An overview of formation and roles of acetaldehyde in winemaking with emphasis on microbiological implications

Shao-Quan Liu & Gordon J. Pilone

Institute of Molecular BioSciences, College of Sciences, Massey University, Palmerston North, New Zealand (Received 1 July 1998; Accepted in revised form 1 December 1998)

Summary

Acetaldehyde is an important aroma compound in wine. This article rst reviews the microbial and chemical formation of acetaldehyde, then the effects of sulphur dioxide on acetaldehyde and effects of acetaldehyde on wine colour and physical stability are described briey. Finally, the microbiological implications of acetaldehyde are emphasized with respect to practical signicance in wine fermentation.
Bacteria, fermentation, phenolics, SO2, yeasts.

Keywords

Introduction

Acetaldehyde (i.e. ethanal, CH3CHO) is a potent volatile avour compound found in many beverages and foods such as apple juice and spirits (Miyake & Shibamoto, 1993), beer (Margalith, 1981; Adams & Moss, 1995), cider and perry (Williams & Tucknott, 1971; Williams, 1975), wine (Dittrich & Barth, 1984), cheese (Urbach, 1995, 1997; Weerkamp et al., 1996; Engels et al., 1997), yoghurt (Bottazzi & Vescovo, 1969; Zourari et al., 1992) and ripened butter (Lindsay et al., 1965). A summary of acetaldehyde levels in alcoholic beverages is given in Table 1. Acetaldehyde at low levels gives a pleasant fruity aroma, but at high concentrations it possesses a pungent irritating odour (Miyake & Shibamoto, 1993). Indeed, excess acetaldehyde produces a green, grassy or apple-like off-avour in beer (Margalith, 1981; Adams & Moss, 1995), cider (Williams, 1974), wine (Henschke & Jiranek, 1993), ripened butter (Lindsay et al., 1965), sour cream and cottage cheese (Rash, 1990). The avour threshold of acetaldehyde varies with
Correspondent: S.-Q. Liu, The New Zealand Dairy Research Institute, Private Bag 11029, Palmerston North, New Zealand. e-mail: SHAO.LIU@nzdri.org.nz

products; for example, approximately 30 mg L1 in cider (Williams, 1974), 2050 mg L1 in beer (Harrison, 1970) and 100125 mg L1 in wine (Berg et al., 1955; Zoecklein et al., 1995). Acetaldehyde is extremely reactive and can react with amino acids to generate various avour compounds (Grifth & Hammond, 1989). Wine is a product of grape juice fermentation. Mature grapes are harvested, crushed, sulphited,

Table 1 Acetaldehyde levels in alcoholic beveragesa Type Red wine White wine Sweet wine Sherry Brandy Cognac Beer Cider Whisky Sak
a

Acetaldehyde (mg L1) 4212 11493 188248 90500 63308 105 512 50 10110 1560

Data summarized from: Amerine & Ough (1980), Castino (1985), Dittrich & Barth (1984), Harrison (1970), Lzaro et al. (1987), Leguerinel et al. (1989), McCloskey & Mahaney (1981), Miyake & Shibamoto (1993), Nykanen (1986), Williams (1974).

claried (white wine), fermented; then racked, aged in barrels or larger vessels, ned, ltered, bottled and bottle aged (Boulton et al., 1996). Yeasts are responsible for the alcoholic fermentation. Sometimes lactic acid bacteria (LAB), either occurring naturally or inoculated, bring about a secondary fermentation (the so-called malolactic fermentation, MLF) usually after the yeast fermentation. This imparts avour complexity, increased microbial stability and deacidication in high acid wine (Kunkee, 1967, 1974; Davis et al., 1985; Wibowo et al., 1985; Henick-Kling, 1993, 1995; Pilone, 1995). Numerous volatile compounds such as aldehydes, alcohols and esters found in wine are primarily responsible for wine aroma (Rapp, 1988). Acetaldehyde is one of the most important sensory carbonyl compounds formed during vinication and constitutes more than 90% of the total aldehyde content in wine (Nykanen, 1986). Various levels of acetaldehyde are found in wine, with average values of about 80 mg L1 for white wine, 30 mg L1 for red wine and 300 mg L1 for sherries (McCloskey & Mahaney, 1981). While high levels of acetaldehyde are generally undesirable in table wines, high concentrations of this volatile compound are considered a unique feature of sherry-type wines (Sponholz, 1993; Cortes et al., 1998). Also, port wine owes its essential features of colour and avour to excess acetaldehyde derived from the brandy used for fortication (Singleton & Guymon, 1963; Singleton et al., 1964; Bakker & Timberlake, 1986). The aim of this article is to give an overview of the formation and roles of acetaldehyde in winemaking, emphasizing its microbiological implications with a view to stimulating further research in this area. References to relevant micro-organisms from other food products, LAB of cheese and yoghurt in particular, are made with regard to acetaldehyde formation and metabolism.

