Cloning and expression of keratinase genes in Escherichia coli BL21(DE3) from Bacillus species

Nguyen Huy Hoang, Nguyen Thu Hien, Hoang Thi Thu Hien, Khuat Huu Thanh


B. licheniformis and B. subtilis are capable to produce enzyme, which hydrolyzed feather highly. Keratinase is a group of commercial enzyme, that has many important applications in industries of waste management, leather, food. Hydrolyzed keratin is used to create some rare amino acids, such as, serine, cysteine and proline or used as animal feed, fertilizer, glue that contribute to reducing environmental pollution. We carried out cloning and expression of recombinant keratinase from Bacillus licheniformis and Bacillus subtilis in Escherichia coli BL21(DE3). Ker 1 from the strain B. licheniformis HT10 was integrated into pET22b vector and Ker 2 from B. subtilis Đ.nd1.2 was integrated into the pET32a vector. In the same expression conditions at 37°C, the 1mM IPTG, two obtained proteins showed a differ in size and activity of the enzyme respectively is 28 kDa, 23.5 U/ml and 36 kDa, 18.5 U/ml.


Keywords: Bacillus lichenniformis, Bacillus subtilis, E. coli, expresssion, keratinase.

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