Influence of pH and ethanol on malolactic fermentation and volatile aroma compound composition in white wines

The present study investigated the influences of pH and ethanol on malolactic fermentation (MLF) and the volatile aroma profile of the subsequent white wines from Riesling and Chardonnay inoculated with two different Oenococcus oeni strains. In all cases MLF was induced after completion of alcoholic fermentation (AF). Partial MLF occurred under low pH 3.2 and high alcohol (118.3 g/L) conditions. In the cases with complete MLF, the time required for each strain varied from 13 to 61 days and was dependent on bacterial culture, cultivar and wine parameter. Chemical properties of each wine were determined after AF, complete and partial MLF. The wines showed significant differences in total higher alcohols, esters and acids that are important for the sensory profile and quality of wine. This work demonstrated that the wine matrix as well as the pH and alcohol concentration affects MLF and the final volatile aroma profile. Results indicate that changes in volatile aroma composition are not necessarily related to complete MLF and that partial MLF already has distinct influences on the wine aroma profile of white wines.     

Malolactic Fermentation and Secondary Metabolite Production by Oenoccocus oeni Strains in Low pH Wines

Abstract: Different wine varieties, including some with low pH, were studied to determine the ability to grow and produce secondary metabolites of a previously selected autochthonous Oenococcus oeni strain (C22L9), compared with a commercial strain. Monitoring of malolactic fermentation (MLF) was carried out by microbiological and chemical analysis of wines. The concentration of some major volatile compounds and biogenic amines in wines before and after malolactic fermentation was also determined. The results showed major differences in MLF duration both between wines and strains, although the differences between strains were slight for most of the analyzed compounds. Statistically significant differences in citric acid degradation were found in all wine varieties and it was confirmed that O. oeni C22L9 is a poor degrader of citric acid; this means that MLF can be prolonged without the risk of producing high concentrations of diacetyl and acetoin. Sensory analysis of wines after MLF showed similar characteristics in wines from both strains. This study thus shows that O. oeni C22L9 possesses even better sensory and fermentation properties than the commercial strain and can be used in wines with different characteristics, which makes it highly valuable for industrial use. Practical Application: The increasingly use of grape varieties of low pH in winemaking and the higher alcohol content of wines, as a consequence of the climatic change, make interesting the study of the behavior during MLF of O. oeni strains in order to determine their ability to grow, when growth conditions are not optimal, and to produce secondary metabolites. A comparative study was conducted using an autochthonous O. oeni strain (C22L9) and a commercial O. oeni strain and 4 wine varieties.     

Impact of malolactic fermentation on the volatile composition of Turkish Kalecik karası red wines

The effects of spontaneous malolactic fermentation (MLF) and a malolactic starter culture (Oenococcus oeni) on the volatile compounds of Kalecik karas? red wines were investigated. The volatile compounds were extracted using liquid–liquid extraction and quantified by GC–MS–flame ionisation detection. Starter cultures of O. oeni had a positive effect in shortening MLF and producing lower volatile acidity. After MLF, the total volatile compounds increased in both the inoculated wines and the spontaneously fermented wine. Ethyl lactate, diethyl succinate and γ‐butyrolactone increased upon inoculation with cultures of O. oeni strains PN4 and VP41. MLF increased the diacetyl concentration in all Kalecik karas? wines. Strain dependent differences were found in the level of diacetyl, with O. oeni PN4 producing more diacetyl than O. oeni VP41. After MLF, the concentrations of acetoin, 4‐ethylguaicol and 4‐ethylphenol increased in spontaneously fermented wine compared with wines inoculated with O. oeni. Accordingly, the use O. oeni PN4 is recommended for a controlled MLF to produce better quality Kalecik karas? wines. © 2018 The Institute of Brewing & Distilling     

Oenococcus oeni and malolactic fermentation – moving into the molecular arena

Malolactic fermentation (MLF) is the bacterial‐driven decarboxylation of L‐malic acid to L‐lactic acid and carbon dioxide, and brings about deacidification, flavour modifications and microbial stability of wine. Pasteur first described the presence of ‘bacteria’ in wine nearly one‐hundred‐and‐fifty years ago and the subsequent elucidation of the bacterial‐driven malolactic reaction was described about fifty years later. Over the following years the occurrence of MLF became apparent in wines worldwide, and eventually Oenococcus oeni was identified as the principal organism involved in the process. O. oeni is remarkable in its ability to tolerate the nutritionally poor and challenging, harsh wine environment; however, it can be a difficult and sometimes unreliable organism to work with in the winery. A greater knowledge of its biology would undoubtedly facilitate the development of strains and practices, with improved performance outcomes. We already know a considerable amount about the biochemistry and physiology of O. oeni, and ironically, although we know little about its genetics, its genome has been sequenced. With this groundwork in place and molecular biology techniques at our disposal we are poised to increase our knowledge and understanding of this organism enormously.     

