Effect of Must Hyperoxygenation on Sensory Expression and Chemical Composition of the Resulting Wines

This paper evaluates the effect of must hyperoxygenation on final wine. Lower concentrations of caftaric acid (0.29 mg·L−1), coutaric acid (1.37 mg·L−1) and Catechin (0.86 mg·L−1) were observed in hyperoxygenated must in contrast to control must (caftaric acid 32.78 mg·L−1, coutaric acid 5.01 mg·L−1 and Catechin 4.45 mg·L−1). In the final wine, hydroxybenzoic acids were found in higher concentrations in the control variant (gallic acid 2.58 mg·L−1, protocatechuic acid 1.02 mg·L−1, vanillic acid 2.05 mg·L−1, syringic acid 2.10 mg·L−1) than in the hyperoxygenated variant (2.01 mg·L−1, 0.86 mg·L−1, 0.98 mg·L−1 and 1.50 mg·L−1 respectively). Higher concentrations of total flavanols (2 mg·L−1 in hyperoxygenated must and 21 mg·L−1 in control must; 7.5 mg·L−1 in hyperoxygenated wine and 19.8 mg·L−1 in control wine) and polyphenols (97 mg·L−1 in hyperoxygenated must and 249 mg·L−1 in control must; 171 mg·L−1 in hyperoxygenated wine and 240 mg·L−1 in control wine) were found in both the must and the control wine. A total of 24 volatiles were determined using gas chromatography mass spectrometry. Statistical differences were achieved for isobutyl alcohol (26.33 mg·L−1 in control wine and 32.84 mg·L−1 in hyperoxygenated wine), or 1-propanol (7.28 mg·L−1 in control wine and 8.51 mg·L−1 in hyperoxygenated wine), while esters such as isoamyl acetate (1534.41 µg·L−1 in control wine and 698.67 µg·L−1 in hyperoxygenated wine), 1-hexyl acetate (136.32 µg·L−1 in control wine and 71.67 µg·L−1 in hyperoxygenated wine) and isobutyl acetate (73.88 µg·L−1 in control wine and 37.27 µg·L−1 in hyperoxygenated wine) had a statistically lower concentration.

Bactrocera oleae (Rossi) (Diptera: Tephritidae) Response to Different Blends of Olive Fruit Fly-Associated Yeast Volatile Compounds as Attractants

The olive fruit fly, Bactrocera oleae (Rossi) is economically the most important olive pest, causing yield losses in all olive growing areas where is detected. Considering that EU requires the reduction of pesticide use by up to 100% by 2050, more effective non-pesticide lures for B. oleae monitoring and/or controlling are needed. This research was aimed at investigating the attractiveness of different blends of olive fruit fly-associated yeast volatiles toward B. oleae. Three blends of olive fruit fly-associated yeast volatiles: isoamyl alcohol and 2-phenethyl alcohol; isoamyl alcohol, 2-phenethyl alcohol and 2-phenethyl acetate; and isoamyl alcohol, 2-phenethyl acetate and isobutyl acetate were selected and tested on yellow sticky traps for attraction of B. oleae in olive orchard. Results showed that traps containing all tested blends of olive fruit fly-associated yeast volatile compounds, in total, were significantly more attractive to B. oleae and were not significantly attractive to green lacewings, compared to the control. Among them, the most promising was the one containing the blend of isoamyl alcohol, 2-phenethyl acetate and isobutyl acetate because its attractiveness was constantly significant during investigation compared to the others. This blend in the future could lead us to the discovery of a new attractant for the monitoring and/or controlling of B. oleae.

