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Metabolic engineering of Escherichia coli for enhanced biosynthesis of poly(3-hydroxybutyrate) based on proteome analysis |
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| Authors: | Seung Hwan Lee Kyoung-Hee Kang Eun Young Kim Tong Un Chae Young Hoon Oh Soon Ho Hong Bong Keun Song Jonggeon Jegals Si Jae Park Sang Yup Lee |
| Affiliation: | 1. Industrial Biochemicals Research Group, Division of Convergence Chemistry, Research Center for Biobased Chemistry, Korea Research Institute of Chemical Technology, P.O. Box 107, 141 Gajeong-ro, Yuseong-gu, Daejeon, 305-600, Republic of Korea 3. Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 Program), Center for Systems and Synthetic Biotechnology, Institute for the BioCentury, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon, 305-701, Republic of Korea 5. School of Chemical Engineering and Bioengineering, University of Ulsan, 93 Daehak-ro, Nam-gu, Ulsan, 680-749, Republic of Korea 2. Department of Environmental Engineering and Energy, Department of Energy and Biotechnology, Myongji University, San 38-2, Nam-dong, Cheoin-gu, Yongin-si, Gyeonggido, 449-728, Republic of Korea 4. Department of Bio and Brain Engineering, Department of Biological Sciences, BioProcess Engineering Research Center, and Bioinformatics Research Center, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon, 305-701, Republic of Korea |
| Abstract: | We have previously analyzed the proteome of recombinant Escherichia coli producing poly(3-hydroxybutyrate) [P(3HB)] and revealed that the expression level of several enzymes in central metabolism are proportional to the amount of P(3HB) accumulated in the cells. Based on these results, the amplification effects of triosephosphate isomerase (TpiA) and fructose-bisphosphate aldolase (FbaA) on P(3HB) synthesis were examined in recombinant E. coli W3110, XL1-Blue, and W lacI mutant strains using glucose, sucrose and xylose as carbon sources. Amplification of TpiA and FbaA significantly increased the P(3HB) contents and concentrations in the three E. coli strains. TpiA amplification in E. coli XL1-Blue lacI increased P(3HB) from 0.4 to 1.6 to g/l from glucose. Thus amplification of glycolytic pathway enzymes is a good strategy for efficient production of P(3HB) by allowing increased glycolytic pathway flux to make more acetyl-CoA available for P(3HB) biosynthesis. |
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