Molecular analysis of Oenococcus oeni population dynamics and the effect of aeration and temperature during alcoholic fermentation on malolactic fermentation

The effects of aeration and temperature during alcoholic fermentation (AF) on spontaneous and inoculated malolactic fermentation (MLF) of wine have been analysed by following the population dynamics of Oenococcus oeni strains with multiplex random amplified polymorphic DNA‐polymerase chain reaction. It has enabled us to follow precisely the proportion of different autochthonous strains and the starter strain through fermentations. Lack of aeration led to delays in AF, which meant that autochthonous lactic acid bacteria could develop earlier and prevented the starter strain from developing correctly. Temperature was not found to lead to any differences. Two strains were isolated in the same spontaneous MLF, suggesting that, in some cases, multiple strains might be responsible for the degradation of malic acid in wine. It can be concluded that delays in the AF can negatively affect the control of MLF and that this can be studied by following the development of the different strains of O. oeni.     

Enhancement of volatile aromatic compounds in black raspberry wines via enzymatic treatment

Robus coreanus Miquel is widely used in the production of Korean black raspberry (KBR) wine owing to its health benefits and commercial value. The effects of three different commercially available glycosidase enzymes on the volatile compounds in KBR wines were investigated with large‐scale fermentation to develop a high‐flavour‐quality wine. Volatile aroma compounds from the wines were analysed using headspace‐solid phase micro‐extraction‐gas chromatography–mass spectrometry (HS‐SPME‐GC–MS) and sensory evaluations were performed to evaluate the flavour characteristics. KBR wines treated with commercial enzymes yielded high concentrations of terpenes and esters compared with the control wine because the odourless non‐volatile glycosides in KBR wines were converted to their corresponding free forms by the enzymes. HS‐SPME‐GC–MS‐SIM analysis showed that the primary monoterpene compounds in KBR wines treated by the enzymes were myrtenol, linalool, citronellol and a significant quantity of compounds such as 2‐phenyl ethanol and ethyl benzoate, which contributed to the flavour of KBR wine, as determined by GC‐FID. KBR wines treated with the enzymes exhibited different sensory characteristics from the control wine owing to higher intensity of floral and fruity aromas. KBR wine treated with CYTOLASE PCL5 on a large scale exhibited the highest sensory preference. Copyright © 2017 The Institute of Brewing & Distilling     

Inhibition and inactivation evaluation of exogenous glycosidases in wines using p‐nitrophenylglycosides

BACKGROUND: Glycosidases are often used to improve the aroma of wines via hydrolysis of glycosidic precursors. The aim of this work was to develop a method to test activity and stability of these enzymes in wine environments using p‐nitrophenylglycosides, avoiding interference by colored substances. RESULTS: The proposed procedure for determination of glycosidase activity in wines involves quantification of the hydrolysis product p‐nitrophenol by HPLC. The method was applied to compare the inhibition of some commercial glycosidases by red and white wines. It was found that inhibition of a β‐D ‐glucosidase by ethanol, glucose and gluconolactone was smaller than that produced by wine. This enzyme was also inhibited by wine fractions rich in phenolic compounds and glycosides. When glycoside fractions were first hydrolyzed, their inhibition was strongly reduced. The stability of glycosidases in red wines was tested, showing a destabilizing effect of β‐glucosidases that was stronger than in white wine. CONCLUSION: The method developed allows measurement of glycosidase activity on p‐nitrophenylglycosides in the presence of colored wine compounds. Comparison of relative glycosidase activity in wines to that in buffer solution under the same conditions can assist in the selection of appropriate aroma‐enhancing enzymes. Copyright © 2009 Society of Chemical Industry     

