Enhanced thermostability and catalytic efficiency of glucose oxidase in Pichia Pastoris

Glucose oxidase (GOD) has many practical applications, but its poor thermostability limits broader use. In this research, three primary mutants of wild-type GOD were designed using rational mutagenesis, and the GODm mutant was constructed by combinatorial design. The expression, purification, enzymatic properties studied. specific enzyme activity 2.10-fold higher than that wild type, (kcat/Km) value increased 1.45-fold. After treatment at 55 ℃ for 3 h, retained 37.5% activity, half-life (t1/2) 65 2.28-fold 3.36-fold respectively. By analyzing three-dimensional structure type mutant, it found T30V formed a new hydrogen bond with FAD strengthened hydrophobic interaction, D315K optimized surface electrostatic A162T improved efficiency electron pathway. Thus, novel catalytic obtained in research.