Truncation of N- and C-terminal regions of Streptococcus mutans dextranase enhances catalytic activity.
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概要
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Multiple forms of native and recombinant endo-dextranases (Dexs) of the glycoside hydrolase family (GH) 66 exist. The GH 66 Dex gene from Streptococcus mutans ATCC 25175 (SmDex) was expressed in Escherichia coli. The recombinant full-size (95.4 kDa) SmDex protein was digested to form an 89.8 kDa isoform (SmDex90). The purified SmDex90 was proteolytically degraded to more than seven polypeptides (23-70 kDa) during long storage. The protease-insensitive protein was desirable for the biochemical analysis and utilization of SmDex. GH 66 Dex was predicted to comprise four regions from the N- to C-termini: N-terminal variable region (N-VR), conserved region (CR), glucan-binding site (GBS), and C-terminal variable region (C-VR). Five truncated SmDexs were generated by deleting N-VR, GBS, and/or C-VR. Two truncation-mutant enzymes devoid of C-VR (TM-NCGΔ) or N-VR/C-VR (TM-ΔCGΔ) were catalytically active, thereby indicating that N-VR and C-VR were not essential for the catalytic activity. TM-ΔCGΔ did not accept any further protease-degradation during long storage. TM-NCGΔ and TM-ΔCGΔ enhanced substrate hydrolysis, suggesting that N-VR and C-VR induce hindered substrate binding to the active site.
- springerの論文
著者
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森 春英
北海道大学大学院農学研究科応用生命科学専攻
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森 春英
北海道大学大学院農学研究院応用生命科学部門
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Kim Young-min
Graduate School Of Agriculture Hokkaido University
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Nakai Hiroyuki
Graduate School Of Agriculture Hokkaido University
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木村 淳夫
北海道大学大学院農学研究院応用生命科学部門
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Matsui Hirokazu
Dept. Biosci. Chem. Fac. Agric. Hokkaido Univ.
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Matsui Hirokazu
Hokkaido Univ. Sapporo Jpn
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Kim Doman
Research Institute For Catalysis Chonnam National University:research Center For New-biomaterials In
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Kim Doman
School Of Biological Sciences And Technology And Institute Of Bioindustrial Technology Jeonnam Natio
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Kobayasi Kazuhiro
Department Of Agriculture Faculty Of Life And Environmental Science Shimane University
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Kimura Atsuo
Hokkaido Univ.
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Kimura Atsuo
Graduate School Of Agriculture Hokkaido University
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Kim Young-min
Hokkaido Univ.
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Kim Young-min
Hokkaido University
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Kim Young-min
Special Business Division Koreaabrasive Ind. Co. Ltd.
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Okuyama Masayuki
Research Faculty Of Agriculture Hokkaido University
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Kimura Atsuo
Hokkaido University
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Funane K
National Food Research Institute
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Matsui Hirokazu
Laboratory Of Biochemistry Research Faculty Of Agriculture Hokkaido University
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Mori Haruhide
Research Faculty Of Agriculture Hokkaido University
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