Abstract
Objectives
To improve the production of 2,3-butanediol (2,3-BD) in Klebsiella pneumoniae, the genes related to the formation of lactic acid, ethanol, and acetic acid were eliminated.
Results
Although the cell growth and 2,3-BD production rates of the K. pneumoniae ΔldhA ΔadhE Δpta-ackA strain were lower than those of the wild-type strain, the mutant produced a higher titer of 2,3-BD and a higher yield in batch fermentation: 91 g 2,3-BD/l with a yield of 0.45 g per g glucose and a productivity of 1.62 g/l.h in fed-batch fermentation. The metabolic characteristics of the mutants were consistent with the results of in silico simulation.
Conclusions
K. pneumoniae knockout mutants developed with an aid of in silico investigation could produce higher amounts of 2,3-BD with increased titer, yield, and productivity.
Similar content being viewed by others
References
Abdel-Hamid AM, Attwood MM, Guest JR (2001) Pyruvate oxidase contributes to the aerobic growth efficiency of Escherichia coli. Microbiology 147:1483–1498
Arasu MV, Kumar V, Ashok S, Hyohak S, Rathnasingh C, Lee HJ, Seung D, Park S (2011) Isolation and characterization of the new Klebsiella pneumoniae J2B strain showing improved growth characteristics with reduced lipopolysaccharide formation. Biotech Bioproc Eng 16:1134–1143
Celinska E, Grajek W (2009) Biotechnological production of 2,3-butanediol: current state and prospects. Biotechnol Adv 27:715–725
Cho JH, Rathnasingh C, Song H, Chung BW, Lee HJ, Seung D (2012) Fermentation and evaluation of Klebsiella pneumoniae and K. oxyotca on the production of 2,3-butanediol. Bioprocess Biosyst Eng 35:1081–1088
Echave P, Tamarit J, Cabiscol E, Ros J (2003) Novel antioxidant role of alcohol dehydrogenase E from Escherichia coli. J Biol Chem 32:30193–30198
Emerson RR, Flickinger MC, Tsao GT (1982) Kinetics of dehydration of aqueous 2,3-butanediol to methyl ethyl ketone. Ind Eng Chem Prod Res Dev 21:473–477
Hansen HG, Henning U (1966) Regulation of pyruvate dehydrogenase activity in Escherichia coli K12. Biochim Biophys Acta 122:355–358
Hugenholtz J, Kleerebezem M (1999) Metabolic engineering of lactic acid bacteria: overview of the approaches and results of pathway rerouting involved in food fermentations. Curr Opin Biotech 10:492–497
Ji XJ, Huang H, Du J, Zhu JG, Ren LJ, Li S, Nie ZK (2009) Development of an industrial medium for economical 2,3-butanediol production through co-fermentation of glucose and xylose by Klebsiella oxytoca. Bioresour Technol 100:5214–5218
Ji XJ, Huang H, Ouyang PK (2011) Microbial 2,3-butanediol production: a state-of-the-art review. Biotechnol Adv 29:351–364
Kim DK, Rathnasingh C, Song H, Lee HJ, Seung D, Chang YK (2013) Metabolic engineering of a novel Klebsiella oxytoca strain for enhanced 2,3-butaneeiol production. J Biosci Bioeng 116:186–192
Kosaric N, Magee RJ, Blaszczyk R (1992) Redox potential measurement for monitoring glucose and xylose conversion by K. pneumoniae. Chem Biochem Eng Q 6:145–152
Liao YC, Huang TW, Chen FC, Charusanti P, Hong JS, Chang HY, Tsai SF, Palsson BO, Hsiung CA (2011) An experimentally validated genome-scale metabolic reconstruction of Klebsiella pneumoniae MGH 78578, iYL1228. J Bacteriol 193:1710–1717
Ma C, Wang A, Qin J, Li L, Ai X, Jiang T, Tang H, Xu P (2009) Enhanced 2,3-butanediol production by Klebsiella pneumoniae SDM. Appl Microbiol Biotechnol 82:49–57
Oliver JW, Machado IM, Yoneda H, Atsumi S (2013) Cyanobacterial conversion of carbon dioxide to 2,3-butanediol. Proc Natl Acad Sci USA 110:1249–1254
Rathnasingh C, Kim DK, Song H, Park S, Lee HJ, Seung D (2012) Isolation and characterization of a new mucoid-free Klebsiella pneumoniae strain for 2,3-butanediol production. Afr J Biotechnol 11:11252–11261
Song H, Lee SY (2006) Production of succinic acid by bacterial fermentation. Enzyme Microb Tech 39:352–361
Winfield ME (1945) The catalytic dehydration of 2,3-butanediol to 1,3-butadiene. J Council Sci Ind Res 18:412–423
Wolfe AJ (2005) The acetate switch. Microbiol Mol Biol Rev 69:12–50
Acknowledgments
This work was supported by the Industrial Strategic Technology Development Program (No. 10050407) funded by the Ministry of Trade, Industry & Energy (MOTIE, Korea) and the Advanced Biomass R&D Center of Global Frontier Project (No. ABC-2010-0029728) funded by the Ministry of Science, ICT and Future Planning (MSIP, Korea).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Rathnasingh, C., Park, J.M., Kim, Dk. et al. Metabolic engineering of Klebsiella pneumoniae and in silico investigation for enhanced 2,3-butanediol production. Biotechnol Lett 38, 975–982 (2016). https://doi.org/10.1007/s10529-016-2062-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-016-2062-y