yeasts; for instance, 0.5286 mg L1 for Saccharomyces cerevisiae and 9.566 mg L1 for Kloeckera apiculata (Table 2). While sugar is the primary substrate of acetaldehyde formation, metabolism of amino acids such as alanine also contributes to the formation of this compound (Henschke & Jiranek, 1993; Boulton et al., 1996). In addition, acetaldehyde is formed from the oxidation of ethanol by lm yeasts (Sponholz, 1993; Zoecklein et al., 1995; Fugelsang, 1997). A considerable number of studies were conducted on the formation of acetaldehyde by lm yeasts in relation to sherry-type wine production over three decades ago (Fornachon, 1953; Amerine, 1958; Ough & Amerine, 1958, 1960; Ough, 1961). Acetaldehyde is excreted mainly during the growth period (Ribreau-Gayon et al., 1956a, 1956b; Amerine & Ough, 1964; Weeks, 1969; Martnez et al., 1997) and can be recatabolized (Fornachon, 1953; Farris et al., 1983). Factors such as temperature, oxygen and SO2 affect the production of acetaldehyde by yeasts (Ough & Amerine, 1958). While Amerine & Ough (1964) reported that fermentation temperature did not inuence the nal total acetaldehyde content in wine, Romano et al. (1994) found increased formation of acetaldehyde at 30 C compared with that formed at 12 C or 24 C. By contrast, Cabranes et al. (1998) observed a signicantly higher acetaldehyde production at 12 C than at 18 C in a cider fermented with S. cerevisiae. SO2

Table 2 Acetaldehyde levels produced by yeasts Yeast Acetaldehyde (mg L1) 0.5286 110350 16683 36125 1033 30 666 0.55 10.528 2340

Acetaldehyde formation in wine

Saccharomyces cerevisiae Saccharomyces uvarum Saccharomyces bayanus Saccharomyces oviformis Saccharomyces fructuum Saccharomyces ludwigii Kloeckera apiculata Torulaspora delbrueckii Hanseniaspora guilliermondii Metschnikowia pulcherrima
a

Formation of acetaldehyde by yeasts Acetaldehyde is considered to be a leakage product of the alcoholic fermentation by yeasts (Margalith, 1981). There are large species and strain differences in acetaldehyde production by

Data summarized from: Cortes et al. (1998), Di Stefano & Ciol (1982), Farris et al. (1983), Fleet & Heard (1993), Herraiz et al. (1989, 1990), Ibeas et al. (1997), Longo et al. (1992), Martnez et al. (1997), Milln & Ortega (1988), Rankine & Pocock (1969), Romano et al. (1994, 1997, 1998), Stratford et al. (1987), Weeks (1969).

induces acetaldehyde formation by yeasts (Rankine & Pocock, 1969; Weeks, 1969; Stratford et al., 1987; Pilkington & Rose, 1988) and wines fermented with SO2 have considerably higher acetaldehyde levels than wines made without SO2 (Herraiz et al., 1989). This SO2-induced production of acetaldehyde appears to be related to SO2 resistance in yeasts (Stratford et al., 1987; Pilkington & Rose, 1988). Anaerobiosis, low pH and/or high sugar content apparently promote acetaldehye production by yeasts (RibreauGayon et al., 1956a). Formation of acetaldehyde by acetic acid bacteria Besides yeasts, acetic acid bacteria (AAB) originating from grapes and winery equipment (Joyeux et al., 1984a; Drysdale & Fleet, 1985) can also produce acetaldehyde. The oenological and practical implications of AAB in winemaking have been reviewed elsewhere (Drysdale & Fleet, 1988). AAB oxidize ethanol to acetaldehyde and acetic acid, and concentrations of up to 250 mg L1 acetaldehyde can be formed (Joyeux et al., 1984b; Lafon-Lafourcade & Ribreau, 1984; Drysdale & Fleet, 1989). This concentration well exceeds the sensory threshold value of 100125 mg L1 for this highly volatile aroma compound, and would adversely affect the organoleptic property of wine. Acetaldehyde tends to accumulate under low oxygen conditions and/or ethanol concentrations higher than 10% (v/v) instead of being oxidized to acetic acid (Zoecklein et al., 1995; Fugelsang, 1997). Formation of acetaldehyde by lactic acid bacteria At present, it is not clear whether wine lactic acid bacteria (LAB) can produce acetaldehyde. In contrast, the formation of acetaldehyde by dairy LAB is well documented, and a great deal of information on this aspect is available. The acetaldehyde producing dairy LAB include lactococci (Harvey, 1960; Bills & Day, 1966; Keenan et al., 1966b; Liu et al., 1997), lactobacilli (Bassette et al., 1967; Keenan & Lindsay, 1967; El-Sayed et al., 1993), leuconostocs (Walsh & Cogan, 1973), pediococci (Keenan et al., 1968)

