Effect of the inactivation of lactate dehydrogenase, ethanol dehydrogenase, and phosphotransacetylase on 2,3-butanediol production in Klebsiella pneumoniae strain

Biotechnology for Biofuels

Xuewu Guo¹  Chunhong Cao¹  Yazhou Wang¹  Chaoqun Li¹  Mingyue Wu¹  Yefu Chen¹  Cuiying Zhang¹  Huadong Pei¹  Dongguang Xiao¹ 

[1] Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education; Tianjin Industrial Microbiology Key Lab, College of Biotechnology, Tianjin University of Science and Technology, Box 08, No. 29, 13ST. TEDA, Tianjin 300457, China

关键词: Klebsiella pneumoniae;    phosphotransacetylase;    ethanol dehydrogenase;    lactate dehydrogenase;    2,3-butanediol;   

Others  :  792998
DOI  :  10.1186/1754-6834-7-44

 received in 2013-11-19, accepted in 2014-03-13,  发布年份 2014

【 摘 要 】

Background

2,3-Butanediol (2,3-BD) is a high-value chemical usually produced petrochemically but which can also be synthesized by some bacteria. To date, Klebsiella pneumoniae is the most powerful 2,3-BD producer which can utilize a wide range of substrates. However, many by-products are also produced by K. pneumoniae, such as ethanol, lactate, and acetate, which negatively regulate the 2,3-BD yield and increase the costs of downstream separation and purification.

Results

In this study, we constructed K. pneumoniae mutants with lactate dehydrogenase (LDH), acetaldehyde dehydrogenase (ADH), and phosphotransacetylase (PTA) deletion individually by suicide vector conjugation.

These mutants showed different behavior of production formation. Knock out of ldhA had little influence on the yield of 2,3-BD, whereas knock out of adhE or pta significantly improved the formation of 2,3-BD. The accumulation of the intermediate of 2,3-BD biosynthesis, acetoin, was decreased in all the mutants. The mutants were then tested in five different carbon sources and increased 2,3-BD was observed. Also a double mutant strain with deletion of adhE and ldhA was constructed which resulted in accelerated fermentation and higher 2,3-BD production. In fed-batch culture this strain achieved more than 100 g/L 2,3-BD from glucose with a relatively high yield of 0.49 g/g.

Conclusion

2,3-BD production was dramatically improved with the inactivation of adhE and pta. The inactivation of ldhA could advance faster cell growth and shorter fermentation time. The double mutant strain with deletion of adhE and ldhA resulted in accelerated fermentation and higher 2,3-BD production. These results provide new insights for industrial production of 2,3-BD by K. pneumoniae.

【 授权许可】

   
2014 Guo et al.; licensee BioMed Central Ltd.

【 图 表 】

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