Recent Advances in

Microalgal Biotechnology
Edited by
www.esciencecentral.org/ebooks Dr. Jin Liu
Dr. Zheng Sun
Dr. Henri Gerken
Ibrahim Hashim
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Recent Advances in Microalgal Biotechnology

Chapter: CO2 Sequestration by Microalgae: Advances and Perspectives

Edited by: Dr. Jin Liu, Dr. Zheng Sun and Dr. Henri Gerken

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CO2 Sequestration by
Microalgae: Advances and
Perspectives
T Chakrabarti*, K Krishnamurthi*, S Saravana Devi and
Abhay B Fulke
Environmental Health Division, CSIR-National Environmental
Engineering Research Institute (CSIR-NEERI), Nagpur, India
*Corresponding authors: T Chakrabarti, Environmental Health
Division, CSIR-National Environmental Engineering Research Institute
(CSIR-NEERI), Nagpur, India, Tel: +91-712-2249885-889; Fax: +91-
712-2249961; E-mail: [email protected]
K Krishnamurthi, Environmental Health Division, CSIR-National
Environmental Engineering Research Institute (CSIR-NEERI), Nagpur,
India, Tel: +91-7122249885-889; Fax: +91-7122249961; E-mail:
[email protected]

Abstract
Carbon dioxide, a Greenhouse Gas (GHG), is the one of the principle pollutant, warming earth. In the past 150 years,
anthropogenic activities have pumped enough carbon dioxide into the atmosphere to raise its levels to 400 ppm, higher than they
have been for hundreds of thousands of years. In the global effort to combat the climate change, several CO2 capture and storage
technologies are deliberated. Because of photosynthetic ability of microalgae, the potential microalgae such as Chlorella vulgaris,
Scenedesmus sp., Chroococcus sp. and Chlamydomonas sp. have been actively used globally in closed and open photobioreactor
for CO2 mitigation. Algae as feed stocks for bio-energy refer to a diverse group of organisms that include microalgae, macroalgae
(seaweed) and cyanobacteria (formerly called blue-green algae). Algae occur in a variety of natural aqueous and terrestrial habitats
in environment. Under certain conditions, some microalgae have the potential to accumulate lipids. Algal biofuel generation imbibes
both carbon sequestration and energy production. Further, special emphasis is required on regulation of biosynthetic genes and its
expression in microalgae under various CO2 stress conditions for sequestration.

Keywords: Algal oil; Bio fuel algae microalgae; Carbon dioxide; Clean technology; GHG; Sequestration
Introduction
Reducing the rate of global climate change which in turn linked to increasing levels of Greenhouse Gases (GHGs) in the atmosphere
is recognized as the worlds biggest environmental challenge [1-3]. In recent years, researchers are focusing on biotechnological routes
which might help to reduce CO2 emissions in the air vis-a-vis sequester more carbon from the air into the ground and oceans. Now,
amply clear that no single technology holds the answer to achieving fuel related GHG emission reductions. Instead, only full use of all
available alternatives such as switching to renewable energy sources, enhancing energy efficiency, curbing emissions from agriculture,
forestry and deploying Carbon Sequestration and Storage (CSS) paradigm will solve the energy and climate challenge. A rational
scientific policy and networking with pockets of excellence conducting their climate change related research in equivalent, elsewhere
in the world, must exists. A strategy for low-carbon escalation and renewable energy systems, especially in the transport sector must
exist in India. The country has three alternatives to high-carbon systems: carbon sequestration, renewable energy and nuclear power,
despite high costs. According to Mr. Sam Pitroda, Chairman of the Knowledge Commission, India has to invent, sometimes reinvent,
model and not to depend on western nations for simulations as western model of consumption, generally, is unsustainable. Biomass
from plants, bagasse, cow dung, agricultural residues etc which could either be anaerobically digested to produce biogas or could be
transformed into gas. A related study was reported by Prajapati et al. [4] and Prajapati et al.[5]. This biomass can be used to generate
steam or power or used as a fuel. Various examples of gasified power plants in India showed that power is generated using rice husk in
Andhra Pradesh State, while several bagasse based plants are present elsewhere in the country. Potential use of biomass as first, second,
third and fourth generations of biofuels are summarized in brief as under:
First generation biofuels: This utilizes food-based feedstocks for example sugar cane, corn or soybean as raw materials, and utilize
processing technologies like fermentation (for ethanol) and trans-esterification for biodiesel [6].
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Second generation biofuels: These exploit non-food feedstocks, for example lingo-cellulosic plant biomass and non-edible
oilseeds through fermentation or thermo-chemical routes. However, the technology has yet not been commercialized.
Third generation biofuels: This purposely utilizes transgenic bioenergy crops to improve biofuel yield. An example is the
development of lignin deficient plants, improve ethanol production, or corn with implanted enzyme cellulase. However, the use of
transgenic plants are still experimented on laboratory scale and their future use in developing countries is questionable because of lack
of monitoring and control strategies to manage transgenic plants under open field conditions. 003
 Fourth generation biofuels: This involves algae (microalgae/macroalgae) production with carbon sequestration and more carbon
neutral to the other generation biofuels [7]. Thus, symbolize Bioenergy with Carbon Storage (BECS).
Literature review revealed that growth of algae is negatively influenced by increasing CO2 [1,8]. Strains that grow well at CO2
concentrations of 5-10% show severe reduction in their growth rate [3,9]. Efficient carbon dioxide capture from industrial flue-gas is an
important development to permit carbon management [1,3,10-14]. Algae have been successfully used to clear outgases from power-plant
[15].Various CO2 mitigation strategies such as chemical absorption, membrane separation, cryogenic fractionation and CO2 adsorption
using molecular sieves have been investigated though their techno-economic feasibility is not so encouraging [16,17]. The key focus for
Government should, therefore, be to support the demonstration projects involving microalgae biomass based biofuel production thereby
enabling the entrepreneurs to go ahead with commercial scale production enabling such cleaner technologies facilitating prosperous
future for everyone.
The key bio-molecules that can be extracted from algal biomass are;
a) Lipids
b) Carbohydrates
c) Proteins
Lipids and carbohydrates can be used as fuel precursors for example, gasoline and biodiesel, while proteins can be used as animal/
fish feeds [18].
In this context, present chapter discusses about the scope of reduction in carbon dioxide emission using microalgae.

