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. 2019 Jul 20;13(3):554–562. doi: 10.1007/s11705-019-1834-z

Construction of a CaHPO4-PGUS1 hybrid nanoflower through protein-inorganic self-assembly, and its application in glycyrrhetinic acid 3-O-mono-β-d-glucuronide preparation

Tian Jiang 1,#, Yuhui Hou 1,#, Tengjiang Zhang 1, Xudong Feng 1,✉, Chun Li 1,✉
PMCID: PMC7089396  PMID: 32215221

Abstract

Glycyrrhetinic acid 3-O-mono-β-d-glucuronide (GAMG), an important pharmaceutical intermediate and functional sweetener, has broad applications in the food and medical industries. A green and cost-effective method for its preparation is highly desired. Using site-directed mutagenesis, we previously obtained a variant of β-glucuronidase from Aspergillus oryzae Li-3 (PGUS1), which can specifically transform glycyrrhizin (GL) into GAMG. In this study, a facile method was established to prepare a CaHPO4-PGUS1 hybrid nanoflower for enzyme immobilization, based on protein-inorganic hybrid self-assembly. Under optimal conditions, 1.2 mg of a CaHPO4-PGUS1 hybrid nanoflower precipitate with 71.2% immobilization efficiency, 35.60 mg·g−1 loading capacity, and 118% relative activity was obtained. Confocal laser scanning microscope and scanning electron microscope results showed that the enzyme was encapsulated in the CaHPO4-PGUS1 hybrid nanoflower. Moreover, the thermostability of the CaHPO4-PGUS1 hybrid nanoflower at 55°C was improved, and its half-life increased by 1.3 folds. Additionally, the CaHPO4-PGUS1 hybrid nanoflower was used for the preparation of GAMG through GL hydrolysis, with the conversion rate of 92% in 8 h, and after eight consecutive runs, it had 60% of its original activity.

Keywords: β-glucuronidase, enzyme-inorganic hybrid nanoflower, biotransformation, glycyrrhizin, glycyrrtinic acid 3-O-mono-β-d-glucuronide

Acknowledgements

This research was funded by the National Natural Science Foundation of China (Grant Nos. 21425624, 21878021, and 21506011).

Footnotes

These authors contributed equally to this work.

Contributor Information

Xudong Feng, Email: xd.feng@bit.edu.cn.

Chun Li, Email: lichun@bit.edu.cn.

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