j.jff.2019.04.008

Journal of Functional Foods 57 (2019) 233–254
Contents lists available at ScienceDirect
Journal of Functional Foods

journal homepage: www.elsevier.com/locate/jff
Sustainable production of natural phenolics for functional food applications T

a,1 a,1 a b a,⁎
Rita Mark , Xiaomei Lyu , Jaslyn J.L. Lee , Roberto Parra-Saldívar , Wei Ning Chen
a
School of Chemical and Biomedical Engineering, College of Engineering, Nanyang Technological University, 62 Nanyang Drive, Singapore 637459, Singapore
b
Tecnologico de Monterrey, School of Engineering and Science, Campus Monterrey, Ave. Eugenio Garza Sada 2501, Monterrey, N.L. CP 64849, Mexico
ARTICLE INFO ABSTRACT
Keywords: Phenolics are a class of plant-derived natural secondary metabolites with good anticancer, anti-inflammatory,
Phenolic compounds antimicrobial and antioxidant activities. In addition to being natural health-benefit compounds abundant in
Metabolic engineering food, phenolic compounds may also have the potential to be ideal food additives and preservatives. Unlike the
Functional food traditional plant extraction and chemical biosynthesis methods, metabolic engineering of microbes provides a
Sustainable production
possible way to sustainably produce phenolic compounds on a large scale, with high commercial availability and
E. coli
S. cerevisiae
low environmental impact. In the last decade, many phenolic compounds have been successfully synthesized and
optimized in the model organisms, especially in Escherichia coli and Saccharomyces cerevisiae. In this review, the
recent efforts on their sustainable biosynthesis of phenolic compounds for functional food applications are
systematically introduced and discussed.
1. Introduction aggression from pathogens or predators (Pandey & Rizvi, 2009). In

recent years, it was discovered that phenolics-enriched diets can help
In recent years, the consumption behavior of the general population prevent a wide-range of diseases, such as cancers, diabetes, heart dis-
has shifted towards a more health-conscious behavior. Healthier life- ease, neurodegenerative and cardiovascular diseases, and aging
style, value-added products, and health-boosting foods and drinks, with (Balasundram, Sundram, & Samman, 2006; Shahidi & Ambigaipalan,
potential to lower the risk of specific illness, are becoming increasingly 2015). For example, curcumin and isoflavonoids could be used for
popular (Bigliardi & Galati, 2013; Siró, Kápolna, Kápolna, & Lugasi, cancer chemotherapy, proanthocyanidins could tackle urinary tract
2008). On the other hand, food quality suffers easily from oxidation and infections, some specific coumarins and flavonoids possess anti-diabetic
microbiological spoilage, which would lead to changes in food color, properties (Shetty, 2004), anthocyanins could relieve the oxidative
odor, texture, and flavor, and give rise to food poisoning. Food pre- stress related to chronic diseases and decrease CVD-related death
servatives provide an effective solution to address this food-safety issue. (Tsuda, 2012), and ferulic acid was officially classified as an anti-
However, current chemical preservatives are associated with potential oxidant in the list of food additives by FDA (Fazary & Ju, 2007). Fur-
health risks, such as allergy and cancer (Etemadi et al., 2017; Varraso & thermore, phenolic compounds are reported to possess hepatoprotec-
Camargo, 2014). Therefore, the search for functional food additives as tive activities to improve liver damage and treatment of digestive
well as natural, effective, and non-toxic food preservatives have problems (Kumar & Pandey, 2013), anti-proliferative, DNA scission
aroused great interest (Shahidi & Ambigaipalan, 2015). inhibitory and anti-obesity activities (Baboota et al., 2013).
Phenolic compounds are a diverse group of plant secondary meta- More interestingly, phenolic compounds are potential natural food
bolites found wide spread in vegetables, fruits, and cereals. In plants, preservatives due to their high antimicrobial and antioxidant activities.
phenolics are synthetized through the pentose phosphate, shikimate, They can donate hydrogen to terminate the free radical production, so
and phenylpropanoid pathways. Based on their carbon skeleton, dif- as to stop the chain reaction which would lead to cell destruction
ferent phenolics can be categorized into the following groups: phenolic during oxidation. This process not only prevents the spoilage of fruits
acids, flavonoids, stilbenes, tannins, lignans, coumarins, curcuminoids, and vegetables to extend the food products’ shelf-life but also can
and quinones, as shown in Fig. 1 (Basli, Belkacem, & Amrani, 2017). protect the human body against the damage of reactive oxygen species
Phenolics play important roles in the growth and reproduction system, (Shahidi & Ambigaipalan, 2015). In addition, numerous in vitro assays
and can also protect the plants against ultraviolet radiation and and food tests have revealed their high antimicrobial properties,
⁎
Corresponding author.
E-mail address: [email protected] (W.N. Chen).
1
These authors contributed equally to this work.
https://doi.org/10.1016/j.jff.2019.04.008
Received 1 February 2019; Received in revised form 2 April 2019; Accepted 2 April 2019
1756-4646/ © 2019 Published by Elsevier Ltd.
R. Mark, et al. Journal of Functional Foods 57 (2019) 233–254
Fig. 1. Biosynthesis paths and classification of phenolic compounds.
especially towards pathogenic strains (Cowan, 1999; Cushnie & Lamb, large amount of chemicals used also causes environmental stress
2005; Puupponen-Pimiä et al., 2001). For example, phenolic acids have (Stéphane, Denis, Céline, & Laurent, 2011). Metabolic engineering of
been used to inhibit the contamination of chicken soup (Stojković et al., microorganisms provides an alternative approach to sustainably pro-
2013). duce phenolics from renewable feedstocks in larger amounts. The ex-
Food consumption is the main way for humans to intake phenolic pression of heterologous enzymes involved in the plant phenolics bio-
compounds. Fruits, vegetables and beverages are all rich in phenolics, synthetic pathway in microorganisms endows them the capacity to
especially coffee, green tea, and wine. The average phenolics intake produce phenolic compounds. In addition, the faster growth rate of
from food is highly dependent on geological locations. Statistically, microorganisms, as compared to plants, makes them more appealing for
each Japanese consume an average of 1492 mg of phenolic compounds commercial use (Pandey, Parajuli, Koffas, & Sohng, 2016). In addition
from food per day, while Brazilians consume 460 mg of phenolic the use of microorganisms potentially results in less environmental
compounds per day. Apart from natural food sources, phenolic com- damage and toxicity to humans compared to chemical biosynthesis
pounds are available from plant extraction and chemical synthesis for methods (Ng, Lyu, Mark, & Chen, 2019).
usage as supplements and food preservatives. In supplements, the total Bacteria Escherichia coli (E. coli) and yeast Saccharomyces cerevisiae
phenolic content can range between 60 and 200 mg/g and fruit extracts (S. cerevisiae) are the most used model organisms for metabolic en-
top the list. However, plants are subjected to long growth cycles and are gineering, due to their genetic tractability and mature high-density
affected by the season and weather. Thus the low concentration of fermentation technologies. S. cerevisiae is especially suitable for food
phenolic compounds obtained from plants, are not able to meet the preservative production, as it is a generally regarded as safe (GRAS)
demands of the food industry (Lyu, Ng, Lee, Mark, & Chen, 2017). For microorganism and has been used in breweries and other food manu-
chemical synthesis, many complex and tedious steps are required. The facture for thousands of years (bread and wine). Yarrowia lipolytica (Y.
234
lipolytica) is another ideal GRAS strain for the production of high-valued

products in the food industry, such as citric acid and α-ketoglutarate. A
lot of pathway assembly tools, such as “YaliBricks” (Wong, Engel, Jin,
Holdridge, & Xu, 2017) and 26 s rDNA-Cre-loxP system (Lv, Edwards,
Zhou, & Xu, 2019), have been developed for Y. lipolytica cell factory
construction. Although great progress has been achieved with meta-
bolic engineering in microorganisms, to the best of our knowledge,
there is no comprehensive review for production of phenolics using
microorganisms. In this work, the recent progress for microbial pro-
duction of individual phenolic compounds, via metabolic engineering
in E. coli and S. cerevisiae, are systematically reviewed, especially me-
tabolic engineering in E. coli, S. cerevisiae and Y. lipolytica.
2. Phenolic acids
2.1. General introduction
Phenolic acids are a class of substances which contain a phenolic

ring and at least one organic carboxylic acid function and are found in
plants. They are the simplest class of phenolics and are the precursors
for the synthesis of many others. As reported in many studies (Birosova,
Mikulasova, & Vaverkova, 2005; Dai & Mumper, 2010; Stich & Rosin,
1984), phenolic acids have antioxidant, antimutagenic, antic-
arcinogenic, anti-inflammatory, antimicrobial, and other biological
properties. For example, phenolic acids are able to inhibit the growth of
many pathogenic bacteria and fungi, such as Staphylococcus aureus, E.
coli, Klebsiella pneumonia, Bacillus cereus, Aspergillus flavus and Asper-
gillus parasiticus, making them attractive food preservatives (Aziz,
Farag, Mousa, & Abo-Zaid, 1998; Shahidi & Ambigaipalan, 2015;
Stojković et al., 2013). Advanced progress on their health beneficial
properties and more details about their antimicrobial properties can be
found in previous reviews by Guzman (2014), Park et al. (2001), and
Sova et al. (2012).
In the phenolic acid biosynthetic pathway chorismate (the ending
product of the shikimate pathway) or cinnamic acid (an intermediate of
the phenylpropanoid pathway) are the substrates, as is shown in Fig. 2.
Depending on the number of carboxylic acids, natural phenolic acids
are classified into two main groups: hydroxybenzoic acids (C6-C1) and
hydroxycinnamic acids (C6-C3), derived from benzoic acid and cin-
namic acid, respectively (Fig. 2). The main hydroxybenzoic acids are:
benzoic acids, salicylic acid, gallic acid, quinic acid, and shikimic acid;
and the main hydroxycinnamic acids are: coumaric acids, ferulic acid
and caffeic acid. The status of the microbial synthesis using E. coli and S.
cerevisiae for the phenolic acids is presented below.
2.2. Production of hydroxybenzoic acids

