Bioconversion de l'acide p-coumarique par Brettanomyces ...

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P-coumaric acid adsorption in wines to avoidethylphenol production by brettanomycesDominique Salameh a,b , Cédric Brandam a , Roger Lteif b andPierre Strehaiano a“Wine active compounds 2008”. Proceedings, Oenoplurimédia Edition, p. 213-216.(a) Laboratoire de Génie Chimique – UMR CNRS 5503 – 5 Rue Paulin Talabot – 31106TOULOUSE – France.(b) Université Saint-Joseph, Faculté des Sciences. Mar Roukoz, Mkallès Po.Box 11514,Beirut 1107-2050, Lebanon.Keywords : p-coumaric acid; adsorption; Brettanomyces; Saccharomyces.1. INTRODUCTIONThe p-coumaric acid is a natural polyphenol of wines, in the hydroxicinnamicacid family. The maceration process brings that hydroxycinnamic acid intogrape juice in the average concentration of 60 mg/L [1]. After fermentation andwine elaboration, the p-coumaric acid concentration cannot exceed 8 mg/L [2].It can be found in a free form, or bounded to other wine components such astartaric acid to form the p-coumaroyltartaric acid or with glucose to form anheteroside.The esterase activity of wine enzymes liberates the p-coumaric acid. In its freeform, it is transformed into 4-vinylphenol by a common enzyme to many winemicroorganisms, the cinnamate decarboxylase. The 4-vinylphenol is associatedto pharmaceutical compounds smell [3]. The problem occurs when wines arecontaminated by brettanomyces yeast. This yeast can turn the 4-vinylphenolinto 4-ethylphenol, due to its specific enzyme presence, the vinylphenolreductase [4]. Nowadays, the horse sweat smell in wines is associated to the 4-ethylphenol molecule. This reaction has a bad organoleptic impact on wine.Therefore, many works studied this bioconversion and proposed treatments toavoid its occurrence. The proposed solutions included many approaches, suchas a high hygienic level to stop brettanomyces contaminations, or physicaltreatments to eliminate the volatile phenol once produced [5].This work presents a new approach based on the p-coumaric acid adsorption onyeasts and oenological components. Its removal from the wine can avoid theethylphenol production. Adsorption on yeast cells mentioned in bibliography[6] was verified. Effects of different factors on the adsorption by two industrialadsorbents were tested.2. MATERIALS AND METHODS2.1. Adsorption tests on Saccharomyces and Brettanomyces cellsDifferent concentrations of Brettanomyces and Saccharomyces yeast wereobtained by fermentations. P-coumaric acid was dissolved in 1ml of pure
ethanol before to be added in different concentrations in a synthetic winemedium. Two minutes after, a sample was taken and centrifuged. The liquidpart was tested on HPLC to detect the p-coumaric acid concentration, and onGC/MS to detect the 4-vinylphenol and 4-ethylphenol content. All the testswere compared with witness samples free of yeast.2.2. Adsorption tests on PVPP and yeast cell wallsPolyvinylpolypyrrolidone (PVPP) and yeast cell walls were used as twooenological adsorbent in the concentration of 3 g/L.The medium used was a synthetic wine medium free of p-coumaric acid. A 2 3factorial experimental design was tested to study the variation of the adsorbedquantity Qe (mol/g) of p-coumaric acid added in different concentrations.Q e = X/m, where m is the adsorbent mass, and X = Ci (initial concentration) - C e(concentration at equilibrium).The factors tested were the pH (3- 3.5 – 4), the temperature (25, 30, 35 o C) andthe ethanol concentration (10- 11.5- 13 % v/v). The agitation was fixed at 250rpm. All the tests were compared with witness samples free of adsorbent. Thep-coumaric acid concentrations tested were 2.5 and 20 mg/L, for each adsorbentand for each level of the experimental design. The contact time for adsorptionwas 5h.3. RESULTS AND DISCUSSION3.1 Adsorption tests in fermentationsResults showed a difference in the adsorption potential of each species. In fact,the brettanomyces sp. showed a better adsorption than the saccharomyces intheir specific conditions (Table 1). There was no 4-vinylphenol or 4-ethylphenol production, corresponding to the lost p-coumaric acid in the contactperiod (2min).Table 1 - Adsorption of p-coumaric acid on Brettanomyces sp. andSaccharomyces sp. in fermentation conditionsInitial concentrationp-coumaric (mg/L)Concentration foundp-coumaric (mg/L)Population(10 6 Cell/ml)Saccharomyces 10 9,83 6010 9,53 12010 9,09 240Brettanomyces 10 9,1 310 8,4 36010 5,2 450These results confirmed the bibliography datas [7]. In fact, the p-coumaric acidis the less adsorbed phenolic acid by the saccharomyces sp. Its adsorption onbrettanomyces cell walls was recently defined [8].3.2 Adsorption tests on PVPP and yeast cell walls
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Bioconversion de l'acide p-coumarique par Brettanomyces ...

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