Evaluation of key odorants in sauvignon blanc wines using three different methodologies.

In this study three different approaches were employed to identify key odorants in Sauvignon blanc wines. First, the concentrations of the odorants were compared to their respective aroma detection thresholds. The resulting odor activity values (OAV) were transformed into a normalized and weighted measure that allows the aroma profiles of different wines to be compared and the contribution of a single aroma in a complex mixture to be evaluated. Based on their OAV, 3-mercaptohexanol and 3-mercaptohexyl acetate were the two most important aroma compounds in many Marlborough Sauvignon blanc wines. Due to limitations with the OAV approach, the study was extended to include aroma extract dilution analysis (AEDA), which revealed that ß-damascenone, together with the varietal thiols, esters, and higher alcohols, are key odorants in Sauvignon blanc wines. The final approach undertaken was aroma reconstitution and omission tests using a deodorized wine base and the creation of a model Marlborough Sauvignon blanc. Single compounds and groups of compounds were omitted from the model to study their impact on the sensory properties of the model wine. Reconstitution and omission confirmed that varietal thiols, esters, terpenes, and ß-damascenone are all important contributors to Sauvignon blanc aroma. The methoxypyrazines showed an important but relatively low impact in all three of the approaches undertaken in this study.

Influence of grape-harvesting steps on varietal thiol aromas in Sauvignon blanc wines.

The intense tropical fruit aroma of Sauvignon blanc wines has been associated with the varietal thiols 3-mercaptohexanol (3MH), derived from odorless precursors in the grape, and 3-mercaptohexyl acetate (3MHA), arising from 3MH during fermentation. Grapes and juice were sourced from five locations in Marlborough, New Zealand, taking hand-picked grapes and samples at four stages during the mechanical harvesting process and pressing, which were then fermented in replicated 750 mL bottles. With each set of juices, the highest concentrations of Cys-3MH and Glut-3MH were found in the juices pressed to 1 bar, but these juices produced wines with lower 3MH and 3MHA concentrations. With three of the juices, there was an increase in varietal thiol content for wines made from juices that had been machine harvested compared to the hand-picked samples, which matched earlier findings of lower 3MH and 3MHA levels in wines made from hand-picked grapes. Juices that were more oxidized, and which showed a higher absorbance at 420 nm, were found to produce wines with lower 3MH and 3MHA concentrations.

The yeast IRC7 gene encodes a ß-lyase responsible for production of the varietal thiol 4-mercapto-4-methylpentan-2-one in wine.

Three varietal thiols are key aroma compounds in Sauvignon Blanc wines: 4-mercapto-4-methylpentan-2-one (4MMP), 3-mercaptohexanol (3MH) and its acetylated derivative 3-mercaptohexyl acetate (3MHA). Screening of Saccharomyces cerevisiae strains identified a clinical isolate with elevated 4MMP production after fermentation. Bulked Segregant Analysis of a cross between this isolate and the laboratory strain revealed a single major locus for 4MMP production near the telomere of chromosome 6. Deletion of the IRC7 gene from this region in YJM450 reduced 4MMP production below detectable levels, but did not affect yields of 3MH, in Sauvignon Blanc wine. Sequencing revealed that the IRC7 gene in YJM450 had been introgressed from a strain of Saccharomyces paradoxus. Most strains of S. cerevisiae, including the laboratory strain S288C, have a 38-bp deletion that inactivates IRC7. Overexpression of a full-length S. cerevisiae allele of IRC7 in a wine yeast, Zymaflore F15, increased 4MMP production in Sauvignon Blanc wine from undetectable levels (<10 ng L(-1)) to concentrations of 1000 ng L(-1), and also increased 3MH and 3MHA. Biochemical analysis of soluble protein extracts showed that both the cerevisiae and paradoxus IRC7 proteins show ß-lyase activity, with a substrate preference for cys-4MMP over cys-3MH.

Ethylene-regulated (methylsulfanyl)alkanoate ester biosynthesis is likely to be modulated by precursor availability in Actinidia chinensis genotypes.

