Enzymatic Production of Chondroitin Oligosaccharides and Its Sulfate Derivatives

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.

Engineering sulfonate group donor regeneration systems to boost biosynthesis of sulfated compounds

“…2d, Supplementary Figs. 3 and 4a) were introduced for sulfonating chondroitin and trehalose to generate chondroitin sulfate A (CSA) 40 and trehalose-2-sulfate, respectively. After modification, the produced CSA and trehalose-2sulfate were pretreated and analyzed by liquid chromatography with tandem mass spectrometry (HPLC-MS/MS, Supplementary Fig.…”

Construction of a Protein Crystalline Inclusion-Based Enzyme Immobilization System for Biosynthesis of PAPS from ATP and Sulfate

“…3′-Phosphoadenosine-5′-phosphosulfate (PAPS) as the universal sulfate donor is used for the biosynthesis of sulfate-containing compounds, such as chondroitin sulfate, , heparin, zosteric acid, sulfated tyrosine, and naringenin 7-sulfate . At present, PAPS is mainly produced by microorganisms and in vitro enzymatic synthesis .…”