Purification and characterization of cold-active endo-1,4-β-glucanase produced by Pseudoalteromonas sp. AN545 from Antarctica |
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引用本文: | 沈继红,阚光锋,史翠娟,雷振环,解秋菊,钱文佳.Purification and characterization of cold-active endo-1,4-β-glucanase produced by Pseudoalteromonas sp. AN545 from Antarctica[J].中国海洋与湖沼学报,2011,29(5):1086-1092. |
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作者姓名: | 沈继红 阚光锋 史翠娟 雷振环 解秋菊 钱文佳 |
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作者单位: | Key Laboratory of Marine Bio-active Substances;State Oceanic Administration;School of the Ocean;Harbin Institute of Technology at Weihai; |
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基金项目: | Supported by the National High Technology Research and Development Program of China (863 Program) (No. 2007AA091905); the Natural Science Foundation of Shandong Province (No. ZR2010DQ010); the Fundamental Research Funds for the Central Universities (No. HIT.IBRSEM.2009148) |
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摘 要: | A bacterium hydrolyzing carboxymethylcellulose, isolated from Antarctic sea ice, was identified as Pseudoalteromonas sp. based on 16S rDNA gene sequences and named as Pseudoalteromonas sp. AN545. The extracellular endo-1,4-β-glucanase AN-1 was purified successively by ammonium sulfate precipitation, DEAE-Sepharose ion exchange chromatography and Sephadex G-75 gel filtration chromatography. The molecular mass of AN-1 was estimated to be 47.5 kDa utilizing SDS-PAGE and gel chromatography analysis. AN-1 could hydrolyze caboxymethylcellulose, avicel and β-glucan, but not cellobiose, xylan and p-Nitrophenyl-β-D-glucopyranoside. The optimal temperature and pH for the β-glucanase activity of AN-1 were determined to be at 30°C and pH 6.0, respectively. AN-1 was stable at acidic solutions of pH 5.0-6.5 and temperatures below 30°C for 1 h. Moreover, the specific activity was enhanced by Ca2+ and Mg2+, and inhibited by Cu2+. The kinetic parameters Michaelis constant (Km) and maximum velocity (Vmax) of AN-1 were 3.96 mg/mL and 6.06×10-2 mg/(min·mL), respectively.
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关 键 词: | Pseudoalteromonas endo-1 4-β-glucanase cold-active enzyme Antarctic sea ice stability |
收稿时间: | 31 March 2005 |
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