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Impact of mixed S. cerevisiae strains on the production of volatiles ...

Impact of mixed S. cerevisiae strains on the production of volatiles ...

728 S. Saberi et al. /

728 S. Saberi et al. / Food Research International 48 (2012) 725–7353.2. Production ong>ofong> volatile compounds by wine yeast ong>strainsong>Eighteen compounds were quantifiable (Table 1) in the headspaceong>ofong> the wines. These included eight higher alcohols, five ethyl esters,three acetate esters, one aldehyde and one organic acid. The concentrationong>ofong> higher alcohols, ethyl esters and acetate esters for the industrial,individual Burgundian and ong>mixedong> Burgundian ong>strainsong> arereported in Tables 2, 3 and 4 respectively. In general, the concentrationong>ofong> the volatile compounds for the individual and ong>mixedong> Burgundianong>strainsong> was mid-range or intermediate, compared to the industrialyeasts. This was exemplified by the concentrations observed for3-methyl-1-butanol for the individual Burgundian (7.007–7.190 mg/L)and ong>mixedong> Burgundian (6.188–7.453mg/L)yeastscomparedtotheindustrialyeast (5.789–9.292 mg/L). This pattern wasobservedforallong>ofong>the compounds, across all classes, with the exception ong>ofong> 2,3-butanediolproduced by M4, which was the lowest ong>ofong> the reported concentrations.Two additional compounds (acetaldehyde, acetic acid) followed asimilar pattern (data not shown), as described above, with the exceptionong>ofong> acetaldehyde concentrations produced by M3 (1.422 mg/L), whichwere at the high end ong>ofong> the range associated with the industrial ong>strainsong>(0.732 mg/L – 1.422 mg/L). Mixed Burgundian yeast ong>strainsong> producedhigher concentrations ong>ofong> higher alcohols such as 2-methyl-1-butanoland lower concentrations ong>ofong> the higher alcohols 2,3-butanediol, propanoland phenylethanol. The Burgundian yeast ong>strainsong> also produced moreethyl hexanoate and ethyl octanoate, and low to moderate concentrationsong>ofong> the acetate esters compared to industrial ong>strainsong>. In contrast,most ong>ofong> the industrial ong>strainsong> produced either very low or very high concentrationsong>ofong> acetate esters.Table 3Mean concentrations 1 (mg/L) ong>ofong> ethyl esters in Chardonnay wines fermented with sixindividual industrial yeast ong>strainsong>, two individual Burgundian and four ong>mixedong> Burgundianyeasts trains, averaged across two fermentation temperatures (n=6). Yeast straineffect 2 is shown for each volatile compound.Yeast strainEthylbutanoateEthylhexanoateEthyloctanoateEthyldecanoateEthyllaurateIndividual industrial ong>strainsong>Blanc 0.200 a 0.036 ab 0.035 a 0.028 ef 0.001 aElegance 0.275 cd 0.045 ab 0.048 bc 0.031 fg 0.005 dFusion 0.299 d 0.059 b 0.059 c 0.034 g 0.010 eCY3079 0.299 d 0.042 ab 0.045 bc 0.025 de 0.003 bcICV-D254 0.361 e 0.030 a 0.030 a 0.022 abcd 0.002 abX16 0.268 bcd 0.039 ab 0.036 ab 0.024 cde 0.004 cdIndividual Burgundian ong>strainsong>C2 0.211 a 0.043ab 0.040 ab 0.017 a 0.002 abC6 0.229 abc 0.044ab 0.040 ab 0.020 abcd 0.002 abMixed Burgundian ong>strainsong>(C2/C6:1/1), 0.246 abcd 0.041 ab 0.042 ab 0.023 bcde 0.001 aM1(C2/C6:1/2), 0.217 ab 0.040 ab 0.042 ab 0.025 de 0.002 abM2(C2/C6:1/3), 0.218 ab 0.040 ab 0.040 ab 0.022 abcd 0.002 abM3(C2/C6:3/2), 0.215 a 0.042 ab 0.041 ab 0.018 ab 0.001 aM4Range 0.200–0.361 0.030–0.059 0.030–0.059 0.017–0.034 0.001–0.011 The mean values ong>ofong> the biological replicates ong>ofong> each yeast strain at two temperaturesare shown (n=6).2 Yeast strain means with different letters are significantly (p≤0.05) different.3.3. Principal component analyses