Inhibition of extracellular proteases and lipases of Candida albicans by hydroxytyrosol and oleuropein

Banović, Franjo (2016) Inhibition of extracellular proteases and lipases of Candida albicans by hydroxytyrosol and oleuropein. Diploma thesis, Faculty of Science > Department of Biology.

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Abstract

Fungal Sap proteases as well as Lip lipases are known as important virulence factors of medically important fungi. It is for this reason that the inhibition of synthesis and activity of these hydrolitic enzymes is an important target in mycosis eradication. The subject of this study was to determine the possibility of modulation of extracellular Sap proteases and Lip lipases of Candida albicans yeast in vitro by hydroxytyrosol and oleuropein. The first step consisted of the cultivation of clinical isolates of C. albicans on the culture medium, the inoculation and incubation followed by induction of proteases and lipases in liquid medium, and the isolation of enzymes from the medium by filtration. Subsequent step was composed of the selection of the enzymatically most productive strains based on short incubation of the filtrate aliquots with matching substrates – BSA (bovine serum albumin) for proteases, DMPTB (2,3-dimercapto-1-propanol tributyrate) for lipases – and the spectrophotometric analysis with corresponding controls. The final step was the examination of the impact that multiple concentrations of hydroxytyrosol and oleuropein have on the activity of these enzymes, by a method similar to the one from previous step. Clinical isolate designated as 781 has proven to be the most productive protease expressor, while it was not possible to determine the most productive lipase expressor. Both substances have shown inhibitory effect on Sap proteases, with the effect of hydroxytyrosol being significantly stronger. It was not possible to confirm their inhibitory effect on Lip lipases since the singling out of the most productive strain was unsuccessful, with the possibility that the induction itself was unsuccessful as well. The attempt to determine the inhibitory effect on the commercially purchased lipase of a related species, C. rugosa, did not yield clear results.

Item Type: Thesis (Diploma thesis)
Keywords: Candida albicans, Sap proteases, Lip lipases, hydroxytyrosol, oleuropein
Supervisor: Kosalec, Ivan
Co-supervisor: Hrenović, Jasna
Date: 2016
Number of Pages: 46
Subjects: NATURAL SCIENCES > Biology
Divisions: Faculty of Science > Department of Biology
Depositing User: Silvana Šehić
Date Deposited: 25 Mar 2016 09:46
Last Modified: 25 Mar 2016 09:46
URI: http://digre.pmf.unizg.hr/id/eprint/4656

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