Malic Acid Fermentation
Malic Acid Fermentation
Malic Acid Fermentation
Malolactic Fermentation
©Copyright Ben Rotter 2002-2009
www.brsquared.org/wine
1 Introduction
Malolactic fermentation (MLF, or "malo") is an important
winemaking process conducted on most red grape wines and some
white grape wines. It is also used with some fruit wines. The
following article gives information concerning the conditions
necessary for MLF, its affects, prevention, progress, suitable wine
type candidates for it and yeast compatibility.
(Cell shape and grouping may depend on the medium in which the
bacteria grow.)
3 Effects of MLF
3.1 General
Bacteria also use only 0.3-2 g/l of sugars yielding about 100 mg/l of
D-lactic acid [Krieger et al., 2000].
3.2 Acidity
MLF affects TA, pH and acid species through the following factors:
3.7 Astringency
3.10 Diacetyl
citric acid metabolisation begins at the same time that malic acid
degradation occurs, but that the degradation of the citric acid is
slower
the higher the initial concentration of citric acid, the more
diacetyl will be produced from MLF
diacetyl concentration increases as citric acid is metabolised by
MLB and decreases again when most of the citric acid has been
consumed
maximum diacetyl concentration tends to coincide with the
exhaustion of malic acid during MLF [Nielsen, 1995]
semi-aerobic (2-4 mg/l oxygen) MLF yielded higher
concentrations of diacetyl, whereas anaerobic (<0.2 mg/l oxygen)
conditions yielded much lower concentrations [Nielsen, 1999]
the higher the pH, the lower the diacetyl production [see
Wibowo et al., 1985]
The diagram shows the main metabolic pathway for citric acid
metabolisation by Leuconostoc oeni.
3.10.4 The breakdown or binding of diacetyl
4 Living conditions
The limiting factors on MLF include total and free sulphur dioxide
(SO2) concentration, alcohol content, pH, and temperature. Each
LAB has it's own limits (and culture manufacturers usually provide
these). The following limitations provide general guidelines (these
are based on Oenococcus oeni unless where otherwise stated).
MLB favour higher pH's. The optimum pH will depend on the strain
and the culturing environment. For most strains, minimal growth
occurs at pH 3.0. Generally, pH's above 3.2 are advised. Optimal
growth for O. oeni occurs at pH 4.2-4.8. Optimal activity occurs
between pH 3.0 and 4.0 and decreases as pH rises - inhibition is
experienced at pH 4.5.
4.4 Temperature
Delay Temperature
slowed by months / essentially no malic acid
10°C (50°F)
decarboxylation
12-13°C (54-
slowed by weeks
55°F)
begins to slow 15°C (59°F)
20-25°C (68-
optimum
77°F)
MLB death >30°C (> 86°F)
4.4 Population
5 LAB development
MLB require a certain population level to be reached before they can
begin MLF. This means that natural/indigenous MLFs (where LAB
have not been inoculated into the must or wine) can have a lag
phase of weeks to several months before they begin MLF.
This lag phase is prolonged the lower the pH. Additionally, the pH
affects which species of LAB will be dominant in the must or wine
[Bousbouras and Kunkee, 1971]. At low pHs Oenococcus oeni is the
primary MLB genus, different strains of which will dominate
throughout MLF. At higher pHs, however, Lactobacillus and
Pediococcus dominate over Leuconostoc [Costello et al., 1983].
During MLF, the population often reaches 1 million cells/ml.
(Leuconostoc oeni in particular requires a population of more than
106-107 CFU/ml to begin MLF.) Their population is usually 10 3-104
CFU/ml following fermentation, and may rise to 10 6-108 CFU/ml
once growth initiates substantially.
After exponential growth, cell populations deline rapidly depending
on the conditions (e.g. elevated tempertures or SO2 will increase
the rate). It is possible that the population of strains of
Lactobacillus and Pediococcus may increase at this point.