Fig. 2. Biosynthetic pathways of diverse phenolic acids and the most effective
strategies for their production improvement. Hydroxycinnamic acids are la-
2.2.1. Benzoic acids: 2,3-DHBA and p-HBA
beled in yellow. Hydroxybenzoic acids are labeled in blue. For improvement of
Important benzoic acids for metabolic engineering are 2,3-dihy-
phenolic acids production, up red arrows indicate the key precursors for en-
droxybenzoic acid (2,3-DHBA) and p-hydroxybenzoic acid (p-HBA), and hancement; black crosses indicate genes for knock-out; black boxes indicate key
they can also be used for production of muconic acid (Mizuno, enzymes for overexpression. The strategies for improving the production of
Yoshikawa, Seki, Mikawa, & Imada, 1988). Muconic acid can be used each specific group of phenolic acids are marked in different colors. Strategies
for bio-plastic production, which offers a sustainable alternative for for production of salicylic acid are marked in purple; the ones for gallic acid are
current non-renewable feedstock based processes (Curran, Leavitt, marked in green; the ones for coumaric acid are marked in yellow; the ones for
Karim, & Alper, 2013). shikimic acid are marked in blue. Enzymes: ppsA, phosphoenolpyruvate syn-
The compound 2,3-DHBA is a native intermediate for the synthesis thase; PST, phosphotransferase system; GalP, galactose permase; GCK, gluco-
of enterobactin in E. coli (Ozenberger, Brickman, & McIntosh, 1989). It kinase; tktA, transketolase 1; aroG, Phe-sensitive phospho-2-dehydro-3-deox-
yheptonate aldolase; aroF, Tyr-sensitive phospho-2-dehydro-3-deoxyheptonate
is catalysed by the enzymes isochorismate synthase (EntC), iso-
aldolase; aroB, 3-dehydroquinate synthase; aroE, shikimate dehydrogenase;
chorismatase (EntB), and 2,3-dihydro-2,3-DHBA dehydrogenase (EntA),
aroK, shikimate kinase 1; aroL, shikimate kinase 2; ydiB, shikimate dehy-
using chorismate as the substrate (Fig. 2). It was reported by Sun et al.
drogenase/quinate dehydrogenase; aroZ, 3-dehydroshikimic acid dehydratase;
(2014), that over-expression of these genes, combined with a knock-out UbiC, chorismate lyase; PobA, p-hydroxybenzoate hydroxylase; ICS, iso-
of the EntE gene, encoding 2,3-dihydroxybenzoate-AMP ligase, in- chorismate synthase; EntA, 2,3-dihydro-2,3-DHBA dehydrogenase; EntB, iso-
hibited the competing consumption of 2,3-DHBA. This resulted in a chorismatase; IPL, isochorismate pyruvate lyase; Irp9, salicylate synthetase;
production of up to 500 mg/L of 2,3-DHBA. On this basis, the avail- pheA, chorismate mutase/prephenate dehydratase; tyrA, prephenate dehy-
ability of chorismate was further increased by expressing the key en- drogenase; PAL, phenylalanine ammonia lyase; TAL, tyrosine ammonia lyase;
zymes in the shikimate pathway (encoded by genes aroL and aroGfbr – C4H, cinnamate 4-hydroxylase; C3H, monooxygenase-p-coumarate 3-hydro-
xylase; COMT, O-methyltransferase. (For interpretation of the references to
235
colour in this figure legend, the reader is referred to the web version of this amount of SA produced (Noda et al., 2016). To achieve this, a pheny-
article.) lalanine-overproducing E. coli strain was constructed by first permitting
the accumulation of intracellular PEP and E4P. This was done by re-
placing the phosphotransferase system (PTS), with a coupled system
feed-back inhibition mutant of aroG) and improving the supply of in-
combining galactose permease (GalP) and glucokinase (GCK) (Fig. 2).
tracellular phosphoenolpyruvate (PEP) and erythrose-4-phosphate
In the second step, the conversion of PEP to pyruvate was inactivated
(E4P) (Fig. 2). This led to a higher titer of 2,3-DHBA (900 mg/L). This is
by the elimination of genes pykF and pykA (encoding pyruvate kinases),
the first report on de novo production of 2,3-DHBA in E. coli (Sun et al.,
while menF gene (expressing menaquinone-specific isochorismate syn-
2014).
thase) from E. coli and pchB gene from Pseudomonas aeruginosa were
In contrast, the pathway for p-HBA biosynthesis is not unique. In
overexpressed.
plants and many microorganisms, chorismic acid is first converted into
phenylpropanoids and then degraded into p-HBA. In E. coli, a shorter
2.2.3. Gallic acid
route was discovered whereby chorismic acid is converted directly into
Gallic acid is widespread in plants, such as gallnuts, sumac, tea
p-HBA, via chorismate lyase (UbiC) (Fig. 2). The first study which de-
leaves and oak bark, and microorganisms such as Phycomyces bla-
monstrated the production of p-HBA in E. coli was reported by Barker
kesleeanus, Enterobacter cloacae, and Aspergillus terreus. Three possible
et al. The UbiC and enzymes involved in the amino acid pathway were
routes have been proposed for the biosynthesis of gallic acid: i) oxi-
over-expressed, which resulted in a production of 12 g/L of p-HBA
dation of 3-dehydroshikimic acid (DHS) followed by spontaneous ar-
under fed-batch fermentation (Barker & Frost, 2001). Metabolic en-
omatization; ii) a two-step catalysis reaction consisting of dehydration
gineering of S. cerevisiae for p-HBA production was also investigated. A
of DHS (by DHS dehydratase AroZ) and hydroxylation of the inter-
total of ∼86 mg/L p-HBA was obtained with the engineered S. cerevisiae
mediate protocatechuic acid (PCA) via p-hydroxybenzoate hydroxylase
strain, expressing the UbiC from E. coli and knocking out of the ARO7
(PobA*, which is mutant enzyme capable of hydroxylating PCA)
gene which encodes for chorismate mutase, thus increasing the accu-
(Fig. 2). Biosynthesis of gallic acid was first reported by Kambourakis
mulation of available chorismate (Krömer et al., 2013).
et al. (2000), in which AroZ from Klebsiella pneumonia and PobA from P.
Based on pathway construction, metabolic engineering for in-
aeruginosa were expressed in E. coli, followed by the elimination of aroE
creasing benzoic acid production was subsequently carried out via
gene (encoding shikimate 5-dehydrogenase) and overexpression of ar-
improving precursor supply and metabolic balance as discussed below.
oFFBR, which is a feedback resistant version of 3-deoxy-7-phospho-
To relieve the metabolic burden caused by genetic modifications in a
heptulonate synthase. This led to the formation of 20 g/L of gallic acid
host cell, a novel dynamic regulation strategy, quorum-sensing circuit
(Kambourakis et al., 2000); iii) a two-step catalysis process which
linked RNA interference, was designed and applied for the production
consisted of Ubic and PobA (Fig. 2). It was designed to eliminate the
of p-HBA. After the growth phase, the key enzymes transketolase
negative impact of these extended pathways from DHS, by using the
(TKL1p), 3-deoxy-7-phosphoheptulonate synthase (ARO4p), and chor-
downstream 4-hydroxybenzoic acid as the substrate. In the study by
ismate lyase (UbiCp)) were overexpressed and the chorismate mutase
Zhenya et al. (2017), protein engineering of PobA were carried out by
(ARO7), anthranilate synthase (TRP3), and pyruvate kinase (CDC19)
rational design, based on structural analysis. A resultant mutant, PobA
were silenced. A higher production of p-HBA was obtained (148 mg/L) Y385F/T294A
, exhibited a 4-fold improvement of activity and a total
(Williams et al., 2015). More recently, a higher production of p-HBA
production of 1.27 g/L of gallic acid (Zhenya et al., 2017).
(2.9 g/L) was achieved in S. cerevisiae by deleting both TRP3 and ARO7,
and expressing of ARO4K229L gene as well as the shikimate kinase, AroL
2.2.4. Shikimic and quinic acids
from E. coli (a major rate-limiting step for formation of aromatic com-
Shikimic acid (SA) is a key intermediate for the synthesis of neur-
pounds), to improve the supply of available precursor chorismate
aminidase inhibitor oseltamivir, an influenza medicine. It is the end
(Averesch, Prima, & Krömer, 2017).
product of the native shikimic acid pathway present in E. coli and S.
cerevisiae (Fig. 2). Quinic acid (QA) is a close relative of shikimic acid. It
2.2.2. Salicylic acid can be obtained from 3-dehydroquinic acid (DHQ), an intermediate of
Salicylic acid (SA) is an important component of many pharma- the shikimic acid pathway, under the catalysis of ydiB (shikimate de-
ceutical products such as aspirin and lamivudine. SA was first synthe- hydrogenase/quinate dehydrogenase) (García et al., 2017). Their bio-
sized in E. coli by Lin et al. (2013), with the aim of producing antic- synthetic pathway and enzymes have been well studied. Recent studies
oagulant precursor 4-hydroxycoumarin. A biosynthesis pathway from focus on ways to improve their productivity and reduce production
Pseudomonas was introduced in E. coli, via over-expression of iso- costs. Taking into consideration that early research work have been
chorismate synthase (ICS) from E. coli (EntC gene) and isochorismate thoroughly reviewed (Bochkov, Sysolyatin, Kalashnikov, &
pyruvate lyase (IPL) from Pseudomonas fluorescence (PchB gene) (Fig. 2). Surmacheva, 2012; Ghosh, Chisti, & Banerjee, 2012; Krämer et al.,
This resulted in a total production of 158.5 mg/L of SA (Lin, Shen, 2003), this review will focus on the research carried out after 2000. The
Yuan, & Yan, 2013). In another study, the Irp9 enzyme from Y. en- major efforts are summarized in Table 1 and the relative strategies are
terocolitica is able to convert chorismate into SA directly (Fig. 2). Upon classified as:
the introduction of this enzyme in combination with a chorismate
mutase/prephenate dehydratase (pheA) and prephenate dehydrogenase 1) Metabolic engineering of the shikimic acid pathway. As mentioned
(tyrA) double deletion, up to 0.9 g/L of SA was achieved by Ahmadi earlier (under the paragraph of p-Coumaric acid), aroG and aroF are
et al. (2016) (Ahmadi, Fawaz, Jones, Zhang, & Pfeifer, 2015; Kerbarh, key genes involved in the shikimate pathway. Shikimate kinase
Ciulli, Howard, & Abell, 2005) (Ahmadi et al., 2016). consumes shikimic acid and produces shikimate 3-phosphate, under
Based on pathway construction, the production of SA was further shikimate kinase, which is encoded by aroK and aroL. Therefore,
enhanced by boosting the availability of chorismate. This was carried over-expression of the feedback-resistant aroGfbr and aroFfbr to-
out by promoting the shikimate pathway (expressing aroL, ppsA, tktA, gether with aroB and/or aroE (the other genes involved in the shi-
and the feedback inhibition resistant mutant of aroG (aroGfbr)) and kimate pathway), and knock-out of aroK and aroL, are the most
disrupting the biosynthesis pathways of phenylalanine and tyrosine (by effective strategies for improving the production of SA (Fig. 2)
knocking out pheA and tyrA) (Fig. 2). As a result, 1.2 g/L of SA was (Chen et al., 2012; Draths, Knop, & Frost, 1999; Escalante et al.,
obtained in the engineered E. coli strain (Lin, Sun, Yuan, & Yan, 2014). 2010; Jian, Kai, Draths, & Frost, 2002; Jian, Draths, Kai, & Frost,
In the study of Noda et al. (2016), an overall 11.5 g/L of salicylic acid 2003; Juminaga et al., 2012; Li et al., 2017; Liu, Lin, Hu, Zhou, &
was finally reached in a batch culture, which was currently the highest Zhu, 2014, 2016; Rodriguez et al., 2013; Yang et al., 2014). Besides,
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R. Mark, et al.
Table 1
Metabolic engineering of E. coli and S. cerevisiae for production of phenolic acids: shikimic, quinic and p-coumaric acid.
Target compound Strategies Host microorganism Yield Reference
Shikimic acid Overexpression of aroB, serA, aroFFBR and areE in the absence of aroL and aroK. E. coli 27.2 g/L Draths et al. (1999)
Quinic acid E. coli 60 g/L
Shikimic acid Decrease of by-product concentration by the repression of shikimate transport. E. coli 52 g/L Knop et al. (2001)
3-dehydroshikimic acid Optimization of phosphoenolpyruvate synthase expression level E. coli 69 g/L Jian et al. (2002)
Shikimic acid Enhancement of phosphoenolpyruvate availability E. coli 87 g/L Chandran et al. (2003)
3-dehydroshikimic acid Optimization of glucose transport system for increased phosphoenolpyruvate availability E. coli 60 g/L Jian et al. (2003)
Shikimic acid Analysis the effects of inactivating aroK, aroL, pykF or pykA and the expression of aroGfbr, tktA, aroB and aroE E. coli 7 g/L Escalante et al. (2010)
Shikimic acid Inactivation of aroK by antisense RNA interference E. coli 1.85 g/L Chen et al. (2012)
Shikimic acid Development of an alternative plasmid-driven constitutive gene expression platform E. coli 43 g/L Rodriguez et al. (2013)
Shikimic acid Targeted proteomics and metabolite profiling in pathway construction and optimization E. coli 750 mg/L Juminaga et al. (2012)
Shikimic acid Genome-wide target gene screening for the pathway flux enhancement, and improvement the enzyme activity of AroG through E. coli 8.9 mg/L Li et al. (2017)
biosensor analysis
Shikimic acid Decrease of byproduct generation and modular expression of aroG, ppsA and tktA genes E. coli 14.6 g/L Chen et al. (2014)
Shikimic acid Optimization of sorbitol as a carbon source E. coli 1077 mg/L Liu et al. (2014)
Shikimic acid Optimization of aerobic fermentation parameters using glycerol as carbon source E. coli 200 mg/L Yang et al. (2014)
Shikimic acid Application of triclosan-induced chromosomal evolution, and analysis the availability of endogenous NADPH E. coli 3.12 g/L Cui et al. (2014)
Shikimic acid Site-specific integration of aroG, aroB, tktA and aroE at ptsHIcrr and ppsA at tyrR. E. coli 27.41 g/L Liu et al. (2016)
237
Shikimic acid Development of a tunable switch for gene regulation E: coli 13.15 g/L Gu, Su, Wang, Liang, and Qi (2016)
Shikimic acid Development of growth phase-dependent control for AroK expression E. coli 5.3 g/L Lee, Hung, & Tsai (2017)
Shikimic acid Enhancement of carbon flux through the implementation of YEASTRACT and OptForce. S. cerevisiae 2 g/L Suástegui et al. (2017)
Quinic acid Investigation of the inactivation or overexpression of ydiB E. coli 7.6 g/L García et al. (2017)
Shikimic acid E. coli 8.2 g/L
p-Coumaric acid Overexpression of R. rubra PAL E. coli Hwang et al. (2003)
p-Coumaric acid Substitution of PAL and C4H for TAL E. coli 2.3 mg/L Watts et al. (2004)
p-Coumaric acid Overexpression of codon-optimized TAL in tyrosine overproducing strain E. coli 974 mg/L Kang et al. (2012)
p-Coumaric acid Determination of TAL and PAL enzyme kinetics and expression in E. coli E. coli 630 µM Vannelli et al. (2007)
p-Coumaric acid Overexpression of TAL, aroGfbr and tktA, in a pheA inactivated E. coli strain, which secrets p-Coa to the media E. coli 22 mg/L Camacho-Zaragoza et al. (2016)
p-Coumaric acid Optimization of yeast extract concentration in a TAL overexpressed E. coli strain E. coli 475 mg/L Chen et al. (2017)
p-Coumaric acid Comparison of TAL enzyme sources E. coli 213 mg/L Santos et al. (2011)
p-Coumaric acid Comparison of TAL, PAL and SeSam8 enzyme sources E. coli 0.54 mM Jendresen et al. (2015)
p-Coumaric acid Mechanism study of carbon allocation into phenylpropanoid metabolism S. cerevisiae Ro and Douglas (2004)
p-Coumaric acid Overexpression of PAL and optimization of precursor feed S. cerevisiae Jiang, Wood, and Morgan (2005)
p-Coumaric acid Determination of TAL and PAL enzyme kinetics and expression in E. coli S. cerevisiae 200 µM Vannelli et al. (2007)
p-Coumaric acid Optimization of phenylalanine feed concentration S. cerevisiae 650 µM Trantas et al. (2009)
p-Coumaric acid Reduction of by-product formation with ARO10 and PDC5 knock-out, overexpression of feedback-resistant DAHP synthase and S. cerevisiae 1.93 g/L Rodriguez et al. (2015) Rodriguez, Chen
chorismate mutase, identification of the shikimate kinases importance in flux-controlling steps Aro2p, optimization of transporters et al. (2017)
p-Coumaric acid Targeted metabolomics study and implementation via global engineering of central metabolism S. cerevisiae Gold et al. (2015)
p-Coumaric acid Identification of new enzymatic bottlenecks by combinatorial metabolic engineering S. cerevisiae 21 mg/L Mao et al. (2017)
repressing shikimate transport can help increase SA, with fewer by- resistant DAHP synthases ARO4K229L and chorismate mutase ARO7G141S
products formed (Knop et al., 2001); were performed to eliminate their feedback inhibition from tyrosine.
2) Metabolic engineering for the biosynthesis of the precursors PEP and The knock-out of phenylpyruvate decarboxylase ARO10 and PDC5
E4P. Over-expression of the tktA gene (encoding transketolase) can eliminated the by-product phenylethanol. These efforts strengthened
enhance the availability of E4P, and consequently increase the titer the supply of tyrosine and obtained more p-Coumaric acid (Gold et al.,
of SA (Knop et al., 2001). Additionally, strengthening the expression 2015; Mao et al., 2017; Rodriguez, Chen et al., 2017; Rodriguez et al.,
of Z. mobilis glf glucose facilitator and Z. mobilis glk glucose kinase in 2015); 3) Discovery and elimination of potential limiting steps by omics
E. coli strain were shown to be another approach with similarly ef- analysis. In the study by Rodriguez, A, seven different key genes (tat1,
fective results (Chandran et al., 2003; Jian et al., 2003). On the tpo1, alp1, agp3, ady2, gal2, and bap2) were identified when com-
other hand, pps gene (encoding PEP synthase) was proven to be a paring metabolic profiling with different S. cerevisiae strains. The var-
critical enzyme for the synthesis of E4P and SA (Chen et al., 2014; ious deletions improved p-coumaric acid production (Rodriguez, Chen
Liu et al., 2016). Instead of using PEP as a substrate, the construc- et al., 2017). For more detailed information, please refer to Table 1.
tion of a novel path from pyruvate, consisting of 2-keto-3-deoxy-6-
phosphogalactonate aldolase, D-erythrose 4-phosphate, 3-dehy- 2.3.2. Caffeic and ferulic acids
droquinate synthase, and 3-dehydroquinate dehydratase, also re- In plants, p-coumaric acid is converted to caffeic acid and ferulic
sulted in substantial production of SA (Ran, Draths, & Frost, 2004). acid by hydroxylation and subsequent methylation reactions, via a
3) Cofactor engineering. In the study by Cui et al., NADPH availability plant-specific cytochrome P450 dependent monooxygenase-p-couma-
has been identified as an important role in the SA biosynthesis, rate 3-hydroxylase (C3H) and O-methyltransferase (COMT), respec-
which could be increased by overexpression of the ge tively (Fig. 2).
4) nes pntAB, encoding NAD(P)+ transhydrogenase, and nadK, en- As the first enzyme, C3Hs belong to P450 enzymes in plants, it is
coding NAD+ kinase(Cui, Ling, Zhang, Huang, & Liu, 2014). difficult to be functionally expressed since they are unstable in bacteria.
5) Condition optimization. The effect of different carbon sources is well To address this issue, recent efforts focus on the expression of microbial
studied. It was concluded that glucose could provide high produc- hydroxylases in E. coli to produce caffeic and ferulic acids. The earliest
tion rates (∼6.8 mg) and glycerol resulted in high yields (∼7 mg) work was reported by Choi et al. (2011), in which 4-coumarate 3-hy-
(Ahn et al., 2008), while phosphate-limited production gave better droxylase (Sam5) from Saccharothrix espanaensis was introduced in E.
product specificity (Johansson & Lidén, 2006) (Louise et al., 2005). coli. As a result, caffeic acid was detected (the titer was not reported)
and ferulic acid was obtained (7.1 mg/L) when COMT from A. thaliana
2.3. Production of hyroxycinnamic acids was expressed (Choi et al., 2011). As reported in another study, ex-
pressing C3H from S. espanaensis and COMT from Medicago sativa re-
2.3.1. p-Coumaric acid sulted in 70 mg/L of caffeic acid and 28.8 mg/L of ferulic acid (Wang
p-Coumaric acid is the precursor for the biosynthesis of the down- et al., 2015). In addition, P450 CYP199A2 from Rhodopseudomonas
stream flavonoids, lignans, stilbenes, condensed tannins, and curcumi- palustris was also found to possess hydroxylation activity of p-coumaric
noids (Fig. 1). It is generated from the compound phenylalanine via acid. By the analysis of its crystal structure and use of site-directed
phenylalanine ammonia lyase (PAL) and cinnamate 4-hydroxylase mutagenesis, a mutant (F185L) with 5.5-fold higher activity was de-
(C4H, a membrane-bound P450 monooxygenase), or produced from veloped. A total of 2.8 g/L of caffeic acid was obtained with whole-cell
tyrosine via tyrosine ammonia lyase (TAL) (Hotze, Schröder, & catalysis process (Furuya, Arai, & Kino, 2012). Besides, recently dis-
Schröder, 1995; Mathias & Schulz, 1999) (Fig. 2). There have been covered hydroxylases are the native E. coli hydroxylase complexes, 4-
many efforts in its synthesis in E. coli and S. cerevisiae, as summarized in hydroxyphenylacetate 3-hydroxylase (4HPA3H), and 4HPA3H from
Table 1. To date, the highest production of p-Coumaric acid in E. coli other bacteria (Pseudomonas aeruginosa and Thermus thermophilus)
was 974 mg/L as reported by Kang et al. (2012), and 1.93 g/L in S. (Takeo et al., 2008).
cerevisiae was reported by Rodriguez et al. (2017). (Rodriguez, Chen The production of caffeic and ferulic acids were further increased by
et al., 2017; Rodriguez, Kildegaard, Li, Borodina, & Nielsen, 2015). improving precursor supply like tyrosine, phenylalanine or p-coumaric
In summary, the main effective strategies (Fig. 2) are: 1) optimizing acid. The most common strategies are over-expression of gene tyrAfbr,
the conversion rate from phenylalanine or tyrosine to p-Coumaric acid, ppsA, tktA and aroGfbr for synthesis of aromatic acids, knock-out of
by gene screening of PAL-4CL and TAL, or gene codon optimization. For metabolic branches consuming tyrosine or phenylalanine, over-ex-
this, PAL from Rhodotorula rubra, A. thaliana, Populus kitakamiensis, R. pression of optimized TAL (Fig. 2) (Kang et al., 2012; Rodrigues,
glutinis were the most popular sources studied, while TAL from Flavo- Araújo, Prather, Kluskens, & Rodrigues, 2015a, 2015b; Santos, Xiao, &
bacterium johnsoniae and Herpetosiphon aurantiacus were found to have Stephanopoulos, 2012; Zhang & Stephanopoulos, 2013). For example,
better performance in converting tyrosine (Watts, Lee, & Schmidt- over-expression of TAL from Rhodobacter capsulatus along with tyrAfbr,
Dannert, 2004) (Jendresen et al., 2015; Kang et al., 2012; Ro & Douglas, ppsA, tktA, and aroGfbr resulted in a production of 50.2 mg/L of caffeic
2004; Santos, Koffas, & Stephanopoulos, 2011; Vannelli, Wei Qi, acid, in the study by Yan et al. (Lin & Yan, 2012). In another study,
Sweigard, Gatenby, & Sariaslani, 2007); 2) Improving the supply of inhibiting pheA (consuming phenylalanine) and tyrA and expressing
precursor phenylalanine or tyrosine, via expression of key enzymes tyrAfbr, ppsA, tktA, and aroGfbr, led to a total of 766 mg/L of caffeic acid
involved in the shikimate pathway, elimination of feedback inhibition, produced from the precursors tyrosine and phenylalanine (Qin, Yuheng,
and knock-out of competing branches. For example, in E. coli, enzy- & Yajun, 2013).
matic catalysis by tyrA, ppsA, tktA, and Aaron were found to be the
limiting steps in the shikimate pathway and aromatic acid biosynthetic 3. Flavonoids
pathway, in which tyrA, aroF, and aroG were feedback inhibited by
tyrosine. As reported in several studies, over-expression of ppsA and 3.1. General introduction
tktA, and introduction of feedback-inhibition-resistant aroGfbr and
tktAfbr led to significant improvement of p-Coumaric acid (Camacho- Flavonoids are a large group of phenolic compounds, consisting of
Zaragoza et al., 2016; Chen, Sun, Li, Yan, & Yuan, 2017; Kang et al., more than 8000 compounds. In plants, flavonoids provide protection
2012; Vannelli et al., 2007). Elimination of phenylalanine consumption, against UV radiation, physical damage, and infections (Forkmann &
via knock-out of gene pheA, provided an effective complementary ap- Martens, 2001). In recent years, it was shown they have antioxidant
proach to further improve its production (Camacho-Zaragoza et al., properties and can lower the risk of cardiovascular, Parkinson's and
2016). Similarly, in S. cerevisiae, over-expression of its feedback- Alzheimer's diseases, cancer and diabetes (Vauzour, Rodriguez-Mateos,
238
Table 2
R. Mark, et al.
Production of flavonone in E. coli and S. cerevisiae.

Target compound Substrate Strategies Host microorganism Yield Reference
Naringenin Tyrosine and Overexpression of three gene clusters with different promoter location E. coli 0.27 µg/L Hwang et al. (2003)
Pinocembrin phenylalanine 0.17 µg/L
Naringenin Tyrosine and Substitution of PAL with TAL E. coli 20.8 mg/L Watts et al. (2004)
phenylalanine
Naringenin Tyrosine Fermentative production of flavanones E. coli 57 mg/L Miyahisa et al. (2005)
Pinocembrin Phenylalanine 58 mg/L
Pinocembrin p-coumaric acid Metabolic engineering of the central metabolism by the enhancement of the intracellular E.coli 710 mg/L Leonard et al. (2007)Leonard et al. (2008)
Naringenin malonyl-CoA pool, analysis of macromolecule complexes, amplification of acetate assimilation 186 mg/L
Eriodictyol pathways, engineering alternative carbon assimilation pathway and inhibition of competitive 54 mg/L
reaction pathways
Naringenin p-coumaric acid Analysis of four‐step metabolic circuit recombinant strains in shake flasks E. coli 33.7 mg/L Yajun et al. (2007)
Liquiritigenin 16.9 mg/L
Pinocembrin Cinnamic acid 28.5 mg/L
7-hydroxyflavanone 1.9 mg/L
Eriodictyol Caffeic acid 5.2 mg/L
Butin 4.2 mg/L
Naringenin p-coumaric acid S. cerevisiae 118.9 mg/L
Liquiritigenin 13.5 mg/L
Pinocembrin Cinnamic acid 57.2 mg/L
Eriodictyol Tyrosine Enhanced intracellular malonyl-CoA and higher cytochrome P450 activity E. coli 107 mg/L Zhu, Wu, Du, Zhou, and Chen (2014)
Eriodictyol Naringenin Optimization of media, induction temperature, induction point and substrate delay time E. coli 62.7 mg/L Jones et al. (2015)
239
Pinocembrin Phenylalanine Overexpression of fatty acid synthases to increase malonyl-CoA availability E. coli 29.9 mg/L Cao et al. (2016)
Pinocembrin Glucose Optimization of metabolic balance by modular and stepwise modular metabolic strategy. E. coli 525 mg/L Wu, Du, et al. (2013), Wu, Zhang, Dong, and Zhou
Identification of ideal gene copy number, promoter strength and pH. Construction of clustered (2016), Wu, Zhang, Zhou, et al. (2016)
regularly interspaced short palindromic repeats interference for malonyl-Coa enhancement.
Naringenin Glucose Optimization of enzyme sources and relative gene expression levels in a tyrosine overproducing E. coli 84 mg/L Santos et al. (2011)
strain
Naringenin p-coumaric acid Enhancement of intracellular malonyl-CoA availability by integrated computational and E. coli 474 mg/L Xu et al. (2011)
experimental approach
Naringenin p-coumaric acid Enhancement of intracellular malonyl-CoA other cofactor availability by a cipher of evolutionary E. coli 215 mg/L Fowler et al. (2009)
Eriodictyol design 114 mg/L
Naringenin p-coumaric acid Development and application of ePathBrick vectors E. coli 68 mg/L Xu et al. (2012)
Naringenin p-coumaric acid Development and application of a CRISPathBrick vector and pCRISPReporter E. coli 18.9 mg/L Cress et al. (2015)
Naringenin Glucose Mutation of targeted locations across the genome and application of sensory proteins for E. coli 61 mg/L Raman et al. (2014)
chemical concentration optimization
Naringenin p-coumaric acid Enhancement of malonyl-Coa by synthetic antisense RNAs E. coli 91.3 mg/L Yang et al. (2015)
Naringenin Glucose Enhancement of malonyl-Coa by clustered regularly interspaced short palindromic repeats E. coli 421 mg/L Wu et al. (2015), Wu, Yu, et al. (2014), Wu, Zhou,
(CRISPR) interference system and antisense RNA et al. (2014)
Optimization of metabolic balance by modifying plasmid gene copy numbers and promoter
strengths
Naringenin Glucose Optimization of co-culture system by IPTG induction timing, inoculation ratio, carbon source and E. coli – E. coli 41.5 mg/L Ganesan et al. (2017)
strain selection
Naringenin Methanol Utilization of methanol as substrate E. coli Whitaker et al. (2017)
Naringenin Tyrosine Overexpression of PAL, 4CL and CHS in a AH22 and pad1 knock out strain S. cerevisiae 7 mg/L Jiang et al. (2005)
Pinocembrin 0.8 mg/L
Naringenin p-coumaric acid Overexpression of C4H, 4CL, CHS and CHI S. cerevisiae 28.3 mg/L Yan, Kohli, and Koffas (2005)
Pinocembrin cinnamic acid 16.3 mg/L
(continued on next page)
Corona, Oruna-Concha, & Spencer, 2010). Their antimicrobial activity

was demonstrated using assays with flavonoid-rich plant extacts and
pure flavonoids, which inhibited Escherichia coli, Salmonella en-
teritidis, Staphylococcus aureus and Micrococcus luteus (Borrás-Linares
et al., 2015). Additionally, quercetin and apigenin were identified as
Rodriguez, Strucko, et al. (2017) potential DNA gyrase inhibitors (Ohemeng, Schwender, Fu, & Barrett,
1993). Some flavonoids have great antifungal activity. For example,
galangin can inhibit Aspergillus tamarii, A. flavus, Cladosporium
sphaerospermum, Penicillium digitatum and Penicillium italicum, and
Koopman et al. (2012)
Trantas et al. (2009)
the viruses HSV and coxsackie B virus (Meyer, Afolayan, Taylor, &
Lehka et al. (2017)
Lyu et al. (2017)
Erasmus, 1997). Additionally, quercetin, dihydroquercetin, dihy-

drofisetin, leucocyanidin, pelargonidin chloride and catechin can be
Reference
helpful against herpes simplex virus, respiratory syncytial virus, po-

liovirus and Sindbis virus (Cushnie & Lamb, 2005).
Flavonoids are synthetized by the phenylpropanoid pathway,
starting from phenylalanine or tryrosine. Through the conversion of p-
15.6 mg/L
1.55 mg/L
5.31 mg/L
108 mg/L
coumaric acid, naringenin is produced, which is at the central branch

8.9 mg/L
90 mg/L
point of the pathway. The main branches are flavones, flavanones,

Yield
flavons, flavanols, anthocyanins, and isoflavanoids (Fig. 1) (Lyu et al.,

2017). The research status regarding the biosynthesis of these above
Host microorganism
compounds in microbes is discussed. Their glycosylated, methylated,

hydroxylated and prenylated forms are not included here but can be
S. cerevisiae
S. cerevisiae
S. cerevisiae
S. cerevisiae
found in reviews of Pandey et al. (2016) and Cao et al. (2015).
3.2. Flavanones
Development of yeast chassis strain with alleviating feedback inhibition and reduced byproduct
Reduction of by-product formation, overexpression of feedback-resistant genes, identification of
Flavanones is the most important flavonoid group, as they are the

precursors for many other flavonoids. They are generally responsible
Application of modified GAL system, enhancement of malonyl-CoA supply, elimination of
for preventing cardiovascular, neurodegenerative and cancer diseases.

Additionally, eriodictyol could reduce the risk of diabetes (Marín et al.,
2017). The main flavanones which are well studied are pinocembrin,
naringenin, eriodictyol, and liquiritigenin (Pandey et al., 2016). The
efforts in the production of these compounds are summarized in Table 2
and discussed below.
feedback inhibition and down-regulation of competing pathways
3.2.1. Pathway construction

Pathway construction is commonly the first step for heterologous
biosynthesis of biochemicals. To date, the pathway for biosynthesis of
Optimization of phenylalanine feed concentration
pinocembrin, naringenin, eriodictyol, and liquiritigenin are well stu-

died (Kaneko, Hwang, Ohnishi, & Horinouchi, 2003; Marín et al., 2017;
Song et al., 2014). Naringenin is synthesized from the phenylpropanoid
pathway, under catalysis of phenylalanine ammonia lyase (PAL), cin-
namate-4-hydroxylase (C4H), 4-coumarate:coenzyme A ligase (4CL),
important flux-controlling genes
chalcone synthase (CHS), and chalcone isomerase (CHI) using pheny-

lalanine as the substrate (Fig. 3). Naringenin can also be synthesized by
tyrosine ammonia lyase (TAL), 4CL, CHS, CHI from tyrosine (Watts
et al., 2004). For the production of pinocembrin, 4CL from Streptomyces
coelicolor was found to convert cinnamic acid into cinnamoyl-CoA,
which bypasses the C4H step for the production of pinocembrin chal-
formation
Strategies
cone (Hwang, Kaneko, Ohnishi, & Horinouchi, 2003). Leguminous

plants were found to have a chalcone reductase (CHR), which can
catalyze the intermediate of the CHS reaction to yield 6′-deox-
ychalcone. Over-expression of CHI from M. sativa, 4CL from P. crispum,
CHS from P.hyvrida or M. sativa and CHR from M. sativa reportedly
Glycerol and sucrose
obtained 16.9 mg/L of liquiritigenin from p-coumaric acid and 5.2 mg/
p-coumaric acid
L eriodictyol from caffeic acid in E. coli, as well as 13.5 mg/L of li-

phenylalanine
quiritigenin and 7.9 mg/L of eriodictyol in S. cerevisiae (Yajun, Lixuan,

Substrate
& Koffas, 2007). For biosynthesis of eriodictyol, F3′H-CPR complex

Glucose
Glucose
(from G. hybrida and C. roseus), as well as E. coli hydroxylase complex

(HpaBC), provide another feasible approach, which uses naringenin as
Table 2 (continued)
the substrate (Fig. 3) (Jones, Collins Shannon, Vernacchio Victoria,

Target compound
Lachance Daniel, & Koffas Mattheos, 2015).

