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Production and Characterization of Keratinolytic Proteases by a Chicken Feather-Degrading Thermophilic Strain, Thermoactinomyces sp YT06

文献类型： 外文期刊

作者： Wang, Lin 1 ; Qian, Yuting 1 ; Cao, Yun 1 ; Huang, Ying 2 ; Chang, Zhizhou 1 ; Huang, Hongying 1 ;

作者机构： 1.Jiangsu Acad Agr Sci, Circular Agr Res Ctr, Nanjing 210014, Jiangsu, Peoples R China

2.Nanjing Inst Agr Sci Jiangsu Hilly Area, Nanjing 210046, Jiangsu, Peoples R China

关键词： Thermophile;keratinase;feather degradation;characterization

期刊名称：JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：2.351； 五年影响因子：2.65 ）

ISSN： 1017-7825

年卷期： 2017 年 27 卷 12 期

页码：

收录情况： SCI

摘要： Thermoactinomyces sp. strain YT06 was isolated from poultry compost and observed to degrade integral chicken feathers completely at 60 degrees C, resulting in the formation of 3.24 mg/ml of free amino acids from 50 ml of culture containing 10 g/l chicken feathers. Strain YT06 could grow and secrete keratinase using feather as the only carbon and nitrogen sources without other supplement, but complementation of 10 g/l sucrose and 4 g/l NaNO3 increased the production of the keratinolytic enzyme. The maximum protease activity obtained was 110 U/ml and for keratinase was 42 U/ml. The keratinase maintained active status over a broad pH (pH 8-11) and temperature (60-75 degrees C). It was inhibited by serine protease inhibitors and most metal ions; however, it could be stimulated by Mn2+ and the surfactant Tween-20. A reductive agent (beta-mercaptoethanol) was observed to cleave the disulfide bond of keratin and improve the access of the enzyme to the keratinaceous substrate. Zymogram analysis showed that strain YT06 primarily secreted keratinase with a molecular mass of approximately 35 kDa. The active band was assessed by MALDI-TOF mass spectrometry and was observed to be completely identical to an alkaline serine protease from Thermoactinomyces sp. Gus2-1. Thermoactinomyces sp. strain YT06 shows great potential as a novel candidate in enzymatic processing of hard-to-degrade proteins into high-value products, such as keratinous wastes.

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