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张锦友,王亚月,曹旭鹏,薛松,张卫,杨艳坤.繁茂膜海绵共附生假单胞菌属菌株DEH138A中脱卤酶的分离纯化及性质表征[J].广西科学,2013,20(2):137-142. [点击复制]
- ZHANG Jin-you,WANG Ya-yue,CAO Xu-peng,XUE Song,ZHANG Wei,YANG Yan-kun.Purification and Characterization of the Dehalogenases from Marine Bacterium Pseudomonas sp. DEH138A Isolated from Marine Sponge Hymeniacidon perlevis[J].Guangxi Sciences,2013,20(2):137-142. [点击复制]
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| 繁茂膜海绵共附生假单胞菌属菌株DEH138A中脱卤酶的分离纯化及性质表征 |
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张锦友1,2, 王亚月1,2, 曹旭鹏1, 薛松1, 张卫3, 杨艳坤4
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| (1.中国科学院大连化学物理研究所海洋生物产品工程组, 辽宁大连 116023;2.中国科学院大学, 北京 100049;3.Flinders大学医学院, Adelaide, SA5042, 澳大利亚;4.郑州大学生物工程系, 河南郑州 450001) |
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| 摘要: |
| 用硫酸铵沉淀,Q-Sepharose HP离子交换层析,Superdex 200凝胶过滤层析和Mono-Q离子交换层析方法,对从大连潮间带繁茂膜海绵中分离得到的1株假单胞菌DEH138A(Pseudomonas sp.DEH138A)中的脱卤酶进行分离纯化和酶学性质表征。结果显示,假单胞菌菌株DEH138A中含有一种能降解L-2-氯丙酸的L-2-卤代酸脱卤酶(L-DEX)。经纯化后,L-DEX亚基分子量为27.8 kDa,全酶分子量为42.5 kDa,推测其为一个二聚体蛋白。L-DEX最适pH值及最适反应温度分别为10.0和30℃,Km值为0.63 mmol/L。L-DEX能够作用于多种2-卤代酸,而且对单溴乙酸的脱卤效果最好。DTT和EDTA对L-DEX无抑制作用,Cu2+和Co2+对L-DEX有明显的抑制作用。L-DEX具有立体选择性等独特的酶学特性,在环境修复、精细化工等领域具有潜在的应用价值。 |
| 关键词: 脱卤酶 假单胞菌 分离纯化 性质表征 |
| DOI： |
| 投稿时间：2013-04-03修订日期：2013-04-27 |
| 基金项目:国家重点基础研究发展计划(973)项目(2009CB724700),中国科学院百人计划项目(A1097),国家自然科学基金项目(31100092)资助。 |
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| Purification and Characterization of the Dehalogenases from Marine Bacterium Pseudomonas sp. DEH138A Isolated from Marine Sponge Hymeniacidon perlevis |
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ZHANG Jin-you1,2, WANG Ya-yue1,2, CAO Xu-peng1, XUE Song1, ZHANG Wei3, YANG Yan-kun4
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| (1.Marine Bioproducts Engineering Group, Dalian Institute of Chemical Physics, CAS, Dalian, Liaoning, 116023, China;2.University of Chinese Academy of Sciences, Beijing, 100049, China;3.School of Medicine, Flinders University, Adelaide, SA5042, Australia;4.Department of Biological Engineering, Zhengzhou University, Zhengzhou, Henan, 450001, China) |
| Abstract: |
| This paper aims at the study of properties of the dehalogenase from bacterium Pseudomonas sp.DEH138A isolated from marine sponge Hymeniacidon perlevis.The dehalogenase from the bacterium Pseudomonas sp.DEH138A was purified by fractional precipitation of (NH4)2SO4,ion exchange chromatography with Q-Sepharose HP,gelfiltration chromatography with Superdex 200 and ion exchange chromatography with Mono-Q.The properties of the L-DEX were also characterized.The results showed that the dehalogenase purified from bacterium Pseudomonas sp.DEH138A was a L-DEX dehalogenase which could specifically dehalogenate L-2-chloropropionate (L-2-CPA).The purified L-DEX was a dimer and its subunit molecular weight was about 27.8 kDa.The optimum pH value and temperature were 10.0 and 30℃,respectively.The Km of the L-DEX was 0.63 mmol/L.The L-DEX could degrade many 2-haloacids and showed the best activity on monobromoacetate.DTT and EDTA could not inhibit the activity of L-DEX,but Cu2+ and Co2+ could inhibit the activity of the L-DEX completely.Considering its promising properties,the dehalogenases from the strain DEH138A may find potential application in environmental and chemical industry. |
| Key words: dehalogenase Pseudomonas sp. DEH138A purification characterization |
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