Engineering robust β-glucuronidase via protein assembly and immobilization for enhanced glycyrrhizin hydrolysis

TY - JOUR

T1 - Engineering robust β-glucuronidase via protein assembly and immobilization for enhanced glycyrrhizin hydrolysis

AU - Wang, Qibin

AU - Yang, Jing

AU - Cao, Weijie

AU - Liu, Hu

AU - Li, Chun

N1 - Publisher Copyright: © 2025

PY - 2025/9

Y1 - 2025/9

N2 - β-Glucuronidase (GUS) plays a crucial role in modifying glycyrrhizin (GL) to enhance its physicochemical properties and biological activities. However, most enzymes suffer from poor thermostability and high production costs, limiting their industrial applications. In this study, we utilized the SpyTag-SpyCatcher pair to assemble tetrameric Aspergillus oryzae GUS (PGUS) into higher-order structures. The thermostability of PGUS was significantly improved by optimizing the assembly sites. Furthermore, we coated an organosilica network (OSN) onto Escherichia coli cells expressing PGUS assemblies using tetraethoxysilane (TEOS) and 3-aminopropyltriethoxysilane (APTES), constructing an immobilized PGUS system. This integrated approach not only enhanced PGUS thermostability without compromising its catalytic activity but also improved the reusability and long-term operational stability, offering a sustainable solution for industrial biocatalysis.

AB - β-Glucuronidase (GUS) plays a crucial role in modifying glycyrrhizin (GL) to enhance its physicochemical properties and biological activities. However, most enzymes suffer from poor thermostability and high production costs, limiting their industrial applications. In this study, we utilized the SpyTag-SpyCatcher pair to assemble tetrameric Aspergillus oryzae GUS (PGUS) into higher-order structures. The thermostability of PGUS was significantly improved by optimizing the assembly sites. Furthermore, we coated an organosilica network (OSN) onto Escherichia coli cells expressing PGUS assemblies using tetraethoxysilane (TEOS) and 3-aminopropyltriethoxysilane (APTES), constructing an immobilized PGUS system. This integrated approach not only enhanced PGUS thermostability without compromising its catalytic activity but also improved the reusability and long-term operational stability, offering a sustainable solution for industrial biocatalysis.

KW - Enzyme assembly

KW - Immobilization

KW - Organosilica network

KW - SpyTag-SpyCatcher

KW - β-Glucuronidase

UR - https://www.scopus.com/pages/publications/105004410122

U2 - 10.1016/j.bej.2025.109772

DO - 10.1016/j.bej.2025.109772

M3 - Article

AN - SCOPUS:105004410122

SN - 1369-703X

VL - 221

JO - Biochemical Engineering Journal

JF - Biochemical Engineering Journal

M1 - 109772

ER -