Journal
BRAZILIAN ARCHIVES OF BIOLOGY AND TECHNOLOGY
Volume 57, Issue 5, Pages 653-662Publisher
INST TECNOLOGIA PARANA
DOI: 10.1590/S1516-8913201402132
Keywords
Fibrinolytic Protease; Production; Agricultural residues; Bacillus subtilis I-2
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Funding
- Indian Council of Medical Research (ICMR Extramural Research Project) [5/4/1/EXM/12-NCD-II]
- Council of Scientific and Industrial Research (CSIR)
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The aim of this work was to study the production of fibrinolytic protease by Bacillus subtilis I-2 on agricultural residues. Molasses substantially enhanced (63%) protease production (652.32 U/mL) than control (398.64 U/mL). Soybean meal supported maximum protease production (797.28 U/mL), followed by malt extract (770.1 U/mL), cotton cake (761.04 U/mL), gelatin (742.92 U/mL) and beef extract (724.8 U/mL). Based on the Plackett-Burman designed experiments, incubation time, soybean meal, mustard cake and molasses were identified as the significant fermentation parameters. Ammonium sulfate precipitation and DEAE sephadex chromatography resulted 4.8-fold purification of protease. Zymography showed the presence of three iso-forms in the partially purified protease preparation, which was confirmed by the SDS-PAGE analysis (42, 48, 60 kDa). Protease exhibited maximum activity at 50 degrees C and at pH 8.0. Significant stability was observed at 30-50 degrees C and at pH 7.0-10.0. Mg2+, Zn2+, Co2+, Ca2+, Mn2+ and Cu2+, EGTA, EDTA and aprotinin severely decreased the enzyme activity.
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