Liquiritigenin
Naringenin
Naringenin
Naringenin
Naringenin
3.2.2. Improving malonyl-CoA supply

Malonyl-CoA is the precursor of flavanones (Fig. 3). Its low con-
centration is the rate-limiting step for flavonoid synthesis. Hwang et al.
240
Fig. 3. Biosynthetic pathways of flavonoids and the most effective strategies for their production improvement. Detailed biosynthesis branches for malonyl-CoA and
relative regulation are present in red box in the bottom right corner. Flavanones are marked with light blue squares, flavones are marked with yellow squares,
flavanols are marked with orange squares, anthocyanidins are marked with green squares, and isoflavonoids are marked with dark blue squares. Up red arrows and
black boxes indicate the key precursors and key enzymes for production of flavonoids respectively, and red crosses indicate the main enzymes, in competing
branches, for the knockout to increase flavonoid accumulation. Enzymes: PAL, phenylalanine ammonia lyase; C4H, cinnamate 4-hydroxylase; TAL, tyrosine ammonia
lyase; 4CL, 4-coumarate:coenzyme A ligase; CHS, chalcone synthase; CHI, chalcone isomerase; CHR, chalcone reductase; IFS, isoflavone synthase; CPR, cytochrome
P450 Reductase; FSI, flavone synthase I; FLS, flavonol synthase; F3H, flavanone 3-hydroxylase; F3′H, flavonoid 3′-hydroxylase; HpaBC, hydroxylase complex; FNS,
flavone synthase; DFR, dihydroflavonol 4-reductase; ANS, anthocyanin synthase; PGK, phosphoglycerate kinase; GAPD, glyceraldehyde 3-phosphate dehydrogenase;
PDH, pyruvate dehydrogenase; ACC, acetyl-CoA carboxylase; fumC, fumarate hydratase; sucC, succinyl-CoA synthetase; adhE, aldehyde-alcohol dehydrogenase;
mdh, malate dehydrogenase; sdhA, succinate dehydrogenase A; fadB, 3-hydroxyacyl-CoA dehydrogenase/enoyl-CoA hydratase; fadD, fatty acyl coenzyme A syn-
thetase; MatB, malonyl-CoA synthetase; MatC, putative dicarboxylate carrier protein. (For interpretation of the references to colour in this figure legend, the reader is
referred to the web version of this article.)
reported that over-expression of acetyl-CoA carboxylase (ACC), which et al., 2016; Santos et al., 2011), or repressing the genes FabD and
catalyse the conversion of acetyl-CoA to malonyl-CoA, led to significant FabB/FabF (encoding for β-ketoacyl-ACP synthase II) responsible for
improvements of the pool of malonyl-CoA (Fig. 3) (Miyahisa et al., fatty acid synthesis via the synthetic antisense RNAs (Leonard et al.,
2005) (Leonard, Lim, Saw, & Koffas, 2007). Another effective approach 2008; Wu, Yu, Du, Zhou, & Chen, 2014; Yang, Lin, Li, Linhardt, & Yan,
was the introduction of a recombinant malonate assimilation pathway 2015). In addition, acetyl-CoA serves as the precursor for malonyl-CoA,
from Rhizobium. trifollii, which enabled the conversion of supplemented which is consumed by the TCA cycle and glycolysis. Simultaneous de-
malonate to malonyl-CoA under the catalysis of matB and matC (Fig. 3) letion or silencing of the related genes eno, mdh, adhE, fumC and sucC
(Wu, Du, Zhou, & Chen, 2013). However, these methods gave rise to an which are involved in the TCA cycle and glycolysis efficiently chan-
extra metabolic burden on the cells, and required supplementation of nelled carbon flux towards malonyl-CoA and led to improved produc-
malonate. Therefore, other strategies focused on genomic regulation. In tion of naringenin or pinocembrin (Wu, Du, Chen, & Zhou, 2015; Wu,
E. coli, fatty acid biosynthesis is the only metabolic process consuming Zhang, Zhou, & Dong, 2016).
malonyl-CoA. This is a bottleneck which prevent efficient flavonoids In addition to macro-control, Xu et al. (2011) displayed a combined
accumulation. Increasing the malonyl-CoA supply have been reported. computational and experimental approach to improve the availability
This was carried out by inhibiting the fatty acid biosynthesis, by sup- of malonyl-CoA in E. coli. It was shown that the OptForce methodology
plementation of the fatty acid synthesis inhibitor cerulenin (Fig. 3) (Cao (Ranganathan, Suthers, & Maranas, 2010) was used to predict a
241
minimal set of genetic interventions for improved yield of malonyl-CoA. fold increase of naringenin production by repressing the phenylethanol
As a result, the knock-out of fumC (encoding fumarate hydratase) and branch (Koopman et al., 2012). Further, the replacement of TSC13
sucC (encoding succinyl-CoA synthetase), and overexpression of target (responsible for fatty acid and phloretic acid production) with ECR from
ACC, PGK (phosphoglycerate kinase), GAPD (glyceraldehyde 3-phos- M. domestica was an effective approach to reduce the production of the
phate dehydrogenase), and PDH (pyruvate dehydrogenase), synergis- side product phloretic acid (Lehka et al., 2017).
tically channeled carbon flux towards malonyl-CoA and obtained a
production of 474 mg/L of naringenin (Xu et al., 2011). Similarly to the 3.3. Flavones
OptForce methodology, a CiED program (Fowler, Gikandi, & Koffas,
2009) was developed by the same group to identify key genetic targets Flavones is another group of flavonoids with beneficial health
involved in malonyl-CoA synthesis. The engineered E. coli strain, which properties for humans. The main compounds well studied are apigenin,
overexpressed ACC and BPL and had deletions of TCA cycle and gly- luteolin, and chrysin. Apigenin has high anticancer and anti-viral ac-
colysis genes, sdhA, adhE, brnQ and cite, produced 215 mg/L naringenin tivity, luteolin possess anti-inflammatory properties, while chrysin has
and 114 mg/L eriodictyol (Fowler et al., 2009). anti-aging activity (Pandey et al., 2016).
Malonyl-CoA biosensors naturally exist in many Gram-positive As reported in several studies, an introduction of flavone synthase
bacteria. They consist of an autogenously regulated FapR transcrip- (FNS) could enable microbes to produce apigenin (Leonard, Yan, Lim, &
tional repressor module and fapO-operator, in which FapR interacts Koffas, 2005; Thuan, Chaudhary, Van Cuong, & Cuong, 2018). For ex-
with the fapO-operator to mediate the repression of downstream fatty ample, expression of FSI from P. crispum along with 4CL from P.
acid synthesis (James & Cronan, 2003). Based on this naturally occur- crispum, CHS and CHI from P. hybrida, resulted in a production of
ring regulon, many malonyl-CoA biosensors have been designed and 415 µg/L of apigenin and 10 µg/L of luteolin, with p-coumaric and
employed for intracellular metabolite monitoring, genetic screening, as caffeic acid as precursors (Leonard, Chemler, Lim, & Koffas, 2006). This
well as to execute feedback control of the expression of designed ge- strain was further optimized by inhibiting fatty acid synthesis. This was
netic circuits for high yield production of aimed products (Johnson carried out by repressing the genes FabB and FabF, which resulted in
et al., 2017). For example, in the study from Li et al. (2015), two novel 5 mg/L of chrysin, 110 mg/L of apigenin and 4 mg/L of luteolin
gene targets for improving malonyl-CoA were identified by combining (Leonard et al., 2008). Similarly, FSI from P. crispum was over-ex-
this sensor with a genome-wide overexpression library (Li, Si, Wang, & pressed in a naringenin and pinocembrin producing strain (Fig. 3)
Zhao, 2015). And 120% improvement of downstream 3-hydro- (Miyahisa et al., 2005), resulted in a production of 13 mg/L of apigenin
xypropionic acid was achieved in the screened recombinant yeast and 9.4 mg/L of chrysin (Lee, Kim, Kim, & Ahn, 2015; Miyahisa et al.,
strain. In the studies from Xu, Li, et al., 2014; Xu, Wang et al., 2014 and 2006).
Liu et al. (2015), novel malonyl-CoA sensors were designed for in vivo
detection of malonyl-CoA concentration based on its fluorescence read- 3.4. Flavanols
out responses and used for dynamic regulation of malonyl-CoA path-
ways towards improving fatty acid production by alleviating over- Flavanols are mainly known for their anticancer properties.
expression-mediated cellular toxicity. These designs might provide Additionally, they can also reduce the risk of coronary artery disease
potential solutions to solve the same problems in increasing flavonoid and prevent age-induced vascular injuries (Vauzour et al., 2010). The
production, caused by insufficient supply/usage of malonyl-CoA (Liu main flavanol compounds are kaempferol, galangin, quercetin, myr-
et al., 2015; Xu, Li, et al., 2014; Xu, Wang et al., 2014). icetin, catechin, afzelechin, and fisetin (Fig. 3). Amongst these,
kaempferol and quercetin are known to reduce the risk of several
3.2.3. Comprehensive regulations chronic diseases, especially cancer, while fisetin has anti-inflammatory
In addition to regulating the precursor supply, studies were per- activity and can reduce the possibility of Alzheimer’s disease (Chen &
formed to further optimize the metabolism of target flavanones. For Chen, 2013; Zheng, Ock, Kwon, & Suk, 2008).
example, a novel sensor-directed evolution of the whole endogenous The first flavanol synthesized in E. coli was kaempferol. This was
pathway of naringenin was performed in E. coli using MAGE for mu- obtained by expression of the flavonol synthase (FLS) from Citrus unshiu
tagenesis (Wang et al., 2009), the ttgR gene (encoding HTH-type tran- grown on naringenin as the substrate (Fig. 3) (Lukačin, Wellmann,
scriptional regulator) was used as the sensor for naringenin, and TolC, Britsch, Martens, & Matern, 2003). The gene F3H is necessary for higher
outer membrane channel was used as the selector (sodium dodecyl- kaempferol production, which works together with FLS to convert more
sulfate was used for positive selection, and colicin E1 was used for naringenin into kaempferol. In the study by Miyahisa et al., over-ex-
negative selection). After 4 rounds of evolution, 36 times more nar- pression of F3H from Citrus sinensis and FLS in a naringenin producing
ingenin was produced as compared to the parental strain (Raman, strain (Miyahisa et al., 2005) resulted in 15 mg/L of kaempferol from
Rogers, Taylor, & Church, 2014). In another study, modular en- tyrosine and 1.1 mg/L of galangin from phenylalanine (Miyahisa et al.,
gineering was employed to balance the different modules, by arranging 2006). Production of kaempferol in a recombinant S. cerevisiae strain
copy numbers and promoters so as to optimize the metabolism of was demonstrated by over-expressing PAL, CPR, C4H, 4CL, CHS and
naringenin (Wu, Zhou, Du, Zhou, & Chen, 2014). More recently, in the CHI, and F3H & FLS (Naesby et al., 2009; Trantas, Panopoulos, &
study by Ganesan et al. (2017), an E. coli - E. coli co-culture system was Ververidis, 2009). In combination with gene screening, the supplement
developed for the production of naringenin. This included a tyrosine of p-coumaric acid and optimization of fermentation conditions,
over-producing strain (Santos, 2010) which expressed TAL from R. 66.3 mg/L of kaempferol was obtained in the engineered S. cerevisiae
glutinis, and a second strain containing the TAL and 4CL genes from P. strain (Duan et al., 2017).
crispum, CHS from P. hybrida and CHI from M. sativa. The co-culture For biosynthesis of quercetin and myricetin, another two enzymes,
system improved the production of naringenin compared to the mono- flavonoid 3′,5′-hydroxylase (F3′5′H) and CPR are required in addition
culture, due to the reduction of metabolic stress and flexible adjustment to the above genes (Fig. 3). Fusion of F3′5′H with CPR, from C. roseus,
between the two engineered strains (Ganesan et al., 2017). along with expression of 4CL from P. crispum, CHS and CHI from P.
The engineering works in S. cerevisiae are fewer as compared to E. hybrida, F3H from Malus domestica and FLS from A. thaliana resulted in
coli. In addition to the over-expression of key enzymes, as well as the 300 µg/L of kaempferol and 50 µg/L of quercetin, when grown on p-
elimination of feedback inhibition from tyrosine, the most effective coumaric acid. With the supplementation of eriodictyol, 1100 µg/L of
strategies for pathway optimization were to inhibit the competing quercetin and 10 µg/L of myricetin were produced (Leonard, Yan, &
branches, such as phenyl ethanol and phloretic acid. The knock-out of Koffas, 2006).
active phenylpyruvate decarboxylase PDC5 and ARO10 resulted in a 3- Catechin and afzelechin synthesis have also been achieved in an
242
engineered E. coli strain. This was carried out by expressing 4CL from P. 6″-O-malonylglucoside. By overexpressing of pgm, galU (encoding
crispum, CHS from P. hybrida, CHI from M. sativa, F3H from M. do- phosphoglucomutase and glucose-1-phosphate uridylyltransferase),
mestica, dihydroflavonol 4-reductase (DFR) from Anthurium andraeanum additionally fusing of 3GT and ANS, 78.9 mg/L of pelargonidin 3-O-
and leucoanthocyanidin reductase (LAR) from Desmodium uncinatum glucoside and 70.7 mg/L of cyanidin 3-O-glucoside were obtained from
(Fig. 3). With culture condition optimization, 8.8 mg/L of catechin was afzelechin and catechin, respectively (Yajun, Zhen, & Koffas, 2008), in
obtained from eriodictyol while 0.74 mg/L of afzelechin was produced which pH was found playing important roles in production of antho-
from naringenin (Leonard et al., 2005). Furthermore, catechin pro- cyanins. More modifications have been performed, including the
duction was further improved by deletion of genes pgi, ppc and pldA, overexpression of transporter protein yadH, knockout of tolC (Leonard
based on a CiED model. By using dihydroquercetin as a substrate, et al., 2008), 3GT and ANS fusion, and overexpression of phosphoglu-
36 mg/L of catechin was produced, with DFR from A. andraeanum and comutase (ycjU), CMP kinase (cmk) and nucleoside-diphosphate kinase
LAR from D. uncinatum (Chemler, Fowler, McHugh, & Koffas, 2010). In (ndk). The last manipulation resulted in 350 mg/L of cyanidin 3-O-
order to enhance catechin production, an ePathBrick system (Xu, glucoside (Lim et al., 2015). Even though production of anthocyanin
Vansiri, Bhan, & Koffas, 2012) was proposed and applied to screen for has been achieved in E. coli as mentioned, they mainly rely on sup-
the best multi-enzymatic pathway construction. The combination of the plementing intermediates as the substrates. Recently, in the study by
following enzymes was created: F3H from C. sinensis, M. domestica and Jones et al, de novo production of anthocyanin was accomplished in E.
P. crispum, DFR from A. andraeanum, C. sinensis and Fragaria ananassa, coli through the development of a polyculture system, which consisted
LAR from C. sinensis and Desmodium uncinatum. The combination with of 4-strains of Escherichia coli expressing 15 exogenous enzymes from
the highest catechin (129–130 mg/L) contained F3H from C. sinensis, diverse plants and microbes. This approach enabled the construction of
LAR from D. unicatum and DFR from C. sinensis/M. domestica, by uti- the whole pathway of anthocyanin while effectively managing the ac-
lizing eriodictyol as the substrate. The strain containing DFR from M. companying metabolic burden. Overall, 9.5 mg/L of pelargoinidin 3-O-
domestica produced the highest afzelechin (39.5 mg/L) from nar- glucoside was produced from glucose (Jones et al., 2017).
ingenin. With further optimization using gene copy-number and an The production of anthocyanins in S. cerevisiae was shown recently.
introduction of scaffold strategies, 910 mg/L of catechin and 68 mg/L A total of 61 mg/L of naringenin was formed via expression of PAL from
afzelechin were obtained in the pgi and ppc knock-out strain (Zhao A. thaliana, C4H from A. majus, native CPR1, 4CL from A. thaliana, CHS
et al., 2015). In addition, the E. coli hydroxylase complex, HpaBC, could from H. androsaemum, and CHI from M. sativa. Next, a series of genes,
also enable the conversion of afzelechin into catechin. With this ap- including F3′H, F3′5′H, F3H, DFR, 3GT, and ANS, were screened in
proach, 34.7 mg/L catechin was obtained (Jones et al., 2015). order to obtain the enzyme with the highest activity. F3′H from P. x
Recently, fisetin was found to be able to prevent Alzheimer’s disease hybrida and F3′5′H from S. lycopersicum were found to be the best
and complications associated with diabetes type I (Zheng et al., 2008). combination amongst the various F3′5′Hs and F3Hs, while all other
However, to date, the native biosynthetic pathway of fisetin in plants is F3Hs performed efficiently. Amongst the five DFRs, DFR from A. an-
still unclear. Rodriguez et al. (2017) proposed a novel approach for the draeanum produced the highest afzelechin, which was then used for
synthesis of fisetin, which was analogous to the pathway leading to pelargonidin 3-O-glucoside production. The DFR from P. trichocarpa
quercetin production. Firstly, p-Coumaroyl-CoA was converted into and I. hollandica were chosen for cyanidin 3-O-glucoside and delpidinin
isoliquiritigenin by CHS and CHR, then subsequently converted into 3-O-glucoside synthesis, respectively. Amongst the nine 3GTs, 3GT
liquiritigenin by CHI. Under catalysis of F3H and FLS, liquiritigenin from D. caryophyllus was the best one for production of pelargonidin 3-
could be converted into resokaempferol thus leading to the production O-glucoside while 3AGT from F. x ananassa was the best one for pro-
of fisetin by FMO and CPR (Fig. 3). With this approach, 0.3 mg/L of duction of cyanidin 3-O-glucoside and delpidinin 3-O-glucoside.
fisetin was detected in the final engineered E. coli (Stahlhut et al., Moreover, 14 ANSs were screened although no large differences were
2015), 2.29 mg/L of fisetin was detected in S. cerevisiae (Rodriguez, displayed. Based on the above efforts, ∼1.25 mg/L of pelargonidin 3-O-
Strucko, et al., 2017). glucoside, ∼1.9 mg/L of cyanidin 3-O-glucoside, and ∼2 mg/L of del-
pidinin 3-O-glucoside were obtained in the recombinant S. cerevisiae
3.5. Anthocyanins strain (Eichenberger, Hansson, Fischer, Dürr, & Naesby, 2018).
Anthocyanins are known to be mainly responsible for the color of 3.6. Isoflavonoids
flowers. Additionally, they have been studied in relation to cardiovas-
cular and neurodegenerative diseases, and anti-cancer or anti-in- Isoflavonoids are a subclass of flavonoids. There are more than 2000
flammatory activities (Vauzour et al., 2010). The main representatives different compounds. The structure consists of having the benzene at-
are pelargonidin, cyandin, and delphinidin (Li, Wang, Luo, Zhao, & tached at C3 instead of the C2 position. They are involved in the de-
Chen, 2017). fence system of plants (Kim, Kim, Jung, & Ahn, 2009), and carry out
Anthocyanins can be generated from naringenin under the catalysis many biological activities in humans, including anticancer, anti-in-
of flavanone 3-hydroxylase (F3H), dihydroflavonol 4-reductase (DFR), flammatory and neurodegenerative properties (Leonard & Koffas,
anthocyanin synthase (ANS), and anthocyanidin 3-O-glucosyl- 2007). The main isoflavonoid compounds which are well studied are
transferase (3GT) (Fig. 3). According to the study by Yan et al. (2005), genistein and daidzein.
5.6 µg/L of pelargonidin 3-O-glucoside and 6 µg/L of cyanidin 3-O- Under the catalysis of isoflavone synthase (IFS), flavanones can be
glucoside were obtained from naringenin and eriodictyol, by introdu- converted into isoflavonoids (Fig. 3). Introduction of IFS from G. max
cing F3H from M. domestica, DFR from A. andraeanum, ANS from M. and Glycyrrhiza echinata enabled S. cerevisiae to produce genistein from
domestica and PGT8 from P. hybrida (Yan, Chemler, Huang, Martens, & chalcone substrates (Katsuyama, Miyahisa, Funa, & Horinouchi, 2007;
Koffas, 2005). In the study, ANS was identified as the key enzyme for Miyahisa et al., 2005; Ralston, Subramanian, Matsuno, & Yu, 2005). In
biosynthesis of anthocyanins. To eliminate this bottleneck, several ANS E. coli, an extra CPR is usually required since it lacks the cytochrome
enzymes, from A. majus, Gerbera hybrida, P. hybrida, and M. domestica, P450 enzymes. 10 mg/L of genistein and 18 mg/L of diadzein were
were screened, amongst which P. hybrida was found to have the highest obtained by fusion of CPR from C. roseus and IFS from G. max (Effendi
activity. The resultant pelargonidin 3-O-glucoside yield was improved Leonard & Koffas, 2007). In another study, with the same strategy, IFS
to 0.98 mg/L, while the cyanidin 3-O-glucoside amount reached from Trifolium pratense and CPR from O. sativa gave 15.1 mg/L of
2.07 mg/L, in combination with cofactor optimization. Furthermore, genistein from naringenin (Kim et al., 2009).
the addition of 3MaT from Dahlia variabilis was found to enable the Instead of using chalcone substrates, production of genistein have
synthesis of pelargonidin 3-O-6″-O-malonylglucoside and cyanidin 3-O- been reported by utilizing phenylalanine or p-coumaric acid as
243
substrates, together with an over-expression of upstream genes PAL overexpression of the malonate synthetase, enzyme fusion (4CL and
C4H, 4CL, CHS, CHI (Trantas et al., 2009). Moreover, gene screening STS) or co-expression with synthetic scaffold, multivariate modular
has been performed with IFS and CPR to obtain enzymes with higher metabolic engineering and fed-batch fermentation, as reported in many
activity. The genes G. max IFS - G. max CPR and T. pratense IFS - C. studies (Li et al., 2015; Lim, Fowler, Hueller, Schaffer, & Koffas, 2011;
roseus were identified to be good combinations. Upon gene screening, Wang et al., 2015; Wang, Chen, & Yu, 2014; Wang & Yu, 2012; Wu, Liu,
an additional introduction of gene HID resulted in higher genistein et al., 2013). Up to date, the highest resveratrol production from glu-
(33–35 mg/L) (Chemler, Lim, Daiss, & Koffas, 2010). cose is 800 mg/L, which was reached by the application of a pull-push-
clock strain engineering strategy. It included the overexpression of the
4. Stilbenes resveratrol biosynthesis pathway, optimization of the electron transfer
to the cytochrome P450 monooxygenase, decrease of the pathway in-
4.1. Resveratrol termediates degradation, an increase of the precursors' supply (Li,
Schneider, Kristensen, Borodina, & Nielsen, 2016). Besides, the co-
Stilbene are synthesized from p-coumaroyl-CoA, together with three culture system was used for the first time for the biosynthesis of re-
malonyl-CoA, which is similar to the synthesis of naringenin (Lin, Jain, sveratrol from glycerol, which could help address toxicity or inhibitory
& Yan, 2014). The main representative and the most widely studied issues due to the intracellular accumulation of intermediaries
stilbene compound is resveratrol (Bhat Krishna & Pezzuto John, 2006; (Camacho-Zaragoza et al., 2016).
Dziggel, Schäfer, & Wink, 2017). It has antioxidant, anti-inflammatory Apart from E. coli and S. cerevisiae, other microorganisms such as
and anti-aggregatory activities, which can be implemented in che- Bacillus licheniformis, were used as hosts for the production of different
motherapies, against cardiovascular or neurodegenerative diseases resveratrol glucosides (Pandey et al., 2014). By using p-coumaric acid
(Giovinazzo, Ingrosso, Paradiso, De Gara, & Santino, 2012). Resveratrol as a precursor, 0.4 mg/L of resveratrol was produced in Streptomyces
and its derivatives can inhibit Bacillus subtilis, Bacillus brevis, Micro- venezuelae (Park et al., 2009), and the production of resveratrol reached
coccus luteus and several Staphylococcus aureus strains, which demon- 158 mg/L in the engineered Corynebacterium glutamicum strain
strate their high antimicrobial properties (Albert, Horbach, Deising, (Kallscheuer et al., 2016). Without supplementation of precursors,
Siewert, & Csuk, 2011), and they also showed high antifungal activity 12 mg/L of resveratrol was generated in Corynebacterium glutamicum
against Trichophyton tonsurans, Trichophyton rubrum, Trichophyton (Braga et al., 2018). In addition, several plants were also studied and
mentagrophytes, Epidermophyton floccosum, and Microsporum gyp- modified for the production of resveratrol with higher efficiency, better
seum (Chan, 2002). Many studies of its biosynthesis in E. coli and S. resistance properties, and better quality. As this topic has been thor-
cerevisiae have been reported, which are summarized in Table 3 and oughly reviewed, this review will not go into the details (Delaunois,
briefly discussed below. Cordelier, Conreux, Clément, & Jeandet, 2009; Donnez, Jeandet,
In 2006, Watts et al. and Beekwilder et al. achieved microbial Clément, & Courot, 2009; Giovinazzo et al., 2012; Jeandet et al., 2017;
production of resveratrol, via overexpression of heterologous 4CL and Jeong et al., 2016; Wang, Chen, & Yu, 2010).
stilbene synthase (STS) and using p-coumaric acid as a precursor in E.
coli and S. cerevisiae (Fig. 4) (Beekwilder et al., 2006; Watts, Lee, & 4.2. Other stilbenes
Schmidt-Dannert, 2006). STS from different origins were screened,
from which STS from R. palmatum displayed good performance (Jeong Apart from resveratrol, there are a few studies on the biosynthesis of
et al., 2015; Wang et al., 2017). Upon the over-expression of 4CL and piceatannol, pterostilbene, pinosylvin, and pinostilbene.
STS, an introduction of TAL or PAL and C4H enabled the production of Resveratrol O-methyltransferase (ROMT) is the common enzyme for
resveratrol from tyrosine and glucose as substrates (Shin, Jung, Kim, the synthesis of pterostilbene and pinostilbene (Fig. 4). ROMT from
Han, & Seo, 2012; Trantas et al., 2009; Wu, Liu, et al., 2013). The Sorghum bicolor was found to have higher activity than that from Vitis
production was further optimized by the overexpression of the feed- riparia, when expressed in E. coli (Jeong et al., 2014). With an addi-
back-insensitive alleles, enhancement of the malonyl-CoA supply by tional expression of 4CL from S. coelicolor and STS from R. palmatum,
Table 3
Production of resveratrol in E. coli and S. cerevisiae.
Substrate Strategies Host microorganism Yield Reference
p-coumaric acid Overexpression of 4CL and STS E. coli 100 mg/L Watts et al. (2006)
p-coumaric acid Overexpression of 4CL and STS E. coli 16 mg/L Beekwilder et al. (2006)
S. cerevisiae 6 mg/L
p-coumaric acid Optimization of stilbene synthases expression efficiency, host strains, promoter and E. coli 2390 mg/L Lim et al. (2011)
construct designs. Enhancement of intracellular malonyl-CoA pool
p-coumaric acid Comparison of STS and ROMT gene sources E. coli 1.9 mg/L Jeong et al. (2015)
p-coumaric acid Comparison of STS genes E. coli 0.187 mg/L Wang et al. (2017)
Tyrosine Development of a single medium fermentation system. Identification and analysis of E. coli 35 mg/L Wu, Liu, et al. (2013)
pathway bottlenecks by multivariate modular metabolic engineering
Glucose and tyrosine Development of biosynthetic enzyme bricks by combinatorial biosynthesis E. coli 114 mg/L Wang et al. (2015)
Glycerol Coculture system of two E. coli strains, each partially expressing the pathway E. coli 22.6 mg/L Camacho-Zaragoza et al.
(2016)
p-coumaric acid Overexpression of 4CL and STS S. cerevisiae 1.45 µg/L Becker et al. (2003)
p-coumaric acid Overexpression of TAL and, 4CL-STS fusion gene and optimization by synthetic S. cerevisiae 14.4 mg/L Zhang et al. (2006)
scaffolds Wang and Yu (2012)
p-coumaric acid Overexpression of 4CL, STS, codon-optimized TAL and araE transporter S. cerevisiae 3.1 mg/L Wang et al. (2011)
Phenylalanine Optimization of phenylalanine feed concentration S. cerevisiae 0.3 mg/L Trantas et al. (2009)
Tyrosine Deletion of PAD1, enhancement of malonyl-CoA pool with the overexpression of ACC1 S. cerevisiae 5.8 mg/L Shin, Han, Park, Kim, and
under GAL1 promoter Seo (2011)
Shin et al. (2012)
Glucose Direct production from glucose, feedback inhibition and enhancement of malonyl-CoA S. cerevisiae 416 mg/L Li et al. (2015)
Ethanol pool 531 mg/L
Phenylalanine Application of pull-push-block strain engineering strategy S. cerevisae 800 mg/L Li et al. (2016)
244
Fig. 4. Biosynthetic pathway for stilbene and the most common strategies for improve their production. Red up arrows and green down arrows indicate the key
precursors and steps for enhancement to improve production of stilbene. Enzymes: PAL, phenylalanine ammonia lyase; C4H, cinnamate 4-hydroxylase; TAL, tyrosine
ammonia lyase; 4CL, 4-coumarate: coenzyme A ligase; STS, stilbene synthase; ROMT, resveratrol O-methyltransferase; HpaBC, hydroxylase complex. (For inter-
pretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
2.4 mg/L of pinostilbene were generated from p-coumaric acid (Jeong thaliana achieved 33.6 mg/L of pterostilbene from tyrosine. When
et al., 2015). The co-expression of TAL from S. espanaensis, 4CL from ROMT from V. vinifera was introduced, 170 mg/L of pterostilbene was
Nicotiana tabacum, and STS from V. vinifera along with ROMT from A. produced from p-coumaric acid in E. coli while 150 mg/L was produced
245
in S. cerevisiae (Wang, Bhuiya, Zhou, & Yu, 2015). In the study by Li ubiquinone 10 (UQ-10) is the most interesting compound with great
et al., the introduction of PAL, C4H, 4CL from A. thaliana, RS from V. health benefits in the treatment of heart disease (Ohara, Kokado,
vinifera and ROMT from S. bicolor and V. vinifera obtained 5.5 mg/L of Yamamoto, Sato, & Yazaki, 2004).
pinostilbene and 35 mg/L of pterostilbene (Li et al., 2016). UQ biosynthesis consists of 10 sequential steps. This includes me-
The over-expression of PAL, 4CL, and STS resulted in successful thylation, decarboxylation, hydroxylation, and isoprenoid transfer. Its
production of pinosylvin (Fig. 4) (Wang et al., 2015). In order to further production followed the process: generation of the benzoquinone
enhance its production in E. coli, several strategies were implemented, frame, sequential condensation of isoprene units with subsequent
including gene screening of PAL, 4CL, and STS, and enhancing pre- transfer of isoprenoid side chain to the benzoquinone frame. In E. coli,
cursor supply. The best gene combination was PAL from P.crispum, 4CL nine genes, ubiA, ispB, ubiB, ubiC, ubiG, ubiH, crtE, COQ1, and COQ2, are
from S. coelicolor, and STS from P. strobus. As a result, 70 mg/L of pi- used for ubiquinone production. The over-expression of these genes
nosylvin was generated from glucose and 91 mg/L was obtained from have led to higher yield of ubiquinone production (UQ 8) in E. coli (Zhu
phenylalanine (van Summeren-Wesenhagen & Marienhagen, 2015). et al., 1995). In UQ biosynthesis, the condensation reaction between 4-
Using a very similar strategy, Liang et al. managed to obtain 47.5 mg/L HB and the isoprenoid side chain is suspected to be the rate-limiting
of pinosylvin. They used a combinatorial bioengineering approach and step. It is catalyzed by 4-HB: polyprenyl diphosphate (4-HB: PPP)
produced pinosylvin straightly in E. coli grown on glycerol. The con- transferase and is encoded by the ubiA gene in E. coli. The introduction
struct included PAL from R. glutinis, 4CL from A. thaliana, and STS from of this enzyme from other sources, such as ppt1 (encoding palmitoyl-
V. vinifera (Liang et al., 2016). In a different approach, the gene enoyl- protein thioesterase I) from Schizosaccharomyces pombe and Coq2 from
reductase III (fabl) expression was repressed. This reduced the tran- yeast and N. tabacum, significantly improved production of UQ (Zhang,
script level of enoyl-reductase I (fabI) which is involved in the fatty acid Shrestha, Niu, Tian, & Tan, 2007) (Ohara et al., 2004). Another key
synthesis. At the same time, the expression of 4CL from S. coelicolor as enzyme is polyprenyl diphosphate (PPP) synthase, which determines
well as STS from V. vinifera, resulted in the engineered strain producing the precise length of the polyprenyl side chain. In E. coli, it exists in the
52.7 mg/L of pinosylvin from cinnamic acid (Salas-Navarrete et al., form of octaprenyl diphosphate synthase (ispB), which bring about UQ
2018). containing C40 octaprenyl tail (UQ10 or CoQ 10). In Agrobacterium tu-
In the study using tyrosine as a substrate, 21.5 mg/L of piceatannol mefaciens, PPP exists in the form of decaprenyl diphosphate synthase
was produced, when TAL, 4CL, and STS, C3H from S. espanaensis were (dps), which lead to the generation of UQ containing C50 octaprenyl tail
expressed (Wang et al., 2015). The amount of piceatannol was almost (UQ8 or CoQ 8). Based on this recognition, over-expression of dps from
double that of Watts et al.’s (2006) earlier study, which introduced 4CL A. tumefaciens/G. suboxydans resulted in production of CoQ 10 in E. coli
from A. thaliana and STS from A. hypogaea into E. coli. The final pro- (Zhang, Li, et al., 2007) (Okada, Kainou, Tanaka, et al., 1998). Simi-
duction of piceatannol was 13 mg/L after caffeic acid was supple- larly, the natural CoQ9 producer, rice, has also been successfully en-
mented to the culture (Watts et al., 2006). Lin and Yan focused on the gineered for CoQ10 production with the introduction of gene ddsA from
conversion of piceatannol from resveratrol in E. coli using a Non‐P450 Gluconobacter suboxydans (Takahashi et al., 2006, 2010).
hydroxylase HpaBC and obtained 1.2 g/L of piceatannol (Lin & Yan, Based on pathway construction, numerous strategies have been
2014). employed for the improvement of CoQ10 production: 1) improving
Apart from E. coli, Streptomyces venezuelae has also been explored as precursor supply, for example, increasing supply of isopentenyl di-
a host microorganism for engineering of stilbenes. The construct in- phosphate (IPP) pool by over-expression of 1-deoxy-d-xylulose-5-
cluded 4CL from S. coelicolor and STS from A. hypogaea. A total of phosphate synthase (DXS) (Harker & Bramley, 1999), 1-deoxy-D-xylu-
0.6 mg/L pinosylvin was produced by the final engineered strain, but lose 5-phosphate reductoisomerase (DXR), isopentenyl-pyrophosphate
which is lower than the production in E coli (Park et al., 2009). delta isomerase (IDI), and 2-C-methyl-D-erythritol 4-phosphate cyti-
dylyltransferase (IspD) (Kim et al., 2006) (Lu et al., 2014), introduction
5. Quinones of a foreign mevalonate pathway (Zahiri et al., 2006), increasing supply
of HBA by overexpression of key enzymes TktA, AroB, AroL, AroA,
Quinones play important roles in the respiratory system of plants. AroC, and UbiC, or strengthening catalysis capacity of the last steps
They are responsible for the electron transport to generate energy, and encoding by UbiE, UbiG, and UbiH (Lu et al., 2015); 2) pathway-
have proven beneficial for human health. Coenzyme Q10 is one of the blocking engineering with competitive branches, for example, repres-
best antioxidants, due to its close location to the oxidation site at the sion of CoQs via deletion of octaprenyl diphosphate synthase gene
membrane. It is already commercially available as food supplement and (ispB) (Cluis et al., 2011) (Choi, Ryu, Park, & Seo, 2009), restraining
has proven to be helpful in cardiovascular disease treatments (Bennett menaquinone production via deletion of the menA gene (Xu et al.,
& San, 2017; Jeya, Moon, Lee, Kim, & Lee, 2010; Kawamukai, 2016). 2014), suppression of the carotenoid pathway (Zhu, Lu, et al., 2017); 3)
Initially, the main approaches to improve quinone production were regulation of the oxygen uptake and the redox potential so as to in-
focused on conventional mutagenesis and fermentation condition op- fluence the NADH/NAD+ ratio, e.g., over-expression of glycer-
timization. The highest production of the quinone, Coenzyme Q10 aldehyde-3-phosphate dehydrogenase type I gene and Vitreoscilla he-
(CoQ10), reached 770 mg/L in a chemically mutant of R. spheroides moglobin gene (Zhu, Ye, et al., 2017). More detailed information is
KY8596. These efforts have been exhaustively reviewed previously displayed in Table 4.
(Choi, Ryu, & Seo, 2005; de Dieu Ndikubwimana & Lee, 2014; Sakato, Apart from CoQ10, anthraquinone (AQ is a quinone compound
1992), and will not be included in this review. Even though some which was also found to be interesting. Studies on this compound
quinones natively exist in plants, yeast, and bacteria, their synthesis is carried out before 2000 have been thoroughly reviewed by Han et al.
quite complex and difficult (de Dieu Ndikubwimana & Lee, 2014; Liu & (Han, Van der Heijden, & Verpoorte, 2001). In the following years, the
Lu, 2016). Related efforts on their pathway discovery and pathway main host organism studied was Rubia cordifolia. As described by Bul-
optimization in microbes will be introduced herein, as summarized in gakov et al., over-expression of rol-genes from A. rhizogenes showed a
Table 4 and discussed below. The focus is on the most effective qui- higher amount of AQ (Bulgakov et al., 2002; Shkryl Yuri et al., 2007).
nones - ubiquinone (UQ) and anthraquinone. Furthermore, calcium‐dependent protein kinases from A. thaliana was
UQ is a lipid component of the electron transport system in mi- found to play an important role in AQ accumulation. Its expression
crobes. It has a quinone structure with isoprenoids side chains. Based on resulted in a 10–12 folds increase in AQ yield, with the highest yield of
the length of isoprenoid side chain, UQ has many different forms. For 523 mg/L (Shkryl, Veremeichik, Bulgakov, & Zhuravlev, 2011; Shkryl
example, in nature, E. coli, S. cerevisiae, and human have ubiquinone et al., 2016).
with 8, 6 and 10-unit side chains, respectively, amongst which
246
Table 4
Metabolic engineering of microbes for production of quinones.
Target compound Substrate Strategies Host microorganism Yield Reference
CoQ10 – Optimization of oxygen supply conditions R. spheroides 770 mg/L Sakato (1992)
Ubiquinone – Overexpression of ubiA and ispB E. coli Zhu et al. (1995)
CoQ10 – Overexpression of dps, coq2, ppt1 and ubiA A. tumrfaciens 30.8 mg/L Zhang, Li, et al. (2007)
CoQ10 CoQ8 Overexpression of ppt1 and application of direct glucose feedback control in E. coli 23 mg/L Zhang, Shrestha, Niu, et al.
high-cell-density fermentation (2007)
CoQ10 CoQ8 Overexpression of ddsA E. coli 25.5 mg/L Park et al. (2005)
CoQ5 – Comparison of different prenyl diphosphate synthases S. cerevisiae 29.6 µg/g Okada, Kainou, Matsuda,
CoQ6 27.7 µg/g and Kawamukai (1998)
CoQ8 29.4 µg/g
CoQ9 31.6 µg/g
CoQ10 12.3 µg/g
CoQ10 CoQ8 Overexpression of DXP synthase and ddsA E. coli 46 mg/L Kim et al. (2006)
CoQ10 CoQ8 Overexpression of ddsA and optimization of pH and mevalonate E. coli 2.4 mg/g Zahiri et al. (2006)
supplementation
CoQ10 CoQ8 Overexpression of dps, ubiC, ubiA and ubiG E. coli 50.3 mg/L Zhang, Shrestha, Li, and Tan
(2007)
CoQ10 CoQ8 Overexpression of dps and dxs, deletion of ispB E. coli 99.4 mg/L Choi et al. (2009)
CoQ10 CoQ8 Replacement of ispB with dps, overexpression of a synthetic mevalonate E. coli 213 µg/L Cluis et al. (2011)
operon, NAP1 and UbiA
CoQ10 CoQ8 Replacement of ispB with ddsA and gapA with gapC, knockout of pykFA, E. coli 10.7 mg/g Huang, Wang, Liu, and Mao
overexpression of pck. Optimization of precursor balance and NADPH (2011)
availability. Chromosomal integration Huang, Chen, and Liu (2014)
CoQ10 CoQ8 Integration of dps into chromosome, deletion of ispB, overexpression of dxs, E. coli 11.7 mg/g Dai et al. (2015)
idi, ubiCA and replacement of PTS with Zymomonas mobilis
CoQ10 – Overexpression of ubiG, ubiE and ubiH. Utilization of trc/tac promoter, LacIq R. sphaeroides 163.5 mg/L Lu et al. (2013)
protein and RBS on dxs, dxr, idi and ispD Lu et al. (2014)
Lu et al. (2015)
Zhu, Ye, et al. (2017)
CoQ10 – Overexpression of gapA-1 and vgb, suppression of carotenoids synthesis R. sphaeroides 73.2 mg/L Zhu, Lu, et al. (2017)
6. The others Usually, one type of PLR converts (+)-pinoresinol to (−)-secoisolar-

