| 4种芽孢杆菌来源木聚糖酶基因在大肠杆菌中的表达及酶活分析 |
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| 引用本文： | 焦志华,徐娥,陆平,李卫芬.4种芽孢杆菌来源木聚糖酶基因在大肠杆菌中的表达及酶活分析[J].饲料工业,2012,33(16):50-54. |
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| 作者姓名： | 焦志华 徐娥 陆平 李卫芬 |
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| 作者单位： | 1. 浙江大学动物科学学院,浙江杭州,310058
2. 贵州大学动物科学学院,贵州贵阳,550025 |
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| 基金项目： | 国家自然科学基金[31072059、30771579]资助 |
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| 摘 要： | 研究旨在比较4种不同来源木聚糖酶基因在大肠杆菌E.coli BL21中表达后的酶活等性质,为生产中筛选高表达量及耐极性木聚糖酶基因提供依据。根据已发表的木聚糖酶基因序列设计引物,分别扩增出浸麻芽孢杆菌(B.macerans)B3、地衣芽孢杆菌(B.licheniformis)B6、蜡样芽孢杆菌(B.cereus)B10和枯草芽孢杆菌(B.subtilis)B12的木聚糖酶基因片段,经克隆表达后获得4种工程菌。经培养和IPTG诱导后进行SDS-PAGE电泳检测及酶特性分析。测序结果表明,4个基因的开放阅读框均为642 bp。其中前3种木聚糖酶基因均为首次报道。4种重组木聚糖酶在55℃下的酶活分别为:36.16、3.85、7.22、98.98 U/ml。4种重组木聚糖酶的最适反应温度均在50~60℃,且枯草芽孢杆菌木聚糖酶最为耐热。结果提示,枯草芽孢杆菌来源的木聚糖酶具有较高的酶活和较好的耐热性。
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| 关 键 词： | 木聚糖酶基因 克隆 表达 酶学性质 |
Expression and enzymatic properties analysis of four sources of bacillus xylanase in Escherichia coli |
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| Jiao Zhihua
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Xu E
,
Lu Ping
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Li Weifen.Expression and enzymatic properties analysis of four sources of bacillus xylanase in Escherichia coli[J].Feed Industry,2012,33(16):50-54. |
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| Authors: | Jiao Zhihua Xu E Lu Ping Li Weifen |
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| Institution: | Jiao Zhihua,Xu E,Lu Ping,Li Weifen |
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| Abstract: | In this study,an enzyme activity comparasion was conducted to find the differences among four kinds of xylanase gene expressions in E.coli BL21,providing the basis for factories to screen the gene with high expression and heat resistance.A pair of primers was designed according to the published nucleotide sequences of putative xylanase genes.With the primers,four target fragments of the β-1,4-endo-xylanase gene was amplified from Bacillus macerans(B.macerans) B3,the lichens Bacillus(B.licheniformis) B6,Bacillus cereus(B.cereus) B10 and Bacillus(B.subtilis) B12,the sequencing results showed that the open reading frame of the four genes were all 642 bp.Three of which were first reported.The results of enzyme characteristics analysis showed that the four recombinant xylanase enzyme activity at 55 ℃ was 36.16,3.85,7.22 and 98.98 U/ml,respectively,in which the bacillus subtilis xylanase enzyme showed the highest enzyme activity.The heat resistance experiment results showed that the bacillus subtilis xylanase had a higher enzyme activity and better heat-resistance. |
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| Keywords: | xylanase gene cloning expression enzymatic properties |
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