ong>ofong> the wine yeast ong>strainsong>Principal component analyses (PCA) ong>ofong> the higher alcohols, ethyl estersand acetate esters in the Chardonnay wines are shown in Fig. 1A-C,respectively. PCA ong>ofong> the higher alcohols (Fig. 1A) accounted for 65.6% ong>ofong>the total variability in the data, with 37% and 28.6% ong>ofong> the varianceexplained by the PC I and PC II, respectively. The industrial ong>strainsong>were scattered throughout the plot, reflecting extremely different patternsong>ofong> production ong>ofong> higher alcohols. This was consistent with concentrationsreported in Table 2. Three yeast ong>strainsong> (Fusion, Elegance andCY3079) were positioned slightly above CY3079 and C2 (Fig. 1A),reflecting much higher concentrations ong>ofong> 3-butanediol (HA-1), hexanol(HA-5), phenylethanol (HA-7) and propanol (HA-8). The X16 and Blancong>strainsong> had very dissimilar patterns ong>ofong> higher alcohol production comparedto all other ong>strainsong>, as shown by their locations, to the far rightand far left ong>ofong> the plot, respectively (Fig. 1A). The ong>mixedong> Burgundianong>strainsong> composed a distinct group, with the exception ong>ofong> M4, whichwas most heavily loaded on negative PC II (Fig. 1A). M4 had lower concentrationsong>ofong> 2,3-butanediol (HA-1) and higher concentrations ong>ofong> HA-6. In contrast, the ong>mixedong> Burgundian ong>strainsong> and C6 had relatively highconcentrations ong>ofong> 2-methyl-1-butanol (HA-2), n-butanol (HA-4) andisobutanol (HA-6) and lower concentrations ong>ofong> 2,3-butanediol (HA-1),n-butanol (HA-4) and propanol (HA-8) (Fig. 1A). This was consistentwith concentrations reported in Table 2. The ong>mixedong> Burgundian ong>strainsong>produced moderate amounts ong>ofong> the higher alcohols compared to X16Table 2Mean concentrations 1 (mg/L) ong>ofong> higher alcohols in Chardonnay wines fermented with six individual industrial yeast ong>strainsong>, two individual Burgundian and four ong>mixedong> Burgundianyeast ong>strainsong>, averaged across two fermentation temperatures (n=6). Strain effect 2 is shown for each volatile compound.Yeast strain 2,3-Butanediol 2-Methyl-1-butanol 3-Methyl-1-butanol n-Butanol 1-Hexanol Isobutanol Phenylethanol PropanolIndividual industrial ong>strainsong>Blanc 1.235 jk 1.150 a 5.789 a 0.049 bc 1.918 de 6.666 a 0.906 cdef 5.034 kElegance 1.065 ghi 1.170 a 7.167 defg 0.046 bc 1.949 ef 9.744 b 1.254 j 2.713 jFusion 1.223 ijk 1.420 def 7.056 cdefg 0.056 cd 2.212 h 7.400 a 1.242 ij 2.111 hiCY3079 1.110 hij 1.181 ab 6.723 abcdefg 0.033 a 1.899 cdef 12.829 f 1.070 h 1.427 aICV-D254 0.864 bcdef 1.249 abcd 6.609 abcdefg 0.101 e 1.811 bcdef 11.825 cdef 0.879 bcde 1.614 abcX16 1.326 k 1.763 f 9.292 h 0.047 bc 1.603 a 15.259 i 1.658 k 2.015 fghiIndividual Burgundian ong>strainsong>C2 1.063 ghi 1.375 cd 7.190 efg 0.072 d 1.993 g 11.999 def 1.023 fgh 2.136 iC6 0.942 defgh 1.291 abcd 7.007 bcdefg 0.039 ab 1.964 fg 19.183 k 1.056 gh 1.941 defghMixed Burgundian ong>strainsong>(C2/C6:1/1), M1 0.975 efgh 1.375 cd 6.188 abcdef 0.056 cd 1.904 cdef 14.892 hi 1.070 h 2.019 ghi(C2/C6:1/2), M2 0.920 cdefg 1.349 bcd 7.203 fg 0.056 cd 1.809 bcdef 14.744 ghi 0.933 efg 1.786 cde(C2/C6:1/3), M3 0.990 fgh 1.572 ef 7.453 g 0.054 bc 1.960 fg 17.327 j 0.916 def 1.960 efghi(C2/C6:3/2), M4 0.545 a 1.291 abcd 6.740 abcdefg 0.053 bc 1.716 ab 12.781 ef 0.474 a 1.664 bcRange 0.545–1.326 1.150–1.763 5.789–9.292 0.033–0.101 1.603–2.212 6.666–19.183 0.474–1.658 1.427–5.0341 The mean values ong>ofong> the biological replicates ong>ofong> each yeast strain at two temperatures are shown (n=6).2 Yeast strain means with different letters are significantly (p≤0.05) different.