Malolactic fermentation of Tempranillo wine: contribution of the lactic acid bacteria inoculation to sensory quality and chemical composition

Malolactic fermentation (MLF) of Tempranillo Rioja wines (Spain) inoculated with two lactic acid bacteria (LAB) strains were studied and compared with spontaneous MLF. Inoculation with selected Oenococcus oeni lyophila shortened MLF duration up to 19 days and lead to wines with more fresh and fruity characters, especially when implantation was 100%. We demonstrated modifications in the concentration of volatile and nitrogenous compounds and a good correlation between analytical and sensory attributes was also noted. In addition, the low initial amino acid concentration and the consumption of these compounds by the inoculated yeast strain during alcoholic fermentation resulted in wines with very low biogenic amines levels (under 3.75 mg L^?1) after MLF and 3 month storage period in all cases. The results showed the significance of choose the most suitable starter to elaborate quality wines and suggest the control of amino acid content in must and wine to prevent the formation of biogenic amines.     

Yeast and bacterial modulation of wine aroma and flavour

Wine is a highly complex mixture of compounds which largely define its appearance, aroma, flavour and mouth‐feel properties. The compounds responsible for those attributes have been derived in turn from three major sources, viz. grapes, microbes and, when used, wood (most commonly, oak). The grape‐derived compounds provide varietal distinction in addition to giving wine its basic structure. Thus, the floral monoterpenes largely define Muscat‐related wines and the fruity volatile thiols define Sauvignon‐related wines; the grape acids and tannins, together with alcohol, contribute the palate and mouth‐feel properties. Yeast fermentation of sugars not only produces ethanol and carbon dioxide but a range of minor but sensorially important volatile metabolites which gives wine its vinous character. These volatile metabolites, which comprise esters, higher alcohols, carbonyls, volatile fatty acids and sulfur compounds, are derived from sugar and amino acid metabolism. The malolactic fermentation, when needed, not only provides deacidification, but can enhance the flavour profile. The aroma and flavour profile of wine is the result of an almost infinite number of variations in production, whether in the vineyard or the winery. In addition to the obvious, such as the grapes selected, the winemaker employs a variety of techniques and tools to produce wines with specific flavour profiles. One of these tools is the choice of microorganism to conduct fermentation. During alcoholic fermentation, the wine yeast Saccharomyces cerevisiae brings forth the major changes between grape must and wine: modifying aroma, flavour, mouth‐feel, colour and chemical complexity. The wine bacterium Oenococcus oeni adds its contribution to wines that undergo malolactic fermentation. Thus flavour‐active yeasts and bacterial strains can produce desirable sensory results by helping to extract compounds from the solids in grape must, by modifying grape‐derived molecules and by producing flavour‐active metabolites. This article reviews some of the most important flavour compounds found in wine, and their microbiological origin.     

Metabolism of SO₂ binding compounds by Oenococcus oeni during and after malolactic fermentation in white wine

Sulfur dioxide SO? is the key additive for the preservation of wines. Carbonyl and keto compounds in wine can bind to SO? and decrease its efficacy, resulting in higher total SO? requirements. Increased consumer demand for low sulfite and organic wines pose production challenges if SO? binders have not been properly managed during vinification. Malolactic fermentation (MLF) has been known to reduce bound SO? levels but detailed time course studies are not available. In this work, the kinetics of major SO? binding compounds and malic acid were followed during MLF in wine with 12 commercially available strains of Oenococcus oeni. Pyruvic acid, acetaldehyde and α-ketoglutaric acid were degraded to various degrees by O. oeni, but galacturonic acid was not. At the time of malic acid depletion, percent degradation of pyruvate, α-ketoglutaric acid and acetaldehyde was 49%, 14% and 30%, respectively. During MLF, the decrease in average bound SO? levels, as calculated from carbonyl metabolism, was 22%. The largest reduction in wine carbonyl content occurred in the week after completion of MLF and was 53% (107 mg/L to 34 mg/L) calculated as bound SO?. Prolonged activity of bacteria in the wines (up to 3 weeks post malic acid depletion) resulted only in reduced additional reductions in bound SO? levels. The results suggest that microbiological wine stabilization one week after malic acid depletion is an effective strategy for maximum removal of SO? binders while reducing the risk of possible post-ML spoilage by O. oeni leading to the production acetic acid and biogenic amines.     

