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Purification and properties of an extracellular lipase from the fungus Botrytis cinerea
Author(s) -
Comménil Pascal,
Belingheri Lionel,
Sancholle Michel,
Dehorter Bertrand
Publication year - 1995
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/bf02536044
Subject(s) - lipase , chromatography , isoelectric point , botrytis cinerea , size exclusion chromatography , chemistry , polyacrylamide gel electrophoresis , esterase , umbelliferone , triolein , hydrolysis , gel electrophoresis , column chromatography , sodium dodecyl sulfate , biochemistry , enzyme , biology , coumarin , organic chemistry , botany
Abstract An extracellular lipase (EC 3.1.1.3) from the fungus Botrytis cinerea has been purified to homogeneity and characterized. The purification included ammonium sulfate fractionation and sequential column chromatography. The purification of the preparation was 31‐fold and recovery yield was 21%. The purified enzyme was associated with esterase activity according to activity staining on polyacrylamide gel. The molecular weight was determined as 60 kDa on sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and estimated at 72 kDa using gel filtration, which suggests that the enzyme may be a monomer. The isoelectric point was 6.5, and optimal activity was obtained at 38°C and pH 6.0. This lipase showed a high specificity for synthetic substrates containing long‐chain unsaturated fatty acids using umbelliferone esters. The effect of β‐cyclodextrin on the hydrolysis of olive oil has been studied. The specific activity was 25 μmole/min/mg in the absence of β‐cyclodextrin and 132 μmole/min/mg in its presence.