Metabolic Bioengineering of Saccharomyces Cerevisiae for the production of beta glucogallin and its Biological activity

Abstract

Natural products or the compounds from nature have been the source for developing new drugs and formulations have inspired for the development of novel derivatives and bioactive compounds. The study of compounds from natural sources have enabled scientists to elucidate and explore various pathways and mechanisms that have helped immensely to develop novel therapeutics. With the advancement of technology and development of various hyphenated analytical techniques, it has enabled us to explore more into the natural sources of compounds to find the bioactive ones. Procurement and extraction of plant material have been a major drawback in the widespread use and supply of such natural products. The increasing unmet global demand for such drugs has led to the development of metabolic engineering which is an amalgamation of biotechnology, bioinformatics and synthetic biology as a tool to engineer microorganisms and enable them as cell factories to produce natural products by altering various metabolic pathways in the cell system. In this thesis, optimization biosynthesis of and#946;-Glucogallin (BGG), which is a gallo-tannin polyphenol compound was undertaken by employing metabolic engineering methods and incorporating the of beta-glucogallin O-galloyl transferase gene in Saccharomyces cerevisiae (S. cerevisiae) by using CRISPR/Cas9 technology and utilizing the HDR mechanism upon DNA break. BGG is a major constituent in amla and other citrus fruits, is a derivative of and#946;-D-glucose and gallic acid. Like other plant polyphenols, BGG has a wide range of bioactive properties viz., antioxidant, anti-inflammatory, antidiabetic, cataract-preventing, anti-glaucoma and UV protectant, etc. To further elucidate its biological properties, its effects on bacterial lipopolysaccharide (LPS) stimulated the cell signaling in RAW 264.7 and mouse peritoneal macrophages was investigated in details at the molecular level......