期刊
METABOLIC ENGINEERING
卷 19, 期 -, 页码 50-56出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymben.2013.05.006
关键词
2-Keto-L-gulonic acid; Pyrroloquinoline quinine; Cofactor engineering; Consortium
资金
- National High Technology Research and Development Program of China (863 Program) [2012AA02A701]
- National Basic Research Program of China (973 Program) [2013CB733601]
- National Natural Science Foundation of China [21020102040]
- NTU
- MOE, Singapore [RG 78/10]
- AcRF Tier-2 Grant, Singapore [MOE2011-T2-2-035, ARC 3/12]
The expression levels of sorbose/sorbosone dehydrogenase genes (sdh and sndh) and the synthesis genes (pqqABCDEN) of the ad joint cofactor pyrroloquinoline quinone (PQQ) were genetically manipulated in Ketogulonigenium vulgare to increase the production of 2-keto-L-gulonic acid (2-KLG), the precursor of vitamin C. in the consortium of K. vulgare and Bacillus cereus. We found that overexpression of sdh sndh alone in K. vulgare could not significantly enhance the production of 2-KLG, revealing the cofactor PQQ was required for the biosynthesis of 2-KLG. Various expression levels of PQQ were achieved by differential expression of pqqA. pqqABCDE and pqqABCDEN, respectively. The combinatorial expression of sdh/sndh and pqqABCDEN in K. vulgare enabled a 20% increase in the production of 2-KLG (79.1 +/- 0.6 g l(-1)) than that of the parental K. vulgare (65.9 +/- 0.4 g l(-1)) in shaking flasks. Our results demonstrated the balanced co-expression of both the key enzymes and the related cofactors was an efficient strategy to increase chemicals' biosynthesis. (C) 2013 Elsevier Inc. All rights reserved.
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