Production of microbial condroitin sulphate with vgb+ Recombinant E. coli strain

In this thesis; it is aimed to produce chondroitin sulfate (CS) which is a product derived from glycosaminoglycan has various applications in medicine, veterinary, pharmaceutical and cosmetics fields due to its unique features, from a new and recombinant microbial source by using reliable biotechnological methods instead of known animal sources. Within the scope of the work; first, the plasmid pUC8:15, carrying the gene vgb, was isolated from E. aerogenes [pUC8:15] strain and replicated. Then, by appropriate procedures, the vgb gene region was transferred to pETM6-PACF plasmid and this new plasmid (pETM6-PACF- pUC8:15) was transferred to the non-pathogenic E. coli (C2987) strain. Molecular structure determination of chondroitin obtained from the recombinant strain of E. coli kfo+-vgb+ was performed by HPLC and NMR analysis. Then, chondroitin sulfate was produced by sulfation process via selective sulfation method. The molecular structure of this product was also confirmed by HPLC and NMR analysis. According to the analysis results the chondroitin and chondroitin sulfate obtained have a molecular weight of 203.0 and 269 Da, respectively. It is known that preparations containing chondroitin and chondroitin sulphate with low MA are absorbed at higher levels after oral intake and are more effective in medical applications. Our results will contribute to overcome the lack of research on this subject in our country. KS obtained from a unique and new strain has a generalizable and preferable value for reliable, high purity KS production. The microbial KS we produce has the potential to be an important source for further experimental studies to investigate the nutritional and pharmaceutical properties and effects.