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Journal of Zhejiang University SCIENCE B 2008 Vol.9 No.9 P.713-720


Purification and characterization of keratinase from a new Bacillus subtilis strain

Author(s):  Cheng-gang CAI, Ji-shuang CHEN, Jiong-jiong QI, Yun YIN, Xiao-dong ZHENG

Affiliation(s):  College of Biology and Environmental Engineering, Zhejiang Shuren University, Hangzhou 310015, China; more

Corresponding email(s):   xdzheng@zju.edu.cn

Key Words:  Ammonium sulfatate, Bacillus subtilis, Characterization, Feather, Keratin, Keratinase, Purification, Reducing agents

Cheng-gang CAI, Ji-shuang CHEN, Jiong-jiong QI, Yun YIN, Xiao-dong ZHENG. Purification and characterization of keratinase from a new Bacillus subtilis strain[J]. Journal of Zhejiang University Science B, 2008, 9(9): 713-720.

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author="Cheng-gang CAI, Ji-shuang CHEN, Jiong-jiong QI, Yun YIN, Xiao-dong ZHENG",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Purification and characterization of keratinase from a new Bacillus subtilis strain
%A Cheng-gang CAI
%A Ji-shuang CHEN
%A Jiong-jiong QI
%A Yun YIN
%A Xiao-dong ZHENG
%J Journal of Zhejiang University SCIENCE B
%V 9
%N 9
%P 713-720
%@ 1673-1581
%D 2008
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B0820128

T1 - Purification and characterization of keratinase from a new Bacillus subtilis strain
A1 - Cheng-gang CAI
A1 - Ji-shuang CHEN
A1 - Jiong-jiong QI
A1 - Yun YIN
A1 - Xiao-dong ZHENG
J0 - Journal of Zhejiang University Science B
VL - 9
IS - 9
SP - 713
EP - 720
%@ 1673-1581
Y1 - 2008
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B0820128

The aim of this study was to purify and characterize a keratinase produced by a new isolated Bacillus subtilis KD-N2 strain. The keratinase produced by the isolate was purified using ammonium sulphate precipitation, Sephadex G-75 and DEAE (diethylaminoethyl)-Sepharose chromatographic techniques. The purified enzyme was shown to have a molecular mass of 30.5 kDa, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The optimum pH at 50 °C was 8.5 and the optimum temperature at pH 8.5 was 55 °C. The keratinase was partially inactivated by some metal ions, organic solvents and serine protease inhibitor phenylmethanesulfonyl fluoride (PMSF). Sodium dodecyl sulfate (SDS) and ethylene diamine tetraacetic acid (EDTA) had positive effect on the keratinase activity. reducing agents including dithiothreitol (DTT), mercaptoethanol, L-cysteine, sodium sulphite, as well as chemicals of SDS, ammonium sulfamate and dimethylsulfoxide (DMSO) stimulated the enzyme activity upon a feather meal substrate. Besides feather keratin, the enzyme is active upon the soluble proteins ovalbumin, bovine serum albumin (BSA), casein and insoluble ones as sheep wool and human hair. Calf hair, silk and collagen could not be hydrolyzed by the keratinase.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


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