NDLI: Directed pathway evolution of the glyoxylate shunt in Escherichia coli for improved aerobic succinate production from glycerol

Content Provider	Springer Nature Link
Author	Li, Ning Zhang, Bo Chen, Tao Wang, Zhiwen Tang, Ya jie Zhao, Xueming
Copyright Year	2013
Abstract	α-Ketoglutarate is accumulated as the main byproduct during the aerobic succinate production from glycerol by Escherichia coli BL21(DE3) in minimal medium. To address this issue, here a strategy of directed pathway evolution was developed to enhance the alternative succinate production route—the glyoxylate shunt. Via the directed pathway evolution, the glyoxylate shunt was recruited as the primary anaplerotic pathway in a ppc mutant, which restored its viability in glycerol minimal medium. Subsequently, the operon sdhCDAB was deleted and the gene ppc was reverted in the evolved strain for succinate production. The resulting strain E2-Δsdh-ppc produced 30 % more succinate and 46 % less α-ketoglutarate than the control strain. A G583T mutation in gene icdA, which significantly decreased the activity of isocitrate dehydrogenase, was identified in the evolved strain as the main mutation responsible for the observed phenotype. Overexpression of α-ketoglutarate dehydrogenase complex in E2-Δsdh-ppc further reduced the amount of byproduct and improved succinate production. The final strain E2-Δsdh-ppc-sucAB produced 366 mM succinate from 1.3 M glycerol in minimal medium in fed-batch fermentation. The maximum and average succinate volumetric productivities were 19.2 and 6.55 mM h−1, respectively, exhibiting potential industrial production capacity from the low-priced substrate.
Starting Page	1461
Ending Page	1475
Page Count	15
File Format	PDF
ISSN	13675435
Journal	Journal of Industrial Microbiology and Biotechnology
Volume Number	40
Issue Number	12
e-ISSN	14765535
Language	English
Publisher	Springer Berlin Heidelberg
Publisher Date	2013-10-02
Publisher Place	Berlin, Heidelberg
Access Restriction	Subscribed
Subject Keyword	Escherichia coli Glycerol Succinate Glyoxylate shunt Isocitrate dehydrogenase Microbiology Biochemistry Inorganic Chemistry Genetic Engineering Biotechnology Bioinformatics
Content Type	Text
Resource Type	Article
Subject	Medicine Bioengineering Applied Microbiology and Biotechnology Biotechnology

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