Engineering poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer composition in E. coli

A strain of Escherichia coli was metabolically engineered to produce poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) of specified composition between 5% and 18% HV. A gene encoding propionyl-CoA synthetase (prpE from S. enterica) was placed under the control of the IPTG-inducible tac promoter (P...

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Bibliographic Details
Published in:Biotechnology and bioengineering 2008-03, Vol.99 (4), p.919-928
Main Authors: Wong, Matthew S, Causey, Thomas B, Mantzaris, Nikos, Bennett, George N, San, Ka-Yiu
Format: Article
Language:English
Subjects:
Acyltransferases - genetics
Acyltransferases - metabolism
Biological and medical sciences
Biotechnology
Coenzyme A Ligases - genetics
Coenzyme A Ligases - metabolism
Copolymers
Correlation analysis
E coli
Escherichia coli
Escherichia coli - physiology
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genetic Enhancement - methods
Molecular weight
PHBV
Polyesters - metabolism
prpE
specified composition
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Summary:A strain of Escherichia coli was metabolically engineered to produce poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) of specified composition between 5% and 18% HV. A gene encoding propionyl-CoA synthetase (prpE from S. enterica) was placed under the control of the IPTG-inducible tac promoter (PtaclacUV₅) while the polyhydroxyalkanoate synthesis operon (phaBCA) from R. eutropha was expressed constitutively. A strain of E. coli harboring both plasmids was grown in defined medium and PHBV was produced with specified hydroxyvalerate (HV) molar content between 5% and 18%. The molecular weight of the copolymer was ~700,000 across various HV contents, and average polydispersity was ~ 1.3. The majority of the PHBV production occurred during the late exponential/stationary phase. The HV content of the copolymer generally peaked early in the incubation before falling to its final value. We found that the time profiles of PrpE activity, propionyl-CoA, and acetyl-CoA were well correlated to the HV content time profile. Despite an abundance of propionyl-CoA, incorporation of HV into the copolymer was inefficient. Therefore, both the PHA operon and conditions affecting the availability of propionyl-CoA must be chosen carefully to achieve the desired HV content. The ability to engineer copolymer composition control into an E. coli strain would be useful in cases where the feedstock composition is not adjustable. Biotechnol. Bioeng. 2008;99: 919-928. © 2007 Wiley Periodicals, Inc.
ISSN:0006-3592
1097-0290
DOI:10.1002/bit.21641