Purification and characterization of two novel halotolerant extracellular proteases from Bacillus subtilis strain FP-133

• Published in: Bioscience, biotechnology, and biochemistry Vol. 70; no. 2; pp. 433 - 440
• Main Authors: Setyorini, E.(Kobe Univ. (Japan)), Takenaka, S, Murakami, S, Aoki, K
• Format: Journal Article
• Language: English
• Published: Tokyo Japan Society for Bioscience, Biotechnology, and Agrochemistry 01.02.2006; Japan Society for Bioscience Biotechnology and Agrochemistry; Oxford University Press
• Subjects: ACTIVIDAD CATALITICA; ACTIVITE CATALYTIQUE; BACILLUS SUBTILIS; Bacillus subtilis - drug effects; Bacillus subtilis - enzymology; Biological and medical sciences; Carbon - pharmacology; Catalysis - drug effects; CATALYTIC ACTIVITY; Cations - chemistry; Culture Media, Conditioned; endoprotease; Enzyme Stability; Fe-containing metalloprotease; FISH PASTES; Fundamental and applied biological sciences. Psychology; halotolerant protease; Hydrogen-Ion Concentration; Metals - chemistry; Metals - pharmacology; Molecular Weight; neutral serine protease; Nitrogen - pharmacology; PASTAS DE PESCADO; PATE DE POISSON; Peptide Hydrolases - isolation & purification; Peptide Hydrolases - metabolism; Peptide Hydrolases - secretion; PROTEASAS; PROTEASE; Protease Inhibitors - pharmacology; PROTEASES; Protein Denaturation; PURIFICACION; PURIFICATION; SALINIDAD; SALINITE; SALINITY; SALT TOLERANCE; Sodium Chloride - pharmacology; Substrate Specificity; Temperature; TOLERANCE AU SEL; TOLERANCIA A LA SAL; Halotolerance; Purification; Serine endopeptidases; Enzyme; Bacillus subtilis; neutral serine protease; Metalloendopeptidases; Bacillaceae; Extracellular; Characterization; Peptidases; Bacillales; endoprotease; Fe-containing metalloprotease; Bacteria; Hydrolases; halotolerant protease
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.70.433

Bacillus subtilis strain FP-133, isolated from a fermented fish paste, synthesized two novel halotolerant extracellular proteases (expro-I and expro-II), showing activity and stability at concentrations of 0-20% (w/v) NaCl. Each protease was purified to homogeneity and characterized. The purified expro-1 was a non-alkaline serine protease with an optimum pH of 7.5, although most serine proteases from Bacillus strains act at the alkaline side. The molecular mass of expro-I was 29 kDa. The purified expro-II was a metalloprotease with a molecular mass of 34 kDa. It was activated by Fesup(2+) which has never been reported as a bacterial protease activator. At a concentration of 7.5% (w/v) NaCl, both proteases preferred animal proteins to vegetable proteins as natural substrates. In addition, under saline conditions, expro-I and II showed high catalytic activity toward gelatin and casein respectively.