WO2009098342A1 - Microorganisme fermenteur producteur de concentrations élevées de glycérol et ses applications dans la production de boissons alcoolisées/de vin - Google Patents

Microorganisme fermenteur producteur de concentrations élevées de glycérol et ses applications dans la production de boissons alcoolisées/de vin Download PDF

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WO2009098342A1
WO2009098342A1 PCT/ES2009/070008 ES2009070008W WO2009098342A1 WO 2009098342 A1 WO2009098342 A1 WO 2009098342A1 ES 2009070008 W ES2009070008 W ES 2009070008W WO 2009098342 A1 WO2009098342 A1 WO 2009098342A1
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wine
ethyl
production
wines
glycerol
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Carmela Belloch Trinidad
José TARÍN
Patricia Martorell Guerola
Ramón GÓMEZ
María Teresa FERNÁNDEZ-ESPINAR GARCÍA
Pablo Ossorio Gonzalez
Amparo QUEROL SIMÓN
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Bodegas Murviedro S A
Consejo Superior de Investigaciones Cientificas CSIC
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Bodegas Murviedro S A
Consejo Superior de Investigaciones Cientificas CSIC
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
    • C12G1/00—Preparation of wine or sparkling wine
    • C12G1/02—Preparation of must from grapes; Must treatment and fermentation
    • C12G1/0203—Preparation of must from grapes; Must treatment and fermentation by microbiological or enzymatic treatment
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
    • C12G3/00—Preparation of other alcoholic beverages
    • C12G3/02—Preparation of other alcoholic beverages by fermentation
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • C12N1/18—Baker's yeast; Brewer's yeast
    • C12N1/185—Saccharomyces isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
    • C12P1/02—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using fungi
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
    • C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
    • C12P7/18—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic polyhydric
    • C12P7/20—Glycerol
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/62—Carboxylic acid esters
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi
    • C12R2001/85—Saccharomyces
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi
    • C12R2001/85—Saccharomyces
    • C12R2001/865—Saccharomyces cerevisiae

Definitions

  • the present invention falls within the Agrifood area and specifically in the winemaking sector.
  • yeast strains of wine origin belonging to the species Saccharomyces bayanus var. uvarum (BM58) and Saccharomyces cerevisiae (BM60), which have the capacity to produce an increase in glycerol concentration in wine of approximately 100%, varying the concentration between 9 and 22 g / L depending on the fermentation temperature and increasing Ia production of secondary aromas, increasing the organoleptic quality of the wine.
  • the chemical composition of the wine is determined by several factors, including the grape variety, the geographical conditions, the microbial ecology of the grapes, the fermentation processes and the vinification practices (CoIe and Noble, 1997).
  • Glycerol is a colorless, odorless and high viscosity triol, commercially known as glycerin. It is the most abundant component in wine after ethanol and is formed during alcoholic fermentation as a result of the oxidation of sugars carried out by yeasts (Scanes et al., 1998).
  • glycerol provides sweetness above about 5.2 g / L in dry white wine.
  • Glycerol concentrations in dry and semi-sweet wines vary between 5 and 14 g / L, although higher concentrations (up to 21 g / L) can be found in white wines made from must whose grapes have been infected by the fungus Botrytis cinerea (Calderone et al., J. Agrie. Food Chem., 52 (19), 5902-5906, 2004).
  • the aroma of the wines is one of the most important characteristics in the assessment of its quality. It can be divided into three main groups: (i) aromas from the grape variety or primary aromas, produced by volatile substances, transferred by the grapes from the grape to the must; (ii) aromas produced during fermentation or secondary aromas, which include two types of major aromatic compounds, higher alcohols and esters; and (iii) the "bouquet", also known as tertiary aroma, produced by the final transformation of the former during aging.