iciresinol while the second one converts (−)-pinoresinol to the (+)
6.1. Tannins enantiomer of secoisolariciresinol (Fujita, Gang, Davin, & Lewis, 1999).
In A. thaliana, PLR was found to possess higher substrate specificity
Tannins are water-soluble phenolic compounds with a molecular (Nakatsubo, Mizutani, Suzuki, Hattori, & Umezawa, 2008). In addition,
weight ranging from 500 to 4000. They have antioxidant, antimicrobial genes for podophyllotoxin production have been identified from Po-
and antitumour activities, and are reported to inhibit HIV (Shahidi & dophyllum hexandrum and expressed in N. benthamiana. Specifically,
Ambigaipalan, 2015). Based on their resistance to hydrolysis, they can PLR, secoisolariciresinol dehydrogenase (SDH), and a type of cyto-
be divided into hydrolyzable tannins and condensed tannins which are chrome P450, CYP719A23 resulted in a very low level of (−)-pluvia-
named as proanthocyanidins (Cai, Luo, Sun, & Corke, 2004; Khoddami, tolide, but increased by 75-fold after supplementation with matair-
Wilkes, & Roberts, 2013). In recent years, metabolic engineering work esinol. Introduction of O-methyltransferases and CYP71CU1 resulted in
have mainly focused on condensed tannins while little information is the production of (−)-yatein, while the overall construct with DIR,
available for hydrolyzable tannins. The biosynthetic pathway of con- PLR, SDH, and CYP719A23 generated 10.3 ng per mg plant dry weight
densed tannins is complex. To date, there are no reports regarding their of (−)-4′-desmethylepipodophyllotoxin when (+)-pinoresinol was
production in microbes. However, reports for plants exists, including used as a substrate (Lau & Sattely, 2015).
the Lotus corniculatus, Medicago and Nicotiana species (Hancock et al., In terms of plant engineering, the Forsythia species is the main focus
2012; Li et al., 2016; Pang et al., 2013; Verdier et al., 2012; Xie, for lignan biosynthesis. In Forsythia koreana, the downregulation of PLR
Sharma, Wright, Wang, & Dixon, 2006) (Fresquet-Corrales et al., 2017) resulted in significant increase in pinoresinol production. The over-
(Li et al., 2016). These findings show that further research is needed to expression of Sesamum CYP81QI increased production of sesamin or
specify the reason condensed tannin production is limited in certain pinoresinol aglycone (Kim et al., 2009; Murata, Matsumoto, Morimoto,
plants, and the transcription factors responsible for these features. Koyama, & Satake, 2015; Ono et al., 2006). On the other hands, PLR
Additionally, the identification of the key enzymes would enable their from Forsythia intermedia was expressed and tested in different wheat
production in microbes. species, and the (+)-secoisolariciresinol diglucoside amounts in the
seeds were measured (Ayella, Trick, & Wang, 2007). In addition, a
comparative study of F. koreana, Forsythia suspense and Forsythia inter-
6.2. Lignans
media has been performed. This resulted in differences in medium
preferences, growth rate and regeneration frequencies, which provided
Lignans have been well studied for their anti-cancer properties
new opportunities for further engineering of Forsythia (Morimoto et al.,
(Ionkova, 2011; Satake et al., 2017; Viladomat & Bastida, 2015).
2011). Apart from Forsythia species, the Linum family has been in-
However, their biosynthesis has not been explored until recently
vestigated as well. In the seeds of Linum utatissimum, the down-
(Marienhagen & Bott, 2013; Satake et al., 2015). In E. coli, only a few
regulation of PLR have led to the loss of (+)-secoisolariciresinol di-
enzymes were expressed. A large gap exists and further research and
glucoside (Renouard et al., 2014), while PLR helped accumulate
investigation is needed. The most interesting compounds of this group
(+)-secoisolariciresinol diglucoside in the seed coat (Hano et al., 2006;
are secoisolariciresinol, matairesinol, and their derivatives pinoresinol,
Renouard et al., 2012). Based on this knowledge about PLR from L.
sesamin, and phyllathin.
utatissimum, a Phyllanthus amarus strain was engineered by RLR ex-
In terms of gene characterization, (+)-pinoresinol/(−)-lariciresinol
pression and resulted in up to 1.23-fold increase of phyllathin (Banerjee
reductase (PLR) isolated from Thuja plicata showed substrate specificity.
247
& Chattopadhyay, 2010). However, the down-regulation of PLR in produced and 9 of those were unnatural (Katsuyama, Hirose, Funa,
Linum perenne hairy roots gave a different result. No differences were Ohnishi, & Horinouchi, 2010). Instead of CUS, a different approach was
found in the amount of justicidin B produced (Hemmati, Schmidt, & employed in E. coli by an introduction of diketide-CoA synthase (DCS)
Fuss, 2007). A third Linum species, Linum corymbulosum was explored as and curcumin synthase (CURS) from Curcuma longa. The engineered
well. Using (+)-pinoresinol as a substrate, (−)-hinokinin was produced strain contained 4CL from A. thaliana and resulted in 70 mg/L curcumin
in the plant's hairy roots and a preferred pathway through (−)-secoi- when grown on ferulic acid as a substrate. When p-coumaric acid was
solariciresinol was discovered (Bayindir, Alfermann, & Fuss, 2008). added as a substrate, bisdemethodycurcumin and demethoxycurcumin
To date, only a few studies have reported the microbial production were produced but in low yield. Furthermore, TAL from R. glutinis, C3H
of lignans. The conversion of (−)-secoisolariciresinol into (−)-ma- from S. espanaensis and CCoAOMT from M. sativa were expressed along
tairesinol has been achieved in E. coli by expression of SDH from F. with the earlier enzymes. The obtained curcumin amount was 3.9 mg/L
intermedia and Podophyllum peltatum (Xia, Costa, Pélissier, Davin, & with caffeic acid as substrate, 0.3 mg/L with p-coumaric acid as a
Lewis, 2001). Additionally, the conversion of (+)-pinoresional to ma- substrate and 0.2 mg/L when grown on tyrosine (Rodrigues et al.,
tairesinol was also achieved in E. coli by the expression of PLR and SDH 2015a, 2015b).
from P. pleianthum, with optimization by protein fusion with PLR (Kuo, In order to increase curcumin production, multiple strategies have
Wei, Lu, Huang, & Lee, 2014). been employed. In the study by Couto et al., fermentation conditions
were optimized, including a one-step fermentation with E. coli species.
6.3. Coumarins In total, 353 mg/L of curcumin was achieved (Couto, Rodrigues, &
Rodrigues, 2017). Furthermore, gene screening, like PAL/TAL (Wang,
Coumarins can be synthesized from cinnamic acid and p-coumaric Zhang, Zhou, Zeng, & Zhan, 2013) (Kim, Cha, Kim, & Ahn, 2017),
acid. However, the exact biosynthetic pathways are still mostly un- protein fusion (Zhang et al., 2016), as well as malonyl-CoA supplement
known (Sun et al., 2015). This limited knowledge makes it difficult for enhancement, were performed to obtain improved production of cur-
metabolic engineers. Hence introduction of the heterologous pathway cumin (Fang, Jones, Zhou, & Koffas, 2017).
have only recently been performed (Marienhagen & Bott, 2013). The
two main metabolites of the group are umbelliferone and scopoletin, 6.5. Violacein
which are beneficial for human health (Chu et al., 2017; Musa,
Cooperwood, & Khan, 2008). Violacein is a purple-colored phenolic compound derived from
The first reported production of umbelliferone and scopoletin were tryptophan, possessing notable antimicrobial, antiviral, anticancer,
performed in E. coli. Introduction of TAL from R. glutini, 4CL from A. antileishmanial, as well as antioxidant properties (Leon, Miranda, De
thaliana, and coumaroyl-CoA 2′-hydroxylase (C2′H) from Ruta grave- Souza, & Durán, 2001; Masuelli et al., 2016; Nakamura, Asada, &
olens led to 4.3 mg/L of umbelliferone from p-coumaric acid, and Sawada, 2003). It is natively produced by various bacteria, such as
2.43 mg/L from tyrosine. Over-expression of TAL from R. glutini, 4CL, Pseudoalteromonas luteoviolacea, Alteromonas luteoviolacea, and Chro-
caffeoyl-CoA O-methyltransferase (CCoAOMT) and caffeoyl-CoA O- mobacterium violaceum (Rahul et al., 2015; Rodrigues et al., 2012; Yada
methyltransferase (F6′H) from A. thaliana, and 4-hydroxyphenylacetate et al., 2008). In the study of August et al. (2000), violacein biosynthetic
3-hydroxylase (4HPA3H) complex obtained 27.8 mg/L scopoletin from pathway in Chromobacterium violaceum was discovered to be encoded
ferulic acid. It reached 3 mg/L when using tyrosine as the substrate by the gene cluster vioABCDE, for converting the precursor tryptophan
(Lin, Sun, Yuan, & Yan, 2013). In another study, a similar strategy, to violacein via multiple enzymatic steps (August et al., 2000). Based on
introducing the above genes from different sources, resulted in higher this discovery, production of violacein has been achieved by the in-
yield of umbelliferone (83 mg/L from coumaric acid) and scopoletin troduction of genes vioABCDE in heterologous hosts like E. coli and Y.
(79.5 mg/L from ferulic acid (Yang, Shim, Kim, & Ahn, 2015) lipolytica. For example, in the study of Ahmetagic et al., improved
Further studies have been conducted on umbelliferone derivatives, production of violacein was obtained in recombinant E. coli, with op-
such as esculatin, skimmin, and herniarin. It was shown that HpaBC timization of plasmids and host strains (Ahmetagic & Pemberton, 2010;
converted umbelliferone to 2.7 g/L (Lin & Yan, 2014), CYP450 BM3 Sarovich & Pemberton, 2007). In a more recent study, enhancement of
from Bacillus megaterium was able to produce 337.10 μM (60.1 mg/L) tryptophan supply and a combination with vioABEDE over-expression
esculetin from umbelliferone, uridine diphosphate glycosyltransferase resulted in 710 mg/L of violacein by E. coli within 12-day fed-batch
YjiC from Bacillus licheniformis resulted in 995.43 μM (322 mg/L) fermentation (Rodrigues et al., 2013). By using YaliBrick vectors, the
skimming, and 37.13 (6.5 mg/L) μM herniarin was obtained with the 12 kb five-gene pathway for violacein production has also been con-
help of an extended acceptor substrate of an O-methyltransferase from structed in Y. lipolytica, which resulted in 31.5 mg/L of violacein (Wong
Streptomyes avermitilis (Chu et al., 2017). et al., 2017). To further improve its production in Y. lipolytica, a sta-
tistical model-based DoE procedure was developed to guide the im-
6.4. Curcuminoids plementation of combinatorial pathway engineering. Firstly, an effi-
cient T7 promoter library was constructed, which covered a broad
Curcuminoids are a small group of phenolic compounds. They have range of gene expression across 1,000-fold transcriptional activity.
antibacterial, anti-inflammatory, antioxidant, anticarcinogenic and Second, the dominant enzyme targets that determine metabolic
anti-HIV activities. Additionally, they can help prevent the develop- pathway efficiency was statistically screened by fractionally sampling
ment of tumors (Shahidi & Ambigaipalan, 2015). Up to now, many the gene expression landscape. Furthermore, an empirical quadratic
metabolic platforms have been already established, with potential for regression model was subsequently used to identify the optimal gene
future implementations (Prasad, Tyagi, & Aggarwal, 2014; Rodrigues, expression patterns of the investigated pathway. This approach finally
Prather, Kluskens, & Rodrigues, 2015). The most popular compounds yielded production of violacein up to 525.4 mg/L in shake flask and
are curcumin, demethoxycurcumin, and bisdemethoxycurcumin, 1.31 g/L in a controlled bench-top bioreactor (Xu, Rizzoni, Sul, &
mainly known as turmeric components. Stephanopoulos, 2017).
Katsuyama et al. (2010) were the first who successfully constructed
the curcuminoid biosynthetic pathway in E. coli. This was done by in- 7. Conclusion
troducing PAL from R. rubra, 4CL from Lithospermum erythrorhizon and
curcuminoid synthase (CUS) from O. sativa. Additionally, they devel- The phenolic compounds’ natural health-beneficial properties as
oped the synthesis of unnatural curcuminoids by supplying two dif- well as antioxidant/antimicrobial activities make them perfect candi-
ferent unnatural carboxylate precursors. In total, 15 curcuminoids were dates for functional food additives and food preservatives. In order to
248
produce them in the most sustainable way, metabolic engineering have Escherichia coli using in silico metabolic predictions. Journal of microbiology and
emerged as an alternative biosynthesis technique. Genetic engineering biotechnology, 18(11), 1773–1784.
Albert, S., Horbach, R., Deising, H. B., Siewert, B., & Csuk, R. (2011). Synthesis and
can be used to increase commercial attractiveness, thereby reducing the antimicrobial activity of (E) stilbene derivatives. Bioorganic & Medicinal Chemistry,
cost of production. In the past decade, big improvements have been 19(17), 5155–5166.
achieved for production of phenolic compounds in both S. cerevisiae and August, P. R., Grossman, T. H., Minor, C., Draper, M. P., MacNeil, I. A., Pemberton, J. M.,
... Osburne, M. S. (2000). Sequence analysis and functional characterization of the
E. coli. Several new techniques were successfully created, demonstrated violacein biosynthetic pathway from Chromobacterium violaceum. Journal of
and established. These techniques have helped to create more efficient Molecular Microbiology and Biotechnology, 2(4), 513–519.
production processes and made it possible to utilize more-cost-effective Averesch, N. J. H., Prima, A., & Krömer, J. O. (2017). Enhanced production of para-
hydroxybenzoic acid by genetically engineered Saccharomyces cerevisiae. Bioprocess
substrates and approaches. Even though the reviewed studies have and Biosystems Engineering, 40(8), 1283–1289.
shown that S. cerevisiae and E. coli are able to efficiently synthesize Ayella, A. K., Trick, H. N., & Wang, W. (2007). Enhancing lignan biosynthesis by over-
different phenolic compounds, none of them had been commercially expressing pinoresinol lariciresinol reductase in transgenic wheat. Molecular Nutrition
& Food Research, 51(12), 1518–1526.
applied. The biggest concern with the commercial application of phe-
Aziz, N. H., Farag, S. E., Mousa, L. A., & Abo-Zaid, M. A. (1998). Comparative anti-
nolic extracts is their safety issues. Currently, many phenolics do not bacterial and antifungal effects of some phenolic compounds. Microbios, 93(374),
have a clear health statement. For example, phenolic extracts in olive 43–54.
oil have been certified as safe (GRAS No. 726), but phenolics’ health Baboota, R. K., Bishnoi, M., Ambalam, P., Kondepudi, K. K., Sarma, S. M., Boparai, R. K.,
& Podili, K. (2013). Functional food ingredients for the management of obesity and
benefits have not yet been approved (EU No. 1924/2006) (EFSA Panel associated co-morbidities – A review. Journal of Functional Foods, 5(3), 997–1012.
on Dietetic Products & Allergies, 2016). In addition, it is worth noting Balasundram, N., Sundram, K., & Samman, S. (2006). Phenolic compounds in plants and
that excessive concentrations of certain natural compounds can also be agri-industrial by-products: Antioxidant activity, occurrence, and potential uses. Food
Chemistry, 99(1), 191–203.
fatal to humans. However, to date, neither the U.S. Food and Drug Banerjee, A., & Chattopadhyay, S. (2010). Effect of over-expression of Linum usita-
Administration (FDA) nor the European Food Safety Authority (EFSA) tissimum PINORESINOL LARICIRESINOL REDUCTASE (LuPLR) gene in transgenic
has identified the ideal dietary intake of phenolic diets. Phenolic Phyllanthus amarus. Plant Cell, Tissue and Organ Culture (PCTOC), 103(3), 315–323.
Barker, J. L., & Frost, J. W. (2001). Microbial synthesis of p-hydroxybenzoic acid from
compounds should be thoroughly tested to determine their safety in glucose. Biotechnology and Bioengineering, 76(4), 376–390.
food and to assess regulatory limits for their daily intake prior to food Basli, A., Belkacem, N., & Amrani, I. (2017). Health benefits of phenolic compounds
use. On the other hand, another limiting factor for commercial use of against cancers. In M. Soto-Hernandez, M. Palma-Tenango, & M.d. R. Garcia-Mateos
(Eds.). Phenolic compounds – Biological activityRijeka: InTech (pp. Ch. 10).
phenolics is the low productivity of microbial cell factories, which is Bayindir, Ü., Alfermann, A. W., & Fuss, E. (2008). Hinokinin biosynthesis in Linum cor-
currently far from industrial applications. Several strains did not utilize ymbulosum Reichenb. The Plant Journal, 55(5), 810–820.
substrate well, or generated unwanted side products. In addition, genes Becker, J. V. W., Armstrong, G. O., van der Merwe, M. J., Lambrechts, M. G., Vivier, M. A.,
& Pretorius, I. S. (2003). Metabolic engineering of Saccharomyces cerevisiae for the
were poorly expressed or the culture conditions were not favourable for
synthesis of the wine-related antioxidant resveratrol. FEMS Yeast Research, 4(1),
the host microorganisms. However, in the last couple of years, the 79–85.
production of phenolic compounds has significantly improved, both in Beekwilder, J., Wolswinkel, R., Jonker, H., Hall, R., de Vos, C. H., & Bovy, A. (2006).
terms of yield and cost-effective culturing conditions. The diversity of Production of resveratrol in recombinant microorganisms. Applied and Environmental
Microbiology, 72.
the demonstrated approaches is wide-ranged. Hopefully, the compar- Bennett, G. N., & San, K.-Y. (2017). Strategies for manipulation of oxygen utilization by
ison and considerations of these findings can lead to a commercially the electron transfer chain in microbes for metabolic engineering purposes. Journal of
applicable strain in the near future. Industrial Microbiology & Biotechnology, 44(4), 647–658.
Bhat Krishna, P. L., & Pezzuto John, M. (2006). Cancer Chemopreventive Activity of
Resveratrol. Annals of the New York Academy of Sciences, 957(1), 210–229.
Acknowledgments Bigliardi, B., & Galati, F. (2013). Innovation trends in the food industry: The case of
functional foods. Trends in Food Science & Technology, 31(2), 118–129.
Birosova, L., Mikulasova, M., & Vaverkova, S. (2005). Antimutagenic effect of phenolic
This work was supported by Nanyang Technological University and acids. Biomedical papers of the Medical Faculty of the University Palacky, Olomouc, Czech
Ministry of Education, Singapore. Republic, 149(2), 489–491.
Bochkov, D. V., Sysolyatin, S. V., Kalashnikov, A. I., & Surmacheva, I. A. (2012). Shikimic
acid: review of its analytical, isolation, and purification techniques from plant and
Author contributions
microbial sources. Journal of Chemical Biology, 5(1), 5–17.
Borrás-Linares, I., Fernández-Arroyo, S., Arráez-Roman, D., Palmeros-Suárez, P. A., Del
Mark and Lyu had researched and compiled prior data. Lee and Val-Díaz, R., Andrade-Gonzáles, I., ... Segura-Carretero, A. (2015). Characterization
of phenolic compounds, anthocyanidin, antioxidant and antimicrobial activity of 25
Parra-Saldívar had interpreted the results. Chen had designed the work
varieties of Mexican Roselle (Hibiscus sabdariffa). Industrial Crops and Products, 69,
and revised the manuscripts. 385–394.
Braga, A., Oliveira, J., Silva, R., Ferreira, P., Rocha, I., Kallscheuer, N., ... Faria, N. (2018).
Conflict of interest Impact of the cultivation strategy on resveratrol production from glucose in en-
gineered Corynebacterium glutamicum. Journal of Biotechnology, 265, 70–75.
Bulgakov, V. P., Tchernoded, G. K., Mischenko, N. P., Khodakovskaya, M. V., Glazunov, V.
The authors declare that they have no conflict of interest. P., Radchenko, S. V., ... Zhuravlev, Y. N. (2002). Effect of salicylic acid, methyl jas-
monate, ethephon and cantharidin on anthraquinone production by Rubia cordifolia
callus cultures transformed with the rolB and rolC genes. Journal of Biotechnology,
Ethical declaration 97(3), 213–221.
Cai, Y., Luo, Q., Sun, M., & Corke, H. (2004). Antioxidant activity and phenolic com-
This research did not include any human subjects and animal ex- pounds of 112 traditional Chinese medicinal plants associated with anticancer. Life
Sciences, 74(17), 2157–2184.
periments. Camacho-Zaragoza, J. M., Hernández-Chávez, G., Moreno-Avitia, F., Ramírez-Iñiguez, R.,
Martínez, A., Bolívar, F., & Gosset, G. (2016). Engineering of a microbial coculture of
References Escherichia coli strains for the biosynthesis of resveratrol. Microbial Cell Factories,
15(1), 163.
Cao, H., Chen, X., Jassbi, A. R., & Xiao, J. (2015). Microbial biotransformation of
Ahmadi, M. K., Fang, L., Moscatello, N., & Pfeifer, B. A. (2016). E. coli metabolic en- bioactive flavonoids. Biotechnology Advances, 33(1), 214–223.
gineering for gram scale production of a plant-based anti-inflammatory agent. Cao, W., Ma, W., Zhang, B., Wang, X., Chen, K., Li, Y., & Ouyang, P. (2016). Improved
Metabolic Engineering, 38, 382–388. pinocembrin production in Escherichia coli by engineering fatty acid synthesis.
Ahmadi, M. K., Fawaz, S., Jones, C. H., Zhang, G., & Pfeifer, B. A. (2015). Total bio- Journal of Industrial Microbiology & Biotechnology, 43(4), 557–566.
synthesis and diverse applications of the nonribosomal peptide-polyketide side- Chan, M. M.-Y. (2002). Antimicrobial effect of resveratrol on dermatophytes and bacterial
rophore yersiniabactin. Applied and Environmental Microbiology, 81(16), 5290–5298. pathogens of the skin. Biochemical Pharmacology, 63(2), 99–104.
Ahmetagic, A., & Pemberton, J. M. (2010). Stable high level expression of the violacein Chandran, S. S., Yi, J., Draths, K. M., Daeniken, R.v., Weber, W., & Frost, J. W. (2003).
indolocarbazole anti-tumour gene cluster and the Streptomyces lividans amyA gene Phosphoenolpyruvate availability and the biosynthesis of shikimic acid. Biotechnology
in E. coli K12. Plasmid, 63(2), 79–85. Progress, 19(3), 808–814.
Ahn, J. O., Lee, H. W., Saha, R., Park, M. S., Jung, J.-K., & Lee, D.-Y. (2008). Exploring the Chemler, J. A., Fowler, Z. L., McHugh, K. P., & Koffas, M. A. G. (2010). Improving NADPH
effects of carbon sources on the metabolic capacity for shikimic acid production in availability for natural product biosynthesis in Escherichia coli by metabolic
249
engineering. Metabolic Engineering, 12(2), 96–104. Fowler, Z. L., Gikandi, W. W., & Koffas, M. A. G. (2009). Increased malonyl coenzyme a
Chemler, J. A., Lim, C. G., Daiss, J. L., & Koffas, M. A. G. (2010). A versatile microbial biosynthesis by tuning the escherichia coli metabolic network and its application to
system for biosynthesis of novel polyphenols with altered estrogen receptor binding flavanone production. Applied and Environmental Microbiology, 75(18), 5831–5839.
activity. Chemistry & Biology, 17(4), 392–401. Fresquet-Corrales, S., Roque, E., Sarrión-Perdigones, A., Rochina, M., López-Gresa, M. P.,
Chen, A. Y., & Chen, Y. C. (2013). A review of the dietary flavonoid, kaempferol on Díaz-Mula, H. M., ... Cañas, L. A. (2017). Metabolic engineering to simultaneously
human health and cancer chemoprevention. Food Chemistry, 138(4), 2099–2107. activate anthocyanin and proanthocyanidin biosynthetic pathways in Nicotiana spp.
Chen, K., Dou, J., Tang, S., Yang, Y., Wang, H., Fang, H., & Zhou, C. (2012). Deletion of PLoS One, 12(9) e0184839.
the aroK gene is essential for high shikimic acid accumulation through the shikimate Fujita, M., Gang, D. R., Davin, L. B., & Lewis, N. G. (1999). Recombinant pinoresinol-
pathway in E. coli. Bioresource Technology, 119, 141–147. lariciresinol reductases from western red cedar (Thuja plicata) catalyze opposite
Chen, X., Li, M., Zhou, L., Shen, W., Algasan, G., Fan, Y., & Wang, Z. (2014). Metabolic enantiospecific conversions. Journal of Biological Chemistry, 274(2), 618–627.
