Acta Scientific Biotechnology

Research Article Volume 1 Issue 9

Genetically Modified Escherichia coli for High Yield Mixed Organic Acid Production on Lignocellulosic Biomass Derived Xylose

Poornima Rao1, Lucy Nainan1, Gunjan Prakash1 and Arvind M Lali2*

1DBT-ICT Centre for Energy Biosciences, Institute of Chemical Technology, A University under Section-3 of UGC Act 1956 and Maharashtra Govt.’s Elite Status and Grade `A’ Centre of Excellence by MHRD, Matunga, Mumbai, India
2Department of Chemical Engineering, Institute of Chemical Technology, A University under Section-3 of UGC Act 1956 and Maharashtra Govt.’s Elite Status and Grade `A’ Centre of Excellence by MHRD, Matunga, Mumbai, India

*Corresponding Author: Arvind M Lali, Department of Chemical Engineering, Institute of Chemical Technology, A University under Section-3 of UGC Act 1956 and Maharashtra Govt.’s Elite Status and Grade `A’ Centre of Excellence by MHRD, Matunga, Mumbai, India.

Received: July 22, 2020; Published: August 31, 2020

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Abstract

  Lignocellulosic biomass deconstruction generates around 20 - 25 w/w% of hemicellulose derived xylose that must be exploited as a sugar platform for production of value-based chemicals. The absence of genes in the metabolic pathways for uptake and utilization of xylose in most microbes restricts its potential applicability to be used as carbon source. Rapidly evolving separation technologies dictate emerging preference for hetero-fermentation with high overall substrate based yields over lower yield homo-fermentations. Escherichia coli is known to metabolize both hexose and pentose sugars, and the present study describes construction and characterization of a double knock-out engineered mutant E. coli BW25113 ΔadhE ΔldhA for hetero-fermentative production of organic acids in > 90% yield on xylose majorly comprising of acetic acid. Fermentation parameters such as aeration, agitation, pH were observed to impact the xylose uptake rate, and consequently the acetate and total organic acid production rate. These investigative studies resulted in acetate yield and productivity of 0.51 ± 0.0027 g/g and 0.33 ± 0.0048 g/L.h; and a total organic acid yields and productivities of 0.92 ± 0.019 g/g and 0.61 ± 0.013 g/L.h on pure xylose. The modified strain demonstrated equivalent acetate yield of 0.51 ± 0.026 g/g and slightly reduced acetate productivity of 0.21 ± 0.011 g/L/h on lignocellulose derived xylose stream containing lignin, nitrate and furfural. Reduction in formate yield was possibly due to disproportion of formate to H2 and CO2 in the presence of nitrate in the lignocellulose derived xylose stream. Recovery of nitrate from the xylose fraction by employing a suitable filtration method can eliminate or reduce nitrate presence and provide higher formate yields. The study provides noteworthy insights on the practical applicability of lignocellulose derived xylose for commercial production of organic acids as important biochemical precursors.

Keywords: Escherichia coli; Lignocellulosic Biomass; Xylose; Organic Acids; Acetic Acid

