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Purification and Biochemical Characterization of Lipase from Ficus carica Latex of Tunisian East Coast Zidi Variety
Author(s) -
LazregAref Houda,
Mosbah Habib,
Fekih Abdelwaheb,
Mars Massoud,
Said Khaled
Publication year - 2012
Publication title -
journal of the american oil chemists' society
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.512
H-Index - 117
eISSN - 1558-9331
pISSN - 0003-021X
DOI - 10.1007/s11746-012-2095-x
Subject(s) - carica , lipase , chemistry , ficus , chromatography , sodium dodecyl sulfate , tributyrin , ripening , pmsf , substrate (aquarium) , enzyme assay , enzyme , food science , biochemistry , botany , biology , ecology
Lipase from Ficus carica L. (Moraceae) latex of the Zidi variety was purified 80.5‐fold with 68.5 % recovery using silica gel chromatography. The molecular weight of the enzyme was 29 kDa as determined by SDS‐PAGE. High lipolytic activity was found in the crude extract during the fruit ripening process. The activity of purified lipase (ZL) seemed to depend strongly on chain length and showed a preference to long chain triacylglycerols. Indeed, ZL specific activity was 370.3 UI/mg using olive oil as a substrate at 45 °C and pH 5.5. In contrast, activity towards short chain triacylglycerols (tributyrin) was 12‐fold lower (32 UI/mg). The enzyme was quite stable in the pH range 4–8, and thermally stable at 60 °C displaying t 1/2 about 90 min using olive oil as a substrate. The values of K m app and V m were found to be 14.3 mM and 294.1 μmol/min/mg, respectively. ZL activity was strongly reduced by Fe 2+ , Mg 2+ and Zn 2+ , while significantly increased by Ca 2+ and Cu 2+ . The enzyme was stimulated by sodium dodecyl sulfate, and Tween‐80, while Triton X‐100 and EDTA had a slight inhibitory effect. No Effect was observed in addition of PMSF and iodoacetic acid.