广西科学2016，Vol.23Issue(3)：228-233,6.DOI:10.13656/j.cnki.gxkx.20160713.003
Shewanella haliotis BP-1海藻酸裂解酶基因的克隆表达
Gene Cloning and Expression of Alginate Lyase from Shewanella haliotis BP-1
摘要
Abstract
Objective]Alginate lyase in Shewanella haliotis BP-1 strains was studied illustrate its biological activity of degrading alginate.[Methods]The gene cloning technology and the Escherichia coli heterologous expression technology were applied to overexpress the alginate lyase;And the enzyme activity was analyzed after the crude enzyme was separated and purified by DEAE Sepharose FF chromatogra-phy.[Results]The alginate lyase gene Alg 1 7S , with a size of 2 1 5 7 bp,was cloned from S. haliotis BP-1 strain genomic DNA and encoded an alginate lyase Alg17S,which belonged to pol-ysaccharide lyase(PL)1 7 family and had a size of 79 726 Da protein(including an N-terminal signal peptide of 26 amino acid signal peptide).Alg17S showed high sequence identity of 5 2% with PL-17 protein sequence Alg17C from Saccharophagus degradans 2-40.Both the purified recombi-nase Alg17S and the △snAlg17S(without the N-terminal signal peptide of 26 amino acids)can degrade alginate,but the enzymatic activity of △snAlg17S revealed a specific activity of 9 635 U/mg,which was more efficient than Alg17S.[Conclusion]The recombinant alginate lyase △s-nAlg17S that has both high-level expression and high enzymatic activity could be a potential en-zyme for further researching on the alginate saccharification and the biofuels production.关键词
海藻酸裂解酶/克隆表达/酶活性测定/Shewanella haliotis BP-1Key words
alginate lyase/cloning and expression/enzyme activity determination/Shewanella haliotis BP-1分类
生物科学引用本文复制引用
黄桂媛,温顺华,李锋,卢明倩,王巧贞,廖威,黄庶识..Shewanella haliotis BP-1海藻酸裂解酶基因的克隆表达[J].广西科学,2016,23(3):228-233,6.基金项目
国家自然科学基金项目(31560017),广西自然科学重点基金项目(2014GXNSFDA118012)和贵州省教育厅自然科学研究重点项目(黔教合KY[2014]286)资助。 （31560017）
