Papers by Volker F Wendisch
Systems Microbiology and Biomanufacturing, Dec 20, 2023
Indole is a signalling molecule produced both by bacteria and plants. In this review its signalli... more Indole is a signalling molecule produced both by bacteria and plants. In this review its signalling role between microbes and in particular in the human gut is discussed. Besides the natural roles, indole also has value for flavour and fragrance applications, for example, in food industry or perfumery. Additionally, indole can be derivatized to several halogenated and oxygenated compounds that can be used as natural colourants or have promising bioactivity with therapeutic potential to treat human diseases. Indole is traditionally obtained from coal tar. Biocatalytic approaches have been developed to convert indole into halogenated and oxygenated derivatives. This review will discuss recent advances in production of indole from glucose or tryptophan by fermentation and the production of derived halogenated and oxygenated derivatives by microbial cell factories.
Processes
Astaxanthin, a highly valuable natural pigment with potent antioxidant properties, is widely used... more Astaxanthin, a highly valuable natural pigment with potent antioxidant properties, is widely used in various industries, including food, pharmaceuticals, and cosmetics. The demand for astaxanthin has led to the development of multiple production methods, including algal, bacterial, and synthetic approaches. Understanding the environmental implications of these production processes is crucial for sustainable decision-making in the astaxanthin industry. This research paper presents a comprehensive Life Cycle Assessment (LCA) comparing the environmental impacts of algal, bacterial, and synthetic astaxanthin production methods. Moreover, a sensitivity analysis is conducted, focusing on the source of energy used during production, and the shift from conventional to renewable energy sources is explored. The environmental importance of natural astaxanthin production is highlighted, considering the potential benefits of reducing dependence on synthetic production methods that rely on non-re...
CRC Press eBooks, Mar 30, 2005
Wendisch VF, Bott M. Phosphorus metabolism of Corynebacterium glutamicum. In: Eggeling L, Bott M,... more Wendisch VF, Bott M. Phosphorus metabolism of Corynebacterium glutamicum. In: Eggeling L, Bott M, eds. Handbook of Corynebacterium glutamicum. Boca Raton, USA: CRC Press; 2005: 377-396
Biospektrum, 2017
Wendisch VF. Journal Club: Calvin-Zyklus-Enzym reguliert Methanol-Stoffwechsel. BIOspektrum. 2017... more Wendisch VF. Journal Club: Calvin-Zyklus-Enzym reguliert Methanol-Stoffwechsel. BIOspektrum. 2017;06.17:666
bioRxiv (Cold Spring Harbor Laboratory), Feb 26, 2021
bioRxiv (Cold Spring Harbor Laboratory), Feb 18, 2021
Fructose utilization in Corynebacterium glutamicum starts with its uptake and concomitant phospho... more Fructose utilization in Corynebacterium glutamicum starts with its uptake and concomitant phosphorylation via the phosphotransferase system (PTS) to yield intracellular fructose 1-phosphate, which enters glycolysis upon ATP dependent phosphorylation to fructose 1,6-bisphosphate by 1-phosphofructokinase. This is known to result in a significantly reduced oxidative pentose phosphate pathway (oxPPP) flux on fructose (~10 %) compared to glucose (~60 %). Consequently, the biosynthesis of NADPH demanding products, e.g. L-lysine, by C. glutamicum is largely decreased, when fructose is the only carbon source. Previous works reported that fructose is partially utilized via the glucose specific PTS presumably generating fructose 6-phosphate. This closer proximity to the entry point of the oxPPP might increase oxPPP flux and consequently NADPH availability. Here, we generated deletion strains either lacking in the fructose-specific PTS or 1-phosphofructokinase activity. We used these strains in short-term evolution experiments on fructose minimal medium and isolated mutant strains, which regained the ability of fast growth on fructose as a sole carbon source. In these fructose mutants, the deletion of the glucose specific PTS, as well as the 6-phosphofructokinase gene, abolished growth, unequivocally showing fructose phosphorylation via glucose specific PTS to fructose 6-phosphate. Gene sequencing revealed three independent amino acid substitutions in PtsG (M260V, M260T, P318S). These three PtsG variants mediated faster fructose uptake and utilization compared to native PtsG. In-depth analysis of the effects of fructose utilization via these PtsG variants revealed significantly increased biomass formation, reduced side-product accumulation, and increased L-lysine production by 50 %.
