Optimization of Media for Mass Production of Xanthine Oxidase from Endophytic Fungus

Abstract

Xanthine Oxidase has been in lot of demand in clinical industry due to its crucial role in purine metabolism pathway. Various ailments like gout, hyperuricemia, myocardial infarction and reperfusion injury are related to the levels of expression of Xanthine Oxidase .A plethora of questions that need to be answered regarding the implication of genes in XO related disorders and mechanism of action and metabolic pathway of oxidoreductase class of enzymes. Xanthine Oxidase can also find application in the development of biosensors for monitoring substrate utilization in cell culture media. #19 NOBASVNP, an Endophytic fungus isolated from Nerium oleander , demonstrated the maximum potential to produce Xanthine Oxidase. The pure strain of this Endophytic fungus was grown on synthetic media to assess the yield and activity of Xanthine Oxidase. The natural media demonstrated higher ability to produce XO than synthetic media. The natural media was optimized by conventional methods optimizing one parameter at a time and by use of design expert version 8. The optimized media gave higher XO activity than unoptimized media. The enzyme activity of crude enzyme precipitate obtained from culture filtrate was assessed by Nitro blue tetrazolium assay. The crude enzyme was purified by gel filtration chromatography to achieve 34% fold purity. The various kinetic parameters of the partially purified enzyme were also deduced.Further studies on full- purification and characterization of enzyme would allow the commercial exploitation of enzyme from Endophytic fungus.