|Table of Contents|

Expression, Characterization and Fermentation Optimization of Keratinase from Bacillus licheniformis in Escherichia coli(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

Issue:
2013 06
Page:
997-1002
Research Field:
Articles
Publishing date:

Info

Title:
Expression, Characterization and Fermentation Optimization of Keratinase from Bacillus licheniformis in Escherichia coli
Author(s):
LIU BaihongZHANG JuanFANG ZhenLIU WentaoDU GuochengCHEN JianLIAO Xiangru
(1Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China) (2National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China) (3Key Laboratory of Carbohydrate and Biotechnology, Jiangnan University, Wuxi 214122, China) (4School of Biotechnology, Jiangnan University, Wuxi 214122, China)
Keywords:
keratinase Bacillus licheniformis Escherichia coli recombinant expression enzymatic property fermentation optimization
CLC:
Q939.9 : Q796
PACS:
DOI:
10.3724/SP.J.1145.2013.00997
DocumentCode:

Abstract:
In order to enhance the production of keratinase in Escherichia coli and study the enzymatic properties, the ker gene (1 140 bp) was cloned from a feather-degrading bacterium of Bacillus licheniformis BBE11-1. After the 1 053 bp fragment of signal peptide-truncated ker gene was inserted into the expression vector pET-22b (+), the recombinant plasmid was transformed into E. coli BL21 (DE3). The recombinant keratinase purified by hydrophobic?interaction?chromatography using HiTrap Phenyl-sepharose Fast Flow was a serine protease most active at 50 ℃ and pH 10. The activity of keratinase was enhanced by Mg2+ and K+, and inhibited by Mn2+. The highest keratinase activity was 460.2 U/mL in growth medium containing inducer (0.05 mmol/L IPTG, 150 mmol/L glycine and 60 g/L yeast extract) at 20 ℃. This report focused on the expression and characterization of recombinant keratinase in E. coli. We found that optimizing culture conditions could preferably enhance production of keratinase in E. coli. The results would be helpful in producing keratinase by gene engineering strain.

References

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Last Update: 2014-01-03