Gupta Sonali and Nigam Arti and Singh Rajni: Purification and characterization of a Bacillus subtilis keratinase and its prospective application in feed industry. In: Acta biologica Szegediensis, (59) 2. pp. 197-204. (2015)
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Abstract
We have isolated a Bacillus subtilis strain (RSE163) from soil and explored for keratinase production. Keratinase was purified using chromatographic methods (Sephadex G-75 and Q Sepharose) resulting in 8.42-fold purification with 3303 U/mg specific activity.The purified enzyme displayed 3 bands in close proximity between 20 to 22 kDa in SDS-PAGE which were apparent to the zone of hydrolysis in gelatin zymogram. Enzyme was stable over a wide pH (7.0-10.0) and temperature (30 °C to 70 °C) range with optimum activity at pH 9.0 and 60 °C. Keratinase activity was stimulated in presence of Mn2+, β-mercaptoethanol and surfactants (Triton-X and Tween-80) and inhibited by Fe3+, Cd2+, K+, PMSF (phenyl methane sulfonyl fluoride) and other chelating and reducing agents. The enzyme efficiently hydrolyzed a variety of complex protein substrates (chicken feather, keratin hydrolyzate and casein) and enzyme kinetics parameters were determined using Lineweaver Burk plot (Km = 6.6 mg/ml, Vmax = 5 U/ml/min). Hydrolyzed feather keratin obtained through fermentation with B. subtilis RSE163 has been explored for its cytotoxicity using liver cell line (HepG2). No cytotoxicity has been determined up to 0.015% concentration of hydrolyzed product indicating its potential applicability as feed supplement.
| Item Type: | Article |
|---|---|
| Heading title: | Articles |
| Journal or Publication Title: | Acta biologica Szegediensis |
| Date: | 2015 |
| Volume: | 59 |
| Number: | 2 |
| ISSN: | 1588-385X |
| Page Range: | pp. 197-204 |
| Language: | English |
| Related URLs: | http://acta.bibl.u-szeged.hu/39273/ |
| Uncontrolled Keywords: | Bacillus subtilis, Mikrobiológia baktériumok |
| Additional Information: | Bibliogr.: p. 203-204. |
| Date Deposited: | 2016. Oct. 17. 10:36 |
| Last Modified: | 2021. Apr. 12. 15:10 |
| URI: | http://acta.bibl.u-szeged.hu/id/eprint/35945 |
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