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Current Microbiology

, Volume 14, Issue 1, pp 7–12 | Cite as

An alkaline proteinase of an alkalophilicBacillus sp.

  • Osamu Tsuchida
  • Yohei Yamagata
  • Takehiko Ishizuka
  • Teruyoshi Arai
  • Jun-Ichi Yamada
  • Michio Takeuchi
  • Eiji Ichishima
Article

Abstract

An alkaline serine proteinase was purfied from the culture broth of an alkalophilicBacillus sp. NKS-21. The molecular weight was estimated to be 22,000 by a gel filtration method and 31,000 by SDS-polyacrylamide gel electrophoresis. The isoelectric point was determined to be 8.2. The amino acid composition and CD spectrum were determined. The alkaline proteinase had a pH optimum at 10–11 for milk casein digestion. The specific activity of the alkaline proteinase was 0.35 katal/kg of protein at pH 10.0 for milk casein hydrolysis.

The substrate specificity of the alkaline proteinase was studied by using the oxidized, insulin B-chain and angiotensin. An initial cleavage site was observed at Leu15-Tyr16, secondary site at Leu11-Val12, and additional sites at Gln4-His5, Tyr26-Thr27, and Asn3-Gln4 in the oxidized insulin B-chain at pH 10.0. In comparison with the subtilisins Carlsberg and Novo, the alkaline proteinase fromBacillus sp. showed a unique specificity toward the oxidized insulin B-chain. Hydrolysis of angiotensin at pH 10.0 with the alkaline proteinase was observed at Tyr4-Ile5. The proteinase has aK m of 0.1 mM andk cat of 3.3 s−1 with angiotensin as substrate.

Keywords

Angiotensin Serine Proteinase Amino Acid Composition Alkaline Proteinase Casein Hydrolysis 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag New York Inc. 1986

Authors and Affiliations

  • Osamu Tsuchida
    • 1
  • Yohei Yamagata
    • 1
  • Takehiko Ishizuka
    • 1
  • Teruyoshi Arai
    • 1
  • Jun-Ichi Yamada
    • 1
  • Michio Takeuchi
    • 1
  • Eiji Ichishima
    • 1
  1. 1.Laboratory of Enzymology and Microbial ChemistryTokyo Noko UniversityFuchu, TokyoJapan

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