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

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal Pub Date : 2025-05-07 DOI:10.1016/j.bej.2025.109772

Qibin Wang , Jing Yang , Weijie Cao , Hu Liu , Chun Li

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引用次数: 0

Abstract

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.

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工程稳健的β-葡萄糖醛酸酶，通过蛋白质组装和固定化，以增强甘草酸水解

β-Glucuronidase （GUS）在修饰甘草酸（glycyrrhizin， GL）以提高其理化性质和生物活性方面起着至关重要的作用。然而，大多数酶的热稳定性差，生产成本高，限制了它们的工业应用。在这项研究中，我们利用SpyTag-SpyCatcher对将四聚体米曲霉GUS （Aspergillus oryzae GUS）组装成高阶结构。通过优化组装位点，PGUS的热稳定性得到了显著提高。此外，我们利用四乙氧基硅烷（TEOS）和3-氨基丙基三乙氧基硅烷（APTES）将有机硅网络（OSN）包被在表达PGUS组装体的大肠杆菌细胞上，构建了固定化PGUS系统。这种集成方法不仅在不影响其催化活性的情况下提高了PGUS的热稳定性，而且提高了可重复使用性和长期运行稳定性，为工业生物催化提供了可持续的解决方案。

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来源期刊

Biochemical Engineering Journal 工程技术-工程：化工

CiteScore

7.10

自引率

5.10%

发文量

380

审稿时长

34 days

期刊介绍： The Biochemical Engineering Journal aims to promote progress in the crucial chemical engineering aspects of the development of biological processes associated with everything from raw materials preparation to product recovery relevant to industries as diverse as medical/healthcare, industrial biotechnology, and environmental biotechnology. The Journal welcomes full length original research papers, short communications, and review papers* in the following research fields: Biocatalysis (enzyme or microbial) and biotransformations, including immobilized biocatalyst preparation and kinetics Biosensors and Biodevices including biofabrication and novel fuel cell development Bioseparations including scale-up and protein refolding/renaturation Environmental Bioengineering including bioconversion, bioremediation, and microbial fuel cells Bioreactor Systems including characterization, optimization and scale-up Bioresources and Biorefinery Engineering including biomass conversion, biofuels, bioenergy, and optimization Industrial Biotechnology including specialty chemicals, platform chemicals and neutraceuticals Biomaterials and Tissue Engineering including bioartificial organs, cell encapsulation, and controlled release Cell Culture Engineering (plant, animal or insect cells) including viral vectors, monoclonal antibodies, recombinant proteins, vaccines, and secondary metabolites Cell Therapies and Stem Cells including pluripotent, mesenchymal and hematopoietic stem cells; immunotherapies; tissue-specific differentiation; and cryopreservation Metabolic Engineering, Systems and Synthetic Biology including OMICS, bioinformatics, in silico biology, and metabolic flux analysis Protein Engineering including enzyme engineering and directed evolution.