Induction and purification by three-phase partitioning of aryl alcohol oxidase (AAO) from Pleurotus ostreatus.
Aryl alcohol oxidase (AAO) is an extracellular flavoenzyme involved in lignin degradation by white rot fungi. Screening of lignolytic and AAO activity from twenty different fungal species were carried out. Among them, seven species showed lignolytic activity and three of them (Pleurotus ostreatus, Pleurotus eous, and Pleurotus platypus) were found to be AAO positive. Maximal AAO activity was observed in batch cultures of P. ostreatus and was found to be induced by aromatic amino acids and aryl alcohols up to a level of 289 U/l. Purification of AAO was carried out by three-phase partitioning (TPP). The 67 kDa enzyme was purified up to 10.19-fold by TPP with an overall recovery of 10.95%. Optimum pH and temperature for P. ostreatus AAO activity was found to be around 6 and 40 °C, respectively. From the LB plot, K (m) value of AAO for oxidizing veratryl alcohol was determined to be 0.6 mM. Results of the study indicate that P. ostreatus is the best producers of AAO, and they could be employed as promising fungal species for biotechnological applications.
Kumar
Vaidyanathan Vinoth
VV
Department of Biotechnology, Kumaraguru College of Technology, Coimbatore, India.
Rapheal
Vinohar Stephen
VS
D000429:Alcohol Oxidoreductases / Q000096:biosynthesis* / Q000302:isolation & purification*; D001709:Biotechnology / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004790:Enzyme Induction; D006863:Hydrogen-Ion Concentration; D008031:Lignin / Q000378:metabolism; D020076:Pleurotus / Q000201:enzymology*; D013696:Temperature
D008031:Lignin; D000429:Alcohol Oxidoreductases; C077759:aryl-alcohol oxidase