提高一个Lytic多糖单氧化酶的酶活性和稳定性
Miesho Hadush Berhe1,2,3,4, Xiangfei Song1,2, Lishan Yao1,2
1Qingdao New Energy Shandong Laboratory, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao 266101, China.
International journal of molecular sciences
|May 27, 2023
概括
蛋白质工程增强了Bacillus amyloliquefaciens LPMO10A (BaLPMO10A) 的活性和热稳定性. 工程 BaLPMO10A 显示改善了纤维素脱聚合,为生物质转化提供了有价值的工具.
科学领域:
- 生物化学和酶学 生物化学和酶学
- 蛋白质工程是指蛋白质工程.
- 生物技术是生物技术.
背景情况:
- 性多糖体单氧化酶 (LPMOs) 是一种关键的依赖铜的酶,用于降解性多糖体,如纤维素和.
- 通过蛋白质工程提高LPMOs的催化效率对于有效的酶生物质转化至关重要.
- 乙氨基面菌LPMO10A (BaLPMO10A) 是优化目标,因为它在多糖分解中的作用.
研究的目的:
- 改造BaLPMO10A的蛋白质序列,以提高其催化活性和稳定性.
- 为了评估工程 BaLPMO10A 变体在各种纤维素基板上的性能.
- 评估工程BaLPMO10A与商业纤维酶的协同效应,以加强生物质降解.
主要方法:
- 使用序列共识方法来优化BaLPMO10A.的蛋白质序列.
- 使用染色基质 2,6-Dimethoxyphenol (2,6-DMP) 量化了酶活性.
- 使用像p-nitrophenyl-β-D-cellobioside (PNPC),carboxymethylcellulose (CMC),酸膨胀纤维素 (PASC),过纸 (FP) 和Avicel.等基质来评估水解和降解潜力. 通过测量明显的化温度来评估热稳定性.
主要成果:
- 与野生类型相比,工程 BaLPMO10A 变种在对 2,6-DMP 的活性上表现出高达 93.7% 的增加.
- BaLPMO10A在PNPC,CMC和PASC上表现出水解活性. 与细胞酶的协同作用导致PASC,FP和Avicel的降解分别增加了2.7倍,2.0倍和1.9倍.
- 突变者表现出增强的热稳定性,明显的化温度比野生类型增加了高达7.5°C.
结论:
- 蛋白质工程成功地提高了BaLPMO10A.的活性和热稳定性.
- 工程 BaLPMO10A 作为一个强大的生物催化剂用于纤维素脱聚合,无论单独还是与纤维素酶协同.
- 这些发现为有效的生物质转化和利用提供了改进的酶工具.
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