Session 2—Introduction to Microbial Catalysis and Engineering

Applied Biochemistry and Biotechnology

, Volume 114, Issue 1, pp 361-372

First online:

Construction of recombinant Escherichia coli strais for production of poly-(3-hydroxybutyrate-co-3-hydroxyvalerate)

  • Kin-Ho LawAffiliated withOpen Laboratory of Chirotechnology of the Institute of Molecular Technology for Drug Discovery & Synthesis, Applied Biology and Chemical Technology, Hong Kong Polytechnic University
  • , Pui-Ling ChanAffiliated withOpen Laboratory of Chirotechnology of the Institute of Molecular Technology for Drug Discovery & Synthesis, Applied Biology and Chemical Technology, Hong Kong Polytechnic University
  • , Wai-Sum LauAffiliated withOpen Laboratory of Chirotechnology of the Institute of Molecular Technology for Drug Discovery & Synthesis, Applied Biology and Chemical Technology, Hong Kong Polytechnic University
  • , Yin-Chung ChengAffiliated withOpen Laboratory of Chirotechnology of the Institute of Molecular Technology for Drug Discovery & Synthesis, Applied Biology and Chemical Technology, Hong Kong Polytechnic University
  • , Yun-Chung LeungAffiliated withOpen Laboratory of Chirotechnology of the Institute of Molecular Technology for Drug Discovery & Synthesis, Applied Biology and Chemical Technology, Hong Kong Polytechnic University
  • , Wai-Hung LoAffiliated withOpen Laboratory of Chirotechnology of the Institute of Molecular Technology for Drug Discovery & Synthesis, Applied Biology and Chemical Technology, Hong Kong Polytechnic University
  • , Hugh LawfordAffiliated withBioengineering Laboratory, Department of Biochemistry, University of Toronto
  • , Hoi-Fu YuAffiliated withOpen Laboratory of Chirotechnology of the Institute of Molecular Technology for Drug Discovery & Synthesis, Applied Biology and Chemical Technology, Hong Kong Polytechnic University Email author 

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Abstract

Plastic wastes constitute a worldwide environmental problem, and the demand for biodegradable plastics has become high. One of the most important characteristics of microbial polyesters is that they are thermoplastic with environmentally degradable properties. In this study, pUC 19/PHA was cloned and transformed into three different Escherichia coli strains. Among the three strains that were successfully expressed in the production of polyhydroxyalkanoates (PHA), E. coli HMS174 had the highest yield in the production of poly-(3-hydroxybutyrate-co-3-hydroxyvalerate) (P[HB-HV]). The cell dry weight and PHA content of recombinant HMS174 reached as high as 10.27 g/L and 43% (w/w), respectively, in fed-batch fermentor culture. The copolymer of PHA, P(HB-HV), was found in the cells, and the biopolymers accumulated were identified and analyzed by gas chromatography, proton nuclear magnetic resonance spectroscopy, and differential scanning calorimetry. We demonstrated clearly that the E. coli host for PHA production has to be carefully selected to obtain a high yield. The results obtained indicated that a superior E. coli with high PHA production can be constructed with a desirable ratio of P(HB-HV), which has potential applications in industry and medicine.

Index Entries

Escherichia coli Polyhydroxyalkanoates fed-batch fermentation nuclear magnetic resonance differential scanning calorimetry