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Download Article Advances in Biological Sciences Research, volume 7 1st International Symposium Innovations in Life Sciences (ISILS 2019) Prospective Trends in Biotechnology for Biofuel Armen Trchounian Department of Biochemistry, Microbiology and Biotechnology Yerevan State University Yerevan, Armenia Department of Biotechnology and Microbiology, Belgorod State National Research University Belgorod, Russian Federation E-mail: [email protected]; ORCID ID: 0000-0002-0901-4893 Abstract—The development of clean alternative biofuel, substrates, including organic waste, and low cost compared including biohydrogen, is of great significant interest. One to thermochemical methods [2-4]. All this is important for a method for biohydrogen is dark and light fermentation when sustainable economy. hydrogenases are responsible enzymes for H2 metabolism. A further development of biotechnology for H2 production can be One method is the production of H2 by bacteria directed in the following ways at least: construction or selection (biohydrogen) that perform dark – mixed acids (Escherichia of appropriate effective bacterial strains (mutants) and coli, Clostridium bejerinckii) and light fermentation cultures; creation of mixed cultures; application of different (Rhodobacter sphaeroides). These bacteria use hydrogenases carbon sources (sugars, alcohols and organic acids) and their (Hyd) and other enzymes for Н metabolism and multiple mixtures for co-fermentation as well as organic wastes (glycerol 2 and lignocellulose waste); control of external technology factors Hyd enzymes have been revealed with E. coli and other (pH, redox potential, substrate concentration), addition of bacteria [2-7]; their mechanisms of action are well known for different compounds and their mixtures, especially some heavy the repeated use of fermentation revisited [8] in the economy. metals (Ni, Fe, Mo, Mg, Cu, etc). Biohydrogen has of interest It can also be produced through biophotolysis of water by when was producing with bioethanol. It can also be applied for microalgae and algae as biocatalysts [9]. enhanced production of biomethane. III. DEVELOPMENT OF BIOTECHNOLOGY FOR HYDROGEN Keywords—biotechnology for biofuel, hydrogen energy, PRODUCTION biomethane, fermentation, bacteria The aim of current development is to improve the contribution of H2 production to renewable energy budget. The research is rapidly increased for last decades, and steps I. INTRODUCTION from lab-scale to large-scale production are putting forward Nowadays the energy supply mainly depends on fossil to H2 production [2-4]. fuels, though significant efforts are being made to use fuels produced from renewable feedstock, as these have less Important experimental evidences have been obtained; greenhouse gas emissions during both fuel production and they have well ground significance for H2 production by using [1-3]. However, due to the current energy crisis and bacteria. Actually, by genetic engineering and selection rising concern over climate change, as well as some limitation approaches, an improved E. coli strain with H2 production -1 -1 with fossil fuels in future, the development of clean rate of 0.68 mmol H2 L h in by-product and new substrate alternative biofuel is of great significant interest. This is of fermentation - glycerol medium has been obtained; this important goal for the global energy strategy to establish a was 20-fold increased H2 production compared with wild low-carbon economy and flexible and adaptive power type [10], and this is not a limitation. Then, sole and mixed systems. culture of dark- and light- fermented bacteria for enhanced H2 There is also one important argument in favor of biofuels production from lignocellulose waste (distillers’ grains, from carbon-containing by-products and waste: this makes it brewery waste) [11-12] provides novel strategic approach for possible to produce decentralized power plants, where power an inexpensive energy generation, as well as to resolve the units can be located in close proximity to carbon resources problem of waste utilization. A 3-7-fold increase can be [4]. In addition, the formation of various contaminants, obtained with E. coli upon glycerol and formate, lactose or including volatile fatty acids, can be significantly reduced or xylose mixture vs. glycerol, formate, lactose or xylose prevented if the production site is next to the power plant. fermentation [13-16]. Prolong production of H2 can be also detected during more than 220 h [15]. II. HYDROGEN AS PERSPECTIVE ENERGY SOURCE Moreover, responsible Hyd enzymes of E. coli have been Hydrogen is an effective, environmentally friendly and revealed with sole and mixed carbon sources, including renewable energy source, since it has 3 times higher energy organic wastes, and