Molecular & Cellular Proteomics, 2005
ABSTRACT The use of Escherichia coli as a model organism has provided a great deal of basic infor... more ABSTRACT The use of Escherichia coli as a model organism has provided a great deal of basic information in biomolecular sciences. Examining trait differences among closely related strains of the same species addresses a fundamental biological question: how much diversity is there at the single species level? The main aim of our research was to identify significant differences in the activities of groups of genes between two laboratory strains of an organism closely related in genome structure. We demonstrate that despite strict and controlled growth conditions, there is high plasticity in the global proteome and genome expression in two closely related E. coli K12 sub-strains (W3110 and MG1655), which differ insignificantly in genome structure. The growth patterns of these two sub-strains were very similar in a well-equipped bioreactor, and their genome structures were shown to be almost identical by DNA microarray. However, detailed profiling of protein and gene expression by 2-dimensional gel electrophoresis and microarray analysis showed many differentially expressed genes and proteins, combinations of which were highly correlated. The differentially regulated genes and proteins belonged to the following functional categories: genes regulated by sigma subunit of RNA polymerase (RpoS), enterobactin-related genes, and genes involved in central metabolism. Genes involved in central cell metabolism - the glycolysis pathway, the tricarboxylic acid cycle and the glyoxylate bypass - were differentially regulated at both the mRNA and proteome levels. The strains differ significantly in central metabolism and thus in the generation of precursor metabolites and energy. This high plasticity probably represents a universal feature of metabolic activities in closely related species, and has the potential to reveal differences in regulatory networks. We suggest that unless care is taken in the choice of strains for any validating experiment, the results might be misleading.
Applied Microbiology and Biotechnology, Jan 9, 2024
This review presents an analysis of formamide, focussing on its occurrence in nature, its functio... more This review presents an analysis of formamide, focussing on its occurrence in nature, its functional roles, and its promising applications in the context of the bioeconomy. We discuss the utilization of formamide as an innovative nitrogen source achieved through metabolic engineering. These approaches underscore formamide's potential in supporting growth and production in biotechnological processes. Furthermore, our review illuminates formamide's role as a nitrogen source capable of safeguarding cultivation systems against contamination in non-sterile conditions. This attribute adds an extra layer of practicality to its application, rendering it an attractive candidate for sustainable and resilient industrial practices. Additionally, the article unveils the versatility of formamide as a potential carbon source that could be combined with formate or CO 2 assimilation pathways. However, its attributes, i.e., enriched nitrogen content and comparatively limited energy content, led to conclude that formamide is more suitable as a co-substrate and that its use as a sole source of carbon for biomass and bio-production is limited. Through our exploration of formamide's properties and its applications, this review underscores the significance of formamide as valuable resource for a large spectrum of industrial applications.
Applied Microbiology and Biotechnology, Mar 26, 2014
Sustainable supply of feedstock has become a key issue in process development in microbial biotec... more Sustainable supply of feedstock has become a key issue in process development in microbial biotechnology. The workhorse of industrial amino acid production Corynebacterium glutamicum has been engineered towards utilization of alternative carbon sources. Utilization of the chitin-derived aminosugar N-acetyl-glucosamine (GlcNAc) for both cultivation and production with C. glutamicum has hitherto not been investigated. Albeit this organism harbors the enzymes N-acetylglucosamine-6-phosphatedeacetylase and glucosamine-6P deaminase of GlcNAc metabolism (encoded by nagA and nagB, respectively) growth of C. glutamicum with GlcNAc as substrate was not observed. This was attributed to the lack of a functional system for GlcNAc uptake. Of the 17 type strains of the genus Corynebacterium tested here for their ability to grow with GlcNAc, only Corynebacterium glycinophilum DSM45794 was able to utilize this substrate. Complementation studies with a GlcNAc-uptake deficient Escherichia coli strain revealed that C. glycinophilum possesses a nagE-encoded EII permease for GlcNAc uptake. Heterologous expression of the C. glycinophilum nagE in C. glutamicum indeed enabled uptake of GlcNAc. For efficient GlcNac utilization in C. glutamicum, improved expression of nagE with concurrent overexpression of the endogenous nagA and nagB genes was found to be necessary. Based on this strategy, C. glutamicum strains for the efficient production of the amino acid L-lysine as well as the carotenoid lycopene from GlcNAc as sole substrate were constructed.
