1. Academic Validation
  2. Structural Stabilization and Activity Enhancement of Glucoamylase via the Machine-Learning Technique and Immobilization

Structural Stabilization and Activity Enhancement of Glucoamylase via the Machine-Learning Technique and Immobilization

  • J Agric Food Chem. 2025 Mar 26;73(12):7347-7363. doi: 10.1021/acs.jafc.4c11907.
Frank Peprah Addai 1 Xinglin Chen 2 Hao Zhu 2 Zongjian Zhen 1 Feng Lin 3 Chengxiang Feng 1 Juan Han 4 Zhirong Wang 5 Yun Wang 1 Yang Zhou 2
Affiliations

Affiliations

  • 1 School of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang, Jiangsu 212013, China.
  • 2 School of Life Sciences, Jiangsu University, Zhenjiang, Jiangsu 212013, China.
  • 3 Key Laboratory of Healthy Freshwater Aquaculture, Ministry of Agriculture, Zhejiang Institute of Freshwater Fisheries, Huzhou, Zhejiang 313001, China.
  • 4 School of Food and Biological Engineering, Jiangsu University, Zhenjiang, Jiangsu 212013, China.
  • 5 Promotion Center for Rural Revitalization of Zhejiang, Hangzhou, Zhejiang 310020, China.
Abstract

Glucoamylases (GLL) hydrolyze starch to glucose syrup without yielding intermediate oligosaccharides, but their lack of stability under industrial conditions poses a major limiting factor. Using consensus- and ancestral-based machine-learning tools, a functional GLL with six mutations (GLLI73l/T130V/N212V/D238G/N327M/S332P) was constructed that exhibited superior hydrolytic activity relative to the wild-type (WT-GLL). An oxidized multi-walled carbon nanotube (oMW-CNT) was used as a solid support to immobilize the WT-GLL with an immobilization capacity of 211.28 mg/g. The specific activity of mutant GLL-6M and GLL@oMW-CNTII was improved by 2.5-fold and 3.9-fold respectively, with both retaining 64.5% residual activity after incubation at 50 °C for 2 h compared to the WT-GLL with 42.6% activity. GLL and GLL-6M were however completely inactivated at 55 °C in 30 min while oMW-CNTII retained ∼43.1% activity. Our results demonstrate that employing a machine-learning approach for Enzyme redesign and immobilization is a practicable alternative for improving Enzyme performance and stability for industrial applications.

Keywords

glucoamylase; machine-learning tools; mutation.

Figures
Products