S. Saberi et al. / Food Research International 48 (2012) 725–735729Table 4Mean concentrations 1 (mg/L) ong>ofong> acetate esters in Chardonnay wines fermented withsix individual industrial ong>strainsong>, two individual Burgundian and four ong>mixedong> Burgundianyeast ong>strainsong>, averaged across two fermentation temperatures (n=6). Yeast strain effect2 is shown for each volatile compound.Yeast strain Ethyl acetate Hexyl acetate Isoamyl acetateIndividual industrial ong>strainsong>Blanc 7.761 ij 0.024 bcde 0.093 defghElegance 6.954 hi 0.027 ef 0.109 hijFusion 5.703 a 0.019 a 0.059 aCY3079 6.635 cdefgh 0.030 f 0.118 jICV-D254 6.245 abcdefgh 0.024 bcde 0.107 ghijX16 8.448 j 0.035 g 0.220 kIndividual Burgundian ong>strainsong>C2 6.617 bcdefgh 0.024 bcde 0.090 cdefghC6 6.663 defgh 0.024 bcde 0.116 ijMixed Burgundian ong>strainsong>(C2/C6:1/1), M1 6.554 abcdefgh 0.025 cde 0.100 efghij(C2/C6:1/2), M2 6.756 efgh 0.027 ef 0.109 hij(C2/C6:1/3), M3 6.917 ghi 0.024 bcde 0.104 fghij(C2/C6:3/2), M4 6.809 fgh 0.026 de 0.085 bcdefRange 5.703–8.448 0.019–0.030 0.059–0.1181 The mean values ong>ofong> the replicates ong>ofong> each yeast strain at two temperatures areshown (n=6).2 Yeast strain means with different letters are significantly (p≤0.05) different.and Blanc as shown in Table 2. In general, the ong>mixedong> Burgundian ong>strainsong>were more similar to one another than to the industrial ong>strainsong>, with theexception ong>ofong> ICV-D254.PCA ong>ofong> the ethyl esters (Fig. 1B) production in the Chardonnaywines described 88.3% ong>ofong> the total variability in the data, with 65%and 23.3% explained by PC I and PC II, respectively. The industrialong>strainsong> were located throughout the plot, reflecting the very differentpatterns ong>ofong> ethyl ester production; Fusion and ICV-D254 yeast ong>strainsong>were most different (Fig. 1B). In contrast, the individual and ong>mixedong>Burgundian ong>strainsong> were grouped together in the center ong>ofong> the plot,with the industrial strain Blanc being most similar to the Burgundianong>strainsong>, as reflected by their close proximity (Fig. 1B). The other threeindustrial ong>strainsong> (X16, CY3079, Elegance) were positioned slightlyabove the PCA center (Fig. 1B) and had similar ethyl ester prong>ofong>ilesto one another. The individual and ong>mixedong> Burgundian ong>strainsong> producedhigher concentrations ong>ofong> ethyl hexanoate (EE-2) and ethyloctanoate (EE-3). In contrast, the Burgundian ong>strainsong> producedlower concentrations ong>ofong> ethyl butanoate (EE-1), while most ong>ofong> theindustrial ong>strainsong> produced higher concentrations ong>ofong> ethyl butanoate,as shown in Table 3. Clearly, the individual and ong>mixedong> Burgundianong>strainsong> were more similar to one another than to the industrialong>strainsong>, with the exception ong>ofong> Blanc.PCA plot ong>ofong> acetate esters (Fig. 1C) accounted for 97.2% ong>ofong> the totalvariability in the data, with 85.8% and 10.4% ong>ofong> the variance explainedby PC I and PC II, respectively. The individual and ong>mixedong> Burgundianyeasts were grouped together forming overlapping clusters in thecentre ong>ofong> the plot, reflecting similar patterns ong>ofong> acetate production.Slightly outside this grouping was ICV-D254, Elegance and CY3079,indicating that these yeasts had very different concentrations ong>ofong>ethyl acetates. In contrast, the industrial ong>strainsong> X16 and Fusionwere positioned to the far right and far left, respectively, reflectingextremely different acetate ester prong>ofong>iles. X16 produced higher concentrationsong>ofong> hexyl acetate (AE-2) and isoamyl acetate (AE-3); consistentwith acetate ester concentrations reported in Table 4. Incontrast, Fusion produced lower