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3. Food Chemistry & Biotechnology 3.1. Lectures

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3. Food Chemistry & Biotechnology 3.1. Lectures Chem. Listy, 102, s265–s1311 (2008) Food Chemistry & Biotechnology 3. FOOD CHEMISTRY been femented for 10 days at 10 °c under semiaerobic condi- & BIOTECHNOLOGY tions or 4–6 weeks at 10 °c under anaerobic conditions. Second cultivation medium: the sterile vinea drinks were inoculated by studied yeast strains and cultuivated at 3.1. Lectures 20 °c for 10 days under semiaerobic conditions. the samples were than sensorially evaluated by a group L01 NONSACCHAROMYCES YEAST IN GRAPE of degustators. the same samples were than analysed by gas MUST – ADVANTAGE OR SPOILAGE? chromatography for the aroma compounds production. each sample was analysed on the Gc MS (Shimadzu QP 2010) JAROSLAVA Kaňuchová PátKováa, EMÍLIA equipment and also on the Gc FiD equipment (Gc 8000 ce Breierováb and INGRID vajcziKová instruments). aInstitute for viticulture and enology of SARC two methods of sample preparation were done: Matuškova 25, 831 01 Bratislava, Slovak republic, the samples (20 ml) were extracted by ether (2 ml), and bInstitute of ChemistrySAS, Dúbravská cesta 9, 845 38 Brati- centrifuged prior to analysis. the etheric extract was used for slava, Slovak republic, analysis (liquid – liquid extraction). this method was used [email protected] for higher alcohols (propanol, isoamylacohol, ethyl ester and higher alcoholos esters) determination Introduction the samples were extracted by tenaq (solid phase mic- the fresh grape must consists of spontaneous microf- roextraction) and than 10 min sampling according to6. this lora formed from 90 to 99 % by yeasts. the most important method was used for monoterpenic compounds determina- genus is without doubt Saccharomyces cerevisiae which is tion.the same column and the same conditions were used by responsible for successive fermentation and good wine qua- both analysis: column: DB WaX 30 m, 0.25 × 0.25, tempe- lity. recently the contribution of non-Saccharomyces yeasts rature programme: 30 °c, hold 2 min, increase by 4 °c min–1 have been widely discussed as there is not definitive opinion up to 230 °c, hold 10 min, 1 ml of sample was injected to on their contribution to the wine quality, especially aroma. injection port at 200 °c, detector temperature 220 °c, carrier Hanseniaspora osmophila and Kloeckera apiculata should gas: helium, injektion mode: split 1 : 100, flow control mode: be considered detrimental yeast species, by higher acetic pressure 70 kPa acid, acetaldehyde, ethyl acetate and acetoin production1. to avoid spoilage it is recommended to inoculate the grape must Results by Saccharomyces cerevisiae immediately after pressing. after inoculation and fermentation of the grape must and on the other hand, the apiculate non-Saccharomyces yeast vinea drink the number of aroma compounds increased sig- is a natural indigenous microflora which contributes to the nificantly, in both cases and more than 60 compounds were wine origin. thus the question wheather to allow the apicu- found. Most of them were recognized as typical fermentation late microflora to start fermentation or not has not yet been products, etc. ethanol, izoamylacohol, propanol, etylester of solved. caproic, caprylic and caprinic acids, ethylacetate, isovaleric the aim of this study work was to determine the aroma acid, pentylacetate, 2,3-butandiol, furfural, 3-hydroxybu- profile of isolated non-Sacaharomyces yeast from the che- tyrate, methionon, 1,4-butandiol. 2-metyl a 3-metylbutanoic mical as well as the sensorial viewpoints. the yeasts of acid, 2-phenyletylacetate, izoamylacetate, cis 3-hexenylace- the genera Rhodotorula, Sporobolomyces, Pichia, Hanse- tate, etylbenzoate, α-terpineol, etyl isobutyrate, etyl butyrate, nula, Issat-chenkia, and Torulospora, were tested from the etyl 2-metylbutyrate, etyl isovalerate, isoamyl acetate, ethyl viewpoint of their contribution to the wine aroma. the results hexanoate, cis-3-hexenol, ethyl octanoate, furfural, linalool, were than exploited and tested in the real wine-making pro- ethyl furoate, ethyl decanoate, ethyl benzoate, α-terpineol, cess. fenylethyl acetate, and geraniol. the increased production of typicall glycolysis products were also confirmed by several Experimental authors2,3,4. ethyl propionate and propyl acetate, characteri- Following yeast strains were isolated from the grape zed the sample of the grape must fermented by Kloeckera must and degraded products: Rhodotorula mucilaginosa apiculata, and 2-propanol and 2-hexanone characterized the (2 strains), Sporobolomyces pararoseus, Pichia membra- sample of the grape must fermented by Pichia membranefa- nefaciens, Pichia anomala (2 strains), Candida intermedi- ciens2. rojas3 studied analysis of non-Saccharomyces yeast ata, Torulospora