| 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 |
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| 引用本文： | 晁雅熙,王淑艳,吴谡琦,魏家强,陈颢.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[J].海洋学报(英文版),2019,38(6):65-74. |
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| 作者姓名： | 晁雅熙 王淑艳 吴谡琦 魏家强 陈颢 |
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| 作者单位： | 自然资源部第一海洋研究所海洋生物活性物质重点实验室,自然资源部第一海洋研究所海洋生物活性物质重点实验室,自然资源部第一海洋研究所海洋生物活性物质重点实验室;海洋科学与技术试点国家实验室海洋生物学与生物技术功能实验室,自然资源部第一海洋研究所海洋生物活性物质重点实验室,自然资源部第一海洋研究所海洋生物活性物质重点实验室;海洋科学与技术试点国家实验室海洋生物学与生物技术功能实验室 |
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| 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. |
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| 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 about62 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°C in the Tris-HCl buffer at pH of 8.9. Moreover, the enzyme activity was enhanced by the metal ions of Na~+, K~+ and Mg~(2+) 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.
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| 关 键 词： | 褐藻胶裂解酶 弧菌 褐藻胶 聚α-L-古洛糖醛酸 寡糖 |
| 收稿时间： | 2018/2/2 0:00:00 |
Cloning and characterization of a new endo-type polyG-specific alginate lyase from bacteria Vibrio sp. QD-5 |
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| Chao Yaxi,Wang Shuyan,Wu Suqi,Wei Jiaqiang and 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. |
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| Authors: | Chao Yaxi Wang Shuyan Wu Suqi Wei Jiaqiang and Chen Hao |
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| Institution: | 1.Key Laboratory of Marine Bioactive Substances, First Institute of Oceanography, Ministry of Natural Resources, Qingdao 266061, China2.Laboratory for Marine Biology and Biotechnology, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266237, China |
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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°C 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. |
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| Keywords: | alginate lyase Vibrio sp alginate poly-α-L-guluronic acid lyase oligosaccharides |
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