Release of glycosidically bound flavour compounds of Chardonnay by Oenococcus oeni during malolactic fermentation

Glycosidases, produced by Oenococcus oeni strain Lalvin EQ54 during malolactic fermentation (MLF) performed in a chemically defined wine (CDW) medium, contributed to the release of volatile aglycons from their glycosylated precursors, present in a Chardonnay wine glycosidic extract. The liberation of wine volatiles during MLF was limited by the low activity of these enzymes in this strain. Six different aglycons examined were 3-hydroxydamascone, alpha-terpineol, vanillin, methyl vanillate, 4-hydroxybenzoic acid and tyrosol. Using p-nitrophenyl synthetic substrates, it was shown that O. oeni Lalvin EQ54 has β-glucosidase, and limited α-l-rhamnopyranosidase and α-l-arabinofuranosidase activities. Release of aglycons from the Chardonnay wine glycosidic extract was increased when purified α-l-rhamnopyranosidase and α-l-arabinofuranosidase were added to the CDW medium together with the malolactic bacteria culture. The results obtained confirmed that O. oeni has the necessary glycosidases for the sequential liberation of the disaccharide sugars from wine glycosides. The rate of MLF proceeded much faster in the CDW supplemented with the Chardonnay wine glycoside extract (8 days compared to 20 days), even though the bacterial growth was unaffected.     

Effects of Fermentation Temperature on Key Aroma Compounds and Sensory Properties of Apple Wine

Fermentation temperature strongly affects yeast metabolism during apple wine making and thus aromatic and quality profiles. In this study, the temperature effect during apple wine making on both the key aroma compounds and sensory properties of apple wine were investigated. The concentration of nine key aroma compounds (ethyl acetate, isobutyl acetate, isopentylacetate, ethyl caprylate, ethyl 4‐hydroxybutanoate, isobutylalcohol, isopentylalcohol, 3‐methylthio‐1‐propanol, and benzeneethanol) in apple wine significantly increased with the increase of fermentation temperature from 17 to 20 °C, and then eight out of the nine key aroma compounds with an exception of ethyl 4‐hydroxybutanoate, decreased when the temperature goes up 20 to 26 °C. Sensory analysis showed that the apple wine fermented at 20 °C had the highest acceptance for consumers. Fermentation at the temperature of 20 °C was therefore considered to be the most suitable condition using the selected yeast strain (Saccharomyces cerevisiae AP05) for apple wine making. Changes in the fermentation temperature can considerably affect the production of key aroma compounds and sensory profiles of apple wine. These results could help apple wine producers make better quality production for consumers at the optimal fermentation temperature.     

Volatile aroma and sensory analysis of black raspberry wines fermented by different yeast strains

Robus coreanus Miquel is a small berry fruit used for Korean black raspberry (KBR) wine‐making. Twelve different yeast strains were investigated by laboratory‐scale fermentation to develop a wine with a high flavour quality. Volatile aroma compounds from the wines were analysed using headspace–solid phase microextraction–gas chromatography–mass spectrometry and sensory evaluation was performed to evaluate the flavour characteristics. The volatile aroma compounds that mostly contributed to the flavour of KBR wines were those related to fruity (esters) and floral (terpenes) aromas. Fifteen out of the 67 identified volatile compounds showed higher odour activity values than other compounds in the wines, and these compounds were considered as important contributors to the final aromas of the wine. Additionally, the KBR wine fermented by the M1 yeast strain had the highest sensory preference because of higher fruity and floral aroma characters compared with other wines. In addition to the M1 strain, the other yeast strains that produced favourable sensory characteristics included Enoferm CSM, Uvaferm VRB, Lalvin ICV GRE, Lalvin ICV Opale and LevureSeche Active. Of these strains, the M1 strain produced a particularly excellent black raspberry wine, and thus could be applied for further large‐scale production of black raspberry wines. It is also expected that this work will expedite research on the production of high‐quality black raspberry wines with beneficial physicochemical properties, functionality and good sensory characteristics. Copyright © 2015 The Institute of Brewing & Distilling     

Improvement of volatile composition of wines by controlled addition of malolactic bacteria

The effect of malolactic fermentation (MLF) on the volatile composition of red wines was studied by inoculation with selected lactic acid bacteria. Four wines were inoculated with different Oenococcus oeni (syn. Leuconostoc oenos) strains, the major malolactic species found in wines, and one was inoculated with a Lactobacillus sp. strain. A non inoculated wine was also analyzed to act as a control. Malolactic fermentation and evolution of non volatile compounds were followed by HPLC and after the depletion of the malic acid present in wine the volatile compounds were extracted and analyzed by gas chromatography with flame ionization and mass spectrometry. Wines which had undergone the MLF showed a significant increment in total higher alcohols, esters and acids that are important in the sensory properties and quality of wine.     

