Enzymatic Production of Chondroitin Oligosaccharides and Its Sulfate Derivatives

Weijiao Zhang; Ruirui Xu; Xuerong Jin; Yang Wang; Litao Hu; Tianmeng Zhang; Guocheng Du; Zhen Kang

doi:10.3389/fbioe.2022.951740

Enzymatic Production of Chondroitin Oligosaccharides and Its Sulfate Derivatives

Front Bioeng Biotechnol. 2022 Jul 13:10:951740. doi: 10.3389/fbioe.2022.951740. eCollection 2022.

Authors

Weijiao Zhang^{1

2

3}, Ruirui Xu^{1

2}, Xuerong Jin^{1

2}, Yang Wang^{1

2}, Litao Hu^{1

2}, Tianmeng Zhang^{1

2}, Guocheng Du^{1

2}, Zhen Kang^{1

2}

Affiliations

¹ The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China.
² The Science Center for Future Foods, Jiangnan University, Wuxi, China.
³ The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China.

Abstract

Chondroitin sulfate (CS) has a wide range of physiological functions and clinical applications. However, the biosynthesis of chondroitin oligosaccharides (o-CHs) and sulfate derivatives with specific length is always challenging. Herein, we report enzymatic strategies for producing homogeneous o-CHs and its sulfate derivatives from microbial sourced chondroitin. Chondroitin disaccharides, tetrasaccharides, hexasaccharides, octasaccharides, and decasaccharides with defined structure were produced by controllably depolymerizing microbial sourced chondroitin with an engineered chondroitinase ABC I. The highest conversion rates of the above corresponding o-CHs were 65.5%, 32.1%, 12.7%, 7.2%, and 16.3%, respectively. A new efficient enzymatic sulfation system that directly initiates from adenosine 5'-triphosphate (ATP) and sulfate was developed and improved the sulfation of chondroitin from 8.3% to 85.8% by optimizing the temperature, sulfate and ATP concentration. o-CHs decasaccharide, octasaccharide, hexasaccharide, tetrasaccharide and disaccharide were modified and the corresponding sulfate derivatives with one sulfate group were prepared. The enzymatic approaches constructed here for preparing o-CHs and its sulfate derivatives pave the way for the study of structure-activity relationship and applications.

Keywords: chondroitin; chondroitin sulfate; chondroitinase ABC I; oligosaccharides; sulfation system.