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222 华 中 农 业 大 学 学 报 第 41 卷

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Optimizing solid fermentation process for chitosanase produced by
Aspergillus tamarii M3b and enzymatic properties of chitosanase

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YAO Yan ,LI Jingjing ,ZHANG Yumeng ,ZHAO Nanpeng ,DING Dezheng ,SU Shuhui ,FU Xing 1
1.College of Food Science and Technology，Huazhong Agricultural University/
National Research and Development Center for Egg Processing，Wuhan 430070，China；
2.Hubei Agricultural Product Quality and Safety Testing Center，Wuhan 430070，China
Abstract A novel fungus strain M3b with high chitosanase activity was screened from the soil of

many places to study the specific chitosanase. The morphological observation and 18S rDNA sequence iden⁃
tification of strain M3b were carried out. The single-factor and orthogonal experiments were used to opti⁃
mize the solid fermentation process for chitosanase produced by M3b. The enzymatic properties of chito⁃
sanase purified were investigated. The results showed that strain M3b was Aspergillus tamarii. The opti⁃
mized fermentation conditions for producing chitosanase produced by M3b based on water as 100% were as
follows：1% colloidal chitosan+1% glucose，0.5% ammonium sulfate，bran∶soybean meal=6∶10，inocu⁃
lation amount 10%，incubation for 6 d，the initial pH 7.0，and the fermentation temperature 32 ℃. Under
these conditions，the activity of chitosanase reached 20.56 U/mL，which was 221.3% of the initial fermenta⁃
tion condition. The results of SDS-PAGE and enzyme profiling showed that the strain M3b produced only
one chitosanase with a molecular weight of 40 ku. The optimum pH and temperature for the enzymatic reac⁃
tion of this enzyme was 5.5 and 60 ℃，with the degree of polymerizing the final product obtained by hydro⁃
lysis of chitosan≥2. It is indicated that producing chitosanase by solid fermentation of Aspergillus tamarii
can significantly improve the activity of chitosanase and reduce the cost of production. The purified chito⁃
sanase AtChito40 has stable enzymatic properties，acid and alkali resistance，and can effectively degrade chi⁃
tosan. It will provide the theoretical and experimental basis for the enzymatic degradation of chitosan and
the preparation of chitosan oligosaccharide in industrial production.
Keywords Aspergillus tamarii；chitosanase；chitallooligosaccharide；solid fermentation；enzyme ac⁃
tivity；purification

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