engineering of Escherichia coli for improving shikimate synthesis from glucose. Furuya, T., Arai, Y., & Kino, K. (2012). Biotechnological production of caffeic acid by
Bioresource Technology, 166, 64–71. bacterial cytochrome P450 CYP199A2. Applied and Environmental Microbiology,
Chen, Z., Sun, X., Li, Y., Yan, Y., & Yuan, Q. (2017). Metabolic engineering of Escherichia 78(17), 6087–6094.
coli for microbial synthesis of monolignols. Metabolic Engineering, 39, 102–109. Ganesan, V., Li, Z., Wang, X., & Zhang, H. (2017). Heterologous biosynthesis of natural
Choi, J.-H., Ryu, Y.-W., Park, Y.-C., & Seo, J.-H. (2009). Synergistic effects of chromo- product naringenin by co-culture engineering. Synthetic and Systems Biotechnology,
somal ispB deletion and dxs overexpression on coenzyme Q10 production in re- 2(3), 236–242.
combinant Escherichia coli expressing Agrobacterium tumefaciens dps gene. Journal García, S., Flores, N., De Anda, R., Hernández, G., Gosset, G., Bolívar, F., & Escalante, A.
of Biotechnology, 144(1), 64–69. (2017). The role of the ydiB gene, which encodes qui-
Choi, J.-H., Ryu, Y.-W., & Seo, J.-H. (2005). Biotechnological production and applications nate/shikimate dehydrogenase, in the production of quinic, dehydroshikimic and
of coenzyme Q10. Applied Microbiology and Biotechnology, 68(1), 9–15. shikimic acids in a PTS - Strain of Escherichia
Choi, O., Wu, C.-Z., Kang, S. Y., Ahn, J. S., Uhm, T.-B., & Hong, Y.-S. (2011). Biosynthesis coli Journal of Molecular Microbiology and Biotechnology, 27(1),
of plant-specific phenylpropanoids by construction of an artificial biosynthetic 11–21.
pathway in Escherichia coli. Journal of Industrial Microbiology & Biotechnology, 38(10), Ghosh, S., Chisti, Y., & Banerjee, U. C. (2012). Production of shikimic acid. Biotechnology
1657–1665. Advances, 30(6), 1425–1431.
Chu, L. L., Pandey, R. P., Lim, H. N., Jung, H. J., Thuan, N. H., Kim, T.-S., & Sohng, J. K. Giovinazzo, G., Ingrosso, I., Paradiso, A., De Gara, L., & Santino, A. (2012). Resveratrol
(2017). Synthesis of umbelliferone derivatives in Escherichia coli and their biological biosynthesis: plant metabolic engineering for nutritional improvement of food. Plant
activities. Journal of Biological Engineering, 11(1), 15. Foods for Human Nutrition, 67(3), 191–199.
Cluis, C. P., Ekins, A., Narcross, L., Jiang, H., Gold, N. D., Burja, A. M., & Martin, V. J. J. Gold, N. D., Gowen, C. M., Lussier, F.-X., Cautha, S. C., Mahadevan, R., & Martin, V. J. J.
(2011). Identification of bottlenecks in Escherichia coli engineered for the production (2015). Metabolic engineering of a tyrosine-overproducing yeast platform using
of CoQ10. Metabolic Engineering, 13(6), 733–744. targeted metabolomics. Microbial Cell Factories, 14(1), 73.
Couto, M. R., Rodrigues, J. L., & Rodrigues, L. R. (2017). Optimization of fermentation Gu, P., Su, T., Wang, Q., Liang, Q., & Qi, Q. (2016). Tunable switch mediated shikimate
conditions for the production of curcumin by engineered Escherichia biosynthesis in an engineered non-auxotrophic Escherichia coli. Retrieved from
coli Journal of The Royal Society Interface, 14(133). Scientific Reports, 6, 29745.
Cowan, M. M. (1999). Plant products as antimicrobial agents. Clinical Microbiology Guzman, J. D. (2014). Natural cinnamic acids, synthetic derivatives and hybrids with
Reviews, 12(4), 564–582. antimicrobial activity. Molecules, 19(12), 19292.
Cress, B. F., Toparlak, Ö. D., Guleria, S., Lebovich, M., Stieglitz, J. T., Englaender, J. A., ... Han, Y.-S., Van der Heijden, R., & Verpoorte, R. (2001). Biosynthesis of anthraquinones in
Koffas, M. A. G. (2015). CRISPathBrick: Modular combinatorial assembly of Type II-A cell cultures of the Rubiaceae. [journal article]. Plant Cell, Tissue and Organ Culture,
CRISPR arrays for dCas9-mediated multiplex transcriptional repression in E. coli. ACS 67(3), 201–220.
Synthetic Biology, 4(9), 987–1000. Hancock, K. R., Collette, V., Fraser, K., Greig, M., Xue, H., Richardson, K., ... Rasmussen,
Cui, Y.-Y., Ling, C., Zhang, Y.-Y., Huang, J., & Liu, J.-Z. (2014). Production of shikimic S. (2012). Expression of the R2R3-MYB transcription factor TaMYB14
acid from Escherichia coli through chemically inducible chromosomal evolution and from Trifolium arvense activates proanthocyanidin biosynthesis in
cofactor metabolic engineering. Microbial Cell Factories, 13(1), 21. the legumes Trifolium repens and medicago sativa < /
Curran, K. A., Leavitt, J. M., Karim, A. S., & Alper, H. S. (2013). Metabolic engineering of em > Plant Physiology, 159(3), 1204–1220.
muconic acid production in Saccharomyces cerevisiae. Metabolic Engineering, 15, Hano, C., Martin, I., Fliniaux, O., Legrand, B., Gutierrez, L., Arroo, R. R. J., ... Lainé, E.
55–66. (2006). Pinoresinol–lariciresinol reductase gene expression and secoisolariciresinol
Cushnie, T. P. T., & Lamb, A. J. (2005). Antimicrobial activity of flavonoids. International diglucoside accumulation in developing flax (Linum usitatissimum) seeds. Planta,
Journal of Antimicrobial Agents, 26(5), 343–356. 224(6), 1291–1301.
Dai, G., Miao, L., Sun, T., Li, Q., Xiao, D., & Zhang, X. (2015). Production of coenzyme Harker, M., & Bramley, P. M. (1999). Expression of prokaryotic 1-deoxy-d-xylulose-5-
Q10 by metabolically engineered Escherichia coli. Sheng wu gong cheng xue bao = phosphatases in Escherichia coli increases carotenoid and ubiquinone biosynthesis.
Chinese Journal of Biotechnology, 31(2), 206–219. FEBS Letters, 448(1), 115–119.
Dai, J., & Mumper, R. J. (2010). Plant phenolics: Extraction, analysis and their anti- Hemmati, S., Schmidt, T. J., & Fuss, E. (2007). (+)-Pinoresinol/(−)-lariciresinol re-
oxidant and anticancer properties. Molecules, 15(10), 7313–7352. ductase from Linum perenne Himmelszelt involved in the biosynthesis of justicidin B.
de Dieu Ndikubwimana, J., & Lee, B. H. (2014). Enhanced production techniques, FEBS Letters, 581(4), 603–610.
properties and uses of coenzyme Q10. Biotechnology Letters, 36(10), 1917–1926. Hotze, M., Schröder, G., & Schröder, J. (1995). Cinnamate 4-hydroxylase from
Delaunois, B., Cordelier, S., Conreux, A., Clément, C., & Jeandet, P. (2009). Molecular Catharanthus roseus and a strategy for the functional expression of plant cytochrome
engineering of resveratrol in plants. Plant Biotechnology Journal, 7(1), 2–12. P450 proteins as translational fusions with P450 reductase in Escherichia coli. FEBS
Donnez, D., Jeandet, P., Clément, C., & Courot, E. (2009). Bioproduction of resveratrol Letters, 374(3), 345–350.
and stilbene derivatives by plant cells and microorganisms. Trends in Biotechnology, Huang, M., Chen, Y., & Liu, J. (2014). Chromosomal engineering of escherichia coli for
27(12), 706–713. efficient production of coenzyme Q10. Chinese Journal of Chemical Engineering, 22(5),
Draths, K. M., Knop, D. R., & Frost, J. W. (1999). Shikimic acid and quinic acid: Replacing 559–569.
isolation from plant sources with recombinant microbial biocatalysis. Journal of the Huang, M., Wang, Y., Liu, J., & Mao, Z. (2011). Multiple strategies for metabolic en-
American Chemical Society, 121(7), 1603–1604. gineering of escherichia coli for efficient production of coenzyme Q10. Chinese
Duan, L., Ding, W., Liu, X., Cheng, X., Cai, J., Hua, E., & Jiang, H. (2017). Biosynthesis Journal of Chemical Engineering, 19(2), 316–326.
and engineering of kaempferol in Saccharomyces cerevisiae. Microbial Cell Factories, Hwang, E. I., Kaneko, M., Ohnishi, Y., & Horinouchi, S. (2003). Production of plant-
16(1), 165. specific flavanones by escherichia coli containing an artificial gene cluster. Applied
Dziggel, C., Schäfer, H., & Wink, M. (2017). Tools of pathway reconstruction and pro- and Environmental Microbiology, 69(5), 2699–2706.
duction of economically relevant plant secondary metabolites in recombinant mi- Ionkova, I. (2011). Anticancer lignans – From discovery to biotechnology. Mini Reviews in
croorganisms. Biotechnology Journal, 12(1), 1600145. Medicinal Chemistry, 11(10), 843–856.
Eichenberger, M., Hansson, A., Fischer, D., Dürr, L., & Naesby, M. (2018). De novo bio- James, E. S., & Cronan, J. E. (2003). Never fat or gaunt. Developmental Cell, 4(5), 610–611.
synthesis of anthocyanins in Saccharomyces cerevisiae. FEMS Yeast Research, 18(4) Jeandet, P., Courot, E., Clément, C., Ricord, S., Crouzet, J., Aziz, A., & Cordelier, S.
foy046-foy046. (2017). Molecular engineering of phytoalexins in plants: Benefits and limitations for
Escalante, A., Calderón, R., Valdivia, A., de Anda, R., Hernández, G., Ramírez, O. T., ... food and agriculture. Journal of Agricultural and Food Chemistry, 65(13), 2643–2644.
Bolívar, F. (2010). Metabolic engineering for the production of shikimic acid in an Jendresen, C. B., Stahlhut, S. G., Li, M., Gaspar, P., Siedler, S., Förster, J., ... Nielsen, A. T.
evolved Escherichia coli strain lacking the phosphoenolpyruvate: carbohydrate (2015). Novel highly active and specific tyrosine ammonia-lyases from diverse ori-
phosphotransferase system. Microbial Cell Factories, 9(1), 21. gins enable enhanced production of aromatic compounds in bacteria and yeast.
Etemadi, A., Sinha, R., Ward, M. H., Graubard, B. I., Inoue-Choi, M., Dawsey, S. M., & Applied and Environmental Microbiology.
Abnet, C. C. (2017). Mortality from different causes associated with meat, heme iron, Jeong, Y. J., An, C. H., Woo, S. G., Jeong, H. J., Kim, Y.-M., Park, S.-J., ... Kim, C. Y.
nitrates, and nitrites in the NIH-AARP diet and health study: Population based cohort (2014). Production of pinostilbene compounds by the expression of resveratrol O-
study. BMJ, 357 j1957. methyltransferase genes in Escherichia coli. Enzyme and Microbial Technology, 54,
Fang, Z., Jones, J. A., Zhou, J., & Koffas, M. A. G. (2017). Engineering escherichia coli co- 8–14.
cultures for production of curcuminoids from glucose. Biotechnology Journal, Jeong, Y. J., An, C. H., Woo, S. G., Park, J. H., Lee, K.-W., Lee, S.-H., ... Kim, C. Y. (2016).
1700576. Enhanced production of resveratrol derivatives in tobacco plants by improving the
Fazary, A. E., & Ju, Y.-H. (2007). Feruloyl esterases as biotechnological tools: Current and metabolic flux of intermediates in the phenylpropanoid pathway. Plant Molecular
future perspectives. Acta Biochimica et Biophysica Sinica, 39(11), 811–828. Biology, 92(1), 117–129.
Forkmann, G., & Martens, S. (2001). Metabolic engineering and applications of flavo- Jeong, Y. J., Woo, S. G., An, C. H., Jeong, H. J., Hong, Y.-S., Kim, Y.-M., ... Kim, C. Y.
noids. Current Opinion in Biotechnology, 12(2), 155–160. (2015). Metabolic engineering for resveratrol derivative biosynthesis in Escherichia
250
coli. Molecules and Cells, 38(4), 318–326. Lau, W., & Sattely, E. S. (2015). Six enzymes from mayapple that complete the biosyn-
Jeya, M., Moon, H.-J., Lee, J.-L., Kim, I.-W., & Lee, J.-K. (2010). Current state of coen- thetic pathway to the etoposide aglycone. Science, 349(6253), 1224–1228.
zyme Q10 production and its applications. Applied Microbiology and Biotechnology, Lee, H., Kim, B. G., Kim, M., & Ahn, J.-H. (2015). Biosynthesis of two flavones, apigenin
85(6), 1653–1663. and genkwanin, in Escherichia coli. Journal of microbiology and biotechnology, 25(9),
Jian, Y., Kai, L., Draths, K. M., & Frost, J. W. (2002). Modulation of phosphoenolpyruvate 1442–1448.
synthase expression increases shikimate pathway product yields in E. coli. Lee, M.-Y., Hung, W.-P., & Tsai, S.-H. (2017). Improvement of shikimic acid production in
Biotechnology Progress, 18(6), 1141–1148. Escherichia coli with growth phase-dependent regulation in the biosynthetic pathway
Jian, Y., Draths, K. M., Kai, L., & Frost, J. W. (2003). Altered glucose transport and shi- from glycerol. World Journal of Microbiology and Biotechnology, 33(2), 25.
kimate pathway product yields in E. coli. Biotechnology Progress, 195, 1450–1459. Lehka, B. J., Eichenberger, M., Bjørn-Yoshimoto, W. E., Vanegas, K. G., Buijs, N., Jensen,
Jiang, H., Wood, K. V., & Morgan, J. A. (2005). Metabolic engineering of the phenyl- N. B., ... Naesby, M. (2017). Improving heterologous production of phenylpropanoids
propanoid pathway in Saccharomyces cerevisiae. Applied and Environmental in Saccharomyces cerevisiae by tackling an unwanted side reaction of Tsc13, an
Microbiology, 71(6), 2962–2969. endogenous double-bond reductase. FEMS Yeast Research, 17(1) fox004-fox004.
Johansson, L., & Lidén, G. (2006). Transcriptome analysis of a shikimic acid producing Leon, L. L., Miranda, C. C., De Souza, A. O., & Durán, N. (2001). Antileishmanial activity
strain of Escherichia coli W3110 grown under carbon- and phosphate-limited con- of the violacein extracted from Chromobacterium violaceum. Journal of Antimicrobial
ditions. Journal of Biotechnology, 126(4), 528–545. Chemotherapy, 48(3), 449–450.
Johnson, A. O., Gonzalez-Villanueva, M., Wong, L., Steinbüchel, A., Tee, K. L., Xu, P., & Leonard, E., Chemler, J., Lim, K. H., & Koffas, M. A. G. (2006). Expression of a soluble
Wong, T. S. (2017). Design and application of genetically-encoded malonyl-CoA flavone synthase allows the biosynthesis of phytoestrogen derivatives in Escherichia
biosensors for metabolic engineering of microbial cell factories. Metabolic Engineering, coli. Applied Microbiology and Biotechnology, 70(1), 85–91.
44, 253–264. Leonard, E., & Koffas, M. A. G. (2007). Engineering of artificial plant cytochrome P450
Jones, J. A., Collins Shannon, M., Vernacchio Victoria, R., Lachance Daniel, M., & Koffas enzymes for synthesis of isoflavones by Escherichia coli. Applied and Environmental
Mattheos, A. G. (2015). Optimization of naringenin and p-coumaric acid hydro- Microbiology, 73(22), 7246–7251.
xylation using the native E. coli hydroxylase complex, HpaBC. Biotechnology Progress, Leonard, E., Lim, K.-H., Saw, P.-N., & Koffas, M. A. G. (2007). Engineering central me-
32(1), 21–25. tabolic pathways for high-level flavonoid production in Escherichia coli. Applied and
Jones, J. A., Vernacchio, V. R., Collins, S. M., Shirke, A. N., Xiu, Y., Englaender, J. A., ... Environmental Microbiology, 73(12), 3877–3886.
Koffas, M. A. G. (2017). Complete biosynthesis of anthocyanins using E. coli poly- Leonard, E., Yan, Y., Fowler, Z. L., Li, Z., Lim, C. G., Lim, K. H., & Koffas, M. A. (2008).
cultures. mBio, 8(3). Strain improvement of recombinant Escherichia coli for efficient production of plant
Juminaga, D., Baidoo, E. E. K., Redding-Johanson, A. M., Batth, T. S., Burd, H., flavonoids. Molecular Pharmaceutics, 5.
Mukhopadhyay, A., ... Keasling, J. D. (2012). Modular engineering of l-tyrosine Leonard, E., Yan, Y., & Koffas, M. A. G. (2006). Functional expression of a P450 flavonoid
production in Escherichia coli. Applied and Environmental Microbiology, 78(1), 89–98. hydroxylase for the biosynthesis of plant-specific hydroxylated flavonols in
Kallscheuer, N., Vogt, M., Stenzel, A., Gätgens, J., Bott, M., & Marienhagen, J. (2016). Escherichia coli. Metabolic Engineering, 8(2), 172–181.
Construction of a corynebacterium glutamicum platform strain for the production of Leonard, E., Yan, Y., Lim, K. H., & Koffas, M. A. G. (2005). Investigation of two distinct
stilbenes and (2S)-flavanones. Metabolic Engineering, 38, 47–55. flavone synthases for plant-specific flavone biosynthesis in Saccharomyces cerevisiae.
Kambourakis, S., Draths, K. M., & Frost, J. W. (2000). Synthesis of gallic acid and pyr- Applied and Environmental Microbiology, 71(12), 8241–8248.
ogallol from glucose: Replacing natural product isolation with microbial catalysis. Li, D., Wang, P., Luo, Y., Zhao, M., & Chen, F. (2017). Health benefits of anthocyanins and
Journal of the American Chemical Society, 122(37), 9042–9043. molecular mechanisms: Update from recent decade. Critical Reviews in Food Science
Kaneko, M., Hwang, E. I., Ohnishi, Y., & Horinouchi, S. (2003). Heterologous production and Nutrition, 57(8), 1729–1741.
of flavanones in Escherichia coli: Potential for combinatorial biosynthesis of flavo- Li, H., Liang, C., Chen, W., Jin, J.-M., Tang, S.-Y., & Tao, Y. (2017). Monitoring in vivo
noids in bacteria. Journal of Industrial Microbiology and Biotechnology, 30(8), 456–461. metabolic flux with a designed whole-cell metabolite biosensor of shikimic acid.
Kang, S.-Y., Choi, O., Lee, J. K., Hwang, B. Y., Uhm, T.-B., & Hong, Y.-S. (2012). Artificial Biosensors and Bioelectronics, 98, 457–465.
biosynthesis of phenylpropanoic acids in a tyrosine overproducing Escherichia coli Li, M., Kildegaard, K. R., Chen, Y., Rodriguez, A., Borodina, I., & Nielsen, J. (2015). De
strain. Microbial Cell Factories, 11(1), 153. novo production of resveratrol from glucose or ethanol by engineered Saccharomyces
Katsuyama, Y., Hirose, Y., Funa, N., Ohnishi, Y., & Horinouchi, S. (2010). Precursor- cerevisiae. Metabolic Engineering, 32, 1–11.
directed biosynthesis of curcumin analogs in Escherichia coli Li, M., Schneider, K., Kristensen, M., Borodina, I., & Nielsen, J. (2016). Engineering yeast
Bioscience, Biotechnology, and Biochemistry, 74(3), 641–645. for high-level production of stilbenoid antioxidants. [Article]. Scientific Reports, 6,
Katsuyama, Y., Miyahisa, I., Funa, N., & Horinouchi, S. (2007). One-pot synthesis of 36827.
genistein from tyrosine by coincubation of genetically engineered Escherichia coli Li, P., Dong, Q., Ge, S., He, X., Verdier, J., Li, D., & Zhao, J. (2016). Metabolic engineering
and Saccharomyces cerevisiae cells. Applied Microbiology and Biotechnology, 73(5), of proanthocyanidin production by repressing the isoflavone pathways and re-
1143–1149. directing anthocyanidin precursor flux in legume. Plant Biotechnology Journal, 14(7),
Kawamukai, M. (2016). Biosynthesis of coenzyme Q in eukaryotes. Bioscience, 1604–1618.
Biotechnology, and Biochemistry, 80(1), 23–33. Li, S., Si, T., Wang, M., & Zhao, H. (2015). Development of a synthetic malonyl-CoA
Kerbarh, O., Ciulli, A., Howard, N. I., & Abell, C. (2005). Salicylate biosynthesis: sensor in saccharomyces cerevisiae for intracellular metabolite monitoring and ge-
Overexpression, purification, and characterization of Irp9, a bifunctional salicylate netic screening. ACS Synthetic Biology, 4(12), 1308–1315.
synthase from Yersinia enterocolitica. Journal of Bacteriology, 187(15), 5061–5066. Liang, J.-L., Guo, L.-Q., Lin, J.-F., He, Z.-Q., Cai, F.-J., & Chen, J.-F. (2016). A novel
Khoddami, A., Wilkes, M., & Roberts, T. (2013). Techniques for analysis of plant phenolic process for obtaining pinosylvin using combinatorial bioengineering in Escherichia
compounds. Molecules, 18(2), 2328. coli. World Journal of Microbiology and Biotechnology, 32(6), 102.
Kim, D. H., Kim, B.-G., Jung, N. R., & Ahn, J.-H. (2009). Production of genistein from Lim, C. G., Fowler, Z. L., Hueller, T., Schaffer, S., & Koffas, M. A. G. (2011). High-yield
naringenin using Escherichia coli containing isoflavone synthase-cytochrome P450 resveratrol production in engineered Escherichia coli. Applied and Environmental
reductase fusion protein. Journal of Microbiology and Biotechnology, 19(12), Microbiology, 77(10), 3451–3460.
1612–1616. Lim, C. G., Wong, L., Bhan, N., Dvora, H., Xu, P., Venkiteswaran, S., & Koffas, M. A. G.
Kim, E. J., Cha, M. N., Kim, B.-G., & Ahn, J.-H. (2017). Production of curcuminoids in (2015). Development of a recombinant Escherichia coli strain for overproduction of
engineered Escherichia coli. Journal of Microbiology and Biotechnology, 27(5), the plant pigment anthocyanin. Applied and Environmental Microbiology, 81(18),
975–982. 6276–6284.
Kim, H. J., Ono, E., Morimoto, K., Yamagaki, T., Okazawa, A., Kobayashi, A., & Satake, H. Lin, Y., Jain, R., & Yan, Y. (2014). Microbial production of antioxidant food ingredients
(2009). Metabolic engineering of lignan biosynthesis in forsythia cell culture. Plant via metabolic engineering. Current Opinion in Biotechnology, 26, 71–78.
and Cell Physiology, 50(12), 2200–2209. Lin, Y., Shen, X., Yuan, Q., & Yan, Y. (2013). Microbial biosynthesis of the anticoagulant
Kim, S.-J., Kim, M.-D., Choi, J.-H., Kim, S.-Y., Ryu, Y.-W., & Seo, J.-H. (2006). precursor 4-hydroxycoumarin. Nature communications, 4, 2603.
Amplification of 1-deoxy-d-xyluose 5-phosphate (DXP) synthase level increases Lin, Y., Sun, X., Yuan, Q., & Yan, Y. (2013). Combinatorial biosynthesis of plant-specific
coenzyme Q10 production in recombinant Escherichia coli. Applied Microbiology and coumarins in bacteria. Metabolic Engineering, 18, 69–77.
Biotechnology, 72(5), 982–985. Lin, Y., Sun, X., Yuan, Q., & Yan, Y. (2014). Extending shikimate pathway for the pro-
Knop, D. R., Draths, K. M., Chandran, S. S., Barker, J. L., von Daeniken, R., Weber, W., & duction of muconic acid and its precursor salicylic acid in Escherichia coli. Metabolic
Frost, J. W. (2001). Hydroaromatic equilibration during biosynthesis of shikimic Engineering, 23, 62–69.
acid. Journal of the American Chemical Society, 123(42), 10173–10182. Lin, Y., & Yan, Y. (2012). Biosynthesis of caffeic acid in Escherichia coli using its en-
Koopman, F., Beekwilder, J., Crimi, B., van Houwelingen, A., Hall, R. D., Bosch, D., ... dogenous hydroxylase complex. Microbial Cell Factories, 11(1), 42.
Daran, J.-M. (2012). De novo production of the flavonoid naringenin in engineered Lin, Y., & Yan, Y. (2014). Biotechnological production of plant-specific hydroxylated
Saccharomyces cerevisiae. Microbial Cell Factories, 11(1), 155. phenylpropanoids. Biotechnology and Bioengineering, 111(9), 1895–1899.
Krämer, M., Bongaerts, J., Bovenberg, R., Kremer, S., Müller, U., Orf, S., ... Raeven, L. Liu, D., Xiao, Y., Evans, B. S., & Zhang, F. (2015). Negative feedback regulation of fatty
(2003). Metabolic engineering for microbial production of shikimic acid. Metabolic acid production based on a malonyl-CoA sensor-actuator. ACS Synthetic Biology, 4(2),
Engineering, 5(4), 277–283. 132–140.
Krömer, J. O., Nunez-Bernal, D., Averesch, N. J. H., Hampe, J., Varela, J., & Varela, C. Liu, M., & Lu, S. (2016). Plastoquinone and ubiquinone in plants: Biosynthesis, physio-
(2013). Production of aromatics in Saccharomyces cerevisiae—A feasibility study. logical function and metabolic engineering. [10.3389/fpls.2016.01898]. Frontiers
Journal of Biotechnology, 163(2), 184–193. Plant Science, 7, 1898.
Kumar, S., & Pandey, A. K. (2013). Chemistry and biological activities of flavonoids: An Liu, X., Lin, J., Hu, H., Zhou, B., & Zhu, B. (2014). Metabolic engineering of Escherichia
overview. The Scientific World Journal, 2013, 16. coli to enhance shikimic acid production from sorbitol. World Journal of Microbiology
Kuo, H.-J., Wei, Z.-Y., Lu, P.-C., Huang, P.-L., & Lee, K.-T. (2014). Bioconversion of pi- and Biotechnology, 30(9), 2543–2550.
noresinol into matairesinol by use of recombinant Escherichia coli. Applied and Liu, X., Lin, J., Hu, H., Zhou, B., & Zhu, B. (2016). Site-specific integration and con-
Environmental Microbiology, 80(9), 2687–2692. stitutive expression of key genes into Escherichia coli chromosome increases shikimic
251
acid yields. Enzyme and Microbial Technology, 82, 96–104. 431(2), 241–244.
Louise, J., Anna, L., Gustav, S., Christian, C., Fog, N. K., & Gunnar, L. (2005). Shikimic Okada, K., Kainou, T., Tanaka, K., Nakagawa, T., Matsuda, H., & Kawamukai, M. (1998).
acid production by a modified strain of E. coli (W3110.shik1) under phosphate- Molecular cloning and mutational analysis of the ddsA gene encoding decaprenyl
limited and carbon-limited conditions. Biotechnology and Bioengineering, 92(5), diphosphate synthase from Gluconobacter suboxydans. European Journal of
541–552. Biochemistry, 255(1), 52–59.
Lu, W., Shi, Y., He, S., Fei, Y., Yu, K., & Yu, H. (2013). Enhanced production of CoQ10 by Ono, E., Nakai, M., Fukui, Y., Tomimori, N., Fukuchi-Mizutani, M., Saito, M., ... Tanaka,
constitutive overexpression of 3-demethyl ubiquinone-9 3-methyltransferase under Y. (2006). Formation of two methylenedioxy bridges by a Sesamum < /