  • aromas from the grape variety or primary aromas produced by volatile substances, transferred by the grapes from the grape to the must
  • aromas produced during fermentation or secondary aromas which include two types of major aromatic compounds, higher alcohols and esters
  • the "bouquet" also known as tertiary aroma
  • glycerol does not have a direct impact on the aromatic characteristics of the wine, it does contribute to the final taste and body of the wine, providing sweetness, hence in some European countries it is used as an indicator of quality in wine. For these reasons, glycerol is sometimes added fraudulently in wine, thus decreasing its quality. Taking into account that the addition of glycerol to wine is a practice not allowed by the regulations of the European Commission (EC Regulation 822/87, appendix IV), it is of great interest to use a yeast strain that, after its addition to the must of grape, produces an increase in glycerol concentration during alcoholic fermentation.
  • Saccharomyces cerevisiae strains whose overexpression in the GPD1 and GPD2 genes results in an overproduction of glycerol (from Barros Lopes et al., 2000).
  • these strains also increase acetic acid concentrations to unacceptable levels.
  • GMO genetically modified microorganisms
  • food containing genetically modified organisms was regulated in 1997 by the European Union, and requires the labeling of products intended for food when they contained legally authorized genetically modified organisms that represent at least 1% of the product.
  • Saccharomyces genus In the must different yeast genera and species are present, but the Saccharomyces genus, and mainly the S. cerevisiae species, is responsible for alcoholic fermentation (Pretorius, 2000). However, there are other species of the Saccharomyces "sensu stricto" complex that have also been found in fermentation processes. Specifically, S. bayanus is typical of wine fermentations carried out at low temperatures, and some strains have fermentative properties better than some S. cerevisiae strains as an increase in glycerol concentration (SS González 2006. Doctoral thesis). Therefore, the use of strains of S. cerevisie or S. bayanus isolated from wine, and that during the alcoholic fermentation produce a Increased glycerol concentration is very interesting for its application in the wine industry.
  • One of these tools consists in improving the composition of the wine, and therefore its quality, by selecting the microorganisms used in the fermentation.
  • the object of the present invention is a microorganism useful for the fermentation of alcoholic beverages, preferably wine, characterized by being a non-genetically modified microorganism, isolated and selected from natural fermentations, which produces high concentrations of glycerol and that generates a series of secondary aromas, improving the organoleptic qualities of the fermentation product.
  • microorganism object of the present invention is a yeast of the genus Saccharomyces comprising the species S. cerevisiae or S. bayanus, and more specifically the yeast strains S. bayanus var. uvarum (BM58) (CECT 13003) or S. cerevisiae (BM60) (CECT 13004).
  • the microorganism object of the present invention is characterized by being isolated and selected from natural fermentations of wines from production areas (denomination of origin DO) controlled Utiel-Requena, Valencia and Alicante, for presenting a majority implantation in wine fermentation , up to 100% with any type of grape variety and characterized by tolerating: low pH, up to 2.8; growth and fermentation temperature both high and low, between 10 and 3O 0 C; osmotic pressure of up to 300 g / l of sugars; and high concentrations of alcohol in the medium, up to 15%.
  • DO production areas
  • Utiel-Requena Valencia and Alicante
  • Another object of the invention consists essentially in that the microorganism of the present invention produces an increase in glycerol concentration greater than 9 g / L, even reaching up to 22.27 g / L, which is equivalent to up to 150% more glycerol.
  • the microorganism object of the present invention is characterized by producing an increase in the production of secondary aromas with respect to a commercial yeast characterized by being a very aromatic yeast that has been used as a control.
  • These secondary aromas may belong to the group of acetates, ethyl esters, and / or alcohols.
  • a final object of the present invention is the use of the microorganism object of the present invention for the production of fermented alcoholic beverages, and particularly in the preparation of any type of wine from grape musts of any variety.
  • yeasts In the market there is a large number of yeasts.
