A method to quantify transesterification 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 but this reaction is reversible, depending on the water content of the reaction medium, via esterification and transesterification. The synthetic activity of lipases can be of major importance in natural ecosystems since it can be involved in carbon stockage in soils or litters. Here, the detection of transesterification activities of lipases in litter is reported for the first time. We used two different litters: litter of Quercus pubescens (QP) and litter of both Q. pubescens and Q. ilex. Different p-nitrophenyl esters and pentanol were used to test transesterification in a reaction medium with an organic solvent (heptane). We showed that these activities were proportional to the amount of litter, the incubation time and the substrate concentration and that they increased with temperature. Furthermore, the lipases from the litters studied were very thermostable since they were still active after 2 h at 70 degrees C. These activities showed common properties of lipases: the highest activities were obtained with a medium acyl-chain substrate p-nitrophenyl caprylate and transesterification activities were correlated to water activity, a(w). The following parameters are recommended to quantify transesterification activities in litter: 10 mM of p-nitrophenyl caprylate, 1 g of litter, 500 microL of pentanol, q.s.p. 4 mL of heptane incubated at 30 degrees C for 2 h.
Equipe Ecologie Microbienne et Biotechnologies, Institut Méditerranéen d'Ecologie et de Paléoécologie, U.M.R. C.N.R.S. I.R.D. 6116, Université Paul Cézanne, Faculté des Sciences et Techniques de St Jérôme, 13397, Marseille, Cedex 20, France.
L
L
Goujard
E
E
Ferre
G
G
Gil
F
F
Ruaudel
A M
AM
Farnet
D002210:Caprylates / Q000378:metabolism*; D004795:Enzyme Stability; D008049:Lipase / Q000032:analysis*; D000439:Pentanols / Q000378:metabolism; D029963:Quercus / Q000201:enzymology*; D013696:Temperature
D002210:Caprylates; D000439:Pentanols; C034106:4-nitrophenyloctanoate; D008049:Lipase