CRC Press eBooks, Aug 25, 2011
Frontiers in Microbiology, Oct 1, 2021
Bacteria respond to pH changes in their environment and use pH homeostasis to keep the intracellu... more Bacteria respond to pH changes in their environment and use pH homeostasis to keep the intracellular pH as constant as possible and within a small range. A change in intracellular pH influences enzyme activity, protein stability, trace element solubilities and proton motive force. Here, the species Corynebacterium glutamicum was chosen as a neutralophilic and moderately alkali-tolerant bacterium capable of maintaining an internal pH of 7.5 ± 0.5 in environments with external pH values ranging between 5.5 and 9. In recent years, the phenotypic response of C. glutamicum to pH changes has been systematically investigated at the bulk population level. A detailed understanding of the C. glutamicum cell response to defined short-term pH perturbations/pulses is missing. In this study, dynamic microfluidic single-cell cultivation (dMSCC) was applied to analyze the physiological growth response of C. glutamicum to precise pH stress pulses at the single-cell level. Analysis by dMSCC of the growth behavior of colonies exposed to single pH stress pulses (pH = 4, 5, 10, 11) revealed a decrease in viability with increasing stress duration w. Colony regrowth was possible for all tested pH values after increasing lag phases for which stress durations w were increased from 5 min to 9 h. Furthermore, single-cell analyses revealed heterogeneous regrowth of cells after pH stress, which can be categorized into three physiological states. Cells in the first physiological state continued to grow without interruption after pH stress pulse. Cells in the second physiological state rested for several hours after pH stress pulse before they started to grow again after this lag phase, and cells in the third physiological state did not divide after the pH stress pulse. This study provides the first insights into single-cell responses to acidic and alkaline pH stress by C. glutamicum.
Bioresource Technology, Dec 1, 2017
Here, a new metabolic pathway for the production of 5-aminovalerate (5AVA) from L-lysine via cada... more Here, a new metabolic pathway for the production of 5-aminovalerate (5AVA) from L-lysine via cadaverine as intermediate was established and this three-step-pathway comprises Llysine decarboxylase (LdcC), putrescine transaminase (PatA) and γ-aminobutyraldehyde dehydrogenase (PatD). Since Corynebacterium glutamicum is used for industrial L-lysine production, the pathway was established in this bacterium. Upon expression of ldcC, patA and patD from Escherichia coli in C. glutamicum wild type, production 5AVA was achieved. Enzyme assays revealed that PatA and PatD also converted cadaverine to 5AVA. Eliminating the by-products cadaverine, N-acetylcadaverine and glutarate in a genome-streamlined Llysine producing strain expressing ldcC, patA and patD improved 5AVA production to a titer of 5.1 g L-1 , a yield of 0.13 g g-1 and a volumetric productivity of 0.12 g L-1 h-1. Moreover, 5AVA production from the alternative feedstocks starch, glucosamine, xylose and arabinose was established.
Metabolic Engineering, Mar 1, 2020
Amino acid fermentation is one of the major pillars of industrial biotechnology. The multi-billio... more Amino acid fermentation is one of the major pillars of industrial biotechnology. The multi-billion USD amino acid market is rising steadily and is diversifying. Metabolic engineering is no longer focused solely on strain development for the bulk amino acids L-glutamate and L-lysine that are produced at the million-ton scale, but targets specialty amino acids. These demands are met by the development and application of new metabolic engineering tools including CRISPR and biosensor technologies as well as production processes by enabling a flexible feedstock concept, co-production and cocultivation schemes. Metabolic engineering advances are exemplified for specialty proteinogenic amino acids, cyclic amino acids, omega-amino acids, and amino acids functionalized by hydroxylation, halogenation and N-methylation.