for different technology conditions content (~ 142 kJ g-1) than fossil fuels such as oil, therefore (Table I); these might open a new way to apply appropriate its conversion efficiency is high; it generates only water and mutant strains for enhanced H2 production. does not contain toxic by-products and can be obtained from biomass [2-4]. Biohydrogen has the advantages of relatively All these approaches can not only enhance H2 production high yield at low temperature (for mesophiles growing best but also decrease in cost of production. at moderate temperatures), cheap and readily available Copyright © 2019, the Authors. Published by Atlantis Press. This is an open access article under the CC BY-NC license (http://creativecommons.org/licenses/by-nc/4.0/). 335 Advances in Biological Sciences Research, volume 7 IV. FURTHER STRATEGY FOR HYDROGEN PRODUCTION well as organic wastes (glycerol and lignocellulose waste) DEVELOPMENT [12-18]; A further development of biotechnology for H2 - control of external technology factors (pH, redox production can be directed in the following ways at least (Fig. potential, substrate concentration), addition of different 1): compounds and their mixtures, especially heavy metals (Ni, Fe, Mo, Mg, Cu, etc) [19-22]. - construction or selection of appropriate effective bacterial strains (mutants) and cultures [4, 10]; creation of In addition, two-step H2 production with dark and light mixed cultures [11]; fermentation when end products of dark mixed acids fermentation (organic acids) can serve as substrates for light - application of different carbon sources (sugars, alcohols fermentation (Fig. 2) is perspective approach to enhance of and organic acids) and their mixtures for co-fermentation as H2 production too [11]. TABLE I. RESPONSIBLE HYD ENZYMES WITH SOLE AND MIXED CARBON SOURCES AND FOR DIFFERENT PH Responsible Hyd enzymes Sole or mixed carbon References pH 7.5 pH 5.5 Hyd-3, Hyd-4 mainly, and Hyd-2 in Glucose (2g L-1) and glycerol (10 g L-1) Hyd-3 [13] some extent Glucose (2 g L-1) and formate (10 mM) Hyd-3 and Hyd-4 Hyd-3 [14] Hyd-3, Hyd-4 mainly, and Hyd-2 in Glycerol (10 g L-1) and formate (10 mM) Hyd-3 [14] some extent Hyd-3, Hyd-4 mainly, and Hyd-2 in Glycerol ((10 g L-1) and lactose (1 g L-1) Hyd-3 and Hyd-4 [15] some extent Glycerol ((10 g L-1) and xylose (4 g L-1) Hyd-3 and Hyd-4 Hyd-3 and Hyd-4 [16] Brewery waste Hyd-3 and Hyd-4 Hyd-3 and Hyd-4 [11-12] Appropriate effective strains (mutants) construction or selection; mixed cultures Control the external technology factors Application of Enhanced H2 like pH, redox different carbon production potential; Addition sources and of various their mixtures, compounds like including cheap single or mixture if and available different metals (Ni, wastes Fe, Mo, etc) etc Fig. 1. Directions to enhanced H2 production by bacteria (E. coli). 336 Advances in Biological Sciences Research, volume 7 H2 H2 Substrates (glycerol, xylose, mixed carbon Dark- organic acids Light- sources) fermentation fermentation ST ND 1 STAGE 2 STAGE Fig. 2. A scheme for two consecutive stages H2 production with dark- and light-fermentation. [8] A. Trchounian, K. Trchounian, “Fermentation revisited: how do V. HYDROGEN AND THE OTHER BIOFUELS PRODUCTION microorganisms survive under energy-limited conditions?”, Trends Biochem. Sci., vol. 44, pp. 391-400, 2019. It is interesting that some bacteria can produce hydrogen [9] L. Gabrielyan, L. Hakobyan, A. Trchounian, “Characterization of with bioethanol from by-products and organic wastes [10, light-dependent hydrogen production by new green microalga 23]. It would be effective to use biohydrogen to improve Parachlorella kessleri in various conditions”, J. Photochem. production of other types of biofuel, especially biomethane. Photobiol. B: Biology, vol. 175, pp. 207-10, 2017. Bioconversion of CO2 evolved during biogas production [10] H. Hu, T. K. Wood, “An evolved Escherichia coli strain for from biomass with biohydrogen to biomethane based on the producing hydrogen and ethanol from glycerol”, Biochem. Biophys. activity of hydrogenotrophic methanogens are being Res. Comm., vol. 391, pp.1033-8, 2010. researched; effectivity and limitations are determined [24- [11] H. Sargsyan, L. Gabrielyan, A. Trchoinian, “Novel approach of ethanol waste utilization: Biohydrogen production by mixed cultures 25]. It was found that the addition of H2 to reactors treating of dark- and photo-fermentative bacteria using distillers’ grains”, Int. food waste showed great potential for enhanced CH4 yield J. Hydrogen Energy, vol. 41, pp. 2377-82, 2016. and biogas upgrade [26]. This creates the possibility to [12] A. Poladyan, K. Trchounian, A. Vasilian, A. Trchouian, “Hydrogen optimize hydrogenotrophic methanogenesis towards production by Escherichia coli
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