Bio-Mitigation of CO2 Emissions with Microalgae

One of the largely considered methods for CO2 mitigation is the use of microalgae in biomass conversion in photo bioreactor [3].
Marine and freshwater microalgae are microscopic photosynthetic organisms. Microalgae namely, Cyanobacterial (Cyanophyceae)
and eukaryotic microalgae, green algae (Chlorophyta) and diatoms (Bacillariophyta) can be used to capture CO2 from three different
sources: atmospheric CO2, CO2 emission from power plants and industrial processes and soluble carbonate [1,19]. In this context,
possibly the transfer of CO2 from the atmosphere to the microalgae through photosynthesis is fundamental route for CO2 capture [3].
However, 360ppm CO2 concentration in atmospheric air makes what economically non-feasible [20]. In contrast, CO2 capture from
flue gas emissions from fossil fuel based power plants achieves better recovery because of, The higher concentration is in exhaust
stream, raceway pond systems for microalgae production, and not in photobioreactor [21]. However, only a small number of algae are
tolerant to the high levels of Sox and NOx present in flue gases. Further, the gases need to be cooled prior to injection into the growth
medium. Some microalgae species can assimilate CO2 from soluble carbonates such as Na2CO3 and NaHCO3 [19] leading to high pH of
the medium because of conversion of carbonate/bicarbonate alkalinity to hydroxyl alkalinity. Such a condition tends to control invasive
species since only a very small number of algae can grow in such extreme conditions [19]. The high cost of process technology and price
competitiveness of biodiesel extraction from microalgae can be offset by bio-mitigation of CO2 emissions which may be simultaneously
exploited to reduce cost [19,22]. The harvested biomass methods, include sun drying, low-pressure shelf drying, drum drying, spray
drying [23,24], fluidized bed drying [25], freeze drying [26] and refractance WindowTM technology drying[27]. Intracellular oils are
extracted more easily from dried biomass [26,28].
Fulke et al. [3] evaluated the photosynthetic ability of different microalgae leading to higher CO2 fixation and calcite formation vis-a-
vis their ability to synthesize biodiesel precursors. Further, Fulke et al.[3]observed the presence of Fatty Acid Methyl Esters (FAME) such
as docosapentaenoic acid (C22:5), palmitic acid (C16:0) and docosahexaenoic acid (C22:6) and superior quality calcite production with
simultaneous CO2 mitigation using Chlorella species. Demirbas, [29] reported that microalgae could covert CO2 into chemical energy
via photosynthesis, subsequently leading to fuels biosynthesis. Thus, the photosynthetic potential of microalgae can be integrated with
advance CO2 sequestration and biodiesel production which is a new area of research interest [1,3].

