Purification and Characterization of a Keratinase from a Feather-degrading Bacterium, Bacillus sp. SH-517
We isolated Bacillus sp. SH-517 from decomposed chicken feathers at a local poultry plant. This strain produced a keratinase that degrades poultry feathers and therefore will be very valuable for industrial use. Most feathers were degraded by the strain within 40 h at 40℃ by shaking culture (180 rpm...
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Veröffentlicht in: | Applied biological chemistry 2010, 53(1), , pp.43-49 |
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Sprache: | eng |
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Zusammenfassung: | We isolated Bacillus sp. SH-517 from decomposed chicken feathers at a local poultry plant. This strain produced a keratinase that degrades poultry feathers and therefore will be very valuable for industrial use. Most feathers were degraded by the strain within 40 h at 40℃ by shaking culture (180 rpm). The keratinase from the culture medium of Bacillus sp. SH-517 was purified by centrifugation, 30-80% ammonium sulfate fractionation, twin-column DEAE-cellulose ion exchange chromatography and Sephadex G-150 gel filtration, to obtain a purified keratinase at 10.82% yield and 14-fold overall purification. The purified enzyme had a specific activity of 825 U/mg. A single protein band was shown on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weight of the keratinase from Bacillus sp. SH-517 was estimated as 51 kDa. The optimum pH and temperature for the enzyme reaction were 7.5 and 40℃, respectively. The enzyme remained stable over the pH range from 4.0 to 9.0 and at temperatures below 50℃. Proteins such as milk casein and chicken feathers were easily hydrolyzed by this enzyme. The enzyme activity was significantly inhibited by Hg²+, Ag²+, ethylene diamine tetraacetic acid (EDTA) and ethylene glycol tetraacetic acid (EGTA), but slightly stimulated by K+ and Na+. |
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ISSN: | 1738-2203 2468-0834 2234-344X 2468-0842 |
DOI: | 10.3839/jksabc.2010.008 |