Limitation of thiamine pyrophosphate supply to growing Escherichia coli switches metabolism to efficient D-lactate formation

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논문

Limitation of thiamine pyrophosphate supply to growing Escherichia coli switches metabolism to efficient D-lactate formation

학술지

Biotechnology and bioengineering

저자명

Tian, Kangming; Niu, Dandan; Liu, Xiaoguang; Prior, Bernard A.; Zhou, Li; Lu, Fuping; Singh, Suren; Wang, Zhengxiang

초록

ABSTRACTEfficient production of d‐lactate by engineered Escherichia coli entails balancing cell growth and product synthesis. To develop a metabolic switch to implement a desirable transition from cell growth to product fermentation, a thiamine auxotroph B0013‐080A was constructed in a highly efficient d‐lactate producer E. coli strain B0013‐070. This was achieved by inactivation of thiE, a gene encoding a thiamine phosphate synthase for biosynthesis of thiamine monophosphate. The resultant mutant B0013‐080A failed to grow on the medium in the absence of thiamine yet growth was restored when exogenous thiamine was provided. A linear relationship between cell mass formation and amount of thiamine supplemented was mathematically determined in a shake flask experiment and confirmed in a 7‐L bioreactor system. This calculation revealed that &sim;95–96 thiamine molecules per cell were required to satisfy cell growth. This relationship was employed to develop a novel fermentation process for d‐lactate production by using thiamine as a limiting condition. A d‐lactate productivity of 4.11 g · L−1 · h−1 from glycerol under microaerobic condition and 3.66 g · L−1 · h−1 from glucose under anaerobic condition was achieved which is 19.1% and 10.2% higher respectively than the parental strain. These results revealed a convenient and reliable method to control cell growth and improve d‐lactate fermentation. This control strategy could be applied to other biotechnological processes that require optimal allocation of carbon between cell growth and product formation. Biotechnol. Bioeng. 2016;113: 182–188. © 2015 Wiley Periodicals, Inc.

발행연도

2016

ISSN

0006-3592

ISSN

1097-0290

권

113

호

페이지

pp.182-188

DOI