[构巢曲霉内切纤维素酶A10在毕赤酵母中的过表达及特性研究]
[Overexpression and characterization of endo-cellulase A10 from Aspergillus nidulans in Pichia pastoris].
作者信息
Li Jianjun, Li Xiaolian, Wang Ziqiang, Li Jianjun
机构信息
National Key Laboratory of Biochemical Engineering, National Engineering Research Center for Biotechnology (Beijing), Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China.
Key Laboratory of Biopharmaceutical Production & Formulation Engineering, The People's Liberation Army, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China.
出版信息
Sheng Wu Gong Cheng Xue Bao. 2017 May 25;33(5):757-765. doi: 10.13345/j.cjb.160442.
Endo-cellulases are important to efficiently hydrolyze cellulose, and widely used in biotechnology. In this study, we overexpressed and characterized an endo-cellulase from Aspergillus nidulans. This endo-cellulase was successfully overexpressed in flasks and fermentor, and its concentration in fermentor reached 0.89 mg/mL. The optimal pH and temperature of the were 4.0 and 80 ℃ respectively, and it was very stable between pH 2.0 and 12.0. It was thermally stable below 60 ℃, whereas it was inactivated very quickly above 70 ℃. Its CMCase activity could be enhanced by Co²⁺, Mn²⁺ and Fe²⁺, whereas it was inhibited by Pb²⁺, Ni²⁺ and Cu²⁺. Therefore, this endo-cellulase exhibited good pH stability and thermostability below 60 ℃, and has the potential as commercial enzymes.
内切纤维素酶对于高效水解纤维素非常重要，并且在生物技术中被广泛应用。在本研究中，我们对构巢曲霉的一种内切纤维素酶进行了过表达及特性分析。该内切纤维素酶在摇瓶和发酵罐中均成功实现了过表达，其在发酵罐中的浓度达到了0.89 mg/mL。其最适pH和温度分别为4.0和80℃，在pH 2.0至12.0之间非常稳定。在60℃以下热稳定，而在70℃以上则很快失活。其羧甲基纤维素酶（CMCase）活性可被Co²⁺、Mn²⁺和Fe²⁺增强，而被Pb²⁺、Ni²⁺和Cu²⁺抑制。因此，这种内切纤维素酶在60℃以下表现出良好的pH稳定性和热稳定性，具有作为商业酶的潜力。
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