Purification and Characterization of Serine Proteinase from a Halophilic Bacterium,<i>Filobacillus</i>sp. RF2-5

• HIRAGA Kazumi

  Department of Applied Biology, Faculty of Textile Science

• NISHIKATA Yasushi

  Department of Applied Biology, Faculty of Textile Science

• NAMWONG Sirilak

  Department of Microbiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University

• TANASUPAWAT Somboon

  Department of Microbiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University

• TAKADA Katsumi

  Peptide Institute Inc.

• ODA Kohei

  Department of Applied Biology, Faculty of Textile Science

書誌事項

タイトル別名

• Purification and Characterization of Serine Proteinase from a Halophilic Bacterium, Filobacillus sp. RF2-5

抄録

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.

収録刊行物

• Ｂｉｏｓｃｉｅｎｃｅ，　Ｂｉｏｔｅｃｈｎｏｌｏｇｙ，　ａｎｄ　Ｂｉｏｃｈｅｍｉｓｔｒｙ

  Ｂｉｏｓｃｉｅｎｃｅ，　Ｂｉｏｔｅｃｈｎｏｌｏｇｙ，　ａｎｄ　Ｂｉｏｃｈｅｍｉｓｔｒｙ 69 (1), 38-44, 2005

  公益社団法人 日本農芸化学会

キーワード

• halophilic bacterium
• <I>Filobacillus</I> sp.
• serine proteinase
• fluorescence resonance energy transfer substrate (FRETS)

詳細情報

• CRID: 1390282681453289856
• NII論文ID: 130000030551
• NII書誌ID: AA10824164
• DOI: 10.1271/bbb.69.38
• ISSN: 13476947
• ISSN: 09168451
• NDL書誌ID: 7237141
• PubMed: 15665465
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I7237141
• Web Site: http://www.tandfonline.com/doi/pdf/10.1271/bbb.69.38
• 本文言語コード: en
• データソース種別:
  • JaLC
  • NDL
  • Crossref
  • PubMed
  • CiNii Articles
• 抄録ライセンスフラグ: 使用不可