The limiting steps of ethylene-dependent (methylsulfanyl)alkanoate ester biosynthesis have been investigated in this study, using closely related Actinidia chinensis genotypes and the commercial cultivar 'Hort16A'. Quantification of methylsulfanyl-compounds from the headspace of ethylene-producing kiwifruits revealed little variation in their volatile composition but remarkable differences in the magnitude of the fruit volatile levels. To test whether the variations in fruit volatile levels can be correlated with the genotype-specific apparent catalytic efficiency, the initial slope of the substrate response curve (V'(Max)K(M)(-1) where V'(Max) is the apparent V(Max) in a crude extract) was evaluated for total alcohol acyltransferase (EC 2.3.1.84) activity. The V'(Max)K(M)(-1) values of different (methylsulfanyl)alkyl-CoAs were in a similar range for most genotypes, which suggests substrate availability as the limiting factor for (methylsulfanyl)alkanoate ester synthesis in these kiwifruit. Furthermore, gene expression analysis of acyltransferase expressed sequence tags points towards the action of multiple isozymes for (methylsulfanyl)alkanoate ester synthesis, emphasizing the central role of substrate levels on final ester concentrations. Volatile levels of the potential precursor methional were increased in ethylene-producing A. chinensis kiwifruit and a close connection between (methylsulfanyl)alkanoate ester formation and ethylene synthesis in plants is proposed. Finally, a possible biosynthetic pathway is presented.

Influence of juice pressing conditions on polyphenols, antioxidants, and varietal aroma of Sauvignon blanc microferments.

The impact of juice press fractions upon the content of varietal thiols in Sauvignon blanc has been examined for wines fermented at the laboratory scale (750 mL). Wines made from pressed juices (taken at 0.25 and 1.0 bar) contained less than half the concentration of 3-mercaptohexan-1-ol (3MH) and 3-mercaptohexyl acetate (3MHA), important contributors to the tropical and passion fruit character of Sauvignon blanc wines, compared to wines made from free run juices. The pressed juices and wines exhibited lower acidity values, more rapid decline in glutathione content, and more advanced polyphenol oxidation. Supplementation of the juices with glutathione (at 67 mg/L) prior to fermentation led to lower varietal thiol concentrations in the finished wines, typically by several percent, whereas treatment with polyvinylpolypyrrolidone (PVPP) made no impact on wine parameters. Pasteurization of pressed juices increased 3MHA content in the finished wines, but also led to a decline in 3MH concentrations.

(Methylsulfanyl)alkanoate ester biosynthesis in Actinidia chinensis kiwifruit and changes during cold storage.

Four 3-(methylsulfanyl)propionate esters, ethyl 3-(methylsulfanyl)prop-2-enoate, two 2-(methylsulfanyl)acetate esters and their possible precursors 2-(methylsulfanyl)ethanol, 3-(methylsulfanyl)propanol and 3-(methylsulfanyl)propanal were quantified from the headspace of Actinidia chinensis 'Hort 16A' kiwifruit pulp by GC-MS-TOF analysis. The majority of these compounds were specific for eating-ripe fruit and their levels increased in parallel with the climacteric rise in ethylene, accumulating towards the very soft end of the eating firmness. No ethylene production could be observed after long-term storage (4-6 months) at 1.5 degrees C and the levels of all methylsulfanyl-volatiles, except methional, declined by 98-100% during that period. This depletion of (methylsulfanyl)alkanoate-esters after prolonged cold storage points towards little flavour impact of these compounds on commercial 'Hort 16A' kiwifruits. However, ethyl 3-(methylsulfanyl)propionate is suggested to be odour active in ripe 'Hort 16A' fruit that has not been stored. Gene expression measured by q-RT PCR of six ripening-specific alcohol acyltransferase (AAT) expressed sequence tags and (methylsulfanyl)alkanoate-ester production of cell-free extracts were also significantly decreased after prolonged cold storage. However, (methylsulfanyl)alkanoate-ester synthesis of cell-free extracts and AAT gene transcript levels could be recovered by ethylene treatment after five months at 1.5 degrees C indicating that the biosynthesis of (methylsulfanyl)alkanoate-esters in 'Hort 16A' kiwifruit is likely to depend on ethylene-regulated AAT-gene expression. That the composition but not the concentration of (methylsulfanyl)alkanoate-esters in fresh fruit could be restored after ethylene treatment suggests that substrate availability might also have an impact on the final levels of these volatiles.