tac promoter in Rhodobacter sphaeroides. Biochemical Engineering Journal, 72, 42–47. em > CYP81Q protein yielding a furofuran lignan, (+)-sesamin. Proceedings of the
Lu, W., Ye, L., Lv, X., Xie, W., Gu, J., Chen, Z., ... Yu, H. (2015). Identification and National Academy of Sciences, 103(26), 10116–10121.
elimination of metabolic bottlenecks in the quinone modification pathway for en- Ozenberger, B. A., Brickman, T. J., & McIntosh, M. A. (1989). Nucleotide sequence of
hanced coenzyme Q10 production in Rhodobacter sphaeroides. Metabolic Engineering, Escherichia coli isochorismate synthetase gene entC and evolutionary relationship of
29, 208–216. isochorismate synthetase and other chorismate-utilizing enzymes. Journal of
Lu, W., Ye, L., Xu, H., Xie, W., Gu, J., & Yu, H. (2014). Enhanced production of coenzyme Bacteriology, 171(2), 775–783.
Q10 by self-regulating the engineered MEP pathway in Rhodobacter sphaeroides. Pandey, K. B., & Rizvi, S. I. (2009). Plant polyphenols as dietary antioxidants in human
Biotechnology and Bioengineering, 111(4), 761–769. health and disease. Oxidative Medicine and Cellular Longevity, 2(5), 270–278.
Lukačin, R., Wellmann, F., Britsch, L., Martens, S., & Matern, U. (2003). Flavonol synthase Pandey, R. P., Parajuli, P., Koffas, M. A. G., & Sohng, J. K. (2016). Microbial production of
from Citrus unshiu is a bifunctional dioxygenase. Phytochemistry, 62(3), 287–292. natural and non-natural flavonoids: Pathway engineering, directed evolution and
Lv, Y., Edwards, H., Zhou, J., & Xu, P. (2019). Combining 26s rDNA and the Cre-loxP systems/synthetic biology. Biotechnology Advances, 34(5), 634–662.
system for iterative gene integration and efficient marker curation in Yarrowia li- Pandey, R. P., Parajuli, P., Shin, J. Y., Lee, J., Lee, S., Hong, Y.-S., ... Sohng, J. K. (2014).
polytica. ACS Synthetic Biology, 8(3), 568–576. Enzymatic biosynthesis of novel resveratrol glucoside and glycoside derivatives.
Lyu, X., Ng, K. R., Lee, J. L., Mark, R., & Chen, W. N. (2017). Enhancement of naringenin Applied and Environmental Microbiology, 80(23), 7235–7243.
biosynthesis from tyrosine by metabolic engineering of Saccharomyces cerevisiae. Pang, Y., Abeysinghe, I. S. B., He, J., He, X., Huhman, D., Mewan, K. M., ... Dixon, R. A.
Journal of Agricultural and Food Chemistry, 65(31), 6638–6646. (2013). Functional characterization of proanthocyanidin pathway enzymes from tea
Mao, J., Liu, Q., Song, X., Wang, H., Feng, H., Xu, H., & Qiao, M. (2017). Combinatorial and their application for metabolic engineering. Plant Physiology, 161(3), 1103–1116.
analysis of enzymatic bottlenecks of l-tyrosine pathway by p-coumaric acid produc- Park, E.-S., Moon, W.-S., Song, M.-J., Kim, M.-N., Chung, K.-H., & Yoon, J.-S. (2001).
tion in Saccharomyces cerevisiae. Biotechnology Letters, 39(7), 977–982. Antimicrobial activity of phenol and benzoic acid derivatives. International
Marienhagen, J., & Bott, M. (2013). Metabolic engineering of microorganisms for the Biodeterioration & Biodegradation, 47(4), 209–214.
synthesis of plant natural products. Journal of Biotechnology, 163(2), 166–178. Park, S. R., Yoon, J. A., Paik, J. H., Park, J. W., Jung, W. S., Ban, Y.-H., ... Yoon, Y. J.
Marín, L., Gutiérrez-del-Río, I., Yagüe, P., Manteca, Á., Villar, C. J., & Lombó, F. (2017). (2009). Engineering of plant-specific phenylpropanoids biosynthesis in Streptomyces
De Novo biosynthesis of apigenin, luteolin, and eriodictyol in the actinomycete venezuelae. Journal of Biotechnology, 141(3), 181–188.
streptomyces albus and production improvement by feeding and spore conditioning. Park, Y.-C., Kim, S.-J., Choi, J.-H., Lee, W.-H., Park, K.-M., Kawamukai, M., ... Seo, J.-H.
[Original Research]. Frontiers in Microbiology, 8(921). (2005). Batch and fed-batch production of coenzyme Q10 in recombinant Escherichia
Masuelli, L., Pantanella, F., La Regina, G., Benvenuto, M., Fantini, M., Mattera, R., ... Bei, coli containing the decaprenyl diphosphate synthase gene from Gluconobacter sub-
R. (2016). Violacein, an indole-derived purple-colored natural pigment produced by oxydans. Applied Microbiology and Biotechnology, 67(2), 192–196.
Janthinobacterium lividum, inhibits the growth of head and neck carcinoma cell lines Prasad, S., Tyagi, A. K., & Aggarwal, B. B. (2014). Recent developments in delivery,
both in vitro and in vivo. Tumor Biology, 37(3), 3705–3717. bioavailability, absorption and metabolism of curcumin: The golden pigment from
Mathias, B., & Schulz, G. E. (1999). Overexpression of a designed 2.2 kb gene of eu- golden spice. Cancer Research and Treatment: Official Journal of Korean Cancer
karyotic phenylalanine ammonia-lyase in Escherichia coli. FEBS Letters, 457(1), Association, 46(1), 2–18.
57–60. Puupponen-Pimiä, R., Nohynek, L., Meier, C., Kähkönen, M., Heinonen, M., Hopia, A., &
Meyer, J. J. M., Afolayan, A. J., Taylor, M. B., & Erasmus, D. (1997). Antiviral activity of Oksman-Caldentey, K.-M. (2001). Antimicrobial properties of phenolic compounds
galangin isolated from the aerial parts of Helichrysum aureonitens. Journal of from berries. Journal of Applied Microbiology, 90(4), 494–507.
Ethnopharmacology, 56(2), 165–169. Qin, H., Yuheng, L., & Yajun, Y. (2013). Caffeic acid production enhancement by en-
Miyahisa, I., Funa, N., Ohnishi, Y., Martens, S., Moriguchi, T., & Horinouchi, S. (2006). gineering a phenylalanine over-producing Escherichia coli strain. Biotechnology and
Combinatorial biosynthesis of flavones and flavonols in Escherichia coli. Applied Bioengineering, 110(12), 3188–3196.
Microbiology and Biotechnology, 71(1), 53–58. Rahul, S., Chandrashekhar, P., Hemant, B., Bipinchandra, S., Mouray, E., Grellier, P., &
Miyahisa, I., Kaneko, M., Funa, N., Kawasaki, H., Kojima, H., Ohnishi, Y., & Horinouchi, S. Satish, P. (2015). In vitro antiparasitic activity of microbial pigments and their
(2005). Efficient production of (2S)-flavanones by Escherichia coli containing an combination with phytosynthesized metal nanoparticles. Parasitology International,
artificial biosynthetic gene cluster. Applied Microbiology and Biotechnology, 68(4), 64(5), 353–356.
498–504. Ralston, L., Subramanian, S., Matsuno, M., & Yu, O. (2005). Partial reconstruction of
Mizuno, S., Yoshikawa, N., Seki, M., Mikawa, T., & Imada, Y. (1988). Microbial pro- flavonoid and isoflavonoid biosynthesis in yeast using soybean Type I and Type II
duction of cis, cis-muconic acid from benzoic acid. Applied Microbiology and chalcone isomerases. Plant Physiology, 137(4), 1375–1388.
Biotechnology, 28(1), 20–25. Raman, S., Rogers, J. K., Taylor, N. D., & Church, G. M. (2014). Evolution-guided opti-
Morimoto, K., Ono, E., Kim, H.-J., Okazawa, A., Kobayashi, A., & Satake, H. (2011). The mization of biosynthetic pathways. Proceedings of the National Academy of Sciences,
construction of transgenic Forsythia plants: Comparative study of 111(50), 17803–17808.
three Forsythia species. Plant Biotechnology, 28(3), 273–280. Ran, N., Draths, K. M., & Frost, J. W. (2004). Creation of a shikimate pathway variant.
Murata, J., Matsumoto, E., Morimoto, K., Koyama, T., & Satake, H. (2015). Generation of Journal of the American Chemical Society, 126(22), 6856–6857.
triple-transgenic forsythia cell cultures as a platform for the efficient, stable, and Ranganathan, S., Suthers, P. F., & Maranas, C. D. (2010). OptForce: An optimization
sustainable production of lignans. PLoS One, 10(12) e0144519. procedure for identifying all genetic manipulations leading to targeted over-
Musa, M. A., Cooperwood, J. S., & Khan, M. O. F. (2008). A review of coumarin deri- productions. PLOS Computational Biology, 6(4) e1000744.
vatives in pharmacotherapy of breast cancer. Current Medicinal Chemistry, 15(26), Renouard, S., Corbin, C., Lopez, T., Montguillon, J., Gutierrez, L., Lamblin, F., ... Hano, C.
2664–2679. (2012). Abscisic acid regulates pinoresinol–lariciresinol reductase gene expression
Naesby, M., Nielsen, S. V., Nielsen, C. A., Green, T., Tange, T. O., Simon, E., ... Titiz, O. and secoisolariciresinol accumulation in developing flax (Linum usitatissimum L.)
(2009). Yeast artificial chromosomes employed for random assembly of biosynthetic seeds. Planta, 235(1), 85–98.
pathways and production of diverse compounds in Saccharomyces cerevisiae. Renouard, S., Tribalatc, M.-A., Lamblin, F., Mongelard, G., Fliniaux, O., Corbin, C., ...
Microbial Cell Factories, 8. Lainé, E. (2014). RNAi-mediated pinoresinol lariciresinol reductase gene silencing in
Nakamura, Y., Asada, C., & Sawada, T. (2003). Production of antibacterial violet pigment flax (Linum usitatissimum L.) seed coat: Consequences on lignans and neolignans
by psychrotropic bacterium RT102 strain. Biotechnology and Bioprocess Engineering, accumulation. Journal of Plant Physiology, 171(15), 1372–1377.
8(1), 37–40. Ro, D.-K., & Douglas, C. J. (2004). Reconstitution of the entry point of plant phenyl-
Nakatsubo, T., Mizutani, M., Suzuki, S., Hattori, T., & Umezawa, T. (2008). propanoid metabolism in yeast (Saccharomyces cerevisiae): Implications for control
Characterization of arabidopsis thaliana pinoresinol reductase, a new type of enzyme of metabolic flux into the phenylpropanoid pathway. Journal of Biological Chemistry,
involved in lignan biosynthesis. Journal of Biological Chemistry, 283(23), 279(4), 2600–2607.
15550–15557. Rodrigues, A. L., Göcke, Y., Bolten, C., Brock, N. L., Dickschat, J. S., & Wittmann, C.
Ng, K. R., Lyu, X., Mark, R., & Chen, W. N. (2019). Antimicrobial and antioxidant ac- (2012). Microbial production of the drugs violacein and deoxyviolacein: analytical
tivities of phenolic metabolites from flavonoid-producing yeast: Potential as natural development and strain comparison. Biotechnology Letters, 34(4), 717–720.
food preservatives. Food Chemistry, 270, 123–129. Rodrigues, A. L., Trachtmann, N., Becker, J., Lohanatha, A. F., Blotenberg, J., Bolten, C.
Noda, S., Shirai, T., Oyama, S., & Kondo, A. (2016). Metabolic design of a platform J., ... Wittmann, C. (2013). Systems metabolic engineering of Escherichia coli for
Escherichia coli strain producing various chorismate derivatives. Metabolic production of the antitumor drugs violacein and deoxyviolacein. Metabolic
Engineering, 33, 119–129. Engineering, 20, 29–41.
Ohara, K., Kokado, Y., Yamamoto, H., Sato, F., & Yazaki, K. (2004). Engineering of ubi- Rodrigues, J. L., Araújo, R. G., Prather, K. L. J., Kluskens, L. D., & Rodrigues, L. R.
quinone biosynthesis using the yeast coq2 gene confers oxidative stress tolerance in (2015a). Heterologous production of caffeic acid from tyrosine in Escherichia coli.
transgenic tobacco. The Plant Journal, 40(5), 734–743. Enzyme and Microbial Technology, 71, 36–44.
Ohemeng, K. A., Schwender, C. F., Fu, K. P., & Barrett, J. F. (1993). DNA gyrase inhibitory Rodrigues, J. L., Araújo, R. G., Prather, K. L. J., Kluskens, L. D., & Rodrigues, L. R.
and antibacterial activity of some flavones(1). Bioorganic & Medicinal Chemistry (2015b). Production of curcuminoids from tyrosine by a metabolically engineered
Letters, 3(2), 225–230. Escherichia coli using caffeic acid as an intermediate. Biotechnology Journal, 10(4),
Okada, K., Kainou, T., Matsuda, H., & Kawamukai, M. (1998). Biological significance of 599–609.
the side chain length of ubiquinone in Saccharomyces cerevisiae. FEBS Letters, Rodrigues, J. L., Prather, K. L. J., Kluskens, L. D., & Rodrigues, L. R. (2015). Heterologous
252
production of curcuminoids. Microbiology and Molecular Biology Reviews, 79(1), Multilevel engineering of the upstream module of aromatic amino acid biosynthesis
39–60. in Saccharomyces cerevisiae for high production of polymer and drug precursors.
Rodriguez, A., Chen, Y., Khoomrung, S., Özdemir, E., Borodina, I., & Nielsen, J. (2017). Metabolic Engineering, 42, 134–144.
Comparison of the metabolic response to over-production of p-coumaric acid in two Sun, X., Lin, Y., Yuan, Q., & Yan, Y. (2014). Biological production of muconic acid via a
yeast strains. Metabolic Engineering, 44, 265–272. prokaryotic 2,3-dihydroxybenzoic acid decarboxylase. ChemSusChem, 7(9),
Rodriguez, A., Kildegaard, K. R., Li, M., Borodina, I., & Nielsen, J. (2015). Establishment 2478–2481.
of a yeast platform strain for production of p-coumaric acid through metabolic en- Sun, X., Shen, X., Jain, R., Lin, Y., Wang, J., Sun, J., ... Yuan, Q. (2015). Synthesis of
gineering of aromatic amino acid biosynthesis. Metabolic Engineering, 31, 181–188. chemicals by metabolic engineering of microbes. [10.1039/C5CS00159E]. Chemical
Rodriguez, A., Martínez, J. A., Báez-Viveros, J. L., Flores, N., Hernández-Chávez, G., Society Reviews, 44(11), 3760–3785.
Ramírez, O. T., ... Bolivar, F. (2013). Constitutive expression of selected genes from Takahashi, S., Ogiyama, Y., Kusano, H., Shimada, H., Kawamukai, M., & Kadowaki, K.-I.
the pentose phosphate and aromatic pathways increases the shikimic acid yield in (2006). Metabolic engineering of coenzyme Q by modification of isoprenoid side
high-glucose batch cultures of an Escherichia coli strain lacking PTS and pykF. chain in plant. FEBS Letters, 580(3), 955–959.
Microbial Cell Factories, 12(1), 86. Takahashi, S., Ohtani, T., Satoh, H., Nakamura, Y., Kawamukai, M., & Kadowaki, K.-I.
Rodriguez, A., Strucko, T., Stahlhut, S. G., Kristensen, M., Svenssen, D. K., Forster, J., ... (2010). Development of coenzyme Q10-enriched rice using sugary and shrunken
Borodina, I. (2017). Metabolic engineering of yeast for fermentative production of mutants. Bioscience, Biotechnology, and Biochemistry, 74(1), 182–184.
flavonoids. Bioresource Technology. Takeo, M., Murakami, M., Niihara, S., Yamamoto, K., Nishimura, M., Kato, D.-I., &
Sakato, K. (1992). Agitation-aeration studies on coenzyme Q10 production using Negoro, S. (2008). Mechanism of 4-nitrophenol oxidation in 
Rhodopseudomonas sphaeroides. Biotechnology and Applied Biochemistry, 16, 19–28. rhodococcus sp. Strain PN1: Characterization of the two-component 4-ni-
Salas-Navarrete, C., Hernández-Chávez, G., Flores, N., Martínez, L. M., Martinez, A., trophenol hydroxylase and regulation of its expression. Journal of Bacteriology,
Bolívar, F., ... Gosset, G. (2018). Increasing pinosylvin production in Escherichia coli 190(22), 7367–7374.
by reducing the expression level of the gene fabI-encoded enoyl-acyl carrier protein Thuan, N. H., Chaudhary, A. K., Van Cuong, D., & Cuong, N. X. (2018). Engineering co-
reductase. Electronic Journal of Biotechnology. culture system for production of apigetrin in Escherichia coli. Journal of Industrial
Santos, C. N. S. (2010). Combinatorial search strategies for the metabolic engineering of mi- Microbiology & Biotechnology, 45(3), 175–185.
croorganisms. PhD thesisCambridge, MA: Massachusetts: Institute of Technology. Trantas, E., Panopoulos, N., & Ververidis, F. (2009). Metabolic engineering of the com-
Santos, C. N. S., Koffas, M., & Stephanopoulos, G. (2011). Optimization of a heterologous plete pathway leading to heterologous biosynthesis of various flavonoids and stil-
pathway for the production of flavonoids from glucose. Metabolic Engineering, 13(4), benoids in Saccharomyces cerevisiae. Metab Eng, 11.
392–400. Tsuda, T. (2012). Dietary anthocyanin-rich plants: Biochemical basis and recent progress
Santos, C. N. S., Xiao, W., & Stephanopoulos, G. (2012). Rational, combinatorial, and in health benefits studies. Molecular Nutrition & Food Research, 56(1), 159–170.
genomic approaches for engineering L-tyrosine production in Escherichia van Summeren-Wesenhagen, P. V., & Marienhagen, J. (2015). Metabolic engineering of
coli Proceedings of the National Academy of Sciences, 109(34), Escherichia coli for the synthesis of the plant polyphenol pinosylvin. Applied and
13538–13543. Environmental Microbiology, 81(3), 840–849.
Sarovich, D. S., & Pemberton, J. M. (2007). pPSX: A novel vector for the cloning and Vannelli, T., Wei Qi, W., Sweigard, J., Gatenby, A. A., & Sariaslani, F. S. (2007).
heterologous expression of antitumor antibiotic gene clusters. Plasmid, 57(3), Production of p-hydroxycinnamic acid from glucose in Saccharomyces cerevisiae and
306–313. Escherichia coli by expression of heterologous genes from plants and fungi. Metabolic
Satake, H., Koyama, T., Bahabadi, S., Matsumoto, E., Ono, E., & Murata, J. (2015). Engineering, 9(2), 142–151.
Essences in metabolic engineering of lignan biosynthesis. Metabolites, 5(2), 270. Varraso, R., & Camargo, C. A. (2014). Processed meat consumption and lung health: More
Satake, H., Shiraishi, A., Koyama, T., Matsumoto, E., Morimoto, K., Bahabadi, S. E., ... evidence for harm. European Respiratory Journal, 43(4), 943–946.
Murata, J. (2017). Chapter 12 - Lignan biosynthesis for food bioengineering A2 - Vauzour, D., Rodriguez-Mateos, A., Corona, G., Oruna-Concha, M. J., & Spencer, J. P. E.
Grumezescu, Alexandru Mihai. In A. M. Holban (Ed.). Food biosynthesis (pp. 351– (2010). Polyphenols and human health: Prevention of disease and mechanisms of
379). Academic Press. action. Nutrients, 2(11), 1106.
Shahidi, F., & Ambigaipalan, P. (2015). Phenolics and polyphenolics in foods, beverages Verdier, J., Zhao, J., Torres-Jerez, I., Ge, S., Liu, C., He, X., ... Udvardi, M. K. (2012).
and spices: Antioxidant activity and health effects – A review. Journal of Functional MtPAR MYB transcription factor acts as an on switch for proanthocyanidin bio-
Foods, 18, 820–897. synthesis in Medicago truncatula Proceedings of the National
Shetty, K. (2004). Role of proline-linked pentose phosphate pathway in biosynthesis of Academy of Sciences, 109(5), 1766–1771.
plant phenolics for functional food and environmental applications: a review. Process Viladomat, F., & Bastida, J. (2015). General overview of plant secondary metabolism. In
Biochemistry, 39(7), 789–804. B. Bahadur, M. Venkat Rajam, L. Sahijram, & K. V. Krishnamurthy (Eds.). Plant
Shin, S.-Y., Han, N. S., Park, Y.-C., Kim, M.-D., & Seo, J.-H. (2011). Production of re- biology and biotechnology: Volume I: Plant diversity, organization, function and im-
sveratrol from p-coumaric acid in recombinant Saccharomyces cerevisiae expressing provement (pp. 539–568). New Delhi: Springer India.
4-coumarate: Coenzyme A ligase and stilbene synthase genes. Enzyme and Microbial Wang, C., Zhi, S., Liu, C., Xu, F., Zhao, A., Wang, X., ... Yu, M. (2017). Characterization of
Technology, 48(1), 48–53. stilbene synthase genes in mulberry (Morus atropurpurea) and metabolic engineering
Shin, S.-Y., Jung, S.-M., Kim, M.-D., Han, N. S., & Seo, J.-H. (2012). Production of re- for the production of resveratrol in Escherichia coli. Journal of Agricultural and Food
sveratrol from tyrosine in metabolically engineered Saccharomyces cerevisiae. Chemistry, 65(8), 1659–1668.
Enzyme and Microbial Technology, 51(4), 211–216. Wang, H. H., Isaacs, F. J., Carr, P. A., Sun, Z. Z., Xu, G., Forest, C. R., & Church, G. M.
Shkryl, Y. N., Veremeichik, G. N., Bulgakov, V. P., & Zhuravlev, Y. N. (2011). Induction of (2009). Programming cells by multiplex genome engineering and accelerated evo-
anthraquinone biosynthesis in Rubia cordifolia cells by heterologous expression of a lution. Nature, 460, 894.
calcium-dependent protein kinase gene. Biotechnology and Bioengineering, 108(7), Wang, S., Zhang, S., Xiao, A., Rasmussen, M., Skidmore, C., & Zhan, J. (2015). Metabolic
1734–1738. engineering of Escherichia coli for the biosynthesis of various phenylpropanoid de-
Shkryl, Y. N., Veremeichik, G. N., Makhazen, D. S., Silantieva, S. A., Mishchenko, N. P., rivatives. Metabolic Engineering, 29, 153–159.
Vasileva, E. A., ... Bulgakov, V. P. (2016). Increase of anthraquinone content in Rubia Wang, S., Zhang, S., Zhou, T., Zeng, J., & Zhan, J. (2013). Design and application of an in
cordifolia cells transformed by native and constitutively active forms of the AtCPK1 vivo reporter assay for phenylalanine ammonia-lyase. Applied Microbiology and
gene. Plant Cell Reports, 35(9), 1907–1916. Biotechnology, 97(17), 7877–7885.
Shkryl Yuri, N., Veremeichik Galina, N., Bulgakov Victor, P., Tchernoded Galina, K., Wang, Y., Bhuiya, M. W., Zhou, R., & Yu, O. (2015). Pterostilbene production by mi-
Mischenko Natalia, P., Fedoreyev Sergei, A., & Zhuravlev Yuri, N. (2007). Individual croorganisms expressing resveratrol O-methyltransferase. Annals of Microbiology,
and combined effects of the rolA, B, and C genes on anthraquinone production in 65(2), 817–826.
Rubia cordifolia transformed calli. Biotechnology and Bioengineering, 100(1), 118–125. Wang, Y., Chen, H., & Yu, O. (2010). Metabolic engineering of resveratrol and other
Siró, I., Kápolna, E., Kápolna, B., & Lugasi, A. (2008). Functional food. Product devel- longevity boosting compounds. BioFactors, 36(5), 394–400.
opment, marketing and consumer acceptance—A review. Appetite, 51(3), 456–467. Wang, Y., Chen, H., & Yu, O. (2014). A plant malonyl-CoA synthetase enhances lipid
Song, M. C., Kim, E. J., Kim, E., Rathwell, K., Nam, S.-J., & Yoon, Y. J. (2014). Microbial content and polyketide yield in yeast cells. Applied Microbiology and Biotechnology,
biosynthesis of medicinally important plant secondary metabolites. [10.1039/ 98(12), 5435–5447.
C4NP00057A]. Natural Product Reports, 31(11), 1497–1509. Wang, Y., Halls, C., Zhang, J., Matsuno, M., Zhang, Y., & Yu, O. (2011). Stepwise increase
Sova, M. (2012). Antioxidant and antimicrobial activities of cinnamic acid derivatives. of resveratrol biosynthesis in yeast Saccharomyces cerevisiae by metabolic en-
Mini Reviews in Medicinal Chemistry, 12(8), 749–767. gineering. Metabolic Engineering, 13(5), 455–463.
Stahlhut, S. G., Siedler, S., Malla, S., Harrison, S. J., Maury, J., Neves, A. R., & Forster, J. Wang, Y., & Yu, O. (2012). Synthetic scaffolds increased resveratrol biosynthesis in en-
(2015). Assembly of a novel biosynthetic pathway for production of the plant fla- gineered yeast cells. Journal of Biotechnology, 157(1), 258–260.
vonoid fisetin in Escherichia coli. Metabolic Engineering, 31, 84–93. Watts, K. T., Lee, P. C., & Schmidt-Dannert, C. (2006). Biosynthesis of plant-specific
Stéphane, Q., Denis, D., Céline, D. C., & Laurent, P. (2011). Plant polyphenols: Chemical stilbene polyketides in metabolically engineered Escherichia coli. BMC Biotechnology,
properties, biological activities, and synthesis. Angewandte Chemie International 6(1), 22.
Edition, 50(3), 586–621. Watts, K. T., Lee, P. C., & Schmidt-Dannert, C. (2004). Exploring recombinant flavonoid
Stich, H. F., & Rosin, M. P. (1984). Naturally occurring phenolics as antimutagenic and biosynthesis in metabolically engineered Escherichia coli. ChemBioChem, 5(4),
anticarcinogenic agents. In M. Friedman (Ed.). Nutritional and toxicological aspects of 500–507.
food safety (pp. 1–29). Boston, MA: Springer, US. Whitaker, W. B., Jones, J. A., Bennett, R. K., Gonzalez, J. E., Vernacchio, V. R., Collins, S.
Stojković, D., Petrović, J., Soković, M., Glamočlija, J., Kukić-Marković, J., & Petrović, S. M., ... Papoutsakis, E. T. (2017). Engineering the biological conversion of methanol to
(2013). In situ antioxidant and antimicrobial activities of naturally occurring caffeic specialty chemicals in Escherichia coli. Metabolic Engineering, 39, 49–59.
acid, p-coumaric acid and rutin, using food systems. Journal of the Science of Food and Williams, T. C., Averesch, N. J. H., Winter, G., Plan, M. R., Vickers, C. E., Nielsen, L. K., &
Agriculture, 93(13), 3205–3208. Krömer, J. O. (2015). Quorum-sensing linked RNA interference for dynamic meta-
Suástegui, M., Yu Ng, C., Chowdhury, A., Sun, W., Cao, M., House, E., ... Shao, Z. (2017). bolic pathway control in Saccharomyces cerevisiae. Metabolic Engineering, 29,
253
124–134. engineering of anthocyanin biosynthesis in Escherichia coli. Applied and

