A modified method based on p-nitrophenol assay to quantify hydrolysis activities of lipases in litters

Lipases are glycerol ester hydrolases (EC 3.1.1.3) produced by a wide range of microorganisms. They catalyse the hydrolysis of different esters depending on the water content of the reaction medium. Here, we developed a simple methodology to quantify lipase hydrolysis activities using two different litters: a litter of Quercus pubescens (QP) and a litter of both Q. pubescens and Q. ilex. Different p-nitrophenyl esters were used to test hydrolysis in a reaction medium with an organic solvent (heptane). We showed that these activities depended on the amount of litter, the incubation time and the substrate concentration and that they increased with temperature. Furthermore, the lipases from the studied litters were still active after 2 h at 70 °C. These activities showed common properties of lipases: the highest activities were obtained with a medium-acyl chain substrate, p-nitrophenyl laurate. Moreover abiotic hydrolysis with short-chain acyl substrates was observable. The following parameters are recommended to quantify hydrolysis activities of lipases in litters: 10 mM of p-nitrophenyl laurate in 2 ml of heptane, 1 g of litter, 2 ml of water incubated at 30 °C for 2 h.