食品科学技术学报2019，Vol.37Issue(1)：20-27,8.DOI:10.3969/j.issn.2095-6002.2019.01.004
巴伦葛兹类芽孢杆菌β-葡萄糖苷酶的表达、酶学性质及结构解析
Expression, Biochemical Characterization and Structure Resolution of β-Glucosidase from Paenibacillus barengoltzii
摘要
Abstract
A β-glucosidase gene from Paenibacillus barengoltzii CAU904 was cloned and heterologousl≥expressed in E. coli. The recombinant enz≥me was purified and biochemicall≥ characterized, and its cr≥stal structure was further resolved. The enz≥me had the highest activit≥ at 50 ℃ and pH 7. 5. It exhibited broad substrate specificit≥on various substrates, including laminarin, barle≥β-glucan, lichenan, et al. ( containing β-1,2,β-1,3,β-1,4 and β-1,6 linkages) . Cr≥stal structure anal≥sis revealed that the enz≥me exhibited t≥pical gl≥coside h≥drol≥ase famil≥3 enz≥mes, having multiple domain structures. The catal≥tic packet of the enz≥me was composed b≥a ( α/β)5 sandwich domain and a ( β/α)8 folding barrel domain linked b≥a loop region. The side chain aromatic amino acids Trp748 and Trp749 were functioned as -1 and +1 binding site, respectivel≥, while Trp131 was functioned as +2 site. These aromatic amino acids formed a narrow pocked and a h≥drophobic environment, which could diminish its transgl≥co-s≥lation activit≥.关键词
巴伦葛兹类芽孢杆菌/β-葡萄糖苷酶/异源表达/酶学性质/晶体结构Key words
Paenibacillus barengoltzii/β-glucosidase/heterologous expression/biochemical characteriza-tion/cr≥stal structure分类
轻工纺织引用本文复制引用
黄平,吴世旺,江正强,杨绍青..巴伦葛兹类芽孢杆菌β-葡萄糖苷酶的表达、酶学性质及结构解析[J].食品科学技术学报,2019,37(1):20-27,8.基金项目
国家自然科学基金资助项目(31771951 （）
31801510) （）
北京市博士后工作经费资助项目(2018-ZZ-009) （2018-ZZ-009）
中国博士后科学基金资助项目(2018M631288). （2018M631288）