6 Why inoculate
The MLF growth (lag) phase associated with spontaneous MLF
(wild/uncultured strains) presents a time of increased risk from
spoilage organisms due to the non-SO2 environment and the
potential production of volatile acidity. Inoculating with a prepared
MLB culture avoids the problems associated with the MLB growth
lag phase by immediately providing the population necessary to
conduct MLF. It is important that the MLB have enough nutrients to
develop. Yeast (Saccharomyces cerevisiae var. "bayanus" in
particular) can reduce the nutrients available to MLB considerably.
Winemakers often add a MLB nutrient when inoculating with MLB to
assist their development.
7 Mixed cultures
Some believe there are advantages in conducting MLF with mixed
MLB cultures.
8.1 Sugars
The chart can then be viewed and the presence of each individual
acid assessed. The background appears blue, with yellowish spots
appearing up the paper. Each acid can only travel a certain distance
up the paper. Tartaric, citric, malic, lactic, and succinic acids are
represented in order at heights progressively up the paper.
13 Preventing MLF
Factors to help in preventing MLF occurring include the following:
(1) The batch is split, one portion goes through full MLF and the
other no MLF. The two portions are then blended back together. The
problem to then overcome is a renewed MLF in the wine, since there
is still malic acid present. The best method in providing stability is
to sterile filter the wine. Alternatively, a thorough clearing and
filtration of the wine coupled with a 0.8 mg/l molecular SO2 level
can be successful in preventing renewed MLF.
(2) The wine undergoes MLF and is closely monitored. When the
winemaker is satisfied with the level of MLF (based on taste and
analytical methods), the MLF is arrested. MLF is terminated by
sterile filtration (<0.45 micron tight filter to remove yeast and lactic
bacteria), a 1.5-2 mg/l molecular SO2 level, keeping wine lees
contact to a minimum and racking when MLF termination is desired
to reduce the MLB population, and possibly the use of lysosyme and
cold temperatures/stabilisation.
Most reds undergo MLF. One of the main reasons for this is to
ensure that the wine will not undergo MLF later in bottle; leading to
lowered acidity, off aromas, and carbon dioxide.
MLF has traditionally been avoided with regard to fruit, flower and
vegetable wines. This is largely because an emphasis is place on
primary ingredient ("fruit" or varietal) character. However, many
non-grape wines may be suitable for MLF. Some winemakers
routinely MLF apple wine, for instance, and reds such as blackberry
are suitable candidates.
Overall, MLF is a stylistic option that will suit some wines and not
others. It is up to the winemaker to assess whether the individual
wine in question will benefit or not from MLF. The above information
hopefully provides clues as to how to discern this.
16 References
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ethanol effects on growth of Leuconostoc oenos. In: Yu, P.-L. (ed.)
Fermentation technologies: Industrial applications, Elsevier, London,
pp 128-131.
Axelsson, L.T. (1993). Lactic acid bacteria: classification and
physiology. In: Salminen S. and von Wright A. (eds.), Lactic acid
bacteria, Marcel Dekker Inc., New York, pp 1-63.
Bartowsky, E.J., Francis, I.L., Bellon, J.R., and Henschke, P.A.
(2002). Is buttery aroma perception in wines predictable from the
diacetyl concentration?, AGJWR, Volume 8, Number 3.
Beelman, R.B., and Kunkee, R.E. (1985). Inducing simultaneous
malolactic-alcoholic fermentation in red table wines. In "Malolactic
Fermentation" (T.H. Lee, ed.), pp. 97-111. Aust. Wine Res. Inst.,
Urrbrae, South Australia.
Bousbouras, G.E., and Kunkee, R.E. (1971). Effect of pH on
malolactic fermentation in wine. Am. J. Enol. Vitic. 22, 121-126.
Buckenh 黶 kes, H.J. (1993). Selection criteria for lactic acid
bacteria to be used as start cultures for various food comodities.
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Costello, P.J., Morrison, R.H., Lee, R.H., and Fleet, G.H.
(1983). Numbers and species of lactic acid bacteria in wines during
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Davis, C.R., Wibowo, D., Eschenbruch, R., Lee, T.H., and
Fleet, G.H. (1985). Practical implications of malolactic
fermentation in wine. J. Appl. Bacteriol. 63, 513-521.
Davis, C.R., Wibowo, D.J., Lee, T.H. and Fleet, G.H. (1986).
