Abstract
A keratin-degrading bacterium of Bacillus licheniformis BBE11-1 was isolated and its ker gene encoding keratinase with native signal peptide was cloned and expressed in Bacillus subtilis WB600 under the strong P HpaII promoter of the pMA0911 vector. In the 3-L fermenter, the recombinant keratinase was secreted with 323 units/mL when non-induced after 24 h at 37 °C. And then, keratinase was concentrated and purified by hydrophobic interaction chromatography using HiTrap Phenyl-Sepharose Fast Flow. The recombinant keratinase had an optimal temperature and the pH at 40 °C and 10.5, respectively, and was stable at 10–50 °C and pH 7–11.5. We found this enzyme can retained 80 % activity after treated 5 h with 1 M H2O2, it was activated by Mg2+, Co2+ and could degraded broad substrates such as degraded feather, bovine serum albumin, casein, gelatin, the keratinase was considered to be a serine protease. Coordinate with Savinase, the keratinase could efficient prevent shrinkage and eliminate fibres of wool, which showed its potential in textile industries and detergent industries.
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Acknowledgments
This project was financially supported by the National Natural Science Foundation of China (No. 30900013), the National High Technology Research and Development Program of China (863 Program, 2011AA100905), the National Key Technology R&D Program in the 12th Five year Plan of China (2011BAK10B03), the Program for Changjiang Scholars and Innovative Research Team in University (No.IRT1135), and the National High Technology Research and Development Program of China (863 Program, 2011AA100901).
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liu, B., Zhang, J., Li, B. et al. Expression and characterization of extreme alkaline, oxidation-resistant keratinase from Bacillus licheniformis in recombinant Bacillus subtilis WB600 expression system and its application in wool fiber processing. World J Microbiol Biotechnol 29, 825–832 (2013). https://doi.org/10.1007/s11274-012-1237-5
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DOI: https://doi.org/10.1007/s11274-012-1237-5