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文章摘要
海洋细菌Catenovulum sp. DP03两个右旋糖酐酶差异分析
Difference Analysis of two Dextranases from Marine Bacterium Catenovulum sp. DP03
投稿时间:2020-09-24  修订日期:2020-10-09
DOI:
中文关键词: Catenovulum sp. DP03  右旋糖酐酶  克隆表达
英文关键词: Catenovulum sp. DP03  dextranase  cloning and expression
基金项目:国家重点研发项目(2018YFC0311106),江苏省研究生科研与实践创新计划项目(SJCX19_1014)
作者单位E-mail
田小鹏 1.江苏省海洋生物资源与环境重点实验室 江苏海洋大学2.江苏省海洋生物技术重点实验室 江苏海洋大学 246562382@qq.com 
邓甜 1.江苏省海洋生物资源与环境重点实验室 江苏海洋大学2.江苏省海洋生物技术重点实验室 江苏海洋大学  
董冬雪 1.江苏省海洋生物资源与环境重点实验室 江苏海洋大学2.江苏省海洋生物技术重点实验室 江苏海洋大学  
吕明生 1.江苏省海洋生物资源与环境重点实验室 江苏海洋大学2.江苏省海洋生物技术重点实验室 江苏海洋大学3.江苏省海洋生物产业技术协同创新中心 江苏海洋大学 246562382@qq.com 
王淑军 1.江苏省海洋生物资源与环境重点实验室 江苏海洋大学2.江苏省海洋生物技术重点实验室 江苏海洋大学3.江苏省海洋生物产业技术协同创新中心 江苏海洋大学  
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中文摘要:
      【目的】右旋糖酐酶能够专一性水解右旋糖酐中的α-(1,6)糖苷键,在食品工业以及龋齿的防治中都有着重要的应用,主要来源于微生物。【方法】将海洋细菌Catenovulum sp. DP03的全基因组测序,从中挖掘编码右旋糖酐酶的基因,并在大肠杆菌中进行异源表达,对重组右旋糖酐酶性质进行比较分析。【结果】海洋细菌Catenovulum sp. DP03中含有两个编码右旋糖酐酶的基因Cadex2870与Cadex2872,其大小分别为2511 bp和2805 bp;Cadex2870与Cadex2872的3D结构与AoDex的结构相似,AoDex的催化区域Q418-D440分别对应Cadex2870的Q431-D453以及Cadex2872的Q425-D447。Cadex2870 与Cadex2872的酶活力分别为16.2 U/mg和 4 U/mg,最适催化温度分别为45℃与30℃,最适催化pH分别为7和8;Cadex2870水解右旋糖酐的产物为异麦芽七糖、异麦芽五糖、异麦芽四糖以及少量异麦芽糖;Cadex2872水解右旋糖酐的产物为异麦芽七糖、异麦芽五糖以及异麦芽四糖。【结论】海洋细菌Catenovulum sp. DP03中含有两个编码右旋糖酐酶的基因,这两个右旋糖酐酶的蛋白质结构以及酶学性质均有差异。
英文摘要:
      [Objective] Dextranase can specifically hydrolyze the α-(1,6) glycosidic bond in dextran, and it has an important applications in industrial production and prevention of dental caries. It is mainly derived from microorganisms. [Methods] The complete genome sequencing of marine bacterium Catenovulum sp. DP03 was completed and dextranase genes were cloned and expressed in Escherichia coli. Then the characterizations of recombinant dextranases were studied. [Results] there were two dextranases genes which was 2511 bp and 2805 bp respectively. The 3D structure of Cadex2870 and Cadex2872 were similar to that of AoDex. The catalytic regions Q418-D440 of AoDex correspond to Q431-D453 of Cadex2870 and Q425-D447 of Cadex2870, respectively. The enzyme activity of Cadex2870 and Cadex2872 were 16.2 U/mg and 4.0 U/mg, respectively. The optimum catalytic temperatures for Cadex 2870 and Cadex 2872 were 45 °C and 30 °C and the optimum catalytic pH were 7 and 8, respectively. The products of Cadex 2870 hydrolyzed dextran were maltoheptaose, maltopentaose, maltotetraose and a small amount of maltose; and Cadex 2872 hydrolyzed dextran were maltoheptaose, maltopentose, and maltotetraose. [Conclusion] The marine bacterium Catenovulum sp. DP03 contains two genes encoding dextranase. There were some differences between the two dextranase in the protein structure and enzymatic properties.
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