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Production, purification and characterization of chitosanase produced by Gongronella sp. JG
Author(s) -
Zhou W.,
Yuan H.,
Wang J.,
Yao J.
Publication year - 2008
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2007.02262.x
Subject(s) - chitosanase , hydrolysis , chromatography , chitin , sodium acetate , chitosan , chemistry , sodium , specific activity , biochemistry , enzyme , organic chemistry
Aims: To optimize the production condition of chitosanases of Gongronella sp. JG and to characterize the major chitosanase. Methods and Results: In the optimized medium and culturing condition, strain JG produced 800 μ mol min −1 l −1 chitosanase activity at 72 h. The major chitosanase – csn1 was purified through three chromatography steps: CM (carboxymethyl)–Sepharose fast flow (FF), Sephacryl S200, SP (sulfopropyl)–Sepharose FF. The molecular weight and the pI value of csn1 were about 90 000 Da and 5·8, respectively. Its specific activity was 82 μ mol min −1 mg −1 . The optimal reaction pH for csn1 was between 4·6 and 4·8. The optimal reaction temperature was 50°C. The half‐life of csn1 at 50°C was estimated to be about 65 min. Mn 2+ was a strong stimulator of csn1 activity, both at 1 and 10 mmol l −1 . csn1 showed its highest activity with chitosan of 85% degree of deacetylation, but did not hydrolyse colloidal chitin and carboxylmethyl cellulose. In 20 mmol l −1 sodium acetate buffer (pH 4·8) and at 50°C, the K m of csn1 was calculated to be 4·5 mg ml −1 . Conclusions: The production condition of chitosanases by Gongronella JG was optimized and the major chitosanase, csn1, was characterized. Significance and Impact of the Study: The present work for the first time reported the production, purification and characterization of chitosanases produced by fungus of Gongronella sp. These results provided us more information on fungal chitosanases.