文献类型：期刊文献

英文题名：Production optimization and expression of pectin releasing enzyme from Aspergillus oryzae PO

作者：Chen, Jia[1];Yang, Rujian[1];Chen, Min[1];Wang, Shaohua[1];Li, Peng[1];Xia, Yuelan[1];Zhou, Li[1];Xie, Jingli[1,2];Wei, Dongzhi[1]

机构：[1]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]E China Univ Sci & Technol, Sch Biotechnol, Dept Food Sci & Technol, Shanghai 200237, Peoples R China

年份：2014

卷号：101

期号：1

起止页码：89

外文期刊名：CARBOHYDRATE POLYMERS

收录：;EI(收录号：20134216850529);WOS:【SCI-EXPANDED(收录号:WOS:000330494800012)】；

基金：This work partially supported by National Natural Science Foundation of China (no. 31201296), "the Fundamental Research Funds for the Central Universities", PR China.

语种：英文

外文关键词：Aspergillus oryzae; Protopectinase; Response surface methodology; Endo-polygalacturonase

摘要：Protopectinase is an enzyme that solubilizes protopectin forming highly polymerized soluble pectin. Protopectinase activity was detected from Aspergillus oryzae PO isolated from soil of persimmon orchard. Response surface methodology of Box-Behnken Design with three fermentation variables (temperature, NaNO3 and apple pomace concentration) was used to optimize protopectinase production of A. oryzae PO, and protopectinase activity was improved to 270.0 U/ml. Endo-polygalacturonase belonged to A-type PPase from A. oryzae PO was cloned and expressed in Pichia pastoris GS115. The endo-polygalacturonase expression was 0.418 mg/ml and the specific activity of purified recombinant endo-polygalacturonase was 7520 U/mg toward polygalacturonic acid. The optimal temperature and pH of recombinant endopolygalacturonase were 45 degrees C and 5.0, respectively. The recombinant endo-polygalacturonase activity was enhanced by the presence of Mg2+, while Ca2+, Ni2+ Mn2+, Cu2+ and SOS strongly inhibited the enzyme activity. The apparent K-m value and V-max value were 5.59 mg/ml and 1.01 mu mol/(min ml), respectively. (C) 2013 Published by Elsevier Ltd.