期刊
METABOLIC ENGINEERING
卷 22, 期 -, 页码 32-39出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymben.2013.12.005
关键词
Acetoin; Candida glabrata; Metabolic engineering; In silico; Cofactor engineering
资金
- Major State Basic Research Development Program of China (973 Program) [2013CB733600]
- Program for Young Talents in China
- Provincial Outstanding Youth Foundation of Jiangsu Province [BK2012002]
- Priority Academic Program Development of Jiangsu Higher Education Institutions
- National Natural Science Foundation of China [31270079]
- Fundamental Research Funds for the Central Universities [JUDCF12027]
Utilization of Candida glabrata overproducing pyruvate is a promising strategy for high-level acetoin production. Based on the known regulatory and metabolic information, acetaldehyde and thiamine were fed to identify the key nodes of carboligase activity reaction (CAR) pathway and provide a direction for engineering C. glabrata. Accordingly, alcohol dehydrogenase, acetaldehyde dehydrogenase, pyruvate decarboxylase, and butanediol dehydrogenase were selected to be manipulated for strengthening the CAR pathway. Following the rational metabolic engineering, the engineered strain exhibited increased acetoin biosynthesis (2.24 g/L). In addition, through in silico simulation and redox balance analysis, NADH was identified as the key factor restricting higher acetoin production. Correspondingly, after introduction of NADH oxidase, the final acetoin production was further increased to 7.33 g/L. By combining the rational metabolic engineering and cofactor engineering, the acetoin-producing C. glabrata was improved stepwise, opening a novel pathway for rational development of microorganisms for bioproduction. (C) 2013 Published by Elsevier Inc. on behalf of International Metabolic Engineering Society.
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