Purification and Characterisation of Keratinase from a Thermotolerant Feather-degrading Bacterium |
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| Authors: | Worapot Suntornsuk Junthip Tongjun Poranee Onnim Hiroshi Oyama Kanok Ratanakanokchai Thanit Kusamran Kohei Oda |
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| Institution: | (1) Department of Microbiology, Faculty of Science, King Mongkut’s University of Technology Thonburi, Bangkok, 10140, Thailand;(2) Department of Applied Biology, Faculty of Textile Science, Kyoto Institute of Technology, Kyoto 606-8585, Japan;(3) Department of Biochemical Technology, School of Bioresources and Technology, King Mongkut’s University of Technology Thonburi, Bangkok, 10140, Thailand;(4) Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand |
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| Abstract: | Summary Isolation and identification of a thermotolerant feather-degrading bacterial strain from Thai soil as well as purification
and properties of its keratinase were investigated. The thermotolerant bacterium was identified as Bacillus licheniformis. The keratinase was purified to homogeneity by three-step chromatography. The purified enzyme exhibited a high specific activity
(218 U mg−1) with 86-fold purification and 25% yield. The enzyme was monomeric and had a molecular mass of 35 kDa. The optimum pH and
temperature for the enzyme were 8.5 and 60 °C, respectively. The enzyme activity was significantly inhibited by PMSF and partly
inhibited by EDTA and iodoacetamide, but was stimulated by metal ions. It hydrolysed soluble proteins with a relative activity
of 4–100% and insoluble proteins, including keratins, with a relative activity of 3–35%. Therefore, the enzyme could improve
the nutritional value of meat- and poultry-processing wastes containing keratins, collagen and gelatin. |
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| Keywords: | Identification purification characterisation keratinase thermotolerant Bacillus |
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