High-efficiency L-lactate production from glycerol by metabolically engineered Escherichia coli.
- Author:
Kangming TIAN
1
;
Guiyang SHI
2
;
Fuping LU
3
;
Suren SINGH
4
;
Zhengxiang WANG
3
Author Information
1. School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China.
2. National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, Jiangsu, China.
3. College of Biotechnology, Tianjin University of Science & Technology, Tianjin 300457, China.
4. Faculty of Applied Sciences, Department of Biotechnology & Food Technology, Durban University of Technology, PO Box 1334, Durban 4001, South Africa.
- Publication Type:Journal Article
- MeSH:
Bacillus;
enzymology;
Cloning, Molecular;
Escherichia coli;
enzymology;
genetics;
metabolism;
Glycerol;
metabolism;
L-Lactate Dehydrogenase;
genetics;
metabolism;
Lactic Acid;
biosynthesis;
Metabolic Engineering;
Promoter Regions, Genetic;
genetics;
Recombinant Proteins;
genetics;
metabolism
- From:
Chinese Journal of Biotechnology
2013;29(9):1268-1277
- CountryChina
- Language:Chinese
-
Abstract:
High-efficient conversion of glycerol to L-lactate is beneficial for the development of both oil hydrolysis industry and biodegradable materials manufacturing industry. In order to construct an L-lactate producer, we first cloned a coding region of gene BcoaLDH encoding an L-lactate dehydrogenase from Bacillus coagulans CICIM B1821 and the promoter sequence (P(ldhA)) of the D-lactate dehydrogenase (LdhA) from Escherichia coli CICIM B0013. Then we assembled these two DNA fragments in vitro and yielded an expression cassette, P(ldhA)-BcoaLDH. Then, the cassette was chromosomally integrated into an ldhA mutant strain, Escherichia coli CICIM B0013-080C, by replacing lldD encoding an FMN-dependent L-lactate dehydrogenase. An L-lactate higher-producer strain, designated as E. coli B0013-090B, possessing genotype of lldD::P(ldhA)-BcoaLDH, deltaack-pta deltapps deltapflB deltadld deltapoxB deltaadhE deltafrdA and deltaldhA, was generated. Under the optimal condition, 132.4 g/L L-lactate was accumulated by B0013-090B with the lactate productivity of 4.90 g/Lh and the yield of 93.7% in 27 h from glycerol. The optical purity of L-lactate in broth is above 99.95%.