and the yoghurt bacteria Streptococcus thermophilus and Lactobacillus bulgaricus (Bottazzi & Dellaglio, 1967; Bottazzi & Vescovo, 1969; Hamdan et al., 1971). The amount of acetaldehyde formed by dairy LAB varies with species and strain, and is usually less than 30 mg L1 (Kneifel et al., 1992). Acetaldehyde is produced by dairy LAB from the metabolism of glucose (Lees & Jago, 1976a), 2-deoxy-D-ribose-5-phosphate (Lees & Jago, 1977) and threonine (Lees & Jago, 1976b; Wilkins et al., 1986a, 1986b; Marshall & Cole, 1983; Marranzini et al., 1989; Rysstad et al., 1990; Grozeva et al., 1994). These acetaldehyde precursors are also present in wine, and it will be of practical value to ascertain if wine LAB can form acetaldehyde from these substrates. Formation of acetaldehyde via auto-oxidation of ethanol and phenolic compounds Wildenradt & Singleton (1974) studied the production of acetaldehyde from ethanol in model solutions as a result of oxidation of phenolic compounds. They found that acetaldehyde production by direct oxidation of ethanol is insignificant, and that oxidation of ethanol to acetaldehyde occurs by a coupled auto-oxidation of certain phenolic compounds. A strong oxidant (H2O2), resulting from phenolic oxidation, oxidizes ethanol to acetaldehyde. This nding was veried in model solutions and red wines by Ribreau-Gayon et al. (1983).
Effect of sulphur dioxide on acetaldehyde

Sulphur dioxide (SO2) is used as an antimicrobial, antioxidative, anti-enzymatic and anti-acetaldehyde (taste improvement) agent in winemaking (Peynaud, 1984). The roles of SO2 in wine fermentation and the binding of SO2 by acetaldehyde and other compounds such as pyruvic acid and -keto-glutaric acid have been reviewed elsewhere (Peynaud, 1984; Romano & Suzzi, 1993; Zoecklein et al., 1995; Boulton et al., 1996; Somers, 1998) and interested readers are referred to these articles for detailed information. Therefore, only a brief summary of SO2 binding by acetaldehyde is given below based on these reviews.

The total SO2 consists of bound and free forms, with the former having a weak antimicrobial function. At wine pH of 34, free SO2 consists mainly of bisulphite anion (HSO31) and a small proportion of molecular SO2 (SO2.H2O) and sulphite anion (SO32). A number of carbonyl compounds (mainly acetaldehyde, pyruvic acid and -keto-glutaric acid) can bind with free SO2 (especially the bisulphite ion) to form a complex compound (bound SO2) which has only weak antimicrobial properties. The bisulphiteacetaldehyde addition product (known as hydroxysulphonate) accounts for the majority of the total SO2 and can interefere with the analysis of free SO2 due to hydrolysis, resulting in an overestimation of the actual free SO2. On the other hand, variations in the concentration of acetaldehyde must be largely responsible for the wide range in free SO2 occurring at similar levels of total SO2 because acetaldehyde has a strong afnity for SO2. The binding of bisulphite ion by acetaldehyde reduces the availability of free SO2, resulting in a reduction in the effectiveness of the antimicrobial efcacy of SO2. However, acetaldehydebound SO2 may be inhibitory to wine LAB (Fornachon, 1963; Lafon-Lafourcade, 1975; Somers & Wescombe, 1982; Hood, 1983; Delni & Morsiani, 1992, 1993), due presumably to free SO2 being released upon bacterial metabolism of the acetaldehyde moiety. Acetaldehyde in the free form is a normal constituent of wines made without use of SO2, and could represent an uncontrollable and undesirable inuence unrelated to wine composition. In spite of problems associated with the use of SO2, stable wine of good quality cannot be made without it (Somers, 1998). Therefore, one of the properties of added SO2 is to limit acetaldehyde formation and to bind acetaldehyde formed so that a wines taste and aroma are protected or improved (Peynaud, 1984; Somers, 1998). However, there must be a balance between the concentrations of these two critical constituents. Dependent on its concentration, SO2 can adversely affect the polymerization reaction between acetaldehyde and phenolics (see below) in addition to its decolourizing effect (Ribreau-Gayon et al., 1983). Just as with SO2 which exists in free and bound forms, acetaldehyde also exists in free and bound forms. This complicates the analysis of acetalde-

