甲酸脱氢酶和分子伴侣基于金属有机框架ZIF-8载体的共固定化研究

Co-immobilization of formate dehydrogenase and molecular chaperone based on metal organic framework ZIF-8 carrier

摘要

酶的固定化技术能够在一定程度上提高酶的稳定性, 且易于实现酶的循环利用, 但在固定化过程中出现的酶活损失问题, 目前仍是酶工程领域的研究难点。本研究以嗜热毁丝菌 Myceliophthora thermophila 来源的甲酸脱氢酶为研究对象, 采用分子伴侣共固定化技术, 成功将该酶基于 ZIF-8 固定化过程的酶活损失从 27.8% 降至 2.4%。另外, 研究结果还表明, 分子伴侣的共固定化又进一步提高了固定化甲酸脱氢酶的热稳定性、贮存稳定性和循环使用性能, 使其在 60 ℃ 孵育 0.5 h、4 ℃ 贮存 30 d 和循环使用 15 次后的剩余活性分别提高了 18.7%、53.3% 和 23.1%。

Abstract

The immobilization of enzymes can improve their stability to a certain extent and facilitate their recycling. However, the issue of enzyme activity loss during immobilization remains a research challenge in the field of enzyme engineering. Focusing on the formate dehydrogenase from Myceliophthora thermophila and using the technology of molecular chaperone co-immobilization, this study successfully reduced the enzyme activity loss based on ZIF-8 immobilization process from 27.8% to 2.4%. The experimental results also showed that the co-immobilization of molecular chaperone further improved the thermal stability, storage stability, and reusability of immobilized formate dehydrogenase, enabling its residual activity increased by 18.7%, 53.3%, and 23.1% respectively after the incubation at 60 ℃ for 0.5 h, storage at 4 ℃ for 30 d, and 15 cycles of use.

关键词

甲酸脱氢酶 / 分子伴侣 / ZIF-8 / 共固定化

Key words

formate dehydrogenase / molecular chaperone / ZIF-8 / co-immobilization

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基金资助

国家自然科学基金项目(21473256)