  • the Yeast selection is carried out taking into account a series of general oenological characteristics such as high fermentative power, low volatile acidity, regularity in fermentation, absence of olfactory effects, non-foaming and that the viability of yeasts does not decrease after the production process of active dry yeast and even that are not very nutritionally demanding (Querol et al., 1992a and Suárez and I ⁇ igo, 2004).
  • We can also talk about other more specific criteria such as aromatic yeasts that degrade malic acid or even increase the concentration of glycerol (Pretorius, 2000).
  • the object of the present invention is a microorganism useful for the fermentation of alcoholic beverages, preferably wine, characterized by being a non-genetically modified microorganism, isolated and selected from natural fermentations, which produces high concentrations of glycerol and that generates other secondary aromas, improving the organoleptic qualities of the fermentation product.
  • strain BM58 produces 22.27 g / L
  • a particular object of the present invention is that the microorganism object of the present invention is a yeast of the genus Saccharomyces comprising the species S. cerevisiae or S. bayanus.
  • yeast of the genus Saccharomyces comprising the species S. cerevisiae or S. bayanus.
  • Saccharomyces genus has been related to the production of alcoholic beverages, among which are: S. cerevisiae, S. bayanus (var. Bayanus and var. Uvarum) and S. pastorianus, among others.
  • the classic taxonomy of yeasts is based on morphological, physiological and biochemical characteristics, criteria with which all these names of yeasts related to alcoholic fermentation arose, although they are often insufficient to establish with precision the phylogeny (ratio of microorganisms in different lineages or strains) of yeasts.
  • phylogeny ratio of microorganisms in different lineages or strains
  • the species S. bayanus includes two varieties, S. bayanus var. bayanus and S. bayanus var. uvarum It has been suggested that the type S. bayanus strain (isolated from beer) could be a hybrid between S. uvarum (wine yeasts included in the taxon S. bayanus) and S. cerevisiae, with which it has been proposed to recover the name specific S. uvarum for non-hybrid strains included in the S. bayanus taxon and mostly isolated from wines (Nguyen et al., 2000).
  • the microorganism object of the present invention is the yeast strain S. bayanus var. uvarum (BM58) (CECT 13003) or S. cerevisiae (BM60) (CECT 13004).
  • Said identification was carried out by means of the PCR amplification of the 5,8S-ITS ribosomal region using the universal primers ITS1 and ITS4 and their subsequent digestion with the restriction enzyme Hae III following the methodology developed by Fernández-Espinar et al. (2000) and comparing with the species patterns already obtained by our group and available in a database accessible through the IATA webb sheet (http://yeast-id.com/).
  • the BM60 strain corresponds to the S. cerevisiae species by giving a restriction pattern of 325, 230, 170, 125 bp and the BM 58 strain is S. bayanus var. uvarum when giving the typical pattern of this species, 495, 230, 125 bp with the enzyme Hae ⁇ .
  • the rapid restriction analysis technique of mtDNA was used for the molecular level characterization of the clone or strain.
  • the analysis of polymorphisms by restriction of mitochondrial DNA has been widely used as a method of characterizing yeast strains of wine origin. This technique allows to determine the molecular profile of each strain in a precise and very reproducible way.
  • the DNA was used extracted according to the protocol described by Querol et al. (1992).
  • the pattern of the strains, BM58 and BM60 is shown in Figure 2 where a totally different band pattern can be observed between both strains indicating that they are different strains.
  • These patterns have been compared with that of other commercial yeasts (Fernández-Espinar et al., 2001), where we have been able to verify that the present pattern of BM58 and BM60 yeasts does not correspond to that of any of the yeasts commonly used by the wineries Spanish.
  • a particular embodiment of the invention consists essentially in that the microorganism object of the present invention is characterized by being isolated and selected from natural fermentations of wines from production areas (designation of origin DO) controlled Utiel-Requena, Valencia and Alicante, to present a majority implementation in wine fermentation, up to 100% with any type of grape variety and characterized by tolerating: low pH, up to 2.8; growth and fermentation temperature both high and low, between 10 and 3O 0 C; osmotic pressure of up to 300 g / l of sugars; and high concentrations of alcohol in the medium, up to 15%.
