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Geranyl diphosphate synthase: An important regulation point in balancing a recombinant monoterpene pathway in Escherichia coli

Paper ID Volume ID Publish Year Pages File Format Full-Text
16968 42627 2015 6 PDF Available
Title
Geranyl diphosphate synthase: An important regulation point in balancing a recombinant monoterpene pathway in Escherichia coli
Abstract

•A monoterpene (geraniol) biosynthesis pathway was re-engineered in E. coli.•GPP synthase is indicated as an important regulation point for geraniol production.•Geraniol production was improved from 183 mg/L to 1119 mg/L.•Non-optimized expression of GPP synthase causes metabolic imbalance and plasmid instability.

The expression level of geranyl diphosphate synthase (GPPS) was suspected to play a key role for geraniol production in recombinant Escherichia coli harboring an entire mevalonate pathway operon and a geraniol synthesis operon. The expression of GPPS was optimized by using ribosomal binding sites (RBSs) designed to have different translation initiation rates (TIRs). The RBS strength in TIR window of 500 arbitrary unit (au)–1400 au for GPPS appears to be suitable for balancing the geraniol biosynthesis pathway in this study. With the TIR of 500 au, the highest production titer of geraniol was obtained at a level of 1119 mg/L, which represented a 6-fold increase in comparison with the previous titer of 183 mg/L. The TIRs of GPPS locating out of range of the optimal window (500–1400 au) caused significant decreases of cell growth and geraniol production. It was suspected to result from metabolic imbalance and plasmid instability in geraniol production by inappropriate expression level of GPP synthase. Our results collectively indicated GPPS as an important regulation point in balancing a recombinant geraniol synthesis pathway. The GPPS-based regulation approach could be applicable for optimizing microbial production of other monoterpenes.

Keywords
Escherichia coli; Geraniol; GPP synthesis; Mevalonate pathway; Plasmid maintenance; RBS strength
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Geranyl diphosphate synthase: An important regulation point in balancing a recombinant monoterpene pathway in Escherichia coli
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 68, January 2015, Pages 50–55
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us