A pH-stable, detergent and chelator resistant type I pullulanase from Bacillus pseudofirmus 703 with high catalytic efficiency.
A pH-stable, detergent and chelator resistant type I pullulanase from Bacillus pseudofirmus 703 with high catalytic efficiency.
复制标题
DOI：
10.1016/j.ijbiomac.2017.11.139
复制
发表时间：
2017-11
影响因子：
8.2
通讯作者：
Zhenghui Lu;Xinling Hu;Panpan Shen;Qinhong Wang;Yuling Zhou;Guimin Zhang;Yanhe Ma
中科院分区：
文献类型：
--
作者：
Zhenghui Lu;Xinling Hu;Panpan Shen;Qinhong Wang;Yuling Zhou;Guimin Zhang;Yanhe Ma
普鲁兰酶具有高效的催化活性，在淀粉冷水解中有着广泛的应用，引起了学术界和工业界的广泛关注。从嗜温菌Bacilluspseudofirmus 703中克隆到一个新的普鲁兰酶基因pul 703。Pul 703是一种I型普鲁兰酶，在45 °C时酶活最高，在25-35 °C下培养72 h，酶活仍保持在70%以上，且具有良好的低温稳定性。pul 703在pH7.0 -8.0范围内酶活最高，在pH5.5 -9.5范围内酶活稳定，12 h后酶活仍保持在80%以上。Pul 703耐EDTA和耐洗涤剂，在10 mM EDTA、10% Triton X-100和Tween 20存在下，相对活性分别为100、99和114.8%。Pul 703能高效地水解普鲁兰多糖，比活为270 U/mg，高于目前报道的所有I型普鲁兰酶。此外，Pul 703可以与α-淀粉酶BLA协同作用，有效地水解支链淀粉。这些结果表明，Pul 703是一个很好的候选人冷淀粉水解。
Pullulanase with high catalytic efficiency has attracted great attention from both the academic and industrial communities for its wide application in cold starch hydrolysis. A novel pullulanase genepul703was cloned from a mesophilic bacteriaBacilluspseudofirmus703. Pul703 was characterized to be a type I pullulanase with maximal activity at 45 °C and good low-temperature stability, more than 70% of activity was detected after incubation at 25–35 °C for 72 h. Pul703 obtained the maximal activity around pH 7.0–8.0, and was highly active and stable over a wide pH range of 5.5–9.5, more than 80% of activity was retained after 12 h incubation in these pHs. Pul703 was EDTA-resistant and detergent-tolerant, with a relative activity of 100, 99, and 114.8% at the presence of 10 mM EDTA, 10% of Triton X-100 and Tween 20, respectively. Pul703 can efficiently hydrolyze pullulan with a specific activity of 270 U/mg, which was higher than all reported type I pullulanases. In addition, Pul703 can act synergistically with α-amylase BLA to efficiently hydrolyze amylopectin. These results suggested that Pul703 was a good candidate for cold starch hydrolysis.