  • DO designation of origin DO
  • Table 2 shows that in the presence of 5, 10, 12 and 15% alcohol, the beer strains belonging to the S. cerevisiae species are more sensitive to growth in the presence of alcohol, and only grow well in the presence of 5% of alcohol, and no growth was observed in the presence of 10, 12 or 15% alcohol.
  • the wine strains belonging to this species are more resistant to alcohol and growth is observed both in the presence of 10 and 12% alcohol; However, not all are able to grow in the presence of 15% alcohol.
  • the present invention are capable of growing at lower temperatures than the wine strains.
  • the BM60 strain showed good ability to grow at low temperatures of 10 and 16 0 C and even an adaptation to ferment at low temperatures better than that shown by the other strains belonging to the S. cerevisiae species analyzed and is more sensitive to growing at 37 0 C than other wine yeasts belonging to species S. cerevisiae analyzed (see Table 2). It should also be noted that strain BM58 belonging to S. bayanus (var. Uvarum) also grew to 1O 0 C.
  • strain BM58 is implanted in the fermentations carried out in the cellar (see example 3).
  • Bayanus strain S. uvarum var (BM58) is overwhelmingly present in the first days of fermentation (6 to day), detecting between 40 and 70% of implantation.
  • This high degree of implantation in an illustrative manner and without limiting the scope of the invention, was observed both in white wines made with Merseguera grape variety (fermenters V22, V34-17 and V35), rosé wines made with Tempranillo grape
  • Another object of the invention consists essentially in that the microorganism of the present invention produces an increase in glycerol concentration greater than 9 g / L, even reaching up to 22.27 g / L, which is equivalent to up to 150% more glycerol.
  • a common commercial yeast produces maximum 9 g / L glycerol 28 0 C, normal temperature fermentation (see Table 1).
  • both BM58 and BM60 produce glycerol levels higher than those of a commercial yeast both at low temperatures and at optimal fermentation temperatures, reaching up to 22.27 g / L at
  • Glycerol contributes to the final taste and body of the wine, providing sweetness in both white, rosé and red wines, hence in some European countries it is used as an indicator of quality in wine.
  • glycerol intervenes in softening the astringent character of polyphenols such as tannins, typical in red wines, thus obtaining softer wines on the palate and facilitating the obtaining of red wines aged in barrels.
  • Another of the needs of the sector is the obtaining of yeasts whose performance in alcohol during alcoholic fermentation is wines with a high alcoholic degree, a need that is currently made more crucial with climate change, since musts with a higher degree of maturity are obtained .
  • Yeasts that are capable of diverting the consumption of sugar to produce glycerol during glycolysis also have the characteristic that decreases the yield in obtaining alcohol (Pretorius, 2000). Therefore, the selection of yeasts that, in addition to meeting the general criteria, are capable of increasing the concentration of glycerol in wines is of great interest to the industries.
  • the microorganism object of the present invention is characterized by producing an increase in the production of secondary aromas of the group of acetates, and more specifically ethyl acetate, isoamyl acetate, isobutyl acetate, acetate hexyl acetate, benzyl acetate and phenyl ethanol acetate (see tables 4, 5 and 6).
  • isoamyl acetate should be noted.
  • This compound is characterized by a banana aroma.
  • Quantified levels in wines between 0.16 and 2.52 mg / L, exceed the detection threshold and are considered as usual concentrations described in wines. In general, higher concentrations have been determined in rosé wines, although this acetate was found in lower concentration in the case of wine made with the Noblesse strain, wines fermented with BM58 had the highest concentrations of isoamyl acetate.
  • concentration of acetate 2 phenylethanol was quantified, which is characterized by providing aromas of rose, honey and tobacco.