Wong, L., Engel, J., Jin, E., Holdridge, B., & Xu, P. (2017). YaliBricks, a versatile genetic Environmental Microbiology, 71.
toolkit for streamlined and rapid pathway engineering in Yarrowia lipolytica. Yan, Y., Kohli, A., & Koffas, M. A. G. (2005). Biosynthesis of Natural Flavanones in
Metabolic Engineering Communications, 5, 68–77. Saccharomyces cerevisiae. Applied and Environmental Microbiology, 71(9), 5610–5613.
Wu, J., Du, G., Chen, J., & Zhou, J. (2015). Enhancing flavonoid production by system- Yang, S.-M., Shim, G. Y., Kim, B.-G., & Ahn, J.-H. (2015). Biological synthesis of cou-
atically tuning the central metabolic pathways based on a CRISPR interference marins in Escherichia coli. Microbial Cell Factories, 14(1), 65.
system in Escherichia coli. Retrieved from Scientific reports, 5, 13477. Yang, Y., Lin, Y., Li, L., Linhardt, R. J., & Yan, Y. (2015). Regulating malonyl-CoA me-
Wu, J., Du, G., Zhou, J., & Chen, J. (2013). Metabolic engineering of Escherichia coli for tabolism via synthetic antisense RNAs for enhanced biosynthesis of natural products.
(2S)-pinocembrin production from glucose by a modular metabolic strategy. Metabolic Engineering, 29, 217–226.
Metabolic Engineering, 16, 48–55. Yang, Y., Yuan, C., Dou, J., Han, X., Wang, H., Fang, H., & Zhou, C. (2014). Recombinant
Wu, J., Liu, P., Fan, Y., Bao, H., Du, G., Zhou, J., & Chen, J. (2013). Multivariate modular expression of glpK and glpD genes improves the accumulation of shikimic acid in E.
metabolic engineering of Escherichia coli to produce resveratrol from l-tyrosine. coli grown on glycerol. World Journal of Microbiology and Biotechnology, 30(12),
Journal of Biotechnology, 167(4), 404–411. 3263–3272.
Wu, J., Yu, O., Du, G., Zhou, J., & Chen, J. (2014). Fine-tuning of the fatty acid pathway Zahiri, H. S., Yoon, S. H., Keasling, J. D., Lee, S. H., Won Kim, S., Yoon, S. C., & Shin, Y. C.
by synthetic antisense RNA for enhanced (2S)-naringenin production from l-tyrosine (2006). Coenzyme Q10 production in recombinant Escherichia coli strains en-
in Escherichia coli. Applied and Environmental Microbiology, 80(23), 7283–7292. gineered with a heterologous decaprenyl diphosphate synthase gene and foreign
Wu, J., Zhang, X., Dong, M., & Zhou, J. (2016). Stepwise modular pathway engineering of mevalonate pathway. Metabolic Engineering, 8(5), 406–416.
Escherichia coli for efficient one-step production of (2S)-pinocembrin. Journal of Zhang, D., Li, Z., Wang, F., Shrestha, B., Tian, P., & Tan, T. (2007). Expression of various
Biotechnology, 231, 183–192. genes to enhance ubiquinone metabolic pathway in Agrobacterium tumefaciens.
Wu, J., Zhang, X., Zhou, J., & Dong, M. (2016). Efficient biosynthesis of (2S)-pinocembrin Enzyme and Microbial Technology, 41(6), 772–779.
from d-glucose by integrating engineering central metabolic pathways with a pH-shift Zhang, D., Shrestha, B., Li, Z., & Tan, T. (2007). Ubiquinone-10 production using
control strategy. Bioresource Technology, 218, 999–1007. Agrobacterium tumefaciens dps gene in Escherichia coli by coexpression system.
Wu, J., Zhou, T., Du, G., Zhou, J., & Chen, J. (2014). Modular optimization of hetero- Molecular Biotechnology, 35(1), 1.
logous pathways for de novo synthesis of (2S)-naringenin in Escherichia coli. PLoS Zhang, D., Shrestha, B., Niu, W., Tian, P., & Tan, T. (2007). Phenotypes and fed-batch
One, 9(7) e101492. fermentation of ubiquinone-overproducing fission yeast using ppt1 gene. Journal of
Xia, Z.-Q., Costa, M. A., Pélissier, H. C., Davin, L. B., & Lewis, N. G. (2001). Biotechnology, 128(1), 120–131.
Secoisolariciresinol dehydrogenase purification, cloning, and functional expression: Zhang, H., & Stephanopoulos, G. (2013). Engineering E. coli for caffeic acid biosynthesis
Implications for human health protection. Journal of Biological Chemistry, 276(16), from renewable sugars. Applied Microbiology and Biotechnology, 97(8), 3333–3341.
12614–12623. Zhang, L., Gao, B., Wang, X., Zhang, Z., Liu, X., Wang, J., ... Tu, P. (2016). Identification
Xie, D. Y., Sharma, S. B., Wright, E., Wang, Z. Y., & Dixon, R. A. (2006). Metabolic en- of a new curcumin synthase from ginger and construction of a curcuminoid-produ-
gineering of proanthocyanidins through co-expression of anthocyanidin reductase cing unnatural fusion protein diketide-CoA synthase::curcumin synthase. [10.1039/
and the PAP1 MYB transcription factor. The Plant Journal, 45(6), 895–907. C5RA23401H]. RSC Advances, 6(15), 12519–12524.
Xu, P., Li, L., Zhang, F., Stephanopoulos, G., & Koffas, M. (2014). Improving fatty acids Zhang, Y., Li, S.-Z., Li, J., Pan, X., Cahoon, R. E., Jaworski, J. G., ... Yu, O. (2006). Using
production by engineering dynamic pathway regulation and metabolic control. unnatural protein fusions to engineer resveratrol biosynthesis in yeast and
Proceedings of the National Academy of Sciences, 111(31), 11299–11304. Mammalian cells. Journal of the American Chemical Society, 128(40), 13030–13031.
Xu, P., Ranganathan, S., Fowler, Z. L., Maranas, C. D., & Koffas, M. A. G. (2011). Genome- Zhao, S., Jones, J. A., Lachance, D. M., Bhan, N., Khalidi, O., Venkataraman, S., ... Koffas,
scale metabolic network modeling results in minimal interventions that cooperatively M. A. G. (2015). Improvement of catechin production in Escherichia coli through
force carbon flux towards malonyl-CoA. Metabolic Engineering, 13(5), 578–587. combinatorial metabolic engineering. Metabolic Engineering, 28, 43–53.
Xu, P., Rizzoni, E. A., Sul, S.-Y., & Stephanopoulos, G. (2017). Improving metabolic Zheng, L. T., Ock, J., Kwon, B.-M., & Suk, K. (2008). Suppressive effects of flavonoid
pathway efficiency by statistical model-based multivariate regulatory metabolic en- fisetin on lipopolysaccharide-induced microglial activation and neurotoxicity.
gineering. ACS Synthetic Biology, 6(1), 148–158. International Immunopharmacology, 8(3), 484–494.
Xu, P., Vansiri, A., Bhan, N., & Koffas, M. A. G. (2012). ePathBrick: A synthetic biology Zhenya, C., Xiaolin, S., Jian, W., Jia, W., Qipeng, Y., & Yajun, Y. (2017). Rational en-
platform for engineering metabolic pathways in E. coli. ACS Synthetic Biology, 1(7), gineering of p-hydroxybenzoate hydroxylase to enable efficient gallic acid synthesis
256–266. via a novel artificial biosynthetic pathway. Biotechnology and Bioengineering, 114(11),
Xu, P., Wang, W., Li, L., Bhan, N., Zhang, F., & Koffas, M. A. G. (2014). Design and kinetic 2571–2580.
analysis of a hybrid promoter-regulator system for malonyl-CoA sensing in Zhu, S., Wu, J., Du, G., Zhou, J., & Chen, J. (2014). Efficient synthesis of eriodictyol from
Escherichia coli. ACS Chemical Biology, 9(2), 451–458. l-tyrosine in Escherichia coli. Applied and Environmental Microbiology, 80(10),
Xu, W., Yang, S., Zhao, J., Su, T., Zhao, L., & Liu, J. (2014). Improving coenzyme Q8 3072–3080.
production in Escherichia coli employing multiple strategies. Journal of Industrial Zhu, X., Yuasa, M., Okada, K., Suzuki, K., Nakagawa, T., Kawamukai, M., & Matsuda, H.
Microbiology & Biotechnology, 41(8), 1297–1303. (1995). Production of ubiquinone in Escherichia coli by expression of various genes
Yada, S., Wang, Y., Zou, Y., Nagasaki, K., Hosokawa, K., Osaka, I., ... Enomoto, K. (2008). responsible for ubiquinone biosynthesis. Journal of Fermentation and Bioengineering,
Isolation and characterization of two groups of novel marine bacteria producing 79(5), 493–495.
violacein. Marine Biotechnology, 10(2), 128–132. Zhu, Y., Lu, W., Ye, L., Chen, Z., Hu, W., Wang, C., ... Yu, H. (2017). Enhanced synthesis of
Yajun, Y., Lixuan, H., & Koffas, Mattheos A. G. (2007). Biosynthesis of 5-deoxyflavanones Coenzyme Q10 by reducing the competitive production of carotenoids in
in microorganisms. Biotechnology Journal, 2(10), 1250–1262. Rhodobacter sphaeroides. Biochemical Engineering Journal, 125, 50–55.
Yajun, Y., Zhen, L., & Koffas, Mattheos A. G. (2008). High-yield anthocyanin biosynthesis Zhu, Y., Ye, L., Chen, Z., Hu, W., Shi, Y., Chen, J., ... Yu, H. (2017). Synergic regulation of
in engineered Escherichia coli. Biotechnology and Bioengineering, 100(1), 126–140. redox potential and oxygen uptake to enhance production of coenzyme Q10 in
Yan, Y., Chemler, J., Huang, L., Martens, S., & Koffas, M. A. (2005). Metabolic Rhodobacter sphaeroides. Enzyme and Microbial Technology, 101, 36–43.
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Journal of Functional Foods 57 (2019) 233–254

