Purification and Characterization of Proteases from Bacillus amyloliquefaciens Isolated from Traditional Soybean Fermentation Starter

Bacillus amyloliquefaciens FSE-68 isolated from meju, a Korean soybean fermentation starter, was identified on the basis of biophysical tests and 16S rRNA gene sequence. A neutral metalloprotease (NPR68) and an alkaline serine-protease (APR68) were purified by ammonium sulfate precipitation and cati...

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Bibliographic Details
Published in:Journal of agricultural and food chemistry 2003-12, Vol.51 (26), p.7664-7670
Main Authors: Cho, Seong-Jun, Oh, Sung-Hoon, Pridmore, R. David, Juillerat, Marcel A, Lee, Cherl-Ho
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Ammonium Sulfate
Animal, plant, fungal and microbial proteins, edible seaweeds and food yeasts
Bacillus - enzymology
Bacillus - genetics
Bacterial Proteins
Biological and medical sciences
Cations
Chemical Precipitation
Chromatography, Ion Exchange
Endopeptidases - chemistry
Endopeptidases - isolation & purification
Endopeptidases - metabolism
Enzyme Stability
Fermentation
Food industries
Fundamental and applied biological sciences. Psychology
Glycine max
Hydrogen-Ion Concentration
Metalloendopeptidases - chemistry
Metalloendopeptidases - metabolism
Metalloproteases - chemistry
Metalloproteases - isolation & purification
Metalloproteases - metabolism
Molecular Sequence Data
Phylogeny
Protease Inhibitors - pharmacology
Serine Endopeptidases - chemistry
Serine Endopeptidases - isolation & purification
Serine Endopeptidases - metabolism
Substrate Specificity
Subtilisin - chemistry
Subtilisin - metabolism
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Summary:Bacillus amyloliquefaciens FSE-68 isolated from meju, a Korean soybean fermentation starter, was identified on the basis of biophysical tests and 16S rRNA gene sequence. A neutral metalloprotease (NPR68) and an alkaline serine-protease (APR68) were purified by ammonium sulfate precipitation and cation exchange chromatography and identified on the basis of their activities at different pH values and the selective protease inhibitors. The molecular weights of NPR68 and APR68 measured with ESI-MS were 32743 (± 0.8) and 27443 (± 0.5) Da, respectively. Against oxidized insulin chains, the NPR68 has a cleavage preference at the site where leucine is located as a P1‘ residue followed by phenylalanine, and the APR68 has broad specificity and favors leucine at the P1 site. These results indicate that the proteases are natural variants of subtilisin and bacillolysin. Keywords: Bacillus amyloliquefaciens; strain identification; protease; specificity; insulin; purification; meju
ISSN:0021-8561
1520-5118
DOI:10.1021/jf0259314