chondroitin-sulfates and adenosine-3--phosphate-5--phosphate

Chondroitin sulfate (CS) extracted from animal tissues has been widely used as nutraceutical and pharmaceutical products for osteoarthritis treatment. Here we developed an efficient sulfation-modification system for large scale preparation of CSA in vitro. First, the expression level of C4ST was improved by 30 times with fusion of the chaperone SUMO. Then, glycerol as a protein stabilizer was found to improve rat AST IV stability during the regeneration of cofactor PAPS. Then peptide linkers or protein scaffolds were employed to assemble AST IV and C4ST into artificial complexes to bring the enzymes and PAPS spatially closer and enhance the catalytic efficiency of chondroitin sulfation. Eventually, the system was scaled up to 1 L system and 15 g chondroitin was converted to CSA in 24 h, with a 98 % conversion. The present study made a step further towards the industrial production of CSA with different sulfation degrees.

Topics: Adenosine Diphosphate; Animals; Arylsulfotransferase; Chondroitin Sulfates; Escherichia coli; Kinetics; Metabolic Engineering; Organisms, Genetically Modified; Plasmids; Rats; Recombinant Proteins; Saccharomyces cerevisiae; Saccharomycetales; Solubility; Sulfotransferases; Synthetic Biology