Journal of Microbiology

  • 제40권2호
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  • Pages.129-133
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  • 2002
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  • 1225-8873(pISSN)
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  • 1976-3794(eISSN)
한국미생물학회 (The Microbiological Society of Korea)
권호 표지

Purification and Characterization of Poly(3-hydroxybutyrate) Depolymerase from a Fungal Isolate, Emericellopsis minima W2

  • Rhee, Young-Ha (Department of Microbiology, Chungnam National University, Daejeon 305-764) ;
  • Kim, Do-Young (Department of Microbiology, Chungnam National University, Daejeon 305-764) ;
  • Yun, Ji-Hye (Department of Microbiology, Chungnam National University, Daejeon 305-764) ;
  • Kim, Hyung-Woo (Institute of Biotechnology, Chungnam National University, Daejeon 305-764) ;
  • Bae, Kyung-Sook (Korea Research Institute of Biotechnology, Daejeon 305-600)
  • 발행 : 2002.06.01
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초록

The fungus, Emericellopsis minima W2, capable of degrading poly(3-hydroxybutyrate) (PHB) was isolated from a waste water sample. Production of the PHB depolymerase from E. minima W2 (PhaZ/ sub Emi/) was significantly repressed in the presence of glucose. PhaZ/ sub Emi/ was purified by column chromatography on Octyl-Sepharose CL-4B and Sephadex G-100. The molecular mass of the PhaZ/ sub Emi/), which consisted of a single polypeptide chain, was estimated to be 48.0 kDa by SDS-PAGE and its pI vague was 4.4. The maximum activity of the PhaZ/ sub Emi/ was observed at pH 9.0 and 55$\^{C}$. It was significantly inactivated by 1mM dithiothreitol, 2mM diisopropyl fluorphosphate, 0.1mM Tween 80, and 0.1 mM Triton X-l00, but insensitive to phenylmethylsulfonyl fluoride and N-ethylmaleimide. The PhaZ/ sub Emi/ efficiently hydrolyzed PHB and its copolyester with 30 mol% 3-hydroxyvalerate, but did not act on poly(3-hydroxyoctanoate). It also hydrolyzed p-nitrophenylacetate and p-nitrophenylbutyrate but hardly affected the longer-chain forms. The main hydrolysis product of PHB was identified as a dimer of 3-hydroxybutyrate.

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