通过在β-板结构中的疏水性相互作用增强β-环氧德克斯糖转移酶的热稳定性
Zihang Jiang1, Xudong Liu1, Yi Ding1
1School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, People's Republic of China.
Journal of agricultural and food chemistry
|July 30, 2025
概括
经过工程设计的酶,β-循环德克斯特林糖酶转移酶 (β-CGTase),显示出增强的热稳定性. 这一突破改善了β-cyclodextrin (β-CD) 的工业生产,提高了产量并降低了成本.
科学领域:
- 生物技术是生物技术.
- 酶工程是什么? 酶工程是什么?
- 工业微生物学 工业微生物学
背景情况:
- β-环极素 (β-CD) 的生产依赖于β-环极素糖酶转移酶 (β-CGTase).
- β-CGTase的有限热稳定性阻碍了高效的工业β-CD生产,增加了成本.
- 提高酶稳定性是提高β-CD产量和经济可行性的关键.
研究的目的:
- 为了提高来自Bacillus circulans STB01.01的β-CGTase的热稳定性.
- 研究特定突变对酶稳定性和催化效率的影响.
- 使用工程β-CGTase变体优化β-CD生产.
主要方法:
- 用于引入疏水性氨基酸替代物,采用了位点导向的突变发生.
- 突变T487 V和T487 V/T500 V是在靠近β-CGTase链的β-片区域创建的.
- 分析了55°C的酶半衰期,催化效率,基质亲和度和β-CD转化率.
主要成果:
- 与野生类型相比,突变T487 V和T487 V/T500 V在55°C时的半衰期 (51.7分钟和68.4分钟) 显着更长 (51.7分钟和68.4分钟).
- 在β-叶片区域的疏水相互作用改善了热稳定性,而不会影响催化效率.
- 双重突变的T487 V/T500 V实现了62.5%的β-CD转化率,反应时间缩短.
结论:
- 通过水性相互作用,局部定向突变成功增强了β-CGTase的热稳定性.
- 工程β-CGTase变体为工业β-CD生产提供了更好的性能.
- 这些发现对具有成本效益和高效的大规模β-CD制造有重大影响.
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