Purification and characterization of microbial protease produced extracellularly from Bacillus subtilis FBL-1

Si, J.-B., Jang, E.-J., Charalampopoulos, D. ORCID: https://orcid.org/0000-0003-1269-8402 and Wee, Y.-J. (2018) Purification and characterization of microbial protease produced extracellularly from Bacillus subtilis FBL-1. Biotechnology and Bioprocess Engineering, 23 (2). pp. 176-182. ISSN 1226-8372 doi: 10.1007/s12257-017-0495-3

Abstract/Summary

An ammonium sulfate precipitation of fermentation broth produced by Bacillus subtilis FBL-1 resulted in 2.9-fold increase of specific protease activity. An eluted protein fraction from the column chromatographies using DEAE-Cellulose and Sephadex G-75 had 94.2- and 94.9-fold higher specific protease activity, respectively. An SDS-PAGE revealed a band of purified protease at approximately 37.6 kDa. Although purified protease showed the highest activity at 45°C and pH 9.0, the activity remained stable in temperature range from 30 to 50°C and pH range from 7.0 to 9.0. Protease activity was activated by metal ions such as Ca2+, Mg2+, Mn2+, Fe2+, Ca2+ and K+, but 10 mM Fe3+ significantly inhibited enzyme activity (53%). Protease activity was inhibited by 2 mM EDTA as a metalloprotease inhibitor, but it showed good stability against surfactants and organic solvents. The preferred substrates for protease activity were found to be casein (100%) and soybean flour (71.6%).

Item Type	Article
URI	https://reading-clone.eprints-hosting.org/id/eprint/78941
Identification Number/DOI	10.1007/s12257-017-0495-3
Refereed	Yes
Divisions	Life Sciences > School of Chemistry, Food and Pharmacy > Department of Food and Nutritional Sciences > Food Research Group
Publisher	Springer
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