RESEARCH ARTICLE

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 ,
  • Yuhui Hou ,
  • Tengjiang Zhang ,
  • Xudong Feng ,
  • Chun Li
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  • Institute for Synthetic Biosystem/Department of Biochemical Engineering, School of Chemistry and Chemical Engineering, Beijing Institute of Technology, Beijing 100081, China

Received date: 06 Jan 2018

Accepted date: 24 Feb 2019

Published date: 15 Sep 2019

Copyright

2019 Higher Education Press and Springer-Verlag GmbH Germany, part of Springer Nature

Abstract

Glycyrrhetinic acid 3-O-mono-b-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.

Cite this article

Tian Jiang , Yuhui Hou , Tengjiang Zhang , Xudong Feng , Chun Li . 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[J]. Frontiers of Chemical Science and Engineering, 2019 , 13(3) : 554 -562 . DOI: 10.1007/s11705-019-1834-z

Acknowledgments

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