Journal of the Korean Wood Science and Technology

The Korean Society of Wood Science & Technology (한국목재공학회)
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Transformation of Terpene Synthase from Polyporus brumalis in Pichia pastoris for Recombinant Enzyme Production

  • An, Ji-Eun (Division of Wood Chemistry, Department of Forest Products, National Institute of Forest Science) ;
  • Lee, Su-Yeon (Forest Biomaterials Research Center, National Institute of Forest Science) ;
  • Ryu, Sun-Hwa (Division of Forest Industry Research, Department of Forest Policy and Economics, National Institute of Forest Science) ;
  • Kim, Myungkil (Division of Global Forestry, Department of Forest Policy and Economics, National Institute of Forest Science)
  • Received : 2018.05.24
  • Accepted : 2018.07.10
  • Published : 2018.07.25
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Abstract

Terpenoids have a wide range of biological functions and have extensive applications in the pharmaceutical, cosmetic, and flavoring industry. The white-rot fungus, Polyporus brumalis, is able to synthesize terpenoids via terpene synthase, which catalyzes an important step that forms a large variety of sesquiterpene products from farnesyl pyrophosphate (FPP). To improve the production of sesquiterpenes, the terpene synthase gene was isolated from Polyporus brumalis and was heterologously transformed into a Pichia pastoris strain. The open reading frame of the isolated gene (approximately 1.2 kb) was inserted into Pichia pastoris to obtain a recombinant enzyme. Five transformants were obtained and the expression of terpene synthase was analyzed at the transcript level by reverse transcription PCR (polymerase chain reaction) and at the protein level by SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis). Expression of the terpene synthase gene product was elevated in the transformants and as expected the molecular weight of the protein was approximately 45 kDa. These recombinant enzymes have potential practical applications and future studies should focus on their functional characterization.

Keywords

References

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