Biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) by recombinant Escherichia coli harboring propionyl-CoA synthase gene (prpE) or propionate permease gene (prpP) |
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| Authors: | Xiao-Wu Liu Hong-Hui Wang Jing-Yu Chen Xiao-Tao Li Guo-Qiang Chen |
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| Affiliation: | 1. Multidisciplinary Research Center, Shantou University, Shantou 515063, China;2. Department of Biological Sciences and Biotechnology, Tsinghua University, Beijng 100084, China;3. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China;1. MOE Key Lab of Bioinformatics, Department of Biological Science and Biotechnology, School of Life Science, Tsinghua-Peking Center for Life Sciences, Tsinghua University, Beijing 100084, China;2. School of Biological Sciences and Biotechnology, Xinjiang University, Urumchi 830046, China;1. Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 05029, South Korea;2. Institute for Ubiquitous Information Technology and Applications (CBRU), Konkuk University, Seoul 05029, South Korea;3. Intelligent Sustainable Materials R&D Group, Korea Institute of Industrial Technology (KITECH), Chonan 31056, South Korea;4. Department of Chemical Engineering, Soongsil University, Seoul 06978, South Korea;1. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, United States;2. Beijing Key Laboratory of Bioprocess, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, China;1. Unidad Microbiología Molecular - Depto. BIOGEM, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Av. Italia 3318, Montevideo, CP11600, Uruguay;2. Laboratorio de Carbohidratos y Glicoconjugados, Instituto de Higiene, Av. Alfredo Navarro 3051, Montevideo, CP11600, Uruguay;3. Nevada City Biolabs,14175 Dirt Rd., Nevada City, CA, 95959, United States |
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| Abstract: | In order to enhance 3-hydroxyvalerate (3HV) fraction in copolyesters of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), the propionate permease gene prpP or the propionyl-CoA synthase gene prpE was transformed into Escherichia coli XL10-Gold with co-expression of PHB operon (phaCAB) from Ralstonia eutropha. The recombinant E. coli strains were cultured on mixed carbon sources composed of glucose and propionic acid to promote PHBV accumulation. It was shown that the over-expression of prpE suppressed 3HV incorporation into PHBV copolymer, which led to reduced 3HV fraction. In contrast, the over-expression of prpP improved the 3HV content from 5.6 to 14.3 mol%, followed by an increased PHBV accumulation up to 62 wt%. The results showed that the expression of prpP stimulated the uptake and utilization of propionic acid and increased the 3HV fraction in PHBV. However, the over-expression of prpE in E. coli did not affect 3HV content in PHBV. Surprisingly, co-expression of prpE and prpP did not lead to any 3HV formation. This study showed the possibility to change the PHBV composition without overdose of propionic acid which is expensive and toxic for the cells. |
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