Growth and metabolism of lactic acid bacteria during and after
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Dharmadhikari, Murli. (2002). Some Issues in Malolactic
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Dittrich, H.H. (1977). Handbuch der Getr 鋘 ketechnologie:
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manual of systematic bacteriology vol 2, Williams and Wilkins,
Baltimore, pp 1071-1075.
Henick-Kling, T. (1993). Malolactic fermentation. In: Fleet, G.H.
(eds.) Wine microbiology and biotechnology, Harwood Academic
Publishers, Chur, pp 289-326.
Henick-Kling, T. (1995). Control of malo-lactic fermentation in
wine: Energetics, flavour modification and methods of starter
culture preparation. J. Appl. Bacteriol. Symp. Suppl. 79, 29S-37S.
Henick-Kling, T., Sandine, W.E. and Heatherbell, D.A. (1989).
Evaluation of malolactic bacteria isolated from Oregon wines. Appl.
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Jackson, R.S. (1994). "Wine science: principles and applications,"
San Diego Academic Press.
Krieger, S., Triolo, G., and Dulau, L. (2000). "Bacteria and Wine
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Liu, S.-Q. (2002). Malolactic fermentation in wine - beyond
deacidification: A review. J. Apll/ Microbiol. 92, 859-601.
Lonvaud-Funel, A. (1995). Microbiology of the malolactic
fermentation: molecular aspects. FEMS Microbiol. Lett. 126, 209-
214.
Lonvaud-Funel, A., Joyeux, A., Desens, C. (1988). Inhibition of
malolactic fermentation of wines by products of yeast metabolism.
Journal of the Science of Food and Agriculture, 44, 183-191.
Martineau, B. and Henick-Kling, T. (1995). Performance and
diacetyl production of commercial strains of malolactic bacteria in
wine. J. Appl. Bacteriol. 78, 526-536.
Martineau, B., Acree, T.E. and Henick-Kling, T. (1995). Effect
of wine type on threshold for diacetyl. Food Research International
28(2).
McDaniel, M., Henderson, L.A., Watson, B.T., Jr., and
Heatherbell, D. (1987). Sensory panel training and screening for
descriptive analysis of the aroma of Pinot noir wine fermented by
several strains of malolactic bacteria. J. Sens. Stud. 2, 149-167.
Nielsen, J.C. and Prahl, C. (1995). Metabolism of citric acid by
Leuconostoc oenos in direct inoculation. Effect on wine flavour.
Results presented at the 5th International Symposium of Enology,
Bordeuax, 15-17th June 1995.
Nielsen, J. C., and Marianne Richelieu. (1999). Control of flavor
development in wine during and after malolactic fermentation by
Oenococcus oeni. Applied and Environmental Microbiology 65(2):
740-745.
Radler, F., and Yannissis, C. (1972). Weins 鋟 reabbau bei Milchs
鋟 rebakterien. Arch. Mikrobiol. 82, 219-239.
Ramos, A. and Santos, H. (1996). Citrate and sugar
cofermentation in Leuconostoc oenos, a 13C nuclear magnetic
resonance study. Appl. Environm. M
Riesen, R. (1999). Modification of Wine Characteristics by
Malolactic Fermentation. http://www.oardc.ohio-
state.edu/grapeweb/vinevan/van0699.htm.
Rib 閞 eau-Gayon, J., Peynaud, E., Rib 閞 eau-Gayon, P., and
Sudraud, P. (eds.) (1975). "Trait?d'Oenologie: Sciences et
Techniques du Vin," Vol. 2. Dunod, Paris.
Sandine, W.E. and Heatherbell, D.A. (1985). Wine preparation
with new strains of Leuconostoc oenos. US Pat. 4,547,373,
application number 521,988.
Salou, P., Loubiere, P. and Pareilleux, A. (1994). Growth and
energetics of Leuconostoc oenos during cometabolism of glucose
with citrate and fructose. Appl. Environ. Microbiol. 60, 1459-1466.
Semon, J.M., Edwards, C.G., Forsyth, D., and Dinn, C. (2001).