Journal of Biotechnology, Feb 1, 2023
Oikawa T, Uesato S, Tamura H, et al. Building an International Network Exchange Program of Educat... more Oikawa T, Uesato S, Tamura H, et al. Building an International Network Exchange Program of Education and Research for Graduate Course Students in Life Science and Biotechnology. Science and Technology Reports of Kansai University. 2008;50:83-94
Caister Academic Press eBooks, 2015
Frontiers in Bioengineering and Biotechnology, May 28, 2021
Fructose utilization in Corynebacterium glutamicum starts with its uptake and concomitant phospho... more Fructose utilization in Corynebacterium glutamicum starts with its uptake and concomitant phosphorylation via the phosphotransferase system (PTS) to yield intracellular fructose 1-phosphate, which enters glycolysis upon ATP-dependent phosphorylation to fructose 1,6-bisphosphate by 1-phosphofructokinase. This is known to result in a significantly reduced oxidative pentose phosphate pathway (oxPPP) flux on fructose (∼10%) compared to glucose (∼60%). Consequently, the biosynthesis of NADPH demanding products, e.g., L-lysine, by C. glutamicum is largely decreased when fructose is the only carbon source. Previous works reported that fructose is partially utilized via the glucose-specific PTS presumably generating fructose 6phosphate. This closer proximity to the entry point of the oxPPP might increase oxPPP flux and, consequently, NADPH availability. Here, we generated deletion strains lacking either the fructose-specific PTS or 1-phosphofructokinase activity. We used these strains in short-term evolution experiments on fructose minimal medium and isolated mutant strains, which regained the ability of fast growth on fructose as a sole carbon source. In these fructose mutants, the deletion of the glucose-specific PTS as well as the 6-phosphofructokinase gene, abolished growth, unequivocally showing fructose phosphorylation via glucose-specific PTS to fructose 6-phosphate. Gene sequencing revealed three independent amino acid substitutions in PtsG (M260V, M260T, and P318S). These three PtsG variants mediated faster fructose uptake and utilization compared to native PtsG. In-depth analysis of the effects of fructose utilization via these PtsG variants revealed significantly increased ODs, reduced side-product accumulation, and increased L-lysine production by 50%.
Current opinion in green and sustainable chemistry, Dec 1, 2018
Lignin is a complex aromatic heteropolymer that harbors great potential to serve as a basis for t... more Lignin is a complex aromatic heteropolymer that harbors great potential to serve as a basis for the production of chemicals, biofuels, and materials. In a bio-economy, lignin valorization through depolymerization to monomers and upgrading of these monomers to targeted chemicals is highly challenging, but attractive and important for the chemical industry. To address current limitations of lignin valorization and further upgrading, we will summarize the recent lignin depolymerization techniques that yield phenolic aldehydes and phenolics such as alkaline oxidation, fast pyrolysis, hydrogenolysis, and hydrolysis. In addition, this review describes novel upgrading approaches via biological and chemo-catalytic pathways from lignin monomers to targeted bulk chemicals, including cis,cis-muconic acid, adipic acid, and terephthalic acid, and to fine chemicals, and then discuss future prospects.
Bioresource Technology, Jul 1, 2018
In the biorefinery concept renewable feedstocks are converted to a multitude of valueadded compou... more In the biorefinery concept renewable feedstocks are converted to a multitude of valueadded compounds irrespective of seasonal or other variations of the complex biomass substrates. Conceptionally, this can be realized by specialized single microbial strains or by co-culturing various strain combinations. In the latter approach strains for substrate conversion and for product formation can be combined. This study addressed the construction of binary microbial consortia based on starch-and sucrose-based production of L-lysine and derived value-added compounds. A commensalism-based synthetic consortium for L-lysine production from sucrose was developed combining an L-lysine auxotrophic, naturally sucrose-negative E. coli strain with a C. glutamicum strain able to produce L-lysine that secretes fructose when grown with sucrose due to deletion of the fructose importer gene ptsF. Mutualistic synthetic consortia with an L-lysine auxotrophic, α-amylase secreting E. coli strain and naturally amylase-negative C. glutamicum strains was implemented for production of valuable fine chemicals from starch.
Bioresource Technology, Dec 1, 2017
The biotechnological production of amino acids occurs at the million-ton scale and annually about... more The biotechnological production of amino acids occurs at the million-ton scale and annually about 6 million tons of L-glutamate and L-lysine are produced by Escherichia coli and Corynebacterium glutamicum strains. L-glutamate and L-lysine production from starch hydrolysates and molasses is very efficient and access to alternative carbon sources and new products has been enabled by metabolic engineering. This review focusses on genetic and metabolic engineering of amino acid producing strains. In particular, rational approaches involving modulation of transcriptional regulators, regulons, and attenuators will be discussed. To address current limitations of metabolic engineering, this article gives insights on recent systems metabolic engineering approaches based on functional tools and methods such as genome reduction, amino acid sensors based on transcriptional regulators and riboswitches, CRISPR interference, small regulatory RNAs, DNA scaffolding, and optogenetic control, and discusses future prospects.
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Papers by Volker F Wendisch