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The journal of microbiology v.43 no.3, 2005년, pp.285 - 294   피인용횟수: 2

Molecular Characterization of Extracellular Medium-chain-length Poly(3-hydroxyalkanoate) Depolymerase Genes from Pseudomonas alcaligenes Strains

Kim Do Young    (Department of Microbiology, School of Bioscience and Biotechnology, Chungnam National University   ); Kim Hyun Chul    (Department of Microbiology, School of Bioscience and Biotechnology, Chungnam National University   ); Kim Sun Young    (Department of Microbiology, School of Bioscience and Biotechnology, Chungnam National University   ); Rhee Young Ha    (Department of Microbiology, School of Bioscience and Biotechnology, Chungnam National University  );
  • 초록

    A bacterial strain M4-7 capable of degrading various polyesters, such as poly $(\varepsilon-caprolactone)$ , poly(3-hydroxybutyrate-co-3-hydroxyvalerate), poly(3-hydroxyoctanoate), and poly(3-hydroxy-5-phenylvalerate), was isolated from a marine environment and identified as Pseudomonas alcaligenes. The relative molecular mass of a purified extracellular medium-chain-length poly(3-hydroxyalkanoate) (MCL-PHA) depolymerase $(PhaZ_{palM4-7})$ from P. alcaligenes M4-7 was 28.0 kDa, as determined by SDS-PAGE. The $PhaZ_{palM4-7}$ was most active in 50 mM glycine-NaOH buffer (pH 9.0) at $35^{\circ}C$ . It was insensitive to dithiothreitol, sodium azide, and iodoacetamide, but susceptible to p-hydroxymercuribenzoic acid, N-bromosuccinimide, acetic anhydride, EDTA, diisopropyl fluorophosphate, phenylmethylsulfonyl fluoride, Tween 80, and Triton X-100. In this study, the genes encoding MCL-PHA depolymerase were cloned, sequenced, and characterized from a soil bacterium, P. alcaligenes LB19 (Kim et al., 2002, Biomacro-molecules 3, 291-296) as well as P. alcaligenes M4-7. The structural gene $(phaZ_{palLB19})$ of MCL-PHA depolymerase of P. alcaligenes LB19 consisted of an 837 bp open reading frame (ORF) encoding a protein of 278 amino acids with a deduced $M_r$ of 30,188 Da. However, the MCL-PHA depolymerase gene $(phaZ_{palM4-7})$ of P. alcaligenes M4-7 was composed of an 834 bp ORF encoding a protein of 277 amino acids with a deduced Mr of 30,323 Da. Amino acid sequence analyses showed that, in the two different polypeptides, a substrate-binding domain and a catalytic domain are located in the N-terminus and in the C-terminus, respectively. The $PhaZ_{palLB19}$ and the $PhaZ_{palM4-7}$ commonly share the lipase box, GISSG, in their catalytic domains, and utilize $^{111}Asn$ and $^{110}Ser$ residues, respectively, as oxyanions that play an important role in transition-state stabilization of hydrolytic reactions.


  • 주제어

    Medium-chain-length polyhydroxyalkanoate .   polyhydroxyalkanoate depolymerase .   Pseudomonas alcaligenes.  

  • 참고문헌 (27)

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  • 이 논문을 인용한 문헌 (2)

    1. 2006. "" Journal of microbiology and biotechnology, 16(12): 1935~1939     
    2. 2007. "" The journal of microbiology, 45(2): 87~97     

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