Glycerol Utilization as Substrate for Ethanol Production in Escherichia coli Recombinant under an Aerobic Condition

Wahyu Suradi Pranata, Prayoga Suryadarma, Djumali Mangunwidjaja

Abstract


The utilization of glycerol as a substrate for ethanol production in E. coli recombinant harboring ethanologenic gens (PDC and ADHB) under aerobic conditions was investigated. This research was conducted by using E. coli BW25113 which compared the growth characteristics on glycerol and glucose as carbon source. E. coli can grow well on both substrates and it consumed glycerol faster than glucose. On glucose, E. coli occurred overflow metabolism that indicated by high acetate accumulation. Meanwhile, on substrate of glycerol the acetate accumulation could be reduced. Using glycerol substrate on E. coli increased piruvate accumulation, comparing with glucose. Therefore, the growth characteristic of glycerol was more effective. In E. coli ∆pta/pHfdh/pTadhB-pdc utilized glycerol substrate was able to accumulate pyruvate as intermediate metabolite for produce 2.18 gL-1 ethanol.

Keywords


Aerobic condition, Escherichia coli, Ethanol, Glucose, Glycerol

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References


International Energy Agency (2008) From 1st to 2nd-generation biofuel technologies. An overview of current Industry and RD and D Activities. Paris Cedex, OECD/IEA.

Ito T, Nakashimada Y, Senba K et al. (2005) Hydrogen and ethanol production from glycerol containing wastes discharged after biodiesel manufacturing process. Journal of Bioscience and Bioengineering 100 (3): 260 – 265.

Dharmadi Y, Murarka A, Gonzalez R (2006) Anaerobic fermentation of glycerol by Escherichia coli: A new platform for metabolic engineering. Biotechnology and Bioengineerin 94 (5): 821 – 829.

Yazdani SS, Gonzalez R (2007) Anaerobic fermentation of glycerol: a path to economic viability for the biofuels industry. Current Opinion in Biotechnology 18 (3): 213 – 219.

Suryadarma P, Ojima Y, Fukuda Y et al. (2012) The rpoS deficiency suppresses acetate accumulation in glucose-enriched culture of Escherichia coli under an aerobic con-dition. Frontiers of Chemical Science and Engineering 6 (2): 152 – 157. doi: 10.1007/s11705-012-1287-0.

Ingram LO, Conway T, Clark DP et al. (1987) Genetic engineering of ethanol production in Escherichia coli. Applied and Environmental Microbiology 53 (10): 2420 – 2425.

Sutapa IDA (1999) Lumpur aktif: Alternatif pengolah limbah cair. Jurnal Studi Pembangunan, Kemasyarakatan dan Lingkungan 1 (3): 25 – 38.

Ojima Y, Kawase D, Nishioka M, Taya M (2009) Functionally undefined gene, yggE, alleviates oxidative stress generated by monoamine oxidase in recombinant Escherichia coli. Biotechnology Letters 31: 139 – 145.

Causey TB, Shanmugam KT, Yomano LP, Ingram LO (2004) Engineering Escherichia coli for efficient conversion of glucose to pyruvate. Proceedings of the National Acad-emy of Sciences of the United States of America 101 (8): 2235 –2240.

Vemuri GN, Altman E, Sangurdekar DP et al. (2006) Over-flow metabolism in Escherichia coli during steady-state growth: Transcriptional regulation and effect of the redox ratio. Applied and Environmental Microbiology 72 (5): 3653 – 3661.

Martinez-Gomez K, Flores N, Castaneda HM et al. (2012) New insights into Escherichia coli metabolism: carbon scavenging, acetate metabolism and carbon recycling re-sponses during growth on glycerol. Microbial Cell Factories 11: 1 – 21. doi: 10.1186/1475-2859-11-46.

Baba T, Ara T, Hasegawa M et al. (2006) Construction of Escherichia coli K-12 in-frame, single gene knockout mu-tants: the keio collection. Molecular Systems Biology 2 (1): 1 – 11. doi: 10.1038/msb4100050.

Murarka A, Dharmadi Y, Yazdani SS, Gonzalez R (2008) Fermentative utilization of glycerol by Escherichia coli and its implications for the production of fuels and chemicals. Applied and Environmental Microbiology 74 (4): 1124 – 1135. doi: 10.1128/AEM.02192-07.

Shah P, Chiu F, Lan JC (2014) Aerobic utilization of crude glycerol by recombinant Escherichia coli for simultaneous production of poly 3-hydroxybutyrate and bioethanol. Journal of Bioscience and Bioengineering 117 (3): 343 – 350. doi: 10.1016/j.jbiosc.2013.08.018.

Durnin G, Clomburg J, Yeates Z et al. (2008) Understand-ng and harnessing the microaerobic metabolism of glycer-ol in Escherichia coli. Biotechnology and Bioengineering 103: 148–161. doi: 10.1002/bit.22246

Nielsen J, Villadsen J, Liden G (2003) Bioreaction engineering principles. New York, Kluwer Academic/Plenum Publishers.

Darbon E, Ito K, Huang HS et al. (1999) Glycerol transport and phosphoenolpyruvate-dependent enzyme I- and Hpr-catalyzed phosphorylation of glycerol kinase in Thermus flavus. Microbiology 145: 3205 – 3212. doi: 10.1099/002212 87-145-11-3205.

Frankel DG (1996) Glycolysis. In: Neidhardt FC (eds) Escherichia coli and Salmonella tiphymurium: Cellular and molecular biology. 2nd edition. Washington DC, ASM Press.

Trinh CT, Srienc F (2009) Metabolic engineering of Escherichia coli for efficient conversion of crude glycerol to ethanol. Applied and Environmental Microbiology 75 (21): 6696 – 6705. doi: 10.1128/AEM.00670-09.




DOI: http://dx.doi.org/10.11594/jtls.08.01.14

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