  • the microorganism object of the present invention is characterized by producing an increase in the production of secondary aromas of the group of ethyl esters, and more specifically ethyl caproate, ethyl caprylate, diethyl succinate, lactate of ethyl and ethyl caprate (see tables 4, 5 and 6).
  • Ethyl caproate and ethyl caprylate are esters related to descriptors of fruity aromas of apple and pineapple. They are compounds that are commonly found in red wines as the base of aroma.
  • Ethyl caprylate ethyl octanoate
  • whites values between 6.87-12.24 mg / L, and in rosés between 5.20-11, 30 mg / L, exceed the normal concentration described (0.82 mg / L and 0.20 mg / L respectively).
  • Red wines have between 2.1 and 4.45 mg /, which are also higher concentrations than usual (0.78 mg / L). It should be noted that in Los Marcos bobal red wines made with BM58, greater production of this compound was observed.
  • Ethyl succinate and ethyl lactate are related to dairy and coffee aromas.
  • ethyl lactate helps to dampen the bitter and acidic edges in taste and according to some authors it contributes to enriching the volume and roundness in the mouth.
  • the wines analyzed showed an extraordinary production of diethyl succinate, which is higher in red wines (8.8-26.5 mg / L). In these cases, it was also appreciated that wines made with BM58 contained more diethyl succinate than the V17 control.
  • White and rosé wines also have high concentrations (0.8-35.8 mg / L). In the case of ethyl lactate, concentrations between 6.8 and 38.80 mg / L have been determined.
  • Ethyl caprate (ethyl decanoate) also has a significant importance in the quality of the final aroma of the wine, providing a fruity, grape and sweet aroma.
  • the detection threshold of this compound is 0.2 mg / L, so the concentrations that have been quantified in the wines exceed this threshold (between 0.8 and 5.23 mg / L).
  • Wines made with BM58 generally have a higher concentration of ethyl caprate than control wines. Some wines have a much higher content than described so far (0.21 mg / L in white wines and 0.42 mg / L in reds). It is worth highlighting the high concentration of ethyl caprate in R-120 Tempranillo and SAUT26 San Antonio bobal and V44 wines all made with BM58.
  • the microorganism object of the present invention is characterized by producing an increase in the production of secondary aromas of the group of alcohols, and more specifically of isobutanol, isoamyl alcohol, 1-hexanol and 2-phenylethanol ( see tables 4, 5 and 6).
  • Isobutanol is a compound that gives rise to aromas reminiscent of wine, solvent or bitter.
  • concentrations between 1, 3 and 6.8 mg / L of isobutanol have been determined, which are below The threshold detection concentration (40 mg / L). These observed values are much lower than the levels described in wines and are between 25.7 and 103 mg / L.
  • the wines analyzed are also characterized by high concentrations of isoamyl alcohol, which provides aromas of whiskey, malt. Concentrations between 143.41 and 347.16 mg / L were quantified, which exceed the usual levels in wines.
  • the detection threshold is 30 mg / L.
  • the 1-hexanol and 2-phenylethanol alcohols are related to descriptors reminiscent of the herb and roses respectively. However, the levels of 1-hexanol in the wines analyzed, between 0.8 and 2.36 mg / L, are below the detection threshold (8 mg / L). On the other hand, it is worth highlighting the high concentration of 2 phenylethanol observed.
  • Another object of the present invention is the use of the microorganism object of the present invention for the production of fermented alcoholic beverages, and particularly in the preparation of any type of wine from grape musts of any variety.
  • the wines made with the BM60 strain as in the BM58 are mostly ethyl esters such as caproate of ethyl, ethyl caprylate, diethyl succinate and ethyl caprate and isoamyl acetate.
  • microorganism object of the present invention is suitable for the production of fermented alcoholic beverages, preferably wine.
  • fermented alcoholic beverages preferably wine.
