新型β-葡萄糖苷酶BglD2异源表达及水解虎杖苷能力  

Heterologous expression of a novel β-glucosidase BglD2 and its application in polydatin-hydrolyzing

作　　者：何成 吴言 孟春雨 肖亚中[1] 方泽民[1] 房伟[1] Cheng He;Yan Wu;Chunyu Meng;Yazhong Xiao;Zemin Fang;Wei Fang(School of Life Sciences,Anhui University,Hefei 230601,Anhui,China)

机构地区：[1]安徽大学生命科学学院,安徽合肥230601

出　　处：《生物工程学报》2021年第2期580-592,共13页Chinese Journal of Biotechnology

基　　金：国家重点研发计划(No.2018YFC0311106);国家自然科学基金(No.31870056)资助。

摘　　要：从海洋细菌Bacillus sp.D1中克隆、重组表达β-葡萄糖苷酶BglD2,研究其酶学性质,并对其水解虎杖苷制备白藜芦醇的能力进行分析。BglD2的最适催化温度和pH分别为45℃和6.5,在30℃和pH 6.5条件下的半衰期约为20 h。BglD2能够水解含β(1→3)、β(1→4)、β(1→6)等键型的多种底物。BglD2具有良好的糖促活特性,100 mmol/L葡萄糖和150 mmol/L木糖分别将酶活力提升2.0倍和2.3倍。BglD2具有较好的乙醇促活及耐受特性,30℃时,10%乙醇使酶活力提升1.2倍,25%乙醇存在时其仍保留60%的酶活力。BglD2具有水解虎杖苷制备白藜芦醇的能力,35℃条件下反应2 h水解率为86%。具有乙醇耐受及抗产物抑制等特性的β-葡萄糖苷酶BglD2在酶法水解虎杖苷制备白藜芦醇方面有应用潜力。A novelβ-glucosidase BglD2 with glucose and ethanol tolerant properties was screened and cloned from the deep-sea bacterium Bacillus sp.D1.The application potential of BglD2 toward polydatin-hydrolyzing was also evaluated.BglD2 exhibited the maximal β-glucosidase activity at 45℃ and pH 6.5.BglD2 maintained approximately 50%of its origin activity after incubation at 30℃ and pH 6.5 for 20 h.BglD2 could hydrolyze a variety of substrates containingβ(1→3),β(1→4),andβ(1→6)bonds.The activity of β-glucosidase was enhanced to 2.0 fold and 2.3 fold by 100 mmol/L glucose and 150 mmol/L xylose,respectively.BglD2 possessed ethanol-stimulated and-tolerant properties.At 30℃,the activity of BglD2 enhanced to 1.2 fold in the presence of 10%ethanol and even remained 60%in 25%ethanol.BglD2 could hydrolyze polydatin to produce resveratrol.At 35℃,BglD2 hydrolyzed 86%polydatin after incubation for 2 h.Thus,BglD2 possessed glucose and ethanol tolerant properties and can be used as the potential candidate of catalyst for the production of resveratrol from polydatin.

关 键 词：Β-葡萄糖苷酶 葡萄糖耐受 乙醇耐受 虎杖苷 白藜芦醇 

分 类 号：R284.1[医药卫生—中药学]