Inducing malolactic fermentation in Chardonnay musts and wines
using different strains of Oenococcus oeni. AGJWR, Vol. 7, No. 1, p
52-59.
Sharpe, M.E. (1981). The genus Lactobacillus. In: Starr, M.P.,
Stolp, H., Tr 黳 er, H.G., Balows, A. and Schegel, H.G. (eds.).
Prokaryotes, vol. 2, Springer-Verlag. Berlin, pp 1653-1679.
Shimazu, Y., Uehara, M., and Watanbe, M. (1985).
Transformation of citric acid to acetic acid, acetoin and diacetyl by
wine making lactic acid bacteria. Agric. Biol. Chem. 49, 2147-2157.
Subramanian, S. and SivaRaman, C. (1984). Deacidification of
musts from the western United States by the calcium double-salt
precipitation process. Am. J. Enol. Vitic. 29, 153-160.
van Vuuren, H.J.J. and Dicks, L.M.T. (1993). Leuconostoc
oenos: A review. Am. J. Enol. Vitic. 44, 99-112.
Wibowo, D., Eschenbruch, R., Davis, C.R., Fleet, G.H., and
Lee, T.H. (1985). Occurrence and growth of lactic acid bacteria in
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www.brsquared.org/wine
Enology Notes
For example, with mature Cabernet Sauvignon, about 8.5% of the total are in
the seeds, 15% in the skins and 77% in the pulp. It would seem that the
separation of the pulp juice from the skins as occurs with bleeding would not
have a large quantitative effect on juice that is removed. However, there is a
significant qualitative influence.
The two amino acids present in the greatest concentration in the fruit are
proline and arginine. Proline cannot be used by the yeast, while arginine can.
Indeed, because it has four atoms of N per molecule (three of which are
believed used by yeasts), arginine is a very good source of fermentable N.
The ratio of these two amino acids is impacted by maturity. Arginine
accumulation in the fruit begins before veraison, continues to increase,
plateaus, and declines. Proline, on the other hand, increases late in the
season and continues to increase with increased hang-time.
High levels of proline are, therefore, associated with increased maturity. High
proline is also associated with plant stress. The reduction in the arginine to
proline ratio can be significant and can negatively impact the amount of
fermentable nitrogen available to the yeast.
The ratio of arginine to proline is much greater in the skins than the pulp. In
other words, pulp juice taken off in bleeding has a relatively high
concentration of proline (approximately 55%) which cannot be used by the
yeast, and a small concentration of the more potent amino acid arginine and
others needed to carry out a healthy fermentation. The lower incidence of
incomplete fermentation in red compared with white musts supports the
concept that the slow release of nitrogen from grape skins during fermentation
is important.
Also as discussed at the volatile sulfur compound workshop, all wines should
undergo sulfite screening prior to bottling. See Enology Notes and Zoecklein
et al. (1999).
Lisa and Patricia will discuss the latest practical information on conducting
healthy fermentations.
I will present our Traminette, Viognier and Cabernet Franc wines produced
from our training system trials: VSP, GDC and Smart-Dyson (Up vs. Down)
from 2003 and 2004.
Cost: $25 per person. The complete registration fee is due NO LATER THAN
August 18, 2005. Checks are to be written payable to Virginia Tech
Foundation and mailed to Terry Rakestraw, Department of Food Science and
Technology (0418), Virginia Tech, Blacksburg, VA 24061. Course fee is non-
refundable.
The earlier the estimation of average berry weights, the more time the
winemaker has to evaluate the crop load, make adjustments, and plan for the
season. There is a relationship between berry weight at veraison and berry
weight at maturity.
Changes in berry weight confound the measurement of degrees Brix, and are
yet another reason why Brix must not be the sole monitor of fruit maturation.
Research indicates that the maximum rate of production of aroma/flavor
compounds in the fruit occurs at about the time when the berry stops
importing water from the phloem or shortly thereafter. Therefore, maximum
aroma/flavor occurs sometime after the berry reaches maximum weight in
most instances. This is the reason why Syrah producers, for example, monitor
the extent of berry shriveling.
To be added to (or removed from) the Enology Notes listserve send an email
message to [email protected] with the word "ADD" or "REMOVE" in the
subject line.
