Purification and characterizations of a novel recombinant Bacillus velezensis endoglucanase by aqueous two-phase system

Bioresources and Bioprocessing

Wensheng Qin¹  Yang Liu¹  Haipeng Guo¹  Yanwen Wu¹ 

[1] Department of Biology, Lakehead University;

关键词: Purification and characterizations;    Endoglucanase;    Bacillus velezensis;    Aqueous two-phase system;   

DOI  :  10.1186/s40643-018-0204-x

来源: DOAJ

【 摘 要 】

Abstract Background Cellulases played an important role in the production of bioenergy and bio-products. Cellulases from bacteria with some special characteristics drew great attention due to its fast growth speed, wide adaption to harsh environment, and production of multi-function cellulases. Results An endoglucanase gene egls from Bacillus velezensis A4 was cloned and expressed in Escherichia coli BL21 (DE3). The recombinant enzyme Egls was partially purified using aqueous two-phase system. The highest recovery rate of the enzyme was 90.39% at PEG 4000 (25% w/w), phosphate buffer 8.08% (w/w) (pH 6.0), and NaCl (5% w/w). The enzyme molecular weight was 55 KD estimated by zymogram. The optimal pH and temperature of recombinant enzyme Egls were pH 6.0 and 55 °C, respectively. The enzyme was stable at pH range of 5.0–7.0 at 55 °C for 60 min. The enzyme exhibited K m, V max, K cat values as 63.38 mg/ml, 55.6 mg/min, and 3.93 × 103/S, respectively. The addition of 10 mM of Mg2+, Mn2+, or 5% (w/w) of Triton-X 100 in the reaction system enhanced the enzyme activity significantly. The enzyme showed both endoglucanase and exoglucanase activity. Conclusions An endoglucanase gene egls from B. velezensis A4 was cloned and expressed in E. coli BL21 (DE3). The recombinant enzyme Egls was purified by aqueous two-phase system and characterized. The enzyme can be applied for the efficient pretreatment of lignocellulosic biomass for bioenergy and bio-products production.

【 授权许可】

Unknown