Non-Conventional Yeasts to Produce Aroma Compounds by Using Agri-Food Waste Materials

Nowadays, biotechnological applications are emphasized to ensure sustainable development by re-utilizing of waste materials to prevent ecological problems and to produce or recover compounds that may have positive effects on health. Yeasts are fascinating microorganisms that play a key role in several traditional and innovative processes. Although Saccharomyces is the most important genus of yeasts, and they are major producers of biotechnological products worldwide, a variety of other yeast genera and species than Saccharomyces, which called ‘non-Saccharomyces’ or ‘non-conventional’ yeasts also have important potential in order to use in biotechnological applications. Some of the non-conventional yeast strains offer a unique potential for biotechnological applications to produce valuable secondary metabolites due to their characteristics of surviving and growing in such extreme conditions, e.g. wide substrate range, rapid growth, thermotolerance, etc. In this review, we aimed to summarize potential biotechnological applications of some non-conventional yeasts (Kluyveromyces spp., Yarrowia spp., Pichia spp., Candida spp., etc.) to produce industrially important aroma compounds (phenylethyl alcohol, phenylethyl acetate, isobutyl acetate, diacetyl, etc.) by re-utilizing agri-food waste materials in order to prevent ecological problems and to produce or recover compounds that may have positive effects on health.

Formaldehyde Analysis in Non-Aqueous Methanol Solutions by Infrared Spectroscopy and Electrospray Ionization

We present the analysis of formaldehyde (HCHO) in anhydrous methanol (CH3OH) as a case study to quantify HCHO in non-aqueous samples. At higher concentrations (C > 0.07 M), we detect a product of HCHO, methoxy methanol (MM, CH3OCH2OH), by Fourier transform infrared spectroscopy, FTIR. Formaldehyde reacts with CH3OH, CD3OH, and CD3OD as shown by FTIR with a characteristic spectral feature around 1,195 cm−1 for CH3OH used for the qualitative detection of MM, a formaldehyde derivative in neat methanol. Ab initio calculations support this assignment. The extinction coefficient for 1,195 cm−1 is in the order of 1.4 × 102 M−1cm−1, which makes the detection limit by FTIR in the order of 0.07 M. For lower concentrations, we performed the quantitative analysis of non-aqueous samples by derivatization with dinitrophenylhydrazine (DNPH). The derivatization uses an aqueous H2SO4 solution to yield the formaldehyde derivatized hydrazone. Ba(OH)2 removes sulfate ions from the derivatized samples and a final extraction with isobutyl acetate to yield a 1:1 methanol: isobutyl acetate solvent for injection for electrospray ionization (ESI). The ESI analysis gave a linear calibration curve for concentrations from 10 to 200 µM with a time-of-flight analyzer (TOF). The detection and quantification limits are 7.8 and 26 μM, respectively, for a linear correlation with R2 > 0.99. We propose that the formaldehyde in CH3OH is in equilibrium with the MM species, without evidence of HCHO in solution. In the presence of water, the peaks for MM become less resolved, as expected from the well-known equilibria of HCHO that favors the formation of methylene glycol and polymeric species. Our results show that HCHO, in methanol does not exist in the aldehyde form as the main chemical species. Still, HCHO is in equilibrium between the production of MM and the formation of hydrated species in the presence of water. We demonstrate the ESI-MS analysis of HCHO from a non-aqueous TiO2 suspension in methanol. Detection of HCHO after illumination of the colloid indicates that methanol photooxidation yields formaldehyde in equilibrium with the solvent.

Probing Specificities of Alcohol Acyltransferases for Designer Ester Biosynthesis with a High-Throughput Microbial Screening Platform

