Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Molecular Structure of PCR Cloned PHA Synthase Genes of Pseudomonas putida KT2440 and Its Utilization for Medium-Chain Length Polyhydroxyalkanoate Production

  • Kim, Tae-Kwon (Department of Genetic Engineering, College of Natural Sciences, Kyungpook National University) ;
  • Shin, Hyun-Dong (Department of Genetic Engineering, College of Natural Sciences, Kyungpook National University) ;
  • Seo, Min-Cheol (Department of Genetic Engineering, College of Natural Sciences, Kyungpook National University) ;
  • Lee, Jin-Nam (Department of Genetic Engineering, College of Natural Sciences, Kyungpook National University) ;
  • Lee, Yong-Hyun (Department of Genetic Engineering, College of Natural Sciences, Kyungpook National University)
  • Published : 2003.04.01
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Abstract

A new phaC gene cluster encoding polyhydroxyalkanoate (PHA) synthase I PHA depolymerase, and PHA synthase II was cloned using the touchdown PCR method, from medium-chain length (mcl-) PHA-producing strain Pseudomonas putida KT2440. The molecular structure of the cloned phaCl gene was analyzed, and the phylogenic relationship was compared with other phaCl genes cloned from Pseudomonas species. The cloned phaCl gene was expressed in a recombinant E. coli to the similar level of PHA synthase in the parent strain P. putida KT2440, but no significant amount of mcl-PHA was accumulated. The isolated phaCl gene was re-introduced into the parent strain P. putida KT2440 to amplify the PHA synthase I activity, and the recombinant P. purida accumulated mcl-PHA more effectively, increasing from 26.6 to $43.5\%$. The monomer compositions of 3-hydroxylalkanoates in mcl-PHA were also modified significantly in the recombinant P. putida enforcing the cloned phaCl gene.

Keywords

References

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