Biofuel from CO2 Sequestered Microalgae Biomass

Microalgae have a higher CO2 fixation ability compared to plants [30] and produce value-added products [1,3,31,32]. Microalgae
oil production was extensively reviewed by Brennan and Owende [7]. Wastewater effluent from the agri-food industry can be used for
microalgae cultivation [33]. However, care should be taken to prevent introduction of wastewater algal predators such as protozoa and
other species.
Three different algae production mechanisms showed photoautotrophic (requires inorganic carbon and light), heterotrophic
(requires organic carbon and light) and mixotrophic (autotrophic photosynthesis and heterotrophic assimilation) production. Algae can
be commercially cultivated in open raceway ponds or closed photo-bioreactors [34]. According to Brennan and Owende [7] raceway
pond consist of a closed loop, oval shaped recirculation channels as shown in (Figure: 1). Photo-bioreactors, on the other hand, consist
of an arrangement of straight glass or plastic tubes as made known in (Figure: 2) by Ugwu et al. [35]. The tubular array captures sunlight,
minimize contamination and favor constant cell growth [12,36]. Ketheesan and Nirmalakhandan [37] studied an airlift-driven raceway
reactor for microalgal cultivation with the highest CO2 consumption. Cheng et al. [38] had showed that increasing retention time of CO2
in photo bioreactor could significantly enhance the CO2 fixation efficiency. Similar result was also reported by Fulke et al. [3] in which
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Chlorella sp. could clearage CO2 gas up to 15% concentration with air lift photobioreactor. Many researchers have studied microalgal CO2
fixation with closed photo-bioreactors [3,38-40]. A variety of microalgae species have been applied to CO2 fixation, including Chlorella
vulgaris, Scenedesmus obliquus, Chlamydomonas reinhardtii, Spirullina sp. And Botrycoccus braunii [1,38,39,41,42]. Thus, algal based
CO2 sequestration is promising technology for carbon dioxide sequestrations vis-a vis biofuel production [1,3,14,43,44].

004
 Figure 1: Plan view of a raceway pond. Algae broth is introduced after the paddlewheel, and completes a cycle while
being mechanically aerated with CO2. It is harvested before the paddlewheel to start the cycle again (adapted from [7,10]).

Figure 2: Basic design of a horizontal tubular photobioreactor (adapted from [7,84]) Two main sections: airlift
system and solar receiver; the airlift systems allow for the transfer of O2 out of the systems and transfer of CO2
into the system as well as providing a means to harvest the biomass. The solar receiver provides a platform for
the algae to grow by giving a high surface area to volume ratio.

