Optimizing the sulfation-modification system for scale preparation of chondroitin sulfate A.
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.
['Jin|Xuerong|X|', 'Li|Qing|Q|', 'Wang|Yang|Y|', 'Zhang|Weijiao|W|', 'Xu|Ruirui|R|', 'Li|Jianghua|J|', 'Du|Guocheng|G|', 'Kang|Zhen|Z|']
D000244:Adenosine Diphosphate D000818:Animals D015239:Arylsulfotransferase D002809:Chondroitin Sulfates D004926:Escherichia coli D007700:Kinetics D060847:Metabolic Engineering D030781:Organisms, Genetically Modified D010957:Plasmids D051381:Rats D011994:Recombinant Proteins D012441:Saccharomyces cerevisiae D004718:Saccharomycetales D012995:Solubility D015238:Sulfotransferases D058615:Synthetic Biology
Sulfation-modification system Chondroitin sulfate Protein scaffolds Aryl sulfotransferase IV Enzymatic transformation Sulfotransferase