| Chao Yaxi,Wang Shuyan,Wu Suqi,Wei Jiaqiang,Chen Hao. 2019. Cloning and characterization of a new endo-type polyG-specific alginate lyase from bacteria Vibrio sp. QD-5. Acta Oceanologica Sinica, 38(6):65－74 |
| Cloning and characterization of a new endo-type polyG-specific alginate lyase from bacteria Vibrio sp. QD-5 |
| 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 |
| Received:February 02, 2018 |
| DOI：10.1007/s13131-018-1226-0 |
| Key words:alginate lyase Vibrio sp. alginate poly-α-L-guluronic acid lyase oligosaccharides |
| 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 |
| 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. |
| Author Name | Affiliation | E-mail | | Chao Yaxi | Key Laboratory of Marine Bioactive Substances, First Institute of Oceanography, Ministry of Natural Resources, Qingdao 266061, China | | | Wang Shuyan | Key Laboratory of Marine Bioactive Substances, First Institute of Oceanography, Ministry of Natural Resources, Qingdao 266061, China | | | Wu Suqi | Key Laboratory of Marine Bioactive Substances, First Institute of Oceanography, Ministry of Natural Resources, Qingdao 266061, China
Laboratory for Marine Biology and Biotechnology, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266237, China | | | Wei Jiaqiang | Key Laboratory of Marine Bioactive Substances, First Institute of Oceanography, Ministry of Natural Resources, Qingdao 266061, China | | | Chen Hao | Key Laboratory of Marine Bioactive Substances, First Institute of Oceanography, Ministry of Natural Resources, Qingdao 266061, China
Laboratory for Marine Biology and Biotechnology, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266237, China | hchen@fio.org.cn |
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| Abstract: |
| 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. |
| 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。 |
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