文献类型：期刊文献

英文题名：An Acid-Adapted Endo-α-1,5-L-arabinanase for Pectin Releasing

作者：Lang, Chong[1,2];Yang, Rujian[1,2];Yang, Ying[1,2];Gao, Bei[1,2];Zhao, Li[1,2];Wei, Wei[1,2];Wang, Hualei[1,2];Matsukawa, Shingo[3];Xie, Jingli[1,2,4];Wei, Dongzhi[1,2,4]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol, Sch Biotechnol, Dept Food Sci & Technol, Shanghai 200237, Peoples R China;[3]Tokyo Univ Marine Sci & Technol, Dept Food Sci & Technol, Tokyo 1088477, Japan;[4]SCICB, Shanghai 200237, Peoples R China

年份：2016

卷号：180

期号：5

起止页码：900

外文期刊名：APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY

收录：;EI(收录号：20162402481904);WOS:【SCI-EXPANDED(收录号:WOS:000387540700007)】；

基金：This work was supported by National Special Fund for State Key Laboratory of Bioreactor Engineering (2060204) and partially supported by the National Natural Science Foundation of China (Nos. 21506057 and 21506057), the National High Technology Research and Development Program of China (No. 2013AA102109), and the Natural Science Foundation of Shanghai (No. 2013ZR1412100).

语种：英文

外文关键词：Endo-1,5-alpha-L-arabinanase; GH43; Acid-adapted; Pectin extraction; Apple pomace

摘要：An arabinanase gene was cloned by overlap-PCR from Penicillium sp. Y702 and expressed in Pichia pastoris. The recombinant enzyme was named AbnC702 with 20 U/mg of endo-arabinanase activity toward linear alpha-1,5-l-arabinan. The optimal pH and temperature of AbnC702 were 5.0 and 50 A degrees C, respectively. The recombinant AbnC702 was highly stable at pH 5.0-7.0 and 50 A degrees C. It could retain about 72.3 % of maximum specific activity at pH 5.0 after incubation for 2.5 h, which indicated AbnC702 was an acid-adapted enzyme. The K (m) and V (max) values were 24.8 +/- 4.7 mg/ml and 88.5 +/- 5.6 U/mg, respectively. A three-dimensional structure of AbnC702 was made by homology modeling, and the counting of acidic/basic amino residues within the region of 10 around the active site, as well the hydrogen bonds within the area of 5 around the active site, might theoretically interpret the acid adaptability of AbnC702. Analysis of hydrolysis products by thin layer chromatography (TLC) combined with high-performance liquid chromatography (HPLC) verified that the recombinant AbnC702 was an endo-1,5-alpha-l-arabinanase, which yielded arabinobiose and arabinotriose as major products. AbnC702 was applied in pectin extraction from apple pomace with synergistic action of alpha-L-arabinofuranosidase.