Cloning and Identification of Keratinase Gene from Deinococcus gobiensis I-0

doi:10.13560/j.cnki.biotech.bull.1985.2018-0976

Abstract

Abstract: In order to further tap the important gene resources of keratinase,improve its hydrolytic activity,and provide theoretical basis for industrial production,this experiment cloned a gene encoding keratinase from Deinococcus gobiensis I-0 isolated from Gobi desert of Xinjiang and named it as Dgker. Prokaryotic expression vector pET-22B-Dgker was constructed and then induced,expressed and purified in vitro,the optimal temperature and pH of the crude enzyme solution were determined through the hydrolysis activity to feathers. Results showed that the first 50 amino acids of N terminal had a great influence on the expression and purification of protein Dgker. The crude enzyme solution of recombinant strain completely decomposed feathers in three days. The transparent circle on milk powder plate appeared more notable in crude enzyme solution of recombinant strain than that of empty strain. Dgker adapted to a wide range of temperatures and pH,among which the optimal temperature was 60℃ and the optimal pH was 5.0. Dgker can degrade feathers and thus will have great application space in the future industrial production and treatment of waste feathers.

Key words: keratin, Deinococcus gobiensis I-0, keratinase, enzyme activity

GENG Xiu-xiu, ZHOU Zheng-fu, LIU Ying-ying, PING Shu-zhen, WANG Jin. Cloning and Identification of Keratinase Gene from Deinococcus gobiensis I-0[J]. Biotechnology Bulletin, 2019, 35(3): 65-70.

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