Journal
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Volume 101, Issue 10, Pages 4327-4337Publisher
SPRINGER
DOI: 10.1007/s00253-017-8200-1
Keywords
Butanol; Clostridium tyrobutyricum; Carbon catabolite repression; Metabolic engineering; Sucrose; Sugarcane juice
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Funding
- National Science Foundation STTR program [IIP-1026648]
- China Scholarship Council [201406350172]
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Clostridium tyrobutyricum is a promising organism for butyrate and n-butanol production, but cannot grow on sucrose. Three genes (scrA, scrB, and scrK) involved in the sucrose catabolic pathway, along with an aldehyde/alcohol dehydrogenase gene, were cloned from Clostridium acetobutylicum and introduced into C. tyrobutyricum (Delta ack) with acetate kinase knockout. In batch fermentation, the engineered strain Ct(Delta ack)-pscrBAK produced 14.8-18.8 g/L butanol, with a high butanol/total solvent ratio of similar to 0.94 (w/w), from sucrose and sugarcane juice. Moreover, stable high butanol production with a high butanol yield of 0.25 g/g and productivity of 0.28 g/La (TM) h was obtained in batch fermentation without using antibiotics for selection pressure, suggesting that Ct(Delta ack)-pscrBAK is genetically stable. Furthermore, sucrose utilization by Ct(Delta ack)-pscrBAK was not inhibited by glucose, which would usually cause carbon catabolite repression on solventogenic clostridia. Ct(Delta ack)-pscrBAK is thus advantageous for use in biobutanol production from sugarcane juice and other sucrose-rich feedstocks.
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