Hydrolysis of glycosidically bound flavour compounds from oak wood by Oenococcus oeni

Malolactic fermentation (MLF), which is conducted by lactic acid bacteria (LAB), has a significant influence on the stability and organoleptic quality of wine. Recent studies have shown that when MLF is carried out in oak wood barrels, LAB were also able to interact with wood and increase volatile compound contents such as vanillin during MLF. The release of these compounds indicates that LAB may convert vanillin precursors present in oak wood. In this work, the effect of commercial glycosidases on the released vanillin was firstly studied. This aldehyde is present in wood extracts in monoglycosidic forms where the major glycones are arabinose and xylose. Other aglycons released during MLF in barrels, syringaldehyde and whisky-lactones, can be considered as other sources of aroma. Secondly, strains selected with high activities toward glycoside substrates could hydrolyse vanillin glycoside precursors from oak wood with the same efficiency as commercial enzymes.     

Interaction of pH, ethanol concentration and wine matrix on induction of malolactic fermentation with commercial 'direct inoculation' starter cultures

The effect of the interaction between pH (2.9–3.5), alcohol concentration (12.5–14.5% v/v) and wine matrix (Chardonnay, Grenache blend and Cabernet Sauvignon) on malolactic fermentation (MLF) induced by two commercially available directly inoculated Oenococcus oeni starter cultures was determined. Wine matrix had greatest impact on the rate of MLF, followed by pH and alcohol; the matrix effect was significantly modified by pH and bacterial culture. The rate of L-malic acid degradation by starter culture A was 3.3–61.3% in the Grenache blend and 3.3–4.5% in the Chardonnay wine compared to the Cabernet Sauvignon wine, for the pH/alcohol ranges 2.9/14.5 to 3.5/12.5. L-malic acid degradation rate by starter culture B was lower but less affected by wine matrix. The rate of L-malic acid degradation was closely related to culture viability, with a high rate of L-malic acid degradation being associated with bacterial growth and vice versa . Treatment of two wines in which MLF had previously failed to complete with various combinations of nutrients, carbon fining and heating, restored MLF, suggesting that a growth-limiting nutrient was absent and/or a toxic substance not related to pH, alcohol or SO₂ concentration was present. This work demonstrated that the wine matrix may affect culture viability more significantly than either pH or alcohol concentration. A high rate of MLF was shown to depend on factors permitting bacterial growth, however, the two strains studied responded differently to the wine matrix, suggesting that malolactic bacteria vary in tolerance to various stresses in the wine environment.     

The effect of successive uses of oak barrels on the extraction of oak-related volatile compounds from wine

Changes in the concentration of furfuryl compounds, guaiacol and 4‐methylguaiacol, cis‐ and trans‐oak lactone, and vanillin were studied during the maturation of wines in oak barrels of different origins (French oak and American oak). The rate at which these volatile compounds accumulated in the wines followed a curve that fits to mathematical models. The changes in concentration were usually consistent with diffusion kinetics. When the barrels are new, the concentration gradient between the wood surface and the wine is high, so diffusion occurs quickly, especially in the case of lactones, the extraction rate of which could be fitted to a power curve. After some time of maturation, the concentration gradient is not so large and furthermore, the wood surface may be exhausted of some compounds so the rate of extraction decreases. During the second fill, the rate of accumulation of these compounds in the wine was almost linear, especially during the first 6 months of storage, and the quantities measured in wine were much lower. The concentration of some compounds, increased during the final 3 months (between days 180 and 270), exhibiting a quadratic curve when plotted. Furfuryl aldehydes behaved differently as they are extracted very rapidly from the first millimetres of wood surface and transformed into furfuryl alcohol, a compound with no organoleptic significance.