[1]鞠酒,张付云,李倩,等.嗜热毁丝霉菌多糖裂解单加氧酶基因的克隆及生物信息学分析[J].黑龙江农业科学,2018,(03):12-18.[doi:10.11942/j.issn1002-2767.2018.03.0012]
　JU Jiu,ZHANG Fu-yun,LI Qian,et al.Cloning of MtLPMO9F Gene from Thermothelomyces thermophila and Bioinformatic Analysis[J].HEILONGJIANG AGRICULTURAL SCIENCES,2018,(03):12-18.[doi:10.11942/j.issn1002-2767.2018.03.0012]
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嗜热毁丝霉菌多糖裂解单加氧酶基因的克隆及生物信息学分析

《黑龙江农业科学》[ISSN:1002-2767/CN:23-1204/S] 卷: 期数: 2018年03期 页码: 12-18 栏目: 出版日期: 2018-03-10
Title:
Cloning of MtLPMO9F Gene from Thermothelomyces thermophila and Bioinformatic Analysis
作者:
鞠酒1; 2; 张付云1; 李倩1; 2; 谭海东2; 尹恒2
1.大连海洋大学 食品科学与工程学院，辽宁 大连 116023；2.中国科学院 大连化学物理研究所/辽宁省碳水化合物重点实验室，辽宁 大连 116023
Author(s):
JU Jiu1; 2; ZHANG Fu-yun1; LI Qian1; 2; TAN Hai-dong2; YIN Heng2
1.College of Food Science and Engineering，Dalian Ocean University，Dalian 116023,China；2.Dalian Institute of Chemical Physics，Chinese Academy of Sciences，Key Laboratory of Carbohydrates in Liaoning Province，Dalian 116023,China
关键词:
纤维素; 多糖裂解单加氧酶; 嗜热毁丝霉菌; 生物信息学分析
Keywords:
cellulose;  Lytic polysaccharide monooxygenases;  Thermothelomyces thermophila;  bioinformatic analysis
DOI:
10.11942/j.issn1002-2767.2018.03.0012
文献标志码:
A
摘要:
多糖裂解单加氧酶(LPMO)是近来发现的一类裂解结晶多糖(如纤维素和几丁质)的氧化酶，它通过单加氧的形式断裂多糖底物之间的糖苷键。为研究LPMO的功能和结构特性，采用基因工程技术从嗜热毁丝霉菌中克隆出LPMO基因(MtLPMO9F)并构建重组表达载体和重组表达菌株，采用生物信息学在线工具和软件分析MtLPMO9F基因编码的蛋白质MtLPMO9F的理化性质、亲/疏水性、信号肽、结构域及空间结构等。结果表明：成功构建了MtLPMO9F真核表达载体及毕赤酵母表达菌株。生物信息学分析结果表明，MtLPMO9F的预测分子量约35.19 kDa，理论等电点为5.13，有4个潜在的N-糖基化位点和16个O-糖基化位点。系统进化分析显示MtLPMO9F与嗜热子囊菌（Thermoascus aurantiacus）的同家族酶TaLPMO9A有亲缘关系，序列相似性为43%。
Abstract:
Lytic polysaccharide monooxygenases (LPMO)，an oxidase,has been recently found to degrade crystalline polysaccharides- such as cellulose and chitin，which cleaves glycosidic bonds of polysaccharide substrates by oxidized reaction.In order to study the function and structural characteristics of the LPMO encoded by MtLPMO9F,the Cloning of MtLPMO9F gene from Myceliophthora thermophila and the construction of recombinant expression vector and Pichia pastoris expression strain were implemented by using genetic engineering.The results showed that the protein consisted of 338 amino acids,its theory molecule weight was predicted as 35.19 kDa,and the isoelectric point was 5.13.There were 4 potential N-glycosylation sites and 16 O-glycosylation sites in the LPMO.The phylogenetic analysis results showed that MtLPMO9F had a most closed phylogenetic relationship with TaLPMO9A from Thermoascus aurantiacus,and the highest sequence identity between MtLPMO9F and TaLPMO9A was 43%.

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备注/Memo

收稿日期：2018-01-26基金项目：国家自然科学基金资助项目（31670803）；中国科学院STS资助项目（KFJ-SW-STS-143）;中国科学院青年创新促进会资助项目（2015144）。第一作者简介：鞠酒（1993-），男，在读硕士，从事微生物和酶工程研究。E-mail：jujiu1993@126.com。

更新日期/Last Update: 2018-05-02