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

  • Baihong liu
  • Juan Zhang
  • Ben Li
  • Xiangru Liao
  • Guocheng Du
  • Jian Chen
Original Paper

DOI: 10.1007/s11274-012-1237-5

Cite this article as:
liu, B., Zhang, J., Li, B. et al. World J Microbiol Biotechnol (2013) 29: 825. doi:10.1007/s11274-012-1237-5

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 PHpaII 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.

Keywords

Bacillus licheniformis Bacillus subtilis WB600 Keratinase Extracellular expression Wool processing 

Copyright information

© Springer Science+Business Media Dordrecht 2012

Authors and Affiliations

  • Baihong liu
    • 1
    • 4
  • Juan Zhang
    • 1
    • 4
  • Ben Li
    • 5
  • Xiangru Liao
    • 1
    • 4
  • Guocheng Du
    • 3
    • 4
  • Jian Chen
    • 2
    • 4
  1. 1.Key Laboratory of Industrial Biotechnology, Ministry of EducationJiangnan UniversityWuxiChina
  2. 2.National Engineering Laboratory for Cereal Fermentation TechnologyJiangnan UniversityWuxiChina
  3. 3.The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of EducationJiangnan UniversityWuxiChina
  4. 4.School of BiotechnologyJiangnan UniversityWuxiChina
  5. 5.Laboratory of Eco-Dyeing and FinishingJiangnan UniversityWuxiChina

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