期刊
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
卷 146, 期 -, 页码 518-523出版社
ELSEVIER
DOI: 10.1016/j.ijbiomac.2020.01.031
关键词
Chitosanase; Chitosan; GH46 family
资金
- National Natural Science Foundation of China [31801574, 31922072]
- National Key Research and Development Program of China [2019YFD0901902]
- Natural Science Foundation of Shandong Province [ZR2017BC095]
- China Agriculture Research System [CARS-48]
- Shandong Province Post-Doctoral Innovation Projects of Special Funds
- Taishan Scholar Project of Shandong Province [tsqn201812020]
Chitosanases play an important role in chitosan degradation, and the enzymatic degradation products of chitosan show various biological activities. Here, a novel glycoside hydrolase family 46 chitosanase (named Csn-BAC) from Bacillus sp. MD-5 was heterologously expressed in Escherichia coli BL21 (DE3). The recombinant enzyme was purified by Ni-NTA affinity chromatography, and its molecular weight was estimated to be 35 kDa by SDS-PAGE. Csn-BAC showed maximal activity toward colloidal chitosan at pH 7 and 40 degrees C. The enzymatic activity of Csn-BAC was enhanced by Mn2+, Cu2+ and Co2+ at 1 mM, and by Mn2+ at 5 mM. Thin-layer chromatography and electrospray ionization-mass spectrometry results demonstrated that Csn-BAC exhibited an endo-type cleavage pattern and hydrolyzed chitosan to yield, mainly, (GlcN)(2) and (GlcN)(3). The enzymatic properties of this chitosanase may make it a good candidate for use in oligosaccharide production-based industries. (C) 2020 Elsevier B.V. All rights reserved.
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