delbruecki, and Issatchenkia orientalis. For fermentation products and found higher acetate content, espe- comparison 3 Saccharomyces cerevisiae strains were also cially 2-phenylacetate and isoamylacetate for Pichia yeasts. used. two of them were isolated from the grape must, the albertazzi4 also described higher levels of phenylacetate by third was a commercial one (Lallemand). other yeast - Pichia pastoris and Kloeckera saturnus (700– consequently the isolated yeast strains were inoculated 1700 mg dm–3). From our results we can confirm incereased to the first cultivation medium: the sterile grape must had ester production by all studied microorganisms, especially increased content of ethyl acetate. s537 Chem. Listy, 102, s265–s1311 (2008) Food Chemistry & Biotechnology however, we have found that some microorganisms table iii produced special compounds, which were not recognised evaluation of vinea fermented under semiaerobic conditions by other yeasts. after the fermentation of the grape must by R. mucilaginosa and Sp. pararoseus, aroma compounds Semiaerobic conditions significantly increased. acetate, hexanal, heptanal, octanal, Vinea 20 °c cyclopentanone, thiazole, decalactone, propyl-3-dimethyl Yeast strain aminopropyl, nonanone, heptanon, and butanediol were aroma Percept formed. P. anomala produced especially isoamyl benzyl C. intermediata acidic + ether. the medium fermented by Sporobolomyces was rich in R. mucilaginosa 3 acethone + sabinylacetate, 3,4-hexandion and eicosane. R. mucilaginosa T. delbruecki yeasty + generated cyklopentanol and, α-cyklogeraniol. S. cerevisiae S. cerevisiae 5 yeastly + produced vericaldehyd and γ-nonalakton. I. orientalis grape must + We have found out that yeasts of the genera Pichia, Rho- S. cerevisiae 8 honey + dotorula, and Sporobolomyces did not produce the linalool P. membranofaciens vinea – acetate, contrary to S. cerevisiae. R. mucilaginosa 11 acethone + the differences in the compound production within P. anomala honey + the same yeast species were also observed. S. cerevisiae Sp. pararoseus acethone + strain 8 produced caproic aldehdyd, trans-pinocamphon, dodecanal, 5-methyl-3-heptanon, izo-menthylacetate, con- tylformiate and 3,4-hexandion. these compounds produced trary to S. cerevisiae strain 5 which did not produce any of strain Rhodotorula mucilaginosa strain 3. these compounds. R. mucilaginosa strain 11 produced higher one of the most important factors in wine proofing is the amounts of 2,3-butandiol, but did not produce any izopen- sensorial evaluation of wine aroma. it is very difficult to esti- mate which from all above mentioned compounds will prevail table i over the other ones and wheather the wine could give positive aroma compounds typically produced by various yeast spe- or negative impression. it is due to the complexity of wine aro- cies and flavoural characterisation of founded compounds mas, the heterogenity of perception and recognition thresholds for each one compound as well as many interactions occuring Yeast strain compound Flavour fragrance within and after fermentation5. cyklopentanol, mint aroma as shown in table ii, under anaerobic conditions only R. mucilaginosa alfacyklogeraniol spice flavour T. delbruecki from apiculate microflora developed a pleasant carnation odour aroma. all the other yeast strains evolved unpleasant, smelly Pichia anomala izoamyacetat banana flavour aroma. however, the situation was radically changed when the Sp. pararoseus sabinylacetate fruity aroma fermentation occured under semi-aerobic conditions. Torulo- S. cerevisiae valericaldehyd, coffee aroma spora delbruecki produced the pleasant aroma, exactly defined γ-nonalakton coconut odour as vanilla. also both strains of P. anomala, Pichia membranefa- ciens, Sp. pararoseus and one strain of R. mucilaginosa evo- ked very pleasant aroma, some of them with honey notes, table ii some of them fruity or increased fermetative impression. Sensorial evaluation of fermented grape must by various the results of better sensorial evaluation under semi- yeast strains, + positive impression, – negative impression aerobic conditions were subsequently tested by real wine- making fermentation. the problem which usually occurs in Anaerobic Semiaerobic real process is that the present microfloras consists of several conditions conditions yeast genera which can possitively or negatively contribute Yeast strain aroma perc. aroma perc. to the wine quality. C.intermediata yeast + acidic – the group of degustators confirmed that the wines were R.mucilaginosa 3 socks smelly – acethone – better evaluated when they were fermented under semiaerobic T.delbruecki pleasant + vanilla + conditions, with the inocoluation by S. cerevisae not directly I.orientalis autolyses – acethone – after fermentation, but several hours after grape pressing. the Pichia anomala acethone –
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