Improvement of catalytic activity and thermostability of glucose oxidase from <i>Aspergillus heteromorphus</i>.

Shanglin YU; Qiao ZHOU; Honghai ZHANG; Yingguo BAI; Huiying LUO; Xiaojun YANG; Bin YAO

Return

Improvement of catalytic activity and thermostability of glucose oxidase from Aspergillus heteromorphus.

Author: Shanglin YU ¹ ; Qiao ZHOU ¹ ; Honghai ZHANG ² ; Yingguo BAI ² ; Huiying LUO ² ; Xiaojun YANG ¹ ; Bin YAO ²
Author Information

1. College of Animal Science and Technology, Northwest A&F University, Yangling 712100, Shaanxi, China.
2. Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
Publication Type:Journal Article
Keywords: catalytic activity; directed evolution; glucose oxidase; rational design; thermostability
MeSH: Glucose Oxidase/chemistry*; Enzyme Stability; Aspergillus/genetics*; Pichia/metabolism*; Temperature; Catalysis; Fungal Proteins/metabolism*; Hot Temperature
From: Chinese Journal of Biotechnology 2025;41(1):296-307
CountryChina
Language:Chinese
Abstract: Glucose oxidase (GOD) is an oxygen-consuming dehydrogenase that can catalyze the production of gluconic acid hydrogen peroxide from glucose, and its specific mechanism of action makes it promising for applications, while the low catalytic activity and poor thermostability have become the main factors limiting the industrial application of this enzyme. In this study, we used the glucose oxidase AtGOD reported with the best thermostability as the source sequence for phylogenetic analysis to obtain the GOD with excellent performance. Six genes were screened and successfully synthesized for functional validation. Among them, the glucose oxidase AhGODB derived from Aspergillus heteromorphus was expressed in Pichia pastoris and showed better thermostability and catalytic activity, with an optimal temperature of 40 ℃, a specific activity of 112.2 U/mg, and a relative activity of 47% after 5 min of treatment at 70 ℃. To improve its activity and thermal stability, we constructed several mutants by directed evolution combined with rational design. Compared with the original enzyme, the mutant T72R/A153P showcased the optimum temperature increasing from 40 to 50 ℃, the specific activity increasing from 112.2 U/mg to 166.1 U/mg, and the relative activity after treatment at 70 ℃ for 30 min increasing from 0% to 33%. In conclusion, the glucose oxidase mutants obtained in this study have improved catalytic activity and thermostability, and have potential for application.