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划定与分子伴奏体共同表达的重组基基酸碳素基因酶活性

Joplin Nongkhlaw¹, Superior Syngkli¹, Miranda Moirangthem¹

¹Department of Zoology, NEHU, Shillong, 793022, India.

The protein journal

|July 11, 2025

概括

分子陪伴者被用来从 cestodes 生产生物活性的重组聚酸碳酸酶 (PEPCK). 这项研究描述了该酶的特征.

科学领域:

生物化学生化学
分子生物学分子生物学
寄生虫学的寄生虫学

背景情况:

烯酸碳酸酶 (PEPCK) 是一种潜在的药物向因其在宿主新陈代谢中的独特作用.
以前试图表达重组PEPCK的结果是包含物体,需要化学添加剂进行重新化.
分子伴侣为稳定蛋白质结构和功能提供了化学方法的替代方案.

研究的目的:

来自Raillietina echinobothrida的重组PEPCK与大肠杆菌中的分子伴侣一起表达.
为了净化和描述生物活性重组PEPCK.
调查伴侣在产生功能性PEPCK中的作用,以便开发潜在的杀虫剂药物.

主要方法:

在pE-SUMO矢量中对rePEPCK进行子克隆.
在大肠杆菌BL21 (DE3) 中,rePEPCK与沙佩龙等离子体 (pG-KJE8,pG-Tf2) 的同时表达.
通过亲和性染色学净化蛋白质,并对酶动力学进行表征.

主要成果:

通过使用分子伴侣,成功地产生了可溶和活性重组PEPCK (rePEPCK).
描述rePEPCK作为75 kDa单体,在pH 7.0下具有最佳活性.
确定了动力参数,包括Vmax,Km,kcat和催化效率,并确定了最佳的辅因子 (Mn2+) 和核酸 (GDP) 度.

结论:

分子陪伴者促进生物活性的骨干PEPCK的产生.
描述的运动性质为理解PEPCK作为甲虫点提供了基础.
这种方法可以增强对酶的生物化学和生物物理性质的研究,用于药物发现.

关键词:

拉伊利提尼尼埃奇诺波特里达 (Raillietina echinobothrida) 是一种类型.酶动力学酶动力学分子的伴侣是分子的伴侣.基酸是碳酸基因酶的一种.

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