Alcohol acyltransferases (AATs) enables microbial biosynthesis of a large space of esters by condensing an alcohol and an acyl CoA. However, substrate promiscuity of AATs prevents microbial biosynthesis of designer esters with high selectivity. Here, we developed a high-throughput microbial screening platform that facilitates rapid identification of AATs for designer ester biosynthesis. First, we established a microplate-based culturing technique with in situ fermentation and extraction of esters. We validated its capability in rapid profiling of the alcohol substrate specificity of 20 chloramphenicol acetyltransferase variants derived from Staphylococcus aureus (CATSa) for microbial biosynthesis of acetate esters with various exogeneous alcohol supply. By coupling the microplate-based culturing technique with a previously established colorimetric assay, we developed a high-throughput microbial screening platform for AATs. We demonstrated that this platform could not only confirm CATSa F97W with enhanced isobutyl acetate synthesis but also identify three ATF1Sc (P348M, P348A, and P348S) variants, derived from Saccharomyces cerevisiae' s AAT and engineered by model-guided protein design, for enhanced butyl acetate production. We anticipate the high-throughput microbial screening platform is a useful tool to identify novel AATs that have important roles in nature and industrial biocatalysis for designer bioester production.

Effects of Simultaneous Co-Fermentation of Five Indigenous Non-Saccharomyces Strains with S. cerevisiae on Vidal Icewine Aroma Quality

In this study, Vidal grape must was fermented using commercial Saccharomyces cerevisiae F33 in pure culture as a control and in mixed culture with five indigenous non-Saccharomyces yeast strains (Hanseniaspora uvarum QTX22, Saccharomycopsis crataegensis YC30, Pichia kluyveri HSP14, Metschnikowia pulcherrima YC12, and Rhodosporidiobolus lusitaniae QTX15) through simultaneous fermentation in a 1:1 ratio. Simultaneous fermentation inhibited the growth of S. cerevisiae F33 and delayed the time to reach the maximum biomass. Compared with pure fermentation, the contents of polyphenols, acetic esters, ethyl esters, other esters, and terpenes were increased by R. lusitaniae QTX15, S. crataegensis YC30, and P. kluyveri HSP14 through simultaneous fermentation. S. crataegensis YC30 produced the highest total aroma activity and the most abundant aroma substances of all the wine samples. The odor activity values of 1 C13-norisoprenoid, 3 terpenes, 6 acetic esters, and 10 ethyl esters improved significantly, and three lactones (δ-decalactone, γ-nonalactone, and γ-decalactone) related to coconut and creamy flavor were only found in this wine. Moreover, this sample showed obvious “floral” and “fruity” note odor due to having the highest amount of ethyl ester aromatic substances and cinnamene, linalool, citronellol, β-damascenone, isoamyl ethanoate, benzylcarbinyl acetate, isobutyl acetate, etc. We suggest that simultaneous fermentation of S. crataegensis YC30 with S. cerevisiae might represent a novel strategy for the future production of Vidal icewine.

The Use of Yeast Mixed Cultures for Deacidification and Improvement of the Composition of Cold Climate Grape Wines

Interest in the use of non-Saccharomyces yeast in mixed cultures is increasing due to the perceived improvement in the quality and complexity of the resulting wines. The aim of the study was to determine the ability of monocultures and mixed yeast cultures for deacidification and improvement of the composition of cold climate grape wines. Fermentation of grape musts with increased total acidity was carried out with the use of monocultures of Saccharomyces cerevisiae MH020215 (Sc), Zygosaccharomyces bailii 749 (Zb) and Metschnikowia pulcherrima MG970690 (Mp), and their mixed cultures, inoculated simultaneously and sequentially. Oenological parameters, organic acids and volatile compounds profiles of obtained wines were characterized. The fermentation kinetics and analytical profiles of the obtained wines showed that the use of mixed yeast cultures contributed to the reduction of volatile acidity and acetic acid content in the wines, as well as obtaining a favorable aromatic profile of the wines. The dominant higher alcohols in all wines were 2-methyl-1-propanol, 3-methyl-1-butanol and 2-methyl-1-butanol. Significantly higher amounts of the first two compounds were found in wines obtained with M. pulcherrima MG070690, both in monoculture and in mixed cultures. The monocultures of M. pulcherrima MG070690 (Mp) compared with Z. bailli 749 (Zb) synthesized higher levels of esters in wines, including ethyl acetate, ethyl propionate, isobutyl acetate, ethyl pyroracemate and isoamyl acetate.