  • S. cerevisiae and S. bayanus are species that develop alcoholic fermentation in industrial processes that give rise to other types of alcoholic beverages other than wine, by way of illustration and without limiting the scope of the invention in beer and cider (see table 2).
  • Figure 1 Analysis by RFLP ' s with the Hae ⁇ enzyme of the ITS-5,8S ribosomal region amplified by PCR of DNA from colonies isolated from BM60 strains corresponding to the S. cerevisiae species, obtaining a restriction pattern of 325, 230, 170, 125 bp, and of the strain
  • BM 58 which is S. bayanus var. uvarum when obtaining the pattern of 495, 230, 125 bp.
  • yeast fermentation was carried out at less than 22 0 C in the Murviedro winery.
  • Yeasts isolated during different fermentations were identified by molecular techniques (Esteve-Zarzoso et al., 1999). Strains belonging to S. cerevisiae, no isolates of S. kudriavzevii, and only 5 isolates of S. bayanus were mostly isolated. This identification was carried out according to the protocol described by Fernández-Espinar (2000).
  • strain BM58 Given a set of criteria for oenological interest such as fermentative capacity 12 and 28 0 C proceeded to the selection of the strain BM58 since it was the strain which grow more quickly at the two temperatures tested and the two concentrations of sugar. In addition, other criteria such as good fermentation rate were also taken into account; good ethanol yield; high glycerol production and low acetic acid. From the total of 5 strains previously isolated, strain BM58 was selected as it was also the one that presented the most appropriate values for alcoholic fermentation, the results of which are shown in Table 1. The Table shows the sugar concentration residual, glycerol, acetic acid and alcohol of the wines obtained after fermenting must of the Tempranillo variety.
  • Table 1 shows the concentration of residual sugar, glycerol, acetic acid and alcohol of the wines obtained after fermenting must of the Tempranillo variety.
  • the BM60 strain takes less days to ferment at both temperatures than the BM58 and the glycerol concentration is also high of 12g / L and 9.5 to 28 0 C and 12 0 C respectively, being at both temperatures at as is the case with strain BM58 higher than normal in wine yeasts (7-9g / L). The rest of the parameters are within the normal ranges in wines.
  • Table 1 Analytical wines obtained with BM58, BM60 and T73 control strains. Fermentation time, reducing sugars, alcohol, glycerol and acetic acid produced by yeasts BM58 and BM60 compared to the commercial strain of S. cerevisiae (T73).
  • BM58 strain adapts well to these stresses, a comparative study was carried out growing the BM58 strain, as well as other wine strains used as controls belonging to the species S. cerevisiae and S. bayanus (see table 2).
  • the different stresses analyzed are low pH (2.8; 3.0; 3.2), both high and low temperature (10, 16, 30, 37 0 C), osmotic (200, 250, 300 g / l of sugars ), high concentrations of alcohol in the medium (5, 10, 12 and 15%).
  • CJ mitochondrial has been widely used as a method of characterizing yeast strains of wine origin (Querol et al., 1992). This technique allows to determine the molecular profile of each strain in a precise and very reproducible way. However, obtaining results requires more time than the PCR technique, since it is previously necessary to isolate the yeasts from the wine using a suitable culture medium.
  • Example 4 Determination of volatile compounds in wines obtained with strain BM58.
  • Example 5 Elaboration of sparkling wines with BM58 and BM60 strains.
  • Fernández-Espinar MT. ; Esteve-Zarzoso, B .; Querol, A. and E. Barrio.
  • Fernández-Espinar M. T .; Barrio, E .; Querol, A. 2003. Analysis of the genetic variability in the species of the Saccharomyces sensu stricto complex. Yeasts, 20: 1213-1226.

