Purification and Characterization of Serine Proteinase from a Halophilic Bacterium, Filobacillus sp. RF2-5
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概要
- 論文の詳細を見る
In order to find a unique proteinase, proteinase-producing bacteria were screened from fish sauce in Thailand. An isolated moderately halophilic bacterium was classified and named Filobacillus sp. RF2-5. The molecular weight of the purified enzyme was estimated to be 49 kDa. The enzyme showed the highest activity at 60 °C and pH 10–11 under 10% NaCl, and was highly stable in the presence of about 25% NaCl. The activity was strongly inhibited by phenylmethane sulfonyl fluoride (PMSF), chymostatin, and α-microbial alkaline proteinase inhibitor (MAPI). Proteinase activity was activated about 2-fold and 2.5-fold by the addition of 5% and 15–25% NaCl respectively using Suc–Ala–Ala–Phe–pNA as a substrate. The N-terminal 15 amino acid sequence of the purified enzyme showed about 67% identity to that of serine proteinase from Bacillus subtilis 168 and Bacillus subtilis (natto). The proteinase was found to prefer Phe, Met, and Thr at the P1 position, and Ile at the P2 position of peptide substrates, respectively. This is the first serine proteinase with a moderately thermophilic, NaCl-stable, and NaCl-activatable, and that has a unique substrate specificity at the P2 position of substrates from moderately halophilic bacteria, Filobacillus sp.
- 社団法人 日本農芸化学会の論文
- 2005-01-23
著者
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Hiraga Kazumi
Department of Applied Biology, Kyoto Institute of Technology
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Takada Katsumi
Peptide Institute, Inc.
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ODA Kohei
Department of Applied Biology, Kyoto Institute of Technology
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TANASUPAWAT Somboon
Department of Microbiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University
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TANASUPAWAT Somboon
Chulalongkon大学 薬学部 微生物学科
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Hiraga Kazumi
Department Of Applied Biology Faculty Of Textile Science Kyoto Institute Of Technology
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Oda K
National Research Inst. Brewing Tokyo Jpn
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Oda Kohei
Department Of Agricultural Chemistry College Of Agriculture University Of Osaka Prefecture
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Tanasupawat Somboon
Department Of Agricultural Chemistry Tokyo University Of Agriculture
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Tanasupawat Somboon
Dep. Of Biochemistry And Microbiology Fac. Of Pharmaceutical Sciences Chulalongkorn Univ.
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Tanasupawat S
Chulalongkorn Univ. Bangkok Tha
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NAMWONG Sirilak
Department of Microbiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University
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NISHIKATA Yasushi
Department of Applied Biology, Faculty of Textile Science
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Namwong Sirilak
Department Of Microbiology Faculty Of Pharmaceutical Sciences Chulalongkorn University
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Hiraga Kazumi
Department Of Applied Biology Kyoto Institute Of Technology Matsugasaki
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Takada Katsumi
Peptide Institute Inc.
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Nishikata Yasushi
Department Of Applied Biology Faculty Of Textile Science
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