Effect of skin contact and pressure on the composition of Sauvignon Blanc must.

Early white winemaking operations are known to affect the extraction of grape skin compounds into the juice fraction, which will dictate their concentration in the resulting wine. Grape skin contact and the amount of pressure applied during grape pressing affect the extraction of varietal aromas located in the skins. Compounds such as the polyphenols and glutathione, with antioxidant properties involved in juice oxidation processes and white wine stability, are also affected. The present study evaluates how grape skin contact and the amount of pressure applied during grape pressing affect the levels of S-(3-hexan-l-ol)cysteine (3MH-S-cys, a key grape-derived precursor to the volatile thiol 3-mercapto-hexanol (3MH), which is reminiscent of passion fruit aroma); 2-methoxy-3-isobutylpyrazine (IBMP, with a capsicum-like descriptor); phenolic compounds; and glutathione in Sauvignon Blanc juice. The study was conducted using grapes obtained from commercial Marlborough (New Zealand) vineyards, using both commercial and laboratory grape-processing procedures. Immobilized metal ion chromatography was used to isolate the 3MH- S-cys precursor from the juices. The isolated precursor was then volatilized by trimethylsilylation and analyzed using gas chromatography/mass spectrometry (GC/MS). IBMP was analyzed by GC/MS after solvent extraction, and a high-performance liquid chromatography method was used for the quantification of phenolic compounds and glutathione. 3MH- S-cys levels were seen to increase in juice fractions obtained from a winery press operating at higher pressures. The increase was attributed to the cumulative effect of longer skin contact time and the amount of pressure applied. The highly water-soluble IBMP was less affected by the amount of pressure applied during commercial grape pressing. Additional information was generated by the specific assessment of skin contact and applied pressure during grape pressing in a laboratory trial. In this trial, a long (32 h) skin contact time resulted in a greater release of varietal aroma compounds, 3MH- S-cys, and IBMP into the juice, and the concentration was further raised by increasing the pressure applied during pressing. However, for both experiments, the extraction of the varietal aroma compounds was offset by a clear increase in the juice oxidative potential, seen by a decline in glutathione content, a natural grape antioxidant, and an increase in particular oxidizeable polyphenol compounds, which may cause the must or wine to brown and lead to a loss of varietal aromas.

Effect of screwcap and cork closures on SO2 levels and aromas in a Sauvignon Blanc wine.

The development of a Sauvignon Blanc wine sealed under screwcap and cork was undertaken using different fill heights and initial levels of free SO2 (20, 25, and 30 mg/L) over 2 years. More SO2 was lost for wines under cork over the first 3 months, corresponding to a higher level of dissolved oxygen at bottling. From this time wines under cork and screwcap lost SO2 at a similar rate and retained dissolved CO2 equally well, indicating that both types of closure presented a similar effective barrier to gas movement. After 2 years in the bottle, the different treatments retained similar levels of the volatile thiols 3-mercaptohexyl acetate (3MHA) and 3-mercaptohexanol (3MH) responsible for fruity aromas, with initial SO2 levels having no effect, but the thiol concentrations were 18-23% lower under cork, which may be due to absorption of volatiles into the cork. Levels of polyphenols such as caftaric acid and the absorbance at 420 nm were the same for wines under cork and screwcap, whereas some indication was given that more oxidation occurred with a lower level of initial free SO2. Although the different treatments were not readily distinguished by a sensory panel, the data for individual wines showed a positive correlation between passion fruit descriptors and levels of 3MHA and 3MH.