In heterotrophic production [45-47], microalgae used not only glucose as organic carbon substrates. Leading to low cost harvesting
systems with higher biomass growth was discussed by Chen and Chen [48]. Li et al. and Miao and Wu [47,49] investigated the
heterotrophic cultivation of Chlorella protothecoides for high lipid content, used for biodiesel production. Chojnacka and Noworyta [50]
compared Spirulina sp. growth in photoautotrophic, heterotrophic and mixotrophic cultures. Akkerman et al.[51] studied importance of
Photosynthetic Efficiency (PE) during photoautrophic growth. Microalgae, therefore, appear to be the most efficient biomass resource,
used for biofuel production due to its lipid content and specific growth rate [52]. Sheehan et al. [11] reported the site specific variation
is the choice for industrial microalgae production. Yoo et al. [32] evaluated three microalgae species, Chlorella vulgaris, Botryococcus
braunii, and Scenedesmus sp. under high level CO2 growth condition for biodiesel production, authors observed that B. braunii is the
most suitable one for biodiesel production while, Scenedesmus sp. is suitable for CO2 mitigation. Chiu et al. and Tang et al. [40,53]
revealed the high concentration of CO2 supplement, stimulate microalgae to grow faster comparison to atmospheric air. Furthermore,
authors found microalga; Haematococcus pluvialis can tolerate 34% of CO2 and could fixed CO2 with fixation rate of 0.14 g-1L-1 d-1
[12,54,55]. Overall summary on recent studies on microalgae CO2 sequestration is presented in (Table 1).
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S.No. Microalgae CO2concentration tolerance (%) CO2 fixation rate (g-1L-1d-1) References
1. Scenedesmus dimorphus 1.27
2. Scenedesmus incrassatulus 0.03% 1.50 Fulke et al. [1]
3. Chroococcus cohaerens 0.78
4. Chlorella sp. 1.62
5. Chlorella sp. 15 0.46 Jin et al. [56] 005
 6. Chlorella sp. 5 0.7 Ryu et al. [57]
7. Chlorella sp. -- 1.38 Zhao et al. [58]
8. Chlorella sp. 15 -- Fulke et al. [3]
9. Chlorella vulgaris 10 0.25 Sydney et al. [59]
10. Chlorella vulgaris 2 0.43 Yeh and Chang [60]
11. Scenedesmus sp. 15 0.61 Jin et al. [56]
12. Spirulina platensis 15 0.92 Kumar et al. [61]
13. Scenedesmus obliquus 10 0.55 Ho et al. [62]
14. Scenedesmus obliquus 10 0.29 Tang et al. [53]
15. Dunaliella sp. 3 0.31 Kishimoto et al. [63]
16. Scenedesmus obliquus 2.5 1.19 Ho et al. [64]
17. Chlorella kessleri
Chlorella vulgaris
Scenedesmus obliquus 18 -- de Morais and Costa [39]
Spirulina sp.
18. Dunaliella tertiolecta 10 0.27 Sydney et al. [59]
19. Haematococcus pluvialis 34 0.14 Huntley and Redalje [35]
20. Chlorella kessleri 18 0.16 de Morais and Costa [65]
Table 1: Recent studies on microalgal CO2 sequestration.
From the table, Haematococcus pluvialis, Spirulina sp., Chlorella sp., Chlorella vulgaris, Scenedesmus incrassatulus, Scenedesmus
obliquus, Scenedesmus dimorphus and Scenedesmus sp. have been recognized as promising microalgae strains, capable to assimilate
much of carbon dioxide, provide a biological tool for reducing GHG emissions from coal-fired power plants and other carbon severe
industrial processes. Whereas, numerous microalgae strains have high lipid content, it is possible to increase the concentration by
optimizing nitrogen level in the feed [66,67], light intensity [68], temperature, salinity [67], CO2 percentage [39,69] and harvesting
procedure [66,69]. However, increases in lipid accumulation will not outcome in lipid productivity as biomass productivity and lipid
accumulation ratios diverges in microalgae.
The algae can be processed into biofuel instead of first extracting oils then processing. This result in reduced costs associated with the
interim process from biomass to biofuel. A degradable alternative liquid called bio-oil, an advantage to enter directly in refinery stream
with some hydro-treating and hydro-cracking, produce a suitable diesel feedstock. Flash pyrolysis technology has high efficiency,
where feedstock is quickly heated to 350 - 500C for 2 seconds. Since an alga exists fundamentally in small units and has no fiber tissue
to deal with, algae have a major advantage over other biomass sources. A significant roadblock in using pyrolysis for algae conversion
is moisture content, and significant dehydration must be performed upstream for the process to work efficiently. The gasification of the
microalgal biomass to liquid fuels during Fischer-Tropsch Synthesis (FTS) or mixed alcohol synthesis results in syngas [70-72]. FTS
tends to require production at a very large scale to make the process efficient overall. However, most significance problem with FTS is
the generation of tars during the gasification process, causes coking of the synthesis catalyst and any other catalysts used in the syngas
cleanup process and must be removed. Liquefaction with the direct hydrothermal (defined as water held in a liquid state above 100oC
by applying pressure) can be employed to convert wet algal biomass to liquid fuels [73]. A maximum oil yield of 64% using Botryococcus
braunii as a feedstock has also been reported that algal biomass was processed by liquefaction [74]. Prajapati et al. [30] studied the novel
integration of phycoremediation and biogas production from algal biomass. Sialve et al. [75] evaluated that microalgae biomass contains
considerable amount of lipids, carbohydrates and proteins. This makes it a favorable substrate to convert into biodiesel, biogas and other
biofuel production along with their integration with CO2 sequestration.

ValueAdded Products from Algal Biomass

Borowitzka [76] examined that some of Chlorophyceae group algal species accumulate high concentrations of carotenoids. Hirata et
al. [77] suggested that phycobiliproteins, phycocyanin and phycoerythrin from algae can be usedincosmetics, pharmacy and nutrition.
Francisco et al. [78] investigated Chlorella vulgaris as for biodiesel production with lipid content of 27.0%. Muller-Feugaet al. [79]
reported bactericides production using Ulvophyceae, Charophyceae and Spirogyra. Lorenz and Cysewski [80] produced astaxanthin
from H.Pluvialis. The schematic diagram of the overall process of CO2 conversion into algal biomass to biofuel and other Value-Added
Products (VAPs) is shown by authors in (Figure 3). In wastewater stabilization ponds, the algae produce oxygen through photosynthesis
[1,81]. Hejazi and Wijffels [82] highlighted the microalgal milking for commercialization of microalgal products.
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CO2 EMISSION

Figure 3: The overall process of CO2 conversion into algal biomass which can be used for production of biofuel
and other value added products. 006
CO2 Sequestration by Microalgae: Recent Advances and Perspectives
Crucial need is integrated CO2 bio-fixation, bio-fuel production and value addition of algal biomass as an expectant substitute to
current CO2 mitigation strategies. However, the option for most frequently quoted microalgae for competent and economical combination
of CO2 biofixation, wastewater treatment and lipid synthesis toward biofuel production should be explored.
Various studies reported CO2 capture technologies have the adequate capability to reduce greenhouse gas emissions and are able to
address the prospects and challenges based on cost and engineering finances of CO2 capture and sequestration [1,3,83].
Special emphasis is required on regulation of biosynthetic genes and its expression in microalgae under various stress conditions
for CO2 sequestration. To increase the photosynthetic efficiency of microalgae, significant genetic modifications in Rubisco gene and its
functions should be investigated.

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