hyde (free and total). Total acetaldehyde can be determined chemically (iodimetry) or enzymatically (aldehyde dehydrogenase), but the chemical method gives results 120% higher than the enzymatic method (Ough & Amerine, 1988). The enzymatic method is considered more accurate and specic, as acetaldehyde is the predominant aldehyde in wine. Free acetaldehyde can be determined by gas chromatography (Ough & Amerine, 1988) or more conveniently by subtracting bound acetaldehyde from total acetaldehyde. Assuming that SO2 present is largely bound to acetaldehyde when the latter is in excess, bound acetaldehyde can be calculated as 0.688 total SO2 (Bakker & Timberlake, 1986).
Effect of acetaldehyde on wine colour and physical stability

Wine contains a variety of phenolic compounds such as anthocyanins, catechins and tannins. While anthocyanins are responsible primarily for the wine colour, the astringency and bitterness of wine can be attributed to catechins, tannins, etc. Further information on the chemistry of phenolics and the relationship between phenolics, wine colour and taste is available elsewhere (Singleton & Esau, 1969; Ribreau-Gayon, 1974; Peynaud, 1984; Singleton, 1988; Somers & Vrette, 1988; Zoecklein et al., 1995; Boulton et al., 1996; Somers, 1998). This section centres on the interactions between acetaldehyde and wine phenolics, and the resultant impact of such interplay on wine colour and physical stability (turbidity). While the colour of new wine is due primarily to its high anthocyanin content, the subsequent colour changes during ageing involves condensation of wine phenolics (Jurd, 1969). Direct condensation between anthocyanins and catechin or tannins without acetaldehyde is very slow. However, rapid polymerization between anthocyanins and catechin or tannins occurs in the presence of acetaldehyde with increased colour intensity and stability, but further reaction with polymerized catechin and tannins leads to instability, precipitation and decreased colour (Somers, 1972; Timberlake & Bridle, 1976; Bakker, 1986). Enhanced colour stability is due presumably to the new compounds formed being partly or wholly resistant to bleaching by SO2

(Bakker & Timberlake, 1997). Although malvidin 3-glucoside (the major anthocyanin in wine) is the only anthocyanin that can slowly condense with acetaldehyde demonstrated in model solutions (Timberlake & Bridle, 1976), other wine anthocyanins would be expected to behave similarly. The condensation of malvidin 3-glucoside and catechin via acetaldehyde forms a dimer consisting of malvidin 3-glucoside linked to catechin by a -CH3CH- bridge (Timberlake & Bridle, 1976, 1977; Baranowski & Nagel, 1983; Bakker et al., 1993). The addition of acetaldehyde to red wine enhances colour intensity, but this is neither legal nor desirable from a avour and health standpoint (Sims & Morris, 1986). However, acetaldehyde plays a key role in the colour increase of port wines, which have higher acetaldehyde levels than normal red wines as a result of fortication with brandy (Singleton & Guymon, 1963; Singleton et al., 1964; Berg & Akiyoshi, 1975; Bakker & Timberlake, 1986). Practices that increase acetaldehyde formation, such as aeration, may promote polymerization of anthocyanins and catechin or tannins, thereby improving colour intensity and stability. Indeed, acetaldehyde arising from a coupled auto-oxidation of ethanol and phenolics provokes co-polymerization of anthocyanins and catechin or tannins (Ribreau-Gayon et al., 1983). From a sensory point of view, this polymerization is desirable since it reduces the tannin level and, hence, alleviates astringency and bitterness (Ribreau-Gayon et al., 1983). Acetaldehyde is also indirectly conducive to wine colour stability by virtue of binding SO2 (see above), a chemical agent known to have a decolourizing effect in wine (Somers, 1975; Timberlake, 1981; RibereauGayon et al., 1983). Furthermore, the anthocyanincatechin and anthocyanintannin polymers formed in the presence of acetaldehyde are resistant to decolourization by SO2 (Zoecklein et al., 1995). The presence of acetaldehyde in red wine can cause turbidity and deposits, presumably due to precipitation of large molecular anthocyanincatechin and anthocyanintannin complexes (Timberlake & Bridle, 1976; Ribreau-Gayon et al., 1983) or formation and precipitation of catechinacetaldehyde colloidal polymers (Saucier