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References

  1. Cheryan M., et al. "Production of acetic acid by Clostridium thermoaceticum”. Advances in Applied Microbiology 43 (1997): 1-33.
  2. Naraian R., et al. “Microbial production of organic acids”. In: Gupta, V. K, Treichel, H., Shapaval, V., de Oliveira, L. A. and Tuohy, M. G. (Edition.), Microbial Functional Foods and Nutraceuticals, 93. John Wiley and Sons Ltd. (2017): 93-121.
  3. Sugaya K., et al. "Production of acetic acid by Clostridium thermoaceticum in batch and continuous fermentations”. Biotechnology and Bioengineering5 (1986): 678-683.
  4. Lali Arvind. "Biofuels for India: what, when and how”. Current Science (2016): 552-555.
  5. Anwar Z., et al. "Agro-industrial lignocellulosic biomass a key to unlock the future bio-energy: a brief review”. Journal of Radiation Research and Applied Sciences2 (2014): 163-173.
  6. Wadekar P and Lali A. “Recovery and downstream chemistry of lignocellulosic lignin”. (2015).
  7. Matsushika A., et al. "Efficient bioethanol production from xylose by recombinant Saccharomyces cerevisiae requires high activity of xylose reductase and moderate xylulokinase activity”. Journal of Bioscience and Bioengineering3 (2008): 306-309.
  8. Deng X., et al. "Xylulokinase activity in various yeasts including Saccharomyces cerevisiae containing the cloned xylulokinase gene”. Applied Biochemistry and Biotechnology1 (1990): 193.
  9. Li Xin., et al. "Comparison of xylose fermentation by two high-performance engineered strains of Saccharomyces cerevisiae”. Biotechnology Reports 9 (2016): 53-56.
  10. Karhumaa K., et al. "Comparison of the xylose reductase-xylitol dehydrogenase and the xylose isomerase pathways for xylose fermentation by recombinant Saccharomyces cerevisiae”. Microbial Cell Factories1 (2007): 5.
  11. Olsson L., et al. "Fermentation of lignocellulosic hydrolysates for ethanol production”. Enzyme and Microbial Technology5 (1996): 312-331.
  12. Margaritis A., et al. "Direct fermentation of D-xylose to ethanol by Kluyveromyces marxianus strains”. Applied and Environmental Microbiology5 (1982): 1039-1041.
  13. Moysés D., et al. "Xylose fermentation by Saccharomyces cerevisiae: challenges and prospects”. International Journal of Molecular Sciences3 (2016): 207.
  14. Busche RM., et al. “Recovery of acetic acid from dilute acetate solution”. Biotechnol. Bioeng. Symp. (United States) (Vol. 12, No. CONF-820580-). EI du Pont de Nemours and Company, Inc., Wilmington, DE. (1982).
  15. Pawar PR., et al. "Organic waste streams as feedstock for the production of high volume-low value products”. Environmental Science and Pollution Research (2020): 1-11.
  16. Rault A., et al. "Fermentation pH influences the physiological-state dynamics of Lactobacillus bulgaricus CFL1 during pH-controlled culture”. Applied and Environmental Microbiology13 (2009): 4374-4381.
  17. Garcı́a-Ochoa., et al. "Oxygen transfer and uptake rates during xanthan gum production”. Enzyme and Microbial Technology9 (2000): 680-690.
  18. Çalik P., et al. "Oxygen transfer effects on recombinant benzaldehyde lyase production”. Chemical Engineering Science22-23 (2004): 5075-5083.
  19. Liu Y-S., et al. "Characterization of oxygen transfer conditions and their effects on Phaffia rhodozyma growth and carotenoid production in shake-flask cultures”. Biochemical Engineering Journal3 (2006): 331-335.
  20. Lali AM and Maurya RR. “Separation of organic acids from mixtures containing ammonium salts of organic acids”. U.S. Patent Application 15/577,912 (2018).
  21. Nainan L and Lali A. “Improved production of acetic acid in Escherichia coli” (2018).
  22. Lali AM., et al. “U.S. Patent No. 8,338,139”. Washington, DC: U.S. Patent and Trademark Office. (2012).
  23. Datsenko K., et al. "One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products”. Proceedings of The National Academy of Sciences12 (2000): 6640-6645.
  24. Kleman L., et al. "Acetate metabolism by Escherichia coli in high-cell-density fermentation”. Applied and Environmental Microbiology11 (1994): 3952-3958.
  25. Zhu J., et al. "Effect of a single-gene knockout on the metabolic regulation in Escherichia coli for D-lactate production under microaerobic condition”. Metabolic Engineering2 (2005): 104-115.
  26. Abbott DA., et al. "Metabolic engineering of Saccharomyces cerevisiae for production of carboxylic acids: current status and challenges”. FEMS Yeast Research8 (2009): 1123-1136.
  27. Luli GW., et al. "Comparison of growth, acetate production, and acetate inhibition of Escherichia coli strains in batch and fed-batch fermentations”. Applied and Environmental Microbiology4 (1990): 1004-1011.
  28. Warnecke T., et al. "Organic acid toxicity, tolerance, and production in Escherichia coli biorefining applications”. Microbial Cell Factories1 (2005): 1-8.
  29. Sawers RG. “Formate and its role in hydrogen production in Escherichia coli”. Biochemical Society Transactions 1 (2005): 42-46.
  30. Zelcbuch L., et al. "Pyruvate formate-lyase enables efficient growth of Escherichia coli on acetate and formate”. Biochemistry17 (2016): 2423-2426.
  31. Tajarudin HA., et al. "Intensive Production of Carboxylic Acids Using butyricum in a Membrane Bioreactor (MBR)”. Fermentation 4.4 (2018): 81.
  32. Causey TB., et al. "Engineering the metabolism of Escherichia coli W3110 for the conversion of sugar to redox-neutral and oxidized products: homoacetate production”. Proceedings of the National Academy of Sciences3 (2003): 825-832.
  33. Yarbrough JM., et al. "Bacterial inhibitory effects of nitrite: inhibition of active transport, but not of group translocation, and of intracellular enzymes”. Applied and Environmental Microbiology4 (1980): 831-834.
  34. Brandberg T., et al. "The fermentation performance of nine strains of Saccharomyces cerevisiae in batch and fed-batch cultures in dilute-acid wood hydrolysate”. Journal of Bioscience and Bioengineering2 (2004): 122-125.
  35. Wu W., et al. "Toward engineering coli with an autoregulatory system for lignin valorization”. Proceedings of the National Academy of Sciences 115.12 (2018): 2970-2975.
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Citation

Citation: Arvind M Lali., et al. “Genetically Modified Escherichia coli for High Yield Mixed Organic Acid Production on Lignocellulosic Biomass Derived Xylose". Acta Scientific Biotechnology 1.9 (2020): 28-38.




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