Expression and characterization of a keratinase encoding gene gm2886 in Streptomyces pactum ACT12 strain.

Yiming MA; Xin KE; Xiaoxia LI; Weixue Shu; Wenhan Yang; Yayong Liu; Xia Yan; Lianghui Jia; Hua Yan

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Expression and characterization of a keratinase encoding gene gm2886 in Streptomyces pactum ACT12 strain.

Author: Yiming MA ¹ ; Xin KE ¹ ; Xiaoxia LI ¹ ; Weixue SHU ¹ ; Wenhan YANG ¹ ; Yayong LIU ² ; Xia YAN ¹ ; Lianghui JIA ¹ ; Hua YAN ¹
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1. College of Life Sciences, Northwest A&F University, Yangling 712100, Shaanxi, China.
2. Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
Publication Type:Journal Article
Keywords: Streptomyces albidoflavus Fea-10; Streptomyces pactum; heterologous expression; keratinase; recombinant enzyme
From: Chinese Journal of Biotechnology 2017;33(12):1968-1978
CountryChina
Language:Chinese
Abstract: By bioinformatics analysis, a putative keratinase gene gm2886 (Accession number: KY368946) was discovered in the genome of a feather-degrading strain, Streptomyces albidoflavus Fea-10. gm2886 was ligated into integrative Escherichia coli-Streptomyces shuttle vector pSET152 under the promoter PermE and added with C-terminal His-tag. The expression vector was transformed into Streptomyces pactum ACT12 by conjugal transfer and the recombinant protein GM2886-His6 was detected in fermentation broth. GM2886-His6 was purified and characterized. Its size was nearly 36 kDa. GM2886-His6 showed proteolytic activity towards a variety of substrates and could even degrade insoluble substrates, such as azure keratin and chicken feathers. The optimal pH and temperature of GM2886-His6 for proteolysis of casein was pH 10.0 and 50 ℃, respectively. The enzyme activity was inhibited by PMSF, but not EDTA, indicating that GM2886-His6 was a serine proteinase. Our results laid the foundation for the research of the molecular biological mechanism on feather-degrading and for the further utilization of Fea-10.