Engineering of a highly efficient Escherichia coli strain for mevalonate fermentation through chromosomal integration

Jilong Wang, Suthamat Niyompanich, Yi Shu Tai, Jingyu Wang, Wenqin Bai, Prithviraj Mahida, Tuo Gao, Kechun Zhang

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Chromosomal integration of heterologous metabolic pathways is optimal for industrially relevant fermentation, as plasmidbased fermentation causes extra metabolic burden and genetic instabilities. In this work, chromosomal integration was adapted for the production of mevalonate, which can be readily converted into β-methyl-δ-valerolactone, a monomer for the production of mechanically tunable polyesters. The mevalonate pathway, driven by a constitutive promoter, was integrated into the chromosome of Escherichia coli to replace the native fermentation gene adhE or ldhA. The engineered strains (CMEV-1 and CMEV-2) did not require inducer or antibiotic and showed slightly higher maximal productivities (0.38 to~0.43 g/liter/h) and yields (67.8 to~71.4% of the maximum theoretical yield) than those of the plasmid-based fermentation. Since the glycolysis pathway is the first module for mevalonate synthesis, atpFH deletion was employed to improve the glycolytic rate and the production rate of mevalonate. Shake flask fermentation results showed that the deletion of atpFH in CMEV-1 resulted in a 2.1- fold increase in the maximum productivity. Furthermore, enhancement of the downstream pathway by integrating two copies of the mevalonate pathway genes into the chromosome further improved the mevalonate yield. Finally, our fedbatch fermentation showed that, with deletion of the atpFH and sucA genes and integration of two copies of the mevalonate pathway genes into the chromosome, the engineered strain CMEV-7 exhibited both high maximal productivity (~1.01 g/liter/h) and high yield (86.1% of the maximum theoretical yield, 30 g/liter mevalonate from 61 g/liter glucose after 48 h in a shake flask).

Original languageEnglish (US)
Pages (from-to)7176-7184
Number of pages9
JournalApplied and environmental microbiology
Volume82
Issue number24
DOIs
StatePublished - Jan 1 2016

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Mevalonic Acid
Fermentation
fermentation
engineering
Escherichia coli
chromosome
chromosomes
gene
liquid state fermentation
genes
productivity
Chromosomes
Genes
polyesters
glycolysis
biochemical pathways
plasmids
plasmid
antibiotics
Polyesters

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Engineering of a highly efficient Escherichia coli strain for mevalonate fermentation through chromosomal integration. / Wang, Jilong; Niyompanich, Suthamat; Tai, Yi Shu; Wang, Jingyu; Bai, Wenqin; Mahida, Prithviraj; Gao, Tuo; Zhang, Kechun.

In: Applied and environmental microbiology, Vol. 82, No. 24, 01.01.2016, p. 7176-7184.

Research output: Contribution to journalArticle

Wang, Jilong ; Niyompanich, Suthamat ; Tai, Yi Shu ; Wang, Jingyu ; Bai, Wenqin ; Mahida, Prithviraj ; Gao, Tuo ; Zhang, Kechun. / Engineering of a highly efficient Escherichia coli strain for mevalonate fermentation through chromosomal integration. In: Applied and environmental microbiology. 2016 ; Vol. 82, No. 24. pp. 7176-7184.
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