EXRACTION OF SULPHATED GLYCOSAMINOGLYCAN FROM ANIMAL SOURCES AND OPTIMIZATION OF THE PROCESS PARAMETERS
Project no. 22497 · Status: Kapanmış · 2020
Summary
<p>Glycosaminoglycan (GAG) is known one kind of unbranced heteropolysaccharide. The natural macromolecule can be extracted from the cartilage of animals. Chondroitin-4-sulfate is known as a sulfated glycosaminoglycan, which is the highest proportion of cartilage tissue. It is generally using as a supplement of functional food for preventing and supporting treatment of arthritis. The aim of this study is to obtain sulfated glycosaminoglycan from chicken sternum and bovine tendon by enzymatic hydrolysis and optimize a suitable hydrolysis condition by statistical experimental design using Response Surface Method. In this thesis, in order to determine the effects of the selected four parameters as enzyme amount, pH value, extraction temperature and time on GAG yield, a trial design created with Central Composite Design was applied. It was aimed to reach the highest GAG% efficiency in the shortest time and by using the least amount of enzyme. The most suitable process conditions for the GAG extraction process from chicken sternum were found to be 0.6% enzyme amount, 6.99 pH value, 62.96°C extraction temperature and 10.79 hours extraction time. Under these conditions, the extraction yield value was predicted to be 26.07%. The most suitable process conditions for the GAG extraction process from bovine tendon were found to be 0.8% enzyme amount, 7.02 pH value, 63.20°C extraction temperature and 10.89 hours extraction time. Under these conditions, the extraction yield value was predicted to be 1.72%. The predicted exraction yield values were confirmed experimentally. In the second stage of the study, some properties of sulphated GAGs produced under optimized process conditions were determined by performing SEM analysis, FTIR analysis, total amino acid analysis, color analysis, thermal analysis molecular weight determination and rheological measurements on sulphated GAG to charactarize. It was determined that the characteristic peaks in the FTIR spectrum of GAG samples obtained from chicken sternum and calf tendon were similar to the spectrum of the chondroitin-4-sulphate standard. As a result of SEM analysis, it was observed that chondrocytes and collagen fibrils were integrated into the GAG structure. It was determined that the color values of the GAG samples were comparable to the chondroitin-4-sulphate standard. While the molecular weight of chondroitin-4-sulfate was determined as 2078 Da, the average molecular weight of the polymers of the GAG sample obtained from chicken sternum and calf tendon were found to be 2597 Da and 5391.5 Da, respectively. The flow behavior of solutions prepared using GAG samples were explained with Newton model. Total amino acid content by weight of GAG samples obtained from chicken sternum and calf tendon was obtained as 23.90% and 37.74%, respectively. The results of the analysis showed that sulfated glycosaminoglycan was successfully obtained from chicken sternum and bovine tendon by enzymatic extraction.</p>