文献类型：期刊文献

英文题名：Structural and biochemical properties of a novel pullulanase of Paenibacillus lautus DSM 3035

作者：Chen, Si-Qi[1];Cai, Xiang-Hai[1];Xie, Jing-Li[1];Wei, Wei[1];Wei, Dong-Zhi[1]

机构：[1]East China Univ Sci & Technol, Newworld Inst Biotechnol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2017

卷号：69

期号：1-2

外文期刊名：STARCH-STARKE

收录：;EI(收录号：20161602243875);WOS:【SCI-EXPANDED(收录号:WOS:000392428600006)】；

基金：This research was financially supported by the National High Technology Research and Development Program of China (No. 2013AA102109, 2012AA022206), the National Natural Science Foundation of China (No. C050203-31200596, 31570795), the National Major Science and Technology Projects of China (No. 2012ZX09304009), and the National Basic Research Program of P. R. China (973, Program No. 2012CB721103).

语种：英文

外文关键词：Enzymatic properties; Functional expression; Hydrolysis products; Paenibacillus lautus; Pullulanase type I

摘要：The pullulanase gene (pul(PL)), encoding a novel type I pullulanase (Pul(PL)), was obtained from a Paenibacillus lautus DSM3035 isolate. The gene has an open reading frame of 2355 bp, After optimizing induction conditions in Escherichia coli, we overexpressed recombinant Pul(PL), purified this enzyme, and assayed its function . The level of functional Pul(PL)-like protein reached its maximum (about 0.28mg/mL, 15% of total protein) after induction for 16h at 20 degrees C. Under these optimized harvesting conditions, Pul(PL) activity was 11.1U/mL. The purified recombinant enzyme with an apparent molecular mass of about 87.9kDa was able to specifically attack the -1,6 linkages in pullulan to generate maltotriose as the major product. The purified Pul(PL) exhibited optimal activity at pH 7.0 and 40 degrees C. The Pul(PL) hydrolyzed pullulan, amylopectin, starch, and glycogen, but not amylose. Substrate specificity observations and reaction products identifications indicate that the purified pullulanase from P. lautus DSM3035 is a type I pullulanase. The present report, to our knowledge, the first to identify type I pullulanase in P.lautus, and detail the enzymatic properties of this enzyme after heterologous expression.