O2 comes into contact with must as just after crushing of the grapes,
which catalyses the activation of oxidation enzymes. These enzymes
oxidize phenolic molecules to their corresponding quinone, as Facts, figures,
discussed in the previous article. When juice or wine is saturated with contact details and
oxygen it contains about 7-8 mg/L of O2, depending on the much more in the
temperature. There are several ways available to the winemaker to 2009/10 Directory
avoid unwanted oxidation. The first entails removing the substrate for
oxidation, the phenolic molecules. Hyper oxidation has thus been
investigated as a means of removing phenolic molecules prior to
fermentation by oxidation, where large amounts of O2 are added to
the must. This can be done by sparging the juice in the tank for about
30 min with O2, racking the juice from one tank to another for a few
times or using O2 in stead of N2 when using flotation to clarify the
juice. According to literate white juice that did not receive skin contact
can be saturated once with O2, which should be sufficient to remove
most phenolic compounds. When skin contact has been given the
juice can be left for 30 min after saturating it and then be sparged
again for 30 min. It is obvious that no SO2 should be added prior to
these operations. If SO2 is added before fermentation it should also be
added after the juice has been racked from the precipitate after
saturation. After fermentation SO2 can also be added.
Phenolic molecules can also be removed from the must and wine by
fining with gelatin, PVPP or activated charcoal. The latter should
however be handled with care, as it can strip a wine from its flavour
compounds if added at too high dosages. Higher phenolics being
extracted into the juice and wine can also be avoided by not applying
too long skin contact periods, handling the grapes with care (thus not
pressing at too high pressures and using small amounts of press
must).
Fig.1: O2 pickup in sampling bottles due to different sampling methods.
taken with chain with container middle of tank. Sapl tap: samples taken
from sampling tap from the tank. Port: samples taken from the port and
transferred into sampling bottles. All data were repeatable within 10%.
When samples are taken from a tank for analysis of SO2, O2 etc. it is
important to realize that wine can quickly take up O2 in the sampling
procedure, which may give a wrong impression of the oxidative state
of the wine. In Fig 1. one can clearly see the effect of this. A 2005
Sauvignon blanc wine at the US Welgevalen cellar was used to test
this. The O2 concentration was measured in the tank and was 0.48
mg/L. Wine was then taken from the top by means of a metal
container attached to a chain and added to 250 mL sampling bottles
in triplicate. In three of the bottles CO2 gas was blown prior to adding
the wine and in another three not. The same procedure was also done
by taking samples from the sampling tap, as well as from the bottom
port of the tank and then adding it in the bottles. The lids of the
bottles were then sealed immediately and the dissolved O2 (DO)
measured after a few seconds. It is clear from Fig 1 that where CO2
was added in the bottle little O2 came into contact with the wine,
especially in the wine that was taken from the sampling tap. The wine
picked up almost 1 mg/L O2 from the sampling tap where no CO2 was
added prior to the bottles. Wine that was first collected from the port
into an open container and then transferred into the bottles pickup up
very large amounts of O2. This should be kept in mind when collecting
wine samples for analyses, or when a winemaker wants to keep a
control sample in a bottle when micro oxygenation will be used in a
tank. It is thus possible to add more O2 to the control sample than the
treated wine if done incorrectly!
Literature cited
Let's assume:
It is mid-September in the Okanagan and you have a well-tended field of grapes getting
close to harvest. You have done all you can to ensure moderate cropping, good canopy
management, and enough, but not too much, water—all factors that affect early and
even ripening.
There are no practical problems to get in the way of selecting the best picking date. For
example:
o The weather is good (days are getting cooler but no rain is coming).
o Pickers are available.
o Shipping or storage is ready.
You know the style of wine you want to make.
Now all you have to do is to decide when the grapes are at their
optimum. This may not be an ideal optimum, just the best you can
expect in this year in this field for this variety of grape and for
the style of wine you want to make.