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  • Distillation Of Fermentation Liquor, Processing Of Alcohols, Vinegar And Beer (AREA)

Abstract

L'invention concerne deux souches de levure, BM58 (CECT13003) et BM60 (CECT13004), appartenant respectivement aux espèces Saccharomyces bayanus (var. uvarum) et S. cerevisiae, non modifiées génétiquement, sélectionnées parmi des fermentations viniques naturelles avec une bonne capacité de fermentation des moûts pour obtenir des boissons alcoolisées, de préférence du vin. Les vins obtenus à partir de cette levure se caractérisent en ce qu'ils contiennent des concentrations élevées de glycérol sans augmenter la teneur en acide acétique et qu'ils accroissent la production de certains arômes secondaires. Ces levures produisent principalement des arômes du type esters éthyliques tels que le caproate d'éthyle, le caprylate d'éthyle, le succinate de diéthyle et le caprate d'éthyle. Enfin, il faut mentionner la capacité élevée d'implantation de ces levures dans des fermentations viniques ainsi que leur capacité de croissance à basses températures (10°C).
PCT/ES2009/070008 2008-02-04 2009-01-27 Microorganisme fermenteur producteur de concentrations élevées de glycérol et ses applications dans la production de boissons alcoolisées/de vin Ceased WO2009098342A1 (fr)

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ES200800286A ES2330709B1 (es) 2008-02-04 2008-02-04 Microorganismo fermentador productor de altas concentraciones de glicerol y sus aplicaciones en la produccion de bebidas alcoholicas/vino.

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CN107177520A (zh) * 2017-05-25 2017-09-19 江南大学 一株高产β‑苯乙醇的酿酒酵母菌株及其应用
WO2018214330A1 (fr) * 2017-05-25 2018-11-29 江南大学 SOUCHE DE SACCHAROMYCES CEREVISIAE À RENDEMENT ÉLEVÉ EN β-PHÉNYLÉTHANOL ET APPLICATION CORRESPONDANTE
CN109576165B (zh) * 2019-01-11 2020-08-04 谭瑛 一种贝酵母菌及其应用
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CN117757650A (zh) * 2024-01-03 2024-03-26 泰山学院 一种酿酒酵母及其在生产低高级醇和/或高乙酸乙酯酒产品中的应用

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CN107164250A (zh) * 2017-05-25 2017-09-15 江南大学 一种增强果酒芳香的酿造方法
CN107177520A (zh) * 2017-05-25 2017-09-19 江南大学 一株高产β‑苯乙醇的酿酒酵母菌株及其应用
WO2018214330A1 (fr) * 2017-05-25 2018-11-29 江南大学 SOUCHE DE SACCHAROMYCES CEREVISIAE À RENDEMENT ÉLEVÉ EN β-PHÉNYLÉTHANOL ET APPLICATION CORRESPONDANTE
CN107177520B (zh) * 2017-05-25 2020-05-08 江南大学 一株高产β-苯乙醇的酿酒酵母菌株及其应用
CN107164250B (zh) * 2017-05-25 2020-07-03 江南大学 一种增强果酒芳香的酿造方法
US10982295B2 (en) 2017-05-25 2021-04-20 Jiangnan University Stain Saccharomyces cerevisiae M 2016785 producing high concentration of β-phenylethanol and application thereof
CN109576165B (zh) * 2019-01-11 2020-08-04 谭瑛 一种贝酵母菌及其应用
CN112266937A (zh) * 2020-12-07 2021-01-26 福建农林大学 一种适用于香灰菌生物发酵合成2-苯乙醇的培养基
CN112266937B (zh) * 2020-12-07 2022-07-05 福建农林大学 一种适用于香灰菌生物发酵合成2-苯乙醇的培养基
CN117757650A (zh) * 2024-01-03 2024-03-26 泰山学院 一种酿酒酵母及其在生产低高级醇和/或高乙酸乙酯酒产品中的应用
CN117757650B (zh) * 2024-01-03 2024-06-04 泰山学院 一种酿酒酵母及其在生产低高级醇和/或高乙酸乙酯酒产品中的应用

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