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Production,characterization and application of a keratinase from Chryseobacterium L99 sp. nov.
Institution:1. Synergetic Innovation Center of Food Safety and Nutrition, Wuxi 214122, China;2. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;3. National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China;4. The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;5. School of Biotechnology, Jiangnan University, Wuxi 214122, China;1. PG & Research Department of Biotechnology, Mahendra Arts and Science College, Kalippatti, Namakkal 637501, Tamil Nadu, India;2. Centre for Biotechnology, Muthayammal College of Arts and Science, Rasipuram, Namakkal 637 408, Tamil Nadu, India;3. Division of Biotechnology, College of Environmental and Bioresource Sciences, Chonbuk National University, Iksan 570752, South Korea;1. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;2. Synergetic Innovation Center of Food Safety and Nutrition, Wuxi 214122, China;3. The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;4. National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China;5. School of Biotechnology, Jiangnan University, Wuxi 214122, China;1. Programa de Pós-Graduação em Ambiente e Tecnologias Sustentáveis, Universidade Federal da Fronteira Sul (UFFS), Campus Cerro Largo, Av. Jacob Reinaldo Haupenthal 1580, 97900-000 Cerro Largo, RS, Brazil;2. Laboratório de Bioquímica e Microbiologia Aplicada, Instituto de Ciência e Tecnologia de Alimentos (ICTA), Universidade Federal do Rio Grande do Sul (UFRGS), 91501-970 Porto Alegre, RS, Brazil;1. Biotechnology Center – Bioinovar, Institute of Microbiology Paulo de Góes, Federal University of Rio de Janeiro, Cidade Universitária, 21941-590 Rio de Janeiro, Brazil;2. Embrapa Food Technology, National Research Center of Agroindustrial Technology of Food, Food Engineering, Biotechnology Laboratory, Guaratiba, 23020-470 Rio de Janeiro, Brazil;3. Federal Institute of Education, Science and Technology of Rio de Janeiro, Rio de Janeiro Campus, Maracanã, 20270-021 Rio de Janeiro, Brazil;4. Embrapa Agroenergy, Biological Station Park, Av. w3 Norte, 70770-901 Brasilia, DF, Brazil;1. Department of Biotechnology, Guru Ghasidas Vishwavidyalaya, Bilaspur, Chattisgarh 495009, India;2. School of Biotechnology, Banaras Hindu University, Varanasi, U.P., India;3. Department of Microbiology, Central University of Rajasthan, N.H.8, Bandersindri Kishangarh, Ajmer, India
Abstract:Keratins are important bioresources for apparels and feedstuffs, but recalcitrant to common enzymes. Now, it is popular and essential to develop keratinolytic enzymes for environmental prevention and improvement of keratin product quality. In the study, the medium optimization, purification, characterization and application of the keratinase from a newly isolated Chryseobacterium L99 sp. nov. were conducted. Exogenous sucrose, malt sugar, glucose, starch, tryptone, Mg2+, Zn2+, Ca2+ and Cu2+ could promote the keratinase production, while exogenous urea, NH4Cl and yeast extract exhibited strong inhibition effects. Response surface methodology predicted a maximum keratinase yield of 213.8 U mL?1, at (g L?1) sucrose 16.8, MgCl2·6H2O 1.9, feather keratin 40.0, NaH2PO4·2H2O 6.0 and K2HPO4·6H2O 1.0, where dry cell weight nearly had a minimum 8.58 g L?1. Then, a serine keratinase about 33 kDa was purified, and its optimal activity was acquired at 40 °C and pH 8.0 with K+, Zn2+or Co2+. Compared with Savinase 16 L and transglutaminase, the L99 keratinase could efficient prevent shrinkage and eliminate directional frictional effect of wool, indicating it as a promising prospect in the biotreatment of wool fibres.
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