concentrations ong>ofong> both these compoundscompared to the Burgundian ong>strainsong>.When comparing Fig. 1A-C, it is evident that the commercialyeasts were more similar to the Burgundian yeasts in production ong>ofong>acetate esters (Fig. 1C) than they were for production ong>ofong> higher alcohols(Fig. 1A) or ethyl esters (Fig. 1B).PCA ong>ofong> all the volatile compounds (Fig. 1D) accounted for 56.1% ong>ofong>the total variability in the data, with 29.4% and 26.7% ong>ofong> the varianceexplained by PC I and PC II, respectively. The individual and ong>mixedong>Burgundian ong>strainsong> composed a grouping in the center ong>ofong> PCA plot,with the exception ong>ofong> M4. It had higher concentrations ong>ofong> n-butanol(HA-5) and isobutanol (HA-6) (Table 2). In contrast, the industrialong>strainsong> were scattered throughout the plot, indicative ong>ofong> yeasts withvery different overall volatile prong>ofong>iles. The industrial strain ICV-D254was the most similar to M4; whereas, the industrial strain Blancwas most similar to the Burgundian ong>strainsong> and M1. Interestingly,the ong>mixedong> Burgundian strain M1 (1:1 ratio ong>ofong> C2:C6) was located inthe plot between the two individual Burgundian ong>strainsong> C2 and C6.In contrast, X16 and Fusion were most dissimilar to the other yeastong>strainsong> (Fig. 1D), as reflected by their location on the plot, with highconcentrations ong>ofong> 3-methyl-1-butanol (HA-3), isobutanol (HA-6),and hexyl acetate (EA-3) and acetaldehyde (ACET) and acetic acid(AA), respectively (Tables 2–4).3.4. Odor prong>ofong>iles, OAVs and estimated sensory prong>ofong>iles ong>ofong> the ChardonnaywinesTable 5 summarizes the volatile compounds, aroma descriptors,water thresholds (Campo et al., 2005; Culleré et al., 2004; Francis &Newton, 2005; Schieberle & Hong>ofong>mann, 1997) and odor active values(OAVs) for the individual and ong>mixedong> Burgundian ong>strainsong>. OAVs greaterthan one, less than one reflect concentrations that are above andTable 5Odor thresholds and odor active values (OAVs) ong>ofong> individual volatile compounds calculatedby dividing the mean odor concentration for the yeast groups by the odorantthreshold (mg/L). The odorant concentrations (mg/L) were determined by GC–MSand averaged between two fermentation temperatures for individual Burgundianyeasts (n=12) and ong>mixedong> Burgundian yeasts (n=24).CompoundAromadescriptorOdorthreshold(mg/L)Mean odor active value aIndividualBurgundianMixedBurgundianHigher alcohols2,3-Butanediol Creamy b 150 b 0.007 0.0062-Methyl-1-butanol Sweet fruit c 0.42 c 3.169 3.3223-Methyl-1-butanol Berry d 30 d 0.237 0.230n-Butanol Nail polish d 150 d 0.0004 0.00041-Hexanol Grass b 8 b 0.247 0.231Isobutanol Fusel b 40 b 0.390 0.373Phenylethanol Rose b 14 b 0.074 0.061Propanol Candy d 306 e 0.007 0.006Ethyl estersEthyl butanoate Strawberry c 0.02 c 11.001 11.200Ethyl hexanoate Green apple c 0.14 c 3.130 2.914Ethyl octanoate Pear d 0.005 c 8.005 8.215Ethyl decanoate Dried fruit c 0.2 b 0.092 0.110Ethyl laurate Waxy d 0.5 d 0.004 0.004Acetate estersEthyl acetate Balsamic c 12 c 0.553 0.563Hexyl acetate Apple b 1.5 b 0.016 0.017Isoamyl acetate Banana b 0.03 b 3.432 3.312AldehydeAcetaldehyde Overripe apple b 0.5 b 1.998 2.307Organic acidsAcetic acid Vinegar d 600 d b0.0001 b0.0001aOdor active values (OAVs) obtained by dividing the mean concentration ong>ofong> the volatilefor the yeast group, by the published absolute threshold in water. Individual Burgundianyeasts (n=12); 2 yeasts ×2 temperatures ×3 replications. Mixed Burgundianyeasts (n=24); 4 yeasts ×2 temperatures × 3 replications.b Campo et al. (2005).c Francis and Newton (2005).d Culleré et al. (2004).e Schieberle and Hong>ofong>mann (1997).

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