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

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Issued Date

2016

Resource Type

Article

File Type

application/pdf

ISSN

992240

DOI

10.1128/AEM.02178-16

Other identifier(s)

2-s2.0-84997173327

Rights Holder(s)

Scopus

Bibliographic Citation

Applied and Environmental Microbiology. Vol 82, No.24 (2016), p.7176-7184

Suggested Citation

Wang J., Niyompanich S., Tai Y.-S., Wang J., Bai W., Mahida P., Gao T., Zhang K. Engineering of a highly efficient Escherichia coli strain for mevalonate fermentation through chromosomal integration. Applied and Environmental Microbiology. Vol 82, No.24 (2016), p.7176-7184. doi:10.1128/AEM.02178-16 Retrieved from: https://hdl.handle.net/20.500.14740/5970

Title

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

Author(s)

Wang J.
 Niyompanich S.
 Tai Y.-S.
 Wang J.
 Bai W.
 Mahida P.
 Gao T.
 Zhang K.

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). © 2016, American Society for Microbiology. All Rights Reserved.

Subject(s)

Bottles
 Escherichia coli
 Genes
 Integration
 Metabolism
 Productivity
 Chromosomal integration
 Constitutive promoters
 Fed-batch fermentation
 Genetic instability
 Maximum productivity
 Metabolic pathways
 Mevalonate pathway
 Shake flask fermentations
 Fermentation
 Bioengineering
 Chromosome
 Fecal coliform
 Fermentation
 Gene expression
 Glucose
 Metabolism
 Plasmid
 Polymer
 Escherichia coli
 Glucose
 Mevalonic acid
 Bacterial chromosome
 Escherichia coli
 Fermentation
 Genetics
 Metabolic engineering
 Metabolism
 Plasmid
 Chromosomes, Bacterial
 Escherichia coli
 Fermentation
 Glucose
 Metabolic Engineering
 Mevalonic Acid
 Plasmids

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URI

https://hdl.handle.net/20.500.14740/5970

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Scopus: Year 1983-2021

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