| Chao Yaxi,Wang Shuyan,Wu Suqi,Wei Jiaqiang,Chen Hao.Cloning and characterization of a new endo-type polyG-specific alginate lyase from bacteria Vibrio sp. QD-5[J].Acta Oceanologica Sinica,2019,38(6):65-74 |
| 一个新的来自于弧菌QD-5并具有Poly-G内切功能的褐藻胶裂解酶的克隆和性质表征 |
| Cloning and characterization of a new endo-type polyG-specific alginate lyase from bacteria Vibrio sp. QD-5 |
| 投稿时间：2018-02-02 |
| DOI：10.1007/s13131-018-1226-0 |
| 中文关键词: 褐藻胶裂解酶 弧菌 褐藻胶 聚α-L-古洛糖醛酸 寡糖 |
| 英文关键词:alginate lyase Vibrio sp. alginate poly-α-L-guluronic acid lyase oligosaccharides |
| 基金项目:The Marine Public Welfare Project of SOA under contract No. 201505032; the Scientific and Technological Innovation Project financially of Qingdao National Laboratory for Marine Science and Technology under contract No. 2016ASKJ14. |
|
| 摘要点击次数: 1498 |
| 全文下载次数: 1217 |
| 中文摘要: |
| 7株具有利用褐藻胶活性的菌株被从腐败的海带中分离得到，对具有较强褐藻胶裂解活性的菌株弧菌QD-5进行了全基因组测序，测序分析结果表明弧菌QD-5的基因组中含有一个褐藻胶裂解酶基因簇。首次对其中的一个基因aly-IV进行了克隆和性质表征，酶的表达和纯化结果表明褐藻胶裂解酶Aly-IV的分子量为62KDa，等电点为5.12，此酶在pH 8.9，温度为350C时最大特异性酶活可达到1256.78 U/mg。Na+， K+ and Mg2+等金属离子可促进酶的活性。研究表明Aly-IV以内切的方式降解褐藻胶中的Poly-G单元，产物主要为单体和二聚体，由于其较高的底物特异性，Aly-IV可被用作生产低聚合度PolyG寡糖的潜在工具酶，也可用于褐藻胶的结果分析。 |
| 英文摘要: |
| Seven bacterial clones with alginate-utilizing activity were isolated from rotten kelp. By activity test, the Vibrio sp. QD-5 with the potential alginate-degrading capability was chosen to carry out the draft genome sequencing, and the result showed that the Vibrio sp. QD-5 containing an alginate lyase gene cluster. One of these genes, aly-IV, was cloned and characterized for the first time. After overexpression, Aly-IV, with a molecular mass of about 62 kDa and a theoretical isoelectric point (pI) of 5.12, was purified to a specific activity of 1 256.78 U/mg and showed highest activity at 35℃ in the Tris-HCl buffer at pH of 8.9. Moreover, the enzyme activity was enhanced by the metal ions of Na+, K+ and Mg2+ under certain concentration. Aly-IV degraded favorably polyG blocks in an endo-type, yielding monomer and dimer as the main products. Due to its high substrate specificity, Aly-IV could be used as a potential tool for production of polyG oligosaccharides with low degree of polymerization (DP) and for determining the fine structure of alginate. |
|
查看全文
查看/发表评论 下载PDF阅读器 |
| 关闭 |