ORIGINAL RESEARCH article
Front. Microbiol.
Sec. Microbiotechnology
Volume 16 - 2025 | doi: 10.3389/fmicb.2025.1641598
This article is part of the Research TopicAdvanced Biocatalysts for Sustainable Chemical SynthesisView all articles
A high effective expression of human D-glucuronyl C5-epimerase with dimer structure in Escherichia coli
Provisionally accepted
LiJian Guo1
QinXia Song2*- 1State Key Laboratory of Aridland Crop Science, Gansu Agricultural University, Lanzhou, 730070, China, LanZhou, China
- 2Center of Anaerobic Microbes, Institute of Biological Research, Gansu Academy of Sciences, Lanzhou, 730000, China, Lanzhou, China
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Heparan sulfate (HS), a linear anionic polysaccharide, participates in many physiological processes and exhibits many pharmacological activities. D-glucuronyl C5-epimerase (Glce) is one of the key enzymes in the biosynthesis of heparan sulfate proteoglycans. In the present study, we established a valid method for heterologous expression in Escherichia coli (E. coli) and subsequent purification of the N-terminal truncated Glce protein using the SUMO-fused expression system. The soluble fraction demonstrated significantly enhanced yield of human Glce 167-617 , thus facilitating the recovery of a high-purity recombination protein. Subsequently, the homogeneity and stability of Glce 167-617 was characterized through size-exclusion chromatography and dynamic light scattering. We found that human Glce 167-617 exists as a dimer in solution.X-ray crystallographic result further confirmed its dimeric assembly while maintaining the integrity of the catalytic domain. In summary, this study successfully overexpressed and purified human Glce protein in E. coli. The purified Glce protein will be applied to chemoenzymatic synthesis of heparin and heparan sulfates in vitro, which facilitating the future bioengineering of pharmaceutical heparins.
Keywords: D-glucuronyl C5-epimerase, Heparan sulfate biosynthesis, Escherichia coli protein expression, two-step protein purification, Dimer structure
Received: 05 Jun 2025; Accepted: 17 Jul 2025.
Copyright: © 2025 Guo and Song. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: QinXia Song, Center of Anaerobic Microbes, Institute of Biological Research, Gansu Academy of Sciences, Lanzhou, 730000, China, Lanzhou, China
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