| 突变酶Y93S对嗜热地衣芽孢杆菌SR01葡聚糖酶热稳定性的影响 |
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| 引用本文： | 韦阳道,易 弋,黎 娅,石征宇,伍时华,梁建荣. 突变酶Y93S对嗜热地衣芽孢杆菌SR01葡聚糖酶热稳定性的影响[J]. 中国酿造, 2017, 36(3): 109. DOI: 10.11882/j.issn.0254-5071.2017.03.023 |
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| 作者姓名： | 韦阳道 易 弋 黎 娅 石征宇 伍时华 梁建荣 |
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| 作者单位： | 1.广西科技大学生物与化学工程学院，广西柳州545006；2.广西糖资源绿色加工重点实验室，广西柳州545006；3.广西高校糖资源加工重点实验室，广西柳州545006；4.柳州市鱼家乐饲料有限公司，广西柳州545002 |
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| 基金项目： | 广西科学基金（2015GXNSFBA139068）；广西柳州市中小企业创新基金（2014F030618） |
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| 摘 要： | 为提高嗜热地衣芽孢杆菌SR01葡聚糖酶的热稳定性，对其相关的氨基酸残基进行定点突变改造。通过对其结构的分析，构建了Y93S突变体，利用分子动力学模拟分析评估后，发现Y93S突变体可能具有较高的耐热能力，利用定点突变技术构建Y93S表达载体并分析温度对表达产物酶活的影响。试验结果表明，突变酶Y93S的最适温度由野生型酶的55 ℃提高至70 ℃；在90 ℃条件下，突变酶Y93S较野生型酶的半衰期由60 min提高到120 min以上；pH及pH稳定性较野生型变化不明显。突变酶Y93S极大的提高了野生型酶的热稳定性，具有潜在的工业应用价值，同时为葡聚糖酶的耐热机理提供有力依据。
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| 关 键 词： | 分子动力学 嗜热地衣芽孢杆菌 葡聚糖酶 热稳定性 |
Effect of mutant enzyme Y93S on the thermal stability of glucanase from thermophilic Bacillus licheniformis SR01 |
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| WEI Yangdao,YI Yi,LI Ya,SHI Zhengyu,WU Shihua,LIANG Jianrong. Effect of mutant enzyme Y93S on the thermal stability of glucanase from thermophilic Bacillus licheniformis SR01[J]. China Brewing, 2017, 36(3): 109. DOI: 10.11882/j.issn.0254-5071.2017.03.023 |
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| Authors: | WEI Yangdao YI Yi LI Ya SHI Zhengyu WU Shihua LIANG Jianrong |
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| Affiliation: | 1.College of Biological and Chemical Engineering, Guangxi University of Science and Technology, Liuzhou 545006, China;
2.Guangxi Key Laboratory of Green Processing of Sugar Resources, Guangxi University of Science and Technology, Liuzhou 545006, China; 3.Key Laboratory for Processing of Sugar Resources of Guangxi Higher Education Institutes, Guangxi University of Science and Technology, Liuzhou 545006, China; 4.Liuzhou City Jiale Fish Feed Limited Company, Liuzhou 545002, China |
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| Abstract: | In order to improve the thermal stability of glucanse from thermophilic Bacillus licheniformis SR01, the related amino acid residues were site-specific mutated. Through the analysis of its structure, the Y93S mutants were constructed. By using molecular dynamics simulation analysis, the Y93S mutants may have higher heat resistance ability. The Y93S expression vector was constructed by site directed mutagenesis technique and the effect of temperature on the activity of the expressed product was analyzed. Results showed that the optimum temperature of the mutant enzyme Y93S increased from 55 ℃ to 70 ℃. At 90 ℃, the half-life period of the mutant enzyme Y93S increased from 60 min to 120 min; pH and pH stability did not change obviously than the wild type. The mutant enzyme Y93S greatly improved the thermal stability of the wild-type enzyme, it had potential application value in industry, and it provided a strong basis for the heat resistance mechanism of the glucanase. |
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| Keywords: | molecular dynamics thermophilic Bacillus licheniformis glucanase thermal stability |
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