Heterologous Expression and Activity Analysis of Collagenase Gene from Bacillus cereus
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    Abstract:

    In this study, Bacillus cereus MBL13-U, which degrades bone collagen, was used as the original strain. The whole genome of B. cereus MBL13-U was sequenced, and the collagenase(ColM13) gene was cloned. The target gene was ligated with the expression vector pET30a of Escherichia coli and transferred into E. coli host strain BL21 to obtain an engineering strain pET30a-ColM13 / BL21 that degrades collagen. The molecular weight of the recombinant enzyme ColM13 expressed in heterologous was determined by SDS-PAGE, and the enzyme activity of the enzyme producing bacteria were studied under the different induction time and induced concentration. Finally, the best enzyme activity conditions of recombinant enzyme ColM13 were as follows: 6‰ IPTG (100 mmol/L), 6 h at 37 ℃, the optimized enzyme activity was 64.99 U/mL, increased 4.25 times than the pre-optimization. And further through the hydrolysis ring test, full-wavelength ultraviolet scanning spectroscopy and scanning electron microscopy were validated experiments, indicating that the successful construction of engineering bacteria, which provided a new approach for the depth development and comprehensive utilization of animals’ bone resources in China.

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  • Online: February 03,2020
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