Executive Summary : | In recent years, bioactive pectic oligosaccharides (POS) have emerged as the most promising candidate for new-generation prebiotics with several health benefits. Compared to conventional prebiotics like fructan (FOS and inulin) and galactan; POS, with 13 different sugar constituents, offers comparatively more regulations on a wide range of probiotics. In comparison to the available methods of POS extraction processes, the enzymatic method provides several advantages, such as product formation based on enzymatic selectivity and specificity, avoiding adverse chemical modification and ensuring the synergetic construction of POS in mild environmental conditions. However, the process bears significant challenges due to the involvement of selective commercial expensive enzymes, end-product inhibition effects, and target to achieve pure oligomers with physiochemical selectivity to become regulatory effective for a wide range of microbiota. To accomplish cost-effective enzymatic production of POS with particular structural and functional properties, emphasis must be given to the development of onsite production of pectinolytic enzymes such as polygalacturonases, pectate lyase, and pectin methylesterase. Of various fruit wastes, jack fruit rind is rich in pectic substances, and thus, the current proposal envisages utilizing jack fruit rind as a potential feedstock for production of pectinases and the use of the same in the hydrolytic production of POS through an ultrafiltration membrane integrated hybrid rector system. Presently, India is the world's largest producer of Jack Fruit, with a record production of 1.4 million tons. It was reported that 70-80% of the fruit, including its outer rind, which is rich in pectin and other carbohydrates, is usually wasted. Pectin extracted from such wastes will be used as an inducer substrate for producing thermophilic pectinases with suitable activity and stability using selective microorganisms. For controlled production of non-digestible oligosaccharides with a lower degree of polymerization through enzymatic hydrolysis, lab-scale shake flask experiments will be initially performed. The essential parameters such as hydrolysis time, temperature, and enzyme concentrations in a cocktail will be optimized, and subsequently, the optimal conditions will be scaled up in a membrane-integrated hybrid reactor system for the large-scale production of POS. The strategic use of membrane-integrated hybrid operation will not only overcome product inhibition, and retention of the free enzyme and its reutilization during hydrolysis, but at the same time, facilitates screening and selective separation of POS based on its size and associated degree of polymerization. Finally, the physicochemical properties, antioxidant activity, and prebiotic potential of POS will be assessed with the help of high-performance anion exchange chromatography, high-performance size exclusion chromatography, FTIR Spectroscopy, MALDI-TOF spectrometry. |