et al., 1997a,b). Acetaldehyde is known to have an impact on haze formation in beer through reacting with catechin and pre-formed complex phenolics (Delcour et al., 1982). Indeed, an appreciable presence of acetaldehyde in the free state (above about 5 mg L1) can destabilize a red wine by inducing a phenolic haze and eventual deposition of condensed pigments, which occurs very rapidly at higher levels of acetaldehyde (Somers, 1998).

Effect of acetaldehyde on wine microorganisms and oenological implications

Effect of acetaldehyde on yeasts and implications for alcoholic fermentation As discussed earlier in this paper, yeasts (predominantly strains of S. cerevisiae) are the primary producers of acetaldehyde in wine. Acetaldehyde is a unique compound in that it is highly reactive and biologically toxic. Acetaldehyde is also very polar and may cause water stress in yeasts (Hallsworth, 1998). It has been suggested that acetaldehyde accumulation (both intracellularly and extracellularly) is one of the central mechanisms of ethanol inhibition of cell growth in yeast ethanol fermentations (Jones, 1989, 1990). Recent evidence shows that acetaldehyde accumulates in fermenting cells of S. cerevisiae to concentrations greatly exceeding extracellular levels (Stanley & Pamment, 1993). However, it remains to be substantiated whether intracellular acetaldehyde accumulation indeed inhibits yeast ethanol fermentations. Nonetheless, inhibition of yeast growth, glucose fermentation and ethanol production by externally added acetaldehyde has been documented. An early study showed that exogenously added acetaldehyde at 1000 mg L1 reduced the fermentation rate of glucose by S. cerevisiae by 30% (Freeman & Donald, 1957). It has been reported that externally added acetaldehyde at 400 mg L1 and above signicantly inhibits cell population, glucose utilization and ethanol productivity, and changes yeast cell morphology (Maiorella et al., 1983). Stanley et al. (1993) found that exogenously added acetaldehyde at 400 mg L1 and above lengthened the lag phase and decreased the exponential specic growth rate of both anaero-

bic and aerobic cultures of S. cerevisiae in a medium lacking ethanol. In contrast to the inhibitory effect, some evidence indicates that low levels of acetaldehyde stimulate yeast growth under certain conditions. Two studies have demonstrated that small amounts of acetaldehyde (up to 580 mg L1) reduce the lag phase and increase the specic growth rate of S. cerevisiae in the presence of 36% (v/v) ethanol under both anaerobic and aerobic conditions, although not in its absence (Walker-Caprioglio & Parks, 1987; Stanley et al., 1993). Therefore, acetaldehyde seems to alleviate ethanol-induced growth inhibition. In addition, low levels of added acetaldehyde (100 mg L1) strongly decrease the lag phase of ethanol- or temperature-stressed S. cerevisiae (Stanley et al., 1997). The reason(s) for acetaldehyde stimulation is not fully understood, but may be ascribable to its role in NAD regeneration and energy generation via glycosis (Stanley et al., 1997). Acetaldehyde inhibition and stimulation of yeast growth have implications for alcoholic fermentations in winemaking. While low levels of acetaldehyde may be stimulatory to yeast growth, high concentrations of this compound (both intracellular and extracellular) may retard or even inhibit yeast ethanol fermentations, resulting in sluggish or stuck alcoholic fermentations sometimes experienced by winemakers. A sluggish alcoholic fermentation is a very delayed or protracted vinication where the rate of sugar utilization is extremely low, whereas a stuck alcoholic fermentation is an incomplete vinication where the vinication ceases before the achievement of a complete alcoholic fermentation so that the residual fermentable sugar level in the wine is too high (greatly exceeding the desirable level of 24 g L1 for dry wine). The end-result of either sluggish or stuck alcoholic fermentations is that the quality of the wine and the economics of wine production are jeopardized. Various factors are implicated in the occurrence of sluggish and stuck alcoholic fermentations. These factors include (a) lack of nutrients such as assimilable nitrogen, minerals and vitamins; (b) lack of oxygen; (c) the presence and production of inhibitory substances such as ethanol, medium fatty acids, sulphites and acetic acid; (d) impact of winemaking practices such as clarication; and (e)

