PRODUCTION OF FRUCTOOLIGOSACCHARIDES BY ENDOINULINASE ENZYME OBTAINED FROM LOCAL FUNGAL ISOLATES
Project no. 20752 · Status: Kapanmış (Yayın bekleniyor) · 2019
Summary
Today, as consumers start to learn about the relationship between health and nutrition, many studies are carried out to develop healthy nutrition products in the food ındustry. In addition to their nutritive effects, prebiotics are at the forefront of functional foods that have a positive effect on health when taken into the body. Prebiotics consist of oligosaccharides and polysaccharides. Oligosaccharides, pharmaceutical supplement and dietary fiber in pharmacy for many years; In the food industry, it is used for different purposes such as sweetener, humectant and leavening agent. In recent years, the importance of prebiotic oligosaccharides for the hosts in the intestinal flora has drawn the attention of researchers to this subject. Fructooligosaccharides (FOS), an important member of the oligosaccharides, are obtained by the endoinulinase enzyme hydrolysis of inulin. Inulin is a heterogeneous mixture of fructose polymers, which is commonly found in nature as the storage carbohydrate of plants. Fructooligosaccharides, which are units of inulin with a low degree of polymerization (2-20), are widely used in the food industry due to their gelling, thickening and sweetening properties. It has been demonstrated by in vivo and in vitro studies that FOS contributes to the development of bifidobacteria species in the intestines. In addition, it has been determined by various studies that fructooligosaccharides added to diets are functional nutritional components that can reduce the risk of diabetes, heart diseases, cancer and osteoporosis. In the study, original isolates isolated from various sources in the "Fungal Culture Collection of Ege University Bioengineering Department" were used. Within the scope of this project, firstly, inulinase enzyme screening was carried out in solid nutrient medium containing commercial inulin on 50 fungal isolates. Aspergillus spp. All isolates grew in synthetic solid nutrient media containing inulin as the sole carbon source. In this study, the final destination is to produce high amounts of FOS from inulin, an important prebiotic, with the endoinulinase enzyme. For this purpose, Aspergillus spp. isolates and Aspergillus niger NRRL 363 cultures of 37 fungal culture were grown in broth containing commercial inulin and reducing sugar was determined to determine the endoinulinase enzyme activity. In addition, the total protein determination and specific activities of the isolates in the production media were determined. Then, 16 Aspergillus sp. isolates and Aspergillus niger NRRL 363 strains determined to have high inulinase activity were grown in nutrient media determined for the production of endoinulinase enzyme, and crude enzyme liquids were obtained from the culture broths. At this stage, both commercial inulin and agricultural inulin (dried Jerusalem artichoke) were used. The components of fructooligosaccharides (D-glucose, D-fructose, sucrose, 1-kestose, nystose) formed as a result of fructooligosaccharide (FOS) synthesis by treating the obtained crude enzyme liquids with inulin were quantitatively determined in HPLC. As a result of the analysis, original isolates producing endoinulinase with the potential for commercialization by patenting were determined, and various amounts of FOS were obtained from inulin. It has been proven that all of the Aspergillus isolates have both endo and exo inulinase enzyme activity, FOS components measured in HPLC and fructose, glucose, and sucrose.According to the HPLC data obtained, it was revealed that Jerusalem artichoke is an important substrate for the bioprocesses to be developed for the synthesis of FOS with the endoinulinase enzyme. With this project, an important step has been taken for the enzymatic production of fructooligosaccharides, which have the potential to be used as prebiotics, with the endoinulinase enzyme of our local fungal isolates.