Contents lists available at ScienceDirect

Journal of Functional Foods

Sustainable production of natural phenolics for functional food applications T

ARTICLE INFO ABSTRACT

1. Introduction aggression from pathogens or predators (Pandey & Rizvi, 2009). In

Fig. 1. Biosynthesis paths and classification of phenolic compounds.

lipolytica) is another ideal GRAS strain for the production of high-valued

2.1. General introduction

Phenolic acids are a class of substances which contain a phenolic

2.2. Production of hydroxybenzoic acids

Production of flavonone in E. coli and S. cerevisiae.

Corona, Oruna-Concha, & Spencer, 2010). Their antimicrobial activity

Lyu et al. (2017)

Erasmus, 1997). Additionally, quercetin, dihydroquercetin, dihy-

helpful against herpes simplex virus, respiratory syncytial virus, po-

coumaric acid, naringenin is produced, which is at the central branch

point of the pathway. The main branches are flavones, flavanones,

flavons, flavanols, anthocyanins, and isoflavanoids (Fig. 1) (Lyu et al.,

compounds in microbes is discussed. Their glycosylated, methylated,

found in reviews of Pandey et al. (2016) and Cao et al. (2015).

Reduction of by-product formation, overexpression of feedback-resistant genes, identification of

Flavanones is the most important flavonoid group, as they are the

for preventing cardiovascular, neurodegenerative and cancer diseases.

3.2.1. Pathway construction

pinocembrin, naringenin, eriodictyol, and liquiritigenin are well stu-

chalcone synthase (CHS), and chalcone isomerase (CHI) using pheny-

cone (Hwang, Kaneko, Ohnishi, & Horinouchi, 2003). Leguminous

L eriodictyol from caffeic acid in E. coli, as well as 13.5 mg/L of li-

quiritigenin and 7.9 mg/L of eriodictyol in S. cerevisiae (Yajun, Lixuan,

& Koffas, 2007). For biosynthesis of eriodictyol, F3′H-CPR complex

(from G. hybrida and C. roseus), as well as E. coli hydroxylase complex

the substrate (Fig. 3) (Jones, Collins Shannon, Vernacchio Victoria,

Lachance Daniel, & Koffas Mattheos, 2015).

3.2.2. Improving malonyl-CoA supply

6. The others Usually, one type of PLR converts (+)-pinoresinol to (−)-secoisolar-

124–134. engineering of anthocyanin biosynthesis in Escherichia coli. Applied and

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