Journal
PROTEIN EXPRESSION AND PURIFICATION
Volume 166, Issue -, Pages -Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.pep.2019.105519
Keywords
Beauveria bassiana; Chitosanase; Pichia pastoris; Purification; Enzymatic characteristics
Categories
Funding
- Fundamental Research Funds for the Central Universities [XDJK2018C066, XDJK2018AA006]
- National Natural Sciences Foundation of China [31570137]
- Chongqing Research Program of Basic Research and Frontier Technology [cstc2019jcyj-msxmX0388, cstc2018jcyjAX0073]
- program for innovation research team of Chongqing University [CXTDX201601012]
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Chitosanase (EC 3.2.1.132) is an important chitosan-degrading enzyme involved in industrial applications. In this study, a chitosanase gene (BbCSN-1) from Beauveria bassiana, an insect fungal pathogen, was cloned and expressed in Pichia pastoris. The amount of BbCSN-1 in the fermentation broth of P. pastoris gradually increased after induction with methanol from one to 6 d, reaching 398 mu g/m1 on the 6th day. The molecular characteristics of BbCSN-1 were measured with colloidal chitosan as a substrate. The purified BbCSN-1 exhibited optimum activity at pH 5 and 30 degrees C and was stable at pH 2-8 and below 40 degrees C. The Km value of BbCSN-1 was approximately 0.8 mg/ml at 30 degrees C (pH 6.0). The activity of BbCSN-1 was significantly enhanced by Mn2+ but inhibited by Co2+ and Cu2+. These results indicated that BbCSN-1 from B. bassiana could be easily expressed in P. pastoris, which provided a basis for further study on its application.
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