[1]吴利洋,梁梅芳,倪辉,等.微泡菌ALW1褐藻胶裂解酶AlgL14的表达及信息学分析[J].集美大学学报(自然版),2018,23(5):341-348.
 WU Liyang,LIANG Meifang,NI Hui,et al.Expression and Bioinformatics Analysis of Alginate Lyase AlgL14 from Microbulbifer sp. ALW1[J].Journal of Jimei University,2018,23(5):341-348.
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微泡菌ALW1褐藻胶裂解酶AlgL14的表达及信息学分析()
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《集美大学学报(自然版)》[ISSN:1007-7405/CN:35-1186/N]

卷:
第23卷
期数:
2018年第5期
页码:
341-348
栏目:
水产、食品与生物技术
出版日期:
2018-09-28

文章信息/Info

Title:
Expression and Bioinformatics Analysis of Alginate Lyase AlgL14 from Microbulbifer sp. ALW1
作者:
吴利洋1梁梅芳1倪辉1234肖安风1234姜泽东1234朱艳冰1234
(1.集美大学食品与生物工程学院,福建 厦门 361021;2.福建省食品微生物与酶工程重点实验室,福建 厦门 361021;3.厦门市食品与生物工程技术研究中心,福建 厦门 361021;4.厦门市南方海洋研究中心经济海藻资源化利用与深加工重点实验室,福建 厦门 361021)
Author(s):
WU Liyang1LIANG Meifang1NI Hui1234XIAO Anfeng1234JIANG Zedong1234ZHU Yanbing1234
(1.College of Food and Biological Engineering,Jimei University,Xiamen 361021,China;2.Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering,Xiamen 361021,China;3.Research Center of Food Biotechnology of Xiamen City,Xiamen 361021,China;4.Key Laboratory of Systemic Utilization and In??????-depth Processing of Economic Seaweed,Xiamen Southern Ocean Technology Center of China,Xiamen 361021,China)
关键词:
微泡菌褐藻胶裂解酶克隆基因表达信息学分析
Keywords:
Microbulbifer spalginate lyasecloninggene expressionbioinformatics analysis
文献标志码:
A
摘要:
以微泡菌(Microbulbifer sp.)ALW1的基因组为模板,使用褐藻胶裂解酶AlgL14特异性引物进行PCR扩增,将扩增产物克隆至pMD18??????-T载体后进行测序,并对该基因编码的蛋白质序列进行生物信息学分析。将目的基因插入pET??????-28α(+)表达载体,转入Escherichia.coli BL21(DE3)中进行诱导表达,并利用亲和层析进行重组蛋白纯化。结果显示,克隆基因的大小为1 350 bp,预测编码含有449个氨基酸残基的蛋白质。该蛋白质序列与其他菌株来源的褐藻胶裂解酶序列具有一定的相似性,预测克隆的目的基因编码褐藻胶裂解酶,归属于PL??????-14家族。褐藻胶裂解酶AlgL14的理论分子质量大小为48.772 ku,理论等电点为6.27。采用同源建模法建立菌株ALW1褐藻胶裂解酶AlgL14的三维结构,富含β-折叠。将目的基因在E.coli BL21(DE3)中进行诱导表达,并纯化获得重组褐藻胶裂解酶。SDS??????-PAGE分析显示,表达的目的蛋白分子质量约为48.8 ku。
Abstract:
The genomic DNA of Microbulbifer sp.ALW1 was used as the template for amplification of alginate lyase AlgL14 gene by using PCR with a pair of specific primers.The amplified products were cloned into pMD18??????-T vector and then sequenced.The deduced protein sequence of this gene was further analyzed by bioinformatics.The target gene was inserted into pET??????-28α(+) expression vector.The recombinant plasmid was transformed into Escherichia coli BL21(DE3) and the target gene was induced to express.The recombinant alginate lyase AlgL14 was purified through affinity chromatography.The results showed that the cloned gene was 1 350 bp,encoding 449 amino acid residues.The target protein sequence shared certain identities with the alginate lyase sequences from other bacterial strains.So the cloned target gene was predicted to encode an alginate lyase belonging to PL??????-14 family.The theoretical molecular weight and pI of the alginate lyase AlgL14 were 48.772 ku and 6.27,respectively.The three??????-dimensional structure of alginate lyase AlgL14 from ALW1 was constructed by homology modeling and presented β???-strands rich structure.The target gene was transformed into Escherichia coli BL21(DE3) and induced to express.Then the recombinant alginate lyase AlgL14 was purified.The molecular weight of the target protein was approximately 48.8 ku through SDS??????-PAGE analysis.

相似文献/References:

[1]吴丽云,刘韩,倪辉,等.Pseudomonas syringae褐藻胶裂解酶基因的克隆及信息学分析[J].集美大学学报(自然版),2015,20(3):179.
 WU Li-yun,LIU Han,NI Hui,et al.Cloning and Bioinformatics Analysis of an Alginate Lyase Gene from Pseudomonas syringae[J].Journal of Jimei University,2015,20(5):179.
[2]陈艳红,杨帆,肖安风,等.褐藻胶裂解酶发酵工艺优化及其中试放大[J].集美大学学报(自然版),2016,21(3):184.
 CHEN Yan-hong,YANG Fan,XIAO An-feng,et al.Optimization and Scale-up of Alginate Lyase Fermentation Processes[J].Journal of Jimei University,2016,21(5):184.

更新日期/Last Update: 2018-11-08