ENHANCE OF INHIBITORY EFFECT OF PEPTIDIC FRACTION OF ARGENTINEAN RED WINE ON ANGIOTENSIN-I CONVERTING ENZYME BY Oenococcus oeni EXOPROTEASE.

AREDES FERNÁNDEZ, PEDRO; MANCA DE NADRA, MARÍA C

Cordoba. Argentina.

Congreso; III Congreso Internacional de Ciencia y Tecnología de los Alimentos. Córdoba.; 2009

Ministerio de Ciencia y Tecnología de la Provincia de Córdoba (MINCYT Cba).

<!-- /* Font Definitions */ @font-face {font-family:"Cambria Math"; panose-1:2 4 5 3 5 4 6 3 2 4; mso-font-charset:0; mso-generic-font-family:roman; mso-font-pitch:variable; mso-font-signature:-1610611985 1107304683 0 0 159 0;} @font-face {font-family:Calibri; panose-1:2 15 5 2 2 2 4 3 2 4; mso-font-charset:0; mso-generic-font-family:swiss; mso-font-pitch:variable; mso-font-signature:-1610611985 1073750139 0 0 159 0;} @font-face {font-family:Garamond; panose-1:2 2 4 4 3 3 1 1 8 3; mso-font-charset:0; mso-generic-font-family:roman; mso-font-pitch:variable; mso-font-signature:647 0 0 0 159 0;} /* Style Definitions */ p.MsoNormal, li.MsoNormal, div.MsoNormal {mso-style-unhide:no; mso-style-qformat:yes; mso-style-parent:""; margin-top:0cm; margin-right:0cm; margin-bottom:10.0pt; margin-left:0cm; line-height:115%; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-fareast-font-family:Calibri; mso-bidi-font-family:"Times New Roman"; mso-ansi-language:EN-US; mso-fareast-language:EN-US;} .MsoChpDefault {mso-style-type:export-only; mso-default-props:yes; font-size:10.0pt; mso-ansi-font-size:10.0pt; mso-bidi-font-size:10.0pt; mso-ascii-font-family:Calibri; mso-fareast-font-family:Calibri; mso-hansi-font-family:Calibri;} @page Section1 {size:612.0pt 792.0pt; margin:70.85pt 3.0cm 70.85pt 3.0cm; mso-header-margin:35.4pt; mso-footer-margin:35.4pt; mso-paper-source:0;} div.Section1 {page:Section1;} --> Hypertension is a risk factor for cardiovascular disease and stroke. Angiotensin-converting enzyme (ACE) hydrolyzes the inactive Angiotensin I and release Angiotensin II, which increase artery pressure. Different peptides with ACE-inhibitory (I-ACE) activity have been isolated from different food sources. Nitrogenous macromolecular fraction (NMF) of wine is comprised principally by peptides and proteins. Oenococcus oeni X2L, from argentinean wine, release an exoprotease that modify the peptidic fraction of wine. Phenolic compounds of wine can also exhibit I-ACE activity. The objective of this research was to determine the effect of O. oeni exoprotease in the I-ACE activity of NMF. Proteolytic activity was obtained from O. oeni culture. O. oeni was grown in grape juice medium pH 4.8. In exponential growth phase, cells were harvested, washed and resuspended in citrate buffer 0.05 M, pH 5.0, 2h at 20ºC for starvation. Cells were removed and supernatant constituted the exoprotease solution. Proteolytic activity was assayed by Cd-ninhydrin method. NMF was obtained from argentinean red wine. Wine was dialyzed 48 h against tap water using a 5000 Da pore size membrane. A portion of wine was clarified with active carbon to remove phenolic compounds. Retentate was vacuum concentrate and lyophilized. NMF and NMF without phenolic compounds (NMF-PC) were dissolved in citrate buffer at different concentrations namely A,B,C,D. Where "A" contains the same concentration that original wine; "B", "C", "D" are 1.5; 2.0; 5.0 fold concentrated respect to wine. Same concentrations of NMF and NMF-PC were dissolved in exoprotease solution and incubated 30°C, 60 min. Then; proteolytic enzyme was inactivated by heating 70°C, 15 min. Total phenolic compounds were determined by Singleton and Rossi method. ACE-inhibitory activity percentage (% I-ACE) was measured by Cushman-Cheung method. Each experience was repeated three times. Experimental data were analyzed by ANOVA. The NMF at A,B,C,D concentrations showed an % I-ACE of 19.4, 45.3, 50.5 and 49.8 respectively. Exoprotease solution shows proteolytic activity on NMF and NMF-PC. NMF treated with exoprotease solution at A,B,C concentrations increase the % I-ACE at 35.5, 54.0 and 65.8. Clarified wines diminish 99.8% the phenolic compounds concentration. Compared with NMF, in NMF-PC at A,B,C,D concentrations, a decrease of % I-ACE at 15.2, 32.4, 34.5 and 37.5 respectively was observed. The NMF-PC treated with exoprotease solution, increase % I-ACE at 45.4, 47.3, 54.3 and 55.9 respectively for A,B,C and D concentrations. Proteolytic enzyme heat-inactivated from exoprotease solution does not produce ACE inhibition. I-ACE activity of argentinean red wine was demonstrated for first time. The increase of I-ACE activity in the NMF and NMF-PC samples treated with O. oeni exoprotease, could be due to either; an increase of the concentration of bioactive peptides or a release of peptides with higher I-ACE activity. Phenolic compounds showed a partial participation in total I-ACE activity.