| ZHU Yanbing,LI Hebin,NI Hui,LIU Jingwen,XIAO Anfeng,CAI Huinong.Purification and biochemical characterization of manganesecontaining superoxide dismutase from deep-sea thermophile Geobacillus sp. EPT3[J].Acta Oceanologica Sinica,2014,33(12):163-169 |
| 深海嗜热菌Geobacillus sp. EPT3锰超氧化物歧化酶的纯化与生物化学鉴定 |
| Purification and biochemical characterization of manganesecontaining superoxide dismutase from deep-sea thermophile Geobacillus sp. EPT3 |
| 投稿时间：2013-03-15 修订日期：2013-09-13 |
| DOI：10.1007/s13131-014-0534-2 |
| 中文关键词: Geobacillus sp. 超氧化物歧化酶 热稳定性 纯化 |
| 英文关键词:Geobacillus sp. superoxide dismutase thermostability purification |
| 基金项目:The National Natural Science Foundation of China under contract No. 31371751; the Science and Technology Program of Xiamen, China under contract No. 201303120001; the Foundation for Innovative Research Team of Jimei University, China under contract No. 2010A006. |
| 作者 | 单位 | E-mail | | 朱艳冰 | 集美大学食品与生物工程学院, 集美大学, 厦门 361021, 中国
福建省食品微生物与酶工程重点实验室, 厦门 361021, 中国
福建省高校食品微生物与酶工程技术研究中心, 厦门 361012, 中国 | | | 李鹤宾 | 厦门医学高等专科学校, 厦门 361008, 中国 | | | 倪辉 | 集美大学食品与生物工程学院, 集美大学, 厦门 361021, 中国
福建省食品微生物与酶工程重点实验室, 厦门 361021, 中国
福建省高校食品微生物与酶工程技术研究中心, 厦门 361012, 中国 | | | 刘静雯 | 集美大学食品与生物工程学院, 集美大学, 厦门 361021, 中国 | | | 肖安风 | 集美大学食品与生物工程学院, 集美大学, 厦门 361021, 中国
福建省食品微生物与酶工程重点实验室, 厦门 361021, 中国
福建省高校食品微生物与酶工程技术研究中心, 厦门 361012, 中国 | | | 蔡慧农 | 集美大学食品与生物工程学院, 集美大学, 厦门 361021, 中国
福建省食品微生物与酶工程重点实验室, 厦门 361021, 中国
福建省高校食品微生物与酶工程技术研究中心, 厦门 361012, 中国 | jmswthermo@hotmail.com |
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| 中文摘要: |
| 热稳定SOD是一种在生物技术应用中有潜力的酶.本研究中,嗜热菌Geobacillus sp. EPT3分离自东太平洋一个深海热液区.经硫酸铵分级沉淀、DEAE-Sepharose层析及Phenyl-Sepharose层析,从该菌株中纯化得到热稳定超氧化物歧化酶(SOD),纯化倍数13.4,回收率11.1%,酶比活力达3354 U/mg.SOD对KCN和H2O2不敏感性,说明来自Geobacillus sp. EPT3的超氧化物歧化酶为Mn-SOD类型.SOD为同聚二聚体,亚基分子量为26.0 kDa.该酶在50℃和60℃下有高的热稳定性.在检测条件下,SOD在一些抑制剂和变性剂存在时相对稳定,例如β-ME、DTT、PMSF、尿素和盐酸胍.Geobacillus sp. EPT3超氧化物歧化酶在pH 5.0~11.0的广范围内有好的稳定性,能耐受极酸和极碱的变性条件,有利于它的工业应用. |
| 英文摘要: |
| Thermostable SOD is a promising enzyme in biotechnological applications. In the present study, thermophile Geobacillus sp. EPT3 was isolated from a deep-sea hydrothermal field in the East Pacific. A thermostable superoxide dismutase (SOD) from this strain was purified to homogeneity by steps of fractional ammonium sulfate precipitation, DEAE-Sepharose chromatography, and Phenyl-Sepharose chromatography. SOD was purified 13.4 fold to homogeneity with a specific activity of 3 354 U/mg and 11.1% recovery. SOD from Geobacillus sp. EPT3 was of the Mn-SOD type, judged by the insensitivity of the enzyme to both KCN and H2O2. SOD was determined to be a homodimer with monomeric molecular mass of 26.0 kDa. It had high thermostability at 50℃ and 60℃. At tested conditions, SOD was relatively stable in the presence of some inhibitors and denaturants, such as β-mercaptoethanol (β-ME), dithiothreitol (DTT), phenylmethylsulfonyl fluoride (PMSF), urea, and guanidine hydrochloride. Geobacillus sp. EPT3 SOD showed striking stability across a wide pH range from 5.0 to 11.0. It could withstand denaturants of extremely acidic and alkaline conditions, which makes it useful in the industrial applications. |
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