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Purification and properties of an extracellular lipase from the fungusBotrytis cinerea

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Lipids

Abstract

An extracellular lipase (EC 3.1.1.3) from the fungusBotrytis 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.

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Abbreviations

DG:

diacylglycerol

EMMEG:

ethylene glycol monoethyl ether

FFA:

free fatty acids

MG:

monoacylglycerol

pI:

isoelectric point

SDS-PAGE:

SDS-polyacrylamide gel electrophoresis

TG:

triacylglycerol

TLC:

thin-layer chromatography

UMB:

umbelliferone

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Authors and Affiliations

  1. Laboratoire de Biologie et Physiologie Végétale, Université de Reims, 51062, Reims Cedex, France

    Pascal Comménil

  2. Laboratoire de Cryptogamie, Université des Sciences et Technologies de Lille, Bâtiment SN 2, 59655, Villeneuve d'Ascq Cedex, France

    Lionel Belingheri, Michel Sancholle & Bertrand Dehorter

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  1. Pascal Comménil
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  2. Lionel Belingheri
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  3. Michel Sancholle
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  4. Bertrand Dehorter
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Comménil, P., Belingheri, L., Sancholle, M. et al. Purification and properties of an extracellular lipase from the fungusBotrytis cinerea . Lipids 30, 351–356 (1995). https://doi.org/10.1007/BF02536044

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  • DOI: https://doi.org/10.1007/BF02536044

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