Optimization of Spectrophotometric and Fluorometric Assays Using Alternative Substrates for the High-Throughput Screening of Lipase Activity

The effects of reaction conditions on the spectrophotometric and fluorometric assays using alternative substrates (p-nitrophenyl palmitate 4-methylumbelliferyl oleate) were investigated to optimize them for high-throughput screening lipase activity from agricultural products. Four model lipases Chromobacterium viscosum, Pseudomonas fluorescens, Sus scrofa pancreas, wheat germ (Triticum aestivum) allowed hydrolyze at different substrate concentrations (1–5 mM), operating pH (5.0–8.0), temperatures (25–55°C). results show that both worked well standard (pH 7.0 30°C) finding a typical lipase, although should be considered detect catalytic lipases, which are applicable more acidic or alkaline circumstances. To validate optimized conditions, was conducted 17 domestic A pileus Pleurotus eryngii showed highest in (633.42 μU/mg) (101.77 assays. this research provide practical information microplates.