the antagonism of yeasts by LAB (LafonLafourcade & Ribreau, 1984; Ingledew & Kunkee, 1985; Kunkee, 1991; Rasmussen et al., 1995; Huang et al., 1996; Alexandre & Charpentier, 1998). By contrast, the potential role of acetaldehyde in sluggish and stuck alcoholic fermentations has, up to the present, been overlooked. As reviewed above, high levels of acetaldehyde may cause growth inhibition of yeasts and the fermentation rate of glucose, and acetaldehyde can accumulate both intracellularly and extracellularly to sufciently high concentrations. However, it remains to be investigated whether acetaldehyde indeed inhibits yeast growth and ethanol fermentation under winemaking conditions. In view of the above-mentioned inhibitory effect of intracellular and extracellular accumulation of acetaldehyde, therefore, it is proposed here that the high accumulation of acetaldehyde both intracellularly and extracellularly may be one of the contributing factors of sluggish and stuck alcoholic fermentations. Further work to verify this hypothesis is warranted. Effect of acetaldehyde on lactic acid bacteria and implications for malolactic fermentation To date, there has been no denitive study on the effect of acetaldehyde on wine LAB, although acetaldehyde consumption and/or depletion during MLF and wine conservation has been observed (Mayer, 1978; Somers & Wescombe, 1982, 1987; Eggenberger, 1988). Some wine LAB, especially heterofermenters such as L. brevis and Oenococcus oeni, can catabolize the acetaldehyde moiety of acetaldehyde-bound SO2, releasing free SO2 that inhibits LAB growth (Fornachon, 1963; Somers & Wescombe, 1982; Hood, 1983; Delni & Morsiani, 1992, 1993). The biological implication of the utilization by wine LAB of acetaldehyde in free or bound form is not yet understood, apart from the liberation of free SO2 from acetaldehyde-bound SO2. By comparison, the effect of free acetaldehyde on dairy LAB especially Leuconostoc mesenteroides subsp. cremoris is well dened. It has been demonstrated that low concentrations (100 mg L1) of acetaldehyde stimulate growth of heterofermentative LAB such as Leuc. mesenteroides subsp. cremoris (Lindsay

et al., 1965; Collins & Speckman, 1974; Schmitt & Divis, 1990), while high levels (100 mg L1) of acetaldehyde inhibit growth of these LAB (ElGendy et al., 1983). It is thought that acetaldehyde acts as a hydrogen acceptor during heterofermentation with the formation of extra energy (ATP) which stimulates growth (Lindsay et al., 1965; Collins & Speckman, 1974). Further study is required to dene the impact of acetaldehyde on wine LAB, heterofermenters such as O. oeni in particular. The utilization of acetaldehyde by wine LAB during MLF is expected to have an impact on wine avour. As mentioned at the beginning of this article, acetaldehyde is a potent, volatile aroma compound. Excessive levels of acetaldehyde impart an undesirable apple-like, green, grassy avour note in alcoholic beverages including cider, beer and table wines. Increases in acetaldehyde concentrations during vinication are generally considered to be adverse in relation to sensory properties and are suggestive of jamminess, staleness and oxidation (Somers, 1998). Winemakers endeavour to minimize acetaldehyde levels when there is an excess, especially when SO2 is not used before and during fermentation. Besides its antimicrobial and antioxidative roles (see above), in the wine industry SO2 is added to limit the formation of acetaldehyde and to protect the varietal character of wine by binding the acetaldehyde formed; this is believed to be the essential function of added SO2 (Somers & Wescombe, 1982; Somers, 1998). However, the paradox is that SO2, itself, induces acetaldehyde formation by yeasts during fermentation, as mentioned earlier (see Formation of acetaldehyde by yeasts). Furthermore, the use of SO2 is undesirable from the viewpoint of colour (bleaching) and organoleptic properties (off-odour) of wine. There are also legal limits of SO2 addition because of health concerns (allergenic). As an alternative to SO2, MLF can be used to minimize acetaldehyde levels in order to eliminate green/grassy avour caused by acetaldehyde. This entails a better understanding of acetaldehyde metabolism by wine LAB. Removal of acetaldehyde via MLF to ameliorate wine avour may be an additional advantage of MLF besides deacidication of high acid wines and enhancement of avour complexity. In contrast, Leuc. dextran-

