| 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 |
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| 引用本文： | 朱艳冰,李鹤宾,倪辉,刘静雯,肖安风,蔡慧农. 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[J]. 海洋学报(英文版), 2014, 33(12): 163-169. DOI: 10.1007/s13131-014-0534-2 |
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| 作者姓名： | 朱艳冰 李鹤宾 倪辉 刘静雯 肖安风 蔡慧农 |
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| 作者单位： | 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;厦门医学高等专科学校, 厦门 361008, 中国;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;集美大学食品与生物工程学院, 集美大学, 厦门 361021, 中国;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;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 |
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| 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. |
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| 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 k Da. It had high thermostability at 50°C and 60°C. 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 p H 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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| 关 键 词： | 锰超氧化物歧化酶 嗜热芽孢杆菌 生化特性 深海 Mn-SOD 净化 热稳定性 工业应用 |
| 收稿时间： | 2013-03-15 |
| 修稿时间： | 2013-09-13 |
Purification and biochemical characterization of manganesecontaining superoxide dismutase from deep-sea thermophile Geobacillus sp. EPT3 |
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| ZHU Yanbing,LI Hebin,NI Hui,LIU Jingwen,XIAO Anfeng and 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. DOI: 10.1007/s13131-014-0534-2 |
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| Authors: | ZHU Yanbing LI Hebin NI Hui LIU Jingwen XIAO Anfeng CAI Huinong |
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| Affiliation: | 1.College of Food and Biological Engineering, Jimei University, Xiamen 361021, China;Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering, Xiamen 361021, China;Research Center of Food Microbiology and Enzyme Engineering Technology of Fujian Province, Xiamen 361021, China2.Xiamen Medical College, Xiamen 361008, China3.College of Food and Biological Engineering, Jimei University, Xiamen 361021, China |
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| Abstract: | 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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| Keywords: | Geobacillus sp. superoxide dismutase thermostability purification |
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