Chinese Journal of Tissue Engineering Research ›› 2019, Vol. 23 ›› Issue (在线): 1-4.

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Cloning and expression of β-glucanase gene from Bacillus subtilis in Escherichia coli

Sun Ke, Wang Ya-nan, Zhang Ya-nan, Zhu Wen-hua, Cheng Dian-lin
  

  1. Qingdao University, Department of Life Science, Shandong, Qingdao 266071
  • Online:2019-03-28 Published:2019-03-28
  • Supported by:

    Promotive research fund for young and middle-aged scientisits of Shandong Province, No.BS2010SW007

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Abstract:

BACKGROUND：β-glucan has been widely used for healthcare and clinical treatment. The molecular weight of the reasonable-sized glucan was obtained By the way of enzymatic hydrolysis which is more and more emphasized, but no research reports has been published in china yet.
OBJECTIVE: The experiment aims to build an engineering strain secreting high-expression β-glucanase of which the activity and thermo-stability are analysed later so as to lay a basis on its clinical application.
METHODS: We got the β-glucanase gene of Bacillus Subtilis，make β-glucanase gene and expression veltor pET28b(+) to recombinant plasmid using molecular biology technology and then structure engineered stain in Escherichia coli, using IPTG to induce the expression of the target potein, purificate it by nickel ion chelating resin.
RESULTS AND CONCLUSION: In this research, Gene encoding β-glucanase in Bacillus subtilis was cloned into Escherichia coli and expressed efficiently. The enzymatic assay has found that it has great activity between 50°C and 60°C and highest in 50°C. The relative high-thermostability of β-glucanase under the temperature of 30°C~40°C. It was preliminarily found that β-glucanase expressed by the engineering strain referred was able to get rid of its substrate. But whether it is suitable for the clinical treatment requires more research. The experiment aims to utilize the strain to improve the level of clinical treatment and provide microbial resources.
Keywords: Bacillus subtilis; Escherichia coli; plasmid; β-glucanase; clinical medicine; healthcare; cloning; expression; enzyme activity; treat