icum and Leuc. mesenteroides subsp. cremoris of dairy origin have been applied successfully in dairy fermentations to remove acetaldehyde to improve avour (Lindsay et al., 1965; Keenan et al., 1966a; Keenan & Lindsay, 1966; Keenan, 1968; Prasad & Srinivas, 1987). The metabolism of acetaldehyde and/or its reaction with wine components such as amino acids may also produce avour compounds that can inuence wine avour. This aspect warrants further research. By comparison, aetaldehyde is known to be converted to acetic acid and ethanol by Leuc. mesenteroides subsp. cremoris of dairy origin (Lindsay et al., 1965; Collins & Speckman, 1974; Lees & Jago, 1976a; Liu et al., 1997). Furthermore, the presence of acetaldehyde (100 mg L1) apparently enhances production of diacetyl and acetoin by several dairy LAB, although acetaldehyde per se is not involved (Collins & Speckman, 1974; El-Gendy et al., 1983). However, high concentrations of acetaldehyde (100 mg L1) inhibit acetoin and diacetyl formation, which is commensurate with growth inhibition (Mizuno & Jezeski, 1959; El-Gendy et al., 1983). MLF usually occurs naturally or is induced at the end of alcoholic fermentations by yeasts, although sometimes winemakers simultaneously induce alcoholic fermentation and MLF (Beelman & Kunkee, 1985, 1987). As a consequence, metabolic interactions such as stimulation and inhibition occur between yeasts and LAB, and the specic type of interaction is dependent upon the compatibility of the yeast and LAB (Fornachon, 1968; Beelman et al., 1982; Lafon-Lafourcade & Ribreau-Gayon, 1984; King & Beelman, 1986; Bisson & Kunkee, 1991; Cannon & Pilone, 1993; Huang et al., 1996). Of particular concern to winemakers is the inhibition of LAB by yeasts, which may lead to sluggish or even stuck MLF. A sluggish MLF takes a few months to complete and sometimes may even stick. A number of mechanisms have been put forward for the inhibition of LAB by yeasts; for instance, competition for nutrients or production of ethanol and SO2 (Fornachon, 1968; Beelman et al., 1982; King & Beelman, 1986; Bisson & Kunkee, 1991). As with sluggish and stuck alcoholic fermentations, the role of acetaldehyde in sluggish and stuck MLF has, to date, been over-

looked. As described above, low levels of acetaldehyde stimulate growth of some dairy LAB, whereas high concentrations of this compound inhibit growth of these LAB. Yeasts produce acetaldehyde mainly during growth and the concentration of acetaldehyde decreases thereafter due to re-adsorption by yeasts (RibreauGayon et al., 1956a, 1956b) and/or reaction with other substances. Wine LAB may well be inhibited by the acetaldehyde produced by yeasts during simultaneous alcoholic fermentation and MLF. High residual levels of acetaldehyde after alcoholic fermentation may also inhibit growth of wine LAB if MLF is induced at the end of yeast fermentation when the residual acetaldehyde level is high. The concentration of acetaldehyde is critical and determines whether stimulation or inhibition occurs.
Conclusion

Acetaldehyde in wine is formed by yeasts, acetic acid bacteria, and coupled auto-oxidation of ethanol and phenolic compounds. The binding of SO2 by acetaldehyde reduces the effectiveness of its antimicrobial activity and its antioxidative effect. The interaction of acetaldehyde with phenolics improves wine colour but may cause turbidity. Microbial growth can be stimulated or inhibited by acetaldehyde, depending on its concentration, and this has implications for alcoholic and malolactic fermentations. Bacterial catabolism of acetaldehyde may generate avour components that can inuence sensory properties of wine.
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