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Conflict of Interest: In relation to this article, we declare that there is no conflict of interest.

Publication history: Received July 8, 2014
Accepted August 19, 2014

This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/bync/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Establishment of a biosynthesis pathway for (R)-3-hydroxyalkanoates in recombinant Escherichia coli

Si Jae Park Seung Hwan Lee¹ Young Hoon Oh¹ Sang Yup Lee^2†

Department of Environmental Engineering and Energy, Myongji University, 116, Myongji-ro, Cheoin-gu, Yongin, Gyeonggy 449-728, Korea ¹Industrial Biochemicals Research Group, Research Center for Biobased Chemistry, Division of Convergence Chemistry, Korea Research Institute of Chemical Technology, P. O. Box 107 141, Gajeong-ro, Yuseong-gu, Daejeon 305-600, Korea ²Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 Plus Program), Center for Systems and Synthetic Biotechnology, and Institute for the BioCentury, KAIST, 291, Daehak-ro, Yuseong-gu, Daejeon 305-701, Korea

Korean Journal of Chemical Engineering, April 2015, 32(4), 702-706(5), 10.1007/s11814-014-0240-y

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Abstract

A biosynthetic pathway for the production of (R)-3-hydroxyalkanoates (R3HAs) through in vivo depolymerization of poly(3-hydroxyalkanoates) [P(3HAs)] was constructed in recombinant Escherichia coli fadA mutant WA101 by introducing the Pseudomonas sp. 61-3 PHA synthase gene (phaC2) and the P. aeruginosa intracellular PHA depolymerase gene (phaZ). When recombinant E. coli WA101 strain expressing the phaC2 gene and the phaZ gene was cultured_x000D_ in Luria-Bertani (LB) medium containing 2 g/L of sodium decanoate, R3HAs could be produced to the concentration of 0.49 g/L. The mole fraction of R3HAs was 7.5mol% of 3-hydroxybutyrate (3HB), 31.6mol% of 3-hydroxyhexanoate (3HHx), 30 mol% of 3-hydroxyoctanoate (3HO), 29.4mol% of 3-hydroxydecanoate (3HD), and 1.5mol% of 3-hydroxydodecanoate (3HDD). When the E. coli 3-ketoacyl-ACP reductase gene (fabG) was overexpressed to provide more (R)-3-hydroxyacyl-CoA (R3HA-CoA), the concentration of R3HAs was increased up to 1.05 g/L. Also, expression of the fabG gene resulted in the mole fraction change of produced R3HAs, in which 3HD fraction was enriched from 29.4mol% to 57.9mol% with the decrease of 3HHx fraction from 31.6mol% to 9.6mol%. Interestingly, the only expression of the fabG gene in E. coli WA101 could produce R3HAs to 0.55 g/L, which suggests that E. coli might have unidentified CoA hydrolases that have substrate specificities toward R3HA-CoA. This study shows the enantiomerically pure RHAs can be efficiently produced by metabolically engineered E. coli with high yield.

Keywords

(R)-3-hydroxyalkanoate Escherichia coli Polyhydroxyalkanoate PHA Depolymerase In vivo Depolymerization

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