EXPRESSION OF CHITINASE GENE FROM Bacillus Licheniformis DSM13 IN E.Coli T7 AND BIOCHEMICAL CHARACTERIZATION OF RECOMBINANT ENZYME

Ngày nhận bài: 26-07-2017

Ngày duyệt đăng: 05-09-2017

Ngày xuất bản: 06-08-2025

DOI: https://doi.org/10.31817/tckhnnvn.2017.15.9.

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Số: Tập 15 Số 9 (2017)

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Anh, N. (2025). EXPRESSION OF CHITINASE GENE FROM Bacillus Licheniformis DSM13 IN E.Coli T7 AND BIOCHEMICAL CHARACTERIZATION OF RECOMBINANT ENZYME. Tạp Chí Khoa học Nông nghiệp Việt Nam, 15(9), 1230–1238. https://doi.org/10.31817/tckhnnvn.2017.15.9.

EXPRESSION OF CHITINASE GENE FROM Bacillus Licheniformis DSM13 IN E.Coli T7 AND BIOCHEMICAL CHARACTERIZATION OF RECOMBINANT ENZYME

Nguyen Hoang Anh (*) 1

  • Tác giả liên hệ: [email protected]
  • 1 Faculty of Food Science and Technology, Vietnam National University of Agriculture

    • Từ khóa

    Bacillus licheniformis DSM13, chitinase, E. coli T7

    Tóm tắt


    In this study, the mature gene encoding for the enzyme endochitinase from the Gram positive bacterium Bacillus licheniformis DSM13 (ATCC 14580) was cloned into the expression vector pET-21d, and then overexpressed in
    E. coli T7. A high expression level was indicated by SDS-PAGE and an enzyme activity assay using pNP-(GlcNAc)2 0.18 mM as the substrate. The activity of the enzyme expressed in E. coli T7 was approximately 8 times higher than that previously expressed in Lactobacillus plantarum. Recombinant chitinase was purified from cell extract and characterized using coloidal chitin 2% as the substrate. The enzyme showed good thermostability (half-lives of 15 days and 8 days at 37 and 50° C, respectively), and good stability in the pH range of 5 - 9. The main product of colloidal chitin hydrolysis as indicated by thin layer chromatography was diacetyl glucosamine. The results demonstrated that this enzyme was promising for chitin waste bioconversion into different chitin-oligosaccharides, such as functional diacetyl glucosamine.

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