Precise Immobilization Strategy Combined with Rational Design to Improve β-Agarase Stability.

Autor: Liu X; The State Key Laboratory of Food Science and Resources, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.; School of Food Science and Technology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.; Collaborative Innovation Center of Food Safety and Quality Control in Jiangsu Province, Jiangnan University, Wuxi 214122, China., Li X; The State Key Laboratory of Food Science and Resources, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.; School of Food Science and Technology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.; Collaborative Innovation Center of Food Safety and Quality Control in Jiangsu Province, Jiangnan University, Wuxi 214122, China., Xie Q; The State Key Laboratory of Food Science and Resources, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.; School of Food Science and Technology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.; Collaborative Innovation Center of Food Safety and Quality Control in Jiangsu Province, Jiangnan University, Wuxi 214122, China., Lu C; The State Key Laboratory of Food Science and Resources, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.; School of Bioengineering, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China., Xie Z; The State Key Laboratory of Food Science and Resources, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.; School of Food Science and Technology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.; Collaborative Innovation Center of Food Safety and Quality Control in Jiangsu Province, Jiangnan University, Wuxi 214122, China., Zhou X; The State Key Laboratory of Food Science and Resources, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.; School of Food Science and Technology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China., Chen L; The State Key Laboratory of Food Science and Resources, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.; School of Food Science and Technology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China., Qiu C; The State Key Laboratory of Food Science and Resources, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.; School of Food Science and Technology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China., Jin Z; The State Key Laboratory of Food Science and Resources, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.; School of Food Science and Technology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.; Collaborative Innovation Center of Food Safety and Quality Control in Jiangsu Province, Jiangnan University, Wuxi 214122, China., Long J; The State Key Laboratory of Food Science and Resources, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.; School of Food Science and Technology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.; Collaborative Innovation Center of Food Safety and Quality Control in Jiangsu Province, Jiangnan University, Wuxi 214122, China.
Jazyk: angličtina
Zdroj: Journal of agricultural and food chemistry [J Agric Food Chem] 2024 Oct 23; Vol. 72 (42), pp. 23366-23378. Date of Electronic Publication: 2024 Oct 11.
DOI: 10.1021/acs.jafc.4c06466
Abstrakt: Recently, the orientational immobilization of enzymes has attracted extensive attention. In this study, we report the development of a strategy combined with rational design to achieve precise site-specific covalent immobilization of β-agarase. We first rationally screened six surface sites that can be mutated to cysteine by combining molecular dynamics simulation and energy calculation. Site-specific immobilization was successfully achieved by Michael addition reaction of mutant enzymes and maleimide-modified magnetic nanoparticles (MAL-MNPs). The enzyme activity retention rate of R66C-MAL-MNPs and K588C-MAL-MNPs was greater than 96%. The thermal deactivation kinetics study revealed that the site-specific immobilization strategy significantly improved the thermal stability of Aga50D, resulting in a substantial increase in its antidenaturation activity at elevated temperatures, and the highest t 1/2 of the immobilized mutant enzymes was increased by an impressive 21.25-fold at 40 °C. The immobilized mutant enzymes also showed significantly enhanced tolerance to metal ions and organic reagents. For instance, all of the immobilized enzymes maintained over 90% of their enzymatic activity in the 50% (v/v) acetone/water solution. The present work may pave the way for the design of precisely immobilized enzymes, which can help promote green manufacturing.
Databáze: MEDLINE
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