文献类型：期刊文献

英文题名：Novel α-L-Arabinofuranosidase from Cellulomonas fimi ATCC 484 and Its Substrate-Specificity Analysis with the Aid of Computer

作者：Yang, Ying[1];Zhang, Lujia[1];Guo, Mingrong[1];Sun, Jiaqi[1];Matsukawa, Shingo[3];Xie, Jingli[1,2];Wei, Dongzhi[1,2]

机构：[1]E China Univ Sci & Technol, Sch Biotechnol, Dept Food Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Shanghai Collaborat Innovat Ctr Biomfg, Shanghai 200237, Peoples R China;[3]Tokyo Univ Marine Sci & Technol, Dept Food Sci & Technol, Tokyo 1088477, Japan

年份：2015

卷号：63

期号：14

起止页码：3725

外文期刊名：JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY

收录：;EI(收录号：20151700772933);WOS:【SCI-EXPANDED(收录号:WOS:000353178100014)】；

基金：This work was supported by the National Special Fund for State Key Laboratory of Bioreactor Engineering (2060204), partially supported by the National Natural Science Foundation of China (no. 31201296) and the Fundamental Research Funds for the Central Universities, People 's Republic of China.

语种：英文

外文关键词：alpha-L,-arabinofuranosidase; substrate specificity; binding energy; docking

摘要：In the process of gene mining for novel alpha-l-arabinofuranosidases (AFs), the gene Celf_3321 from Cellulomonas fimi ATCC 484 encodes an AF, termed as AbfCelf, with potent activity, 19.4 U/mg under the optimum condition, pH 6.0 and 40 degrees C. AbfCelf can hydrolyze alpha-1,5-linked oligosaccharides, sugar beet arabinan, linear 1,5-alpha-arabinan, and wheat flour arabinoxylan, which is partly different from some previously well-characterized GH 51 AFs. The traditional substrate-specificity analysis for AFs is labor-consuming and money costing, because the substrates include over 30 kinds of various 4-nitrophenol (PNP)-glycosides, oligosaccharides, and polysaccharides. Hence, a preliminary structure and mechanism based method was applied for substrate-specificity analysis. The binding energy (Delta G, kcal/mol) obtained by docking suggested the reaction possibility and coincided with the experimental results. AbfA crystal 1QW9 was used to test the rationality of docking method in simulating the interaction between enzyme and substrate, as well the credibility of the substrate-specificity analysis method in silico.