| 一株曲霉的一种中温糖化酶的纯化及其部分酶学特性 |
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| 引用本文： | 莫德姣,刘君梁,冼亮,段承杰,冯家勋.一株曲霉的一种中温糖化酶的纯化及其部分酶学特性[J].广西农业生物科学,2010(3):434-440. |
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| 作者姓名： | 莫德姣 刘君梁 冼亮 段承杰 冯家勋 |
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| 作者单位： | [1]广西大学生命科学与技术学院,南宁530005 [2]广西亚热带生物资源保护利用重点实验室,南宁530005 [3]广西微生物及植物遗传工程教育部重点实验室,南宁530005 |
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| 基金项目： | 国家科技支撑计划（2007BAD75B05）; 国家863计划（2007AA021307）共同资助 |
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| 摘 要： | 本研究从广西花坪自然保护区采集的土壤中筛选获得了一株产糖化酶丝状真菌菌株57-45,通过形态学观察和真菌内转录间隔区（internal transcribed spacer,ITS）序列比对分析,将其初步鉴定为曲霉属（Aspergillus sp.）的一个种。纯化真菌57-45所产的一种胞外糖化酶经过硫酸铵分级沉淀、疏水层析和阴离子交换层析三步蛋白质纯化步骤,得到在SDS-PAGE上约60kD的单一蛋白质条带,薄层层析分析表明该纯化的蛋白质水解可溶性淀粉的产物只有葡萄糖,证明该纯化的蛋白质为糖化酶。纯化的糖化酶Km值为1.9mg/mL,Vmax为4148μmol/（min·mg）,最适作用pH5.5,最适作用温度50℃,在同步糖化发酵中有应用的潜力。金属离子Fe3＋、Zn2＋、Cu2＋对酶活有较强的抑制作用,EDTA对酶活有较强的促进作用。本文结果将为进一步研究曲霉糖化酶的酶学特性提供新的材料。
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| 关 键 词： | 曲霉 糖化酶 蛋白质纯化 酶学特性 |
Purification and Partial Characterization of a Mesophilic Glucoamylase Produced by an Aspergillus Strain |
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| Mo Dejiao,Liu Junliang,Xian Liang,Duan Chengjie,Feng Jiaxun.Purification and Partial Characterization of a Mesophilic Glucoamylase Produced by an Aspergillus Strain[J].Journal of Guangxi Agricultural and Biological Science,2010(3):434-440. |
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| Authors: | Mo Dejiao Liu Junliang Xian Liang Duan Chengjie Feng Jiaxun |
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| Institution: | 1,2,3 1 College of Life Science and Technology,Guangxi University,Nanning,530005;2 Guangxi Key Laboratory of Ministry of Education for Microbial and Plant Genetic Engineering,Nanning,530005;3 Guangxi Key Laboratory of Subtropical Bioresources Conservation and Utilization,Guangxi University,Nanning,530005 |
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| Abstract: | In this research,filamentous fungal strain 57-45 which produced glucoamylase was isolated from a soil sample collected from the Huaping Nature Reserve of Guangxi.Fungus 57-45 was identified as Aspergillus sp.by morphological and internal transcribed spacer sequence analysis.One extracellular glucoamylase Produced by this fungus 57-45 was purified in a 3-steps procedure involving ammonium sulfate precipitation,hydrophobic interaction chromatography and anion exchange chromatography.The purified protein showed a single band of about 60 kD molecular mass on sodium dodecyl sulfate-polyacrylamide gel electrophoresis.The thin layer chromatography analysis showed that the purified protein released glucose as sole product from soluble starch,confirming that is a glucoamylase.The Km and Vmaxare 1.9 mg/mL and 4 148 μmol/（min·mg）,respectively.The optimum pH and temperature of the purified enzyme toward soluble starch were 5.5 and 50℃,respectively.This enzyme has potential application in the simultaneous saccharification and fermentation.Enzyme activity was significantly inhibited by Fe3＋ Zn2＋ Cu2＋,but activated by EDTA.The results of this work produced by would provide a novel material for investigating the enzyme properties of glucoamylase Aspergillus. |
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| Keywords: | Aspergillus Glucoamylase Purification Enzymatic properties |
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