Metabolic pathway engineering for production of 1,2-propanediol and 1-propanol by Corynebacterium glutamicum

被引:72
作者
Siebert, Daniel
Wendisch, Volker F. [1 ]
机构
[1] Univ Bielefeld, Fac Biol, Chair Genet Prokaryotes, D-33615 Bielefeld, Germany
关键词
Corynebacterium glutamicum; Metabolic engineering; 1-propanol; 1,2-propanediol; ESCHERICHIA-COLI; AMINO-ACIDS; BIOTECHNOLOGICAL PRODUCTION; MICROBIAL-PRODUCTION; ETHANOL-PRODUCTION; LYSINE PRODUCTION; PYRUVATE-KINASE; ORGANIC-ACIDS; EXPRESSION; GENE;
D O I
10.1186/s13068-015-0269-0
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Background: Production of the versatile bulk chemical 1,2-propanediol and the potential biofuel 1-propanol is still dependent on petroleum, but some approaches to establish bio-based production from renewable feed stocks and to avoid toxic intermediates have been described. The biotechnological workhorse Corynebacterium glutamicum has also been shown to be able to overproduce 1,2-propanediol by metabolic engineering. Additionally, C. glutamicum has previously been engineered for production of the biofuels ethanol and isobutanol but not for 1-propanol. Results: In this study, the improved production of 1,2-propanediol by C. glutamicum is presented. The product yield of a C. glutamicum strain expressing the heterologous genes gldA and mgsA from Escherichia coli that encode methylglyoxal synthase gene and glycerol dehydrogenase, respectively, was improved by additional expression of alcohol dehydrogenase gene yqhD from E. coli leading to a yield of 0.131 mol/mol glucose. Deletion of the endogenous genes hdpA and ldh encoding dihydroxyacetone phosphate phosphatase and lactate dehydrogenase, respectively, prevented formation of glycerol and lactate as by-products and improved the yield to 0.343 mol/mol glucose. To construct a 1-propanol producer, the operon ppdABC from Klebsiella oxytoca encoding diol dehydratase was expressed in the improved 1,2-propanediol producing strain ending up with 12 mM 1-propanol and up to 60 mM unconverted 1,2-propanediol. Thus, B-12-dependent diol dehydratase activity may be limiting 1-propanol production. Conclusions: Production of 1,2-propanediol by C. glutamicum was improved by metabolic engineering targeting endogenous enzymes. Furthermore, to the best of our knowledge, production of 1-propanol by recombinant C. glutamicum was demonstrated for the first time.
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页数:13
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