1. Appearance.
2. Numbers (sugar and acid levels).
3. Taste.
Unless you are lucky (and in the Okanagan you are not likely to be
lucky very often), you will discover that appearance, numbers, and
taste do not point to the same picking time. You have to make some
compromises. In the recent past, the compromises were usually biased
in favor of sugar and acid numbers. Today, top winemakers are more
likely to prefer taste and almost ignore numbers. But any bias or
compromise will affect how you make your wine and the eventual
quality of this wine.
Picking on Appearance
1. Flowering
o Early flowering is preferred as long as the threat of cold is past—best vintages
are often associated with an early start and an early harvest.
o Weather can interfere with complete and even pollination.
2. Green berry growth
o Berries use their own chlorophyll to grow and accumulate acid and sugar.
o Canes and roots also grow rapidly.
3. Arrest of green berry growth
o Berries have reached more than half their final size, but are still green and
hard.
o Berries have all the basic acid they will ever have, but only a small proportion
of the sugar.
o Roots and shoots also stop growing. This is usually the recommended time for
pruning of vines for canopy management and for thinning of clusters.
4. Beginning of véraison
o Berries soften and the color changes (véraison means turning, ripening). The
color change happens dramatically in individual berries (in 24 hours), but
unevenly among and within clusters. Even under ideal conditions it will take
two weeks for a vineyard to turn completely. The date of véraison is based on
an estimate of when half the grapes have turned.
o Véraison comes earlier with a low leaf-to-fruit ratio and with warm, sunny, dry
weather—best vineyards tend to start véraison early
5. Sugar/water accumulation
o Berries gain size and weight, adding somewhere between 25% (a dry season)
and 75% (wet season) of their green berry weight. A 40-50% increase is
normal for good quality grapes.
o Sucrose is manufactured by the leaves and transported to the berries where it
is hydrolyzed and stored as glucose and fructose.
o Acidity declines because of grape growth and the dilution of the existing store
of acid. Malic acid also declines because it is used in grape respiration.
o Flavors begin to accumulate, although most varietal flavor development comes
later in the season.
o Grapes typically do best at about 25°C—temperatures below 15°C and above
35°C will stop the plant from working. Warm (not hot), sunny, dry ripening
seasons tend to produce the best vintages.
o Berries ripen unevenly among and within clusters. Within clusters, berries at
the top (next to the stem) are riper (more sugar, less acid) than those at the
tip. Larger berries have more seeds, and these ripen more slowly than small
berries of the same variety.
6. Arrest of phloem transport
o Plants "shut down" and berries stop accumulating water and sugar—this
happens naturally at the "end" of the season but can happen earlier from
stress.
o Berries soften noticeably and are easily "deformed" with a squeeze. It is easier
to separate seeds and pulp and to detach grapes from their stems
o Stems go brown (in many varieties) and seeds go from green to brown.
o Flavors continue to develop, because most flavors are synthesized in the
grape.
7. Dehydration
o Berries can lose 10% or more of their weight through water loss over a period
of a couple of weeks.
o Sugar levels rise because of water loss. The rise in sugar can suggest that the
plants are still working when they aren't.
o Acidity is reduced. Tartaric acid level can rise a bit because of dehydration, but
malic acid continues to be lost (grapes can lose almost all their remaining
malic acid during this period). Potassium activity in the grapes can continue to
reduce pH even though TA may not decline much.
o Flavor synthesis comes to an end and and flavor deterioration begins (these
periods overlap in ways not well understood).
8. Raisining
o Berry flavors continue to deteriorate.
In Napa and Sonoma you can drop off a sample of 100 berries at the
ETS lab in St. Helena before 2:00 pm, pay $110 (US), and by early
evening have the results for the following tests:
°Brix (% by weight of soluble solids, mainly fermentable sugar). During the time they are
testing, ETS says this commonly varies between 19 and 30, which means people start
testing only well along in the growing season. For another $60 you can get a precise
measure of fermentable sugar.
Various acid measurements:
o pH—commonly varies between 2.90 and 4.20
o Titratable Acid or TA (g/100ml)—commonly varies between 0.35 and 1.20
o Tartaric Acid (g/L)—commonly varies between 1 and 11
o Malic Acid (g/L)—commonly varies between 0.5 and 11
o Potassium (mg/L)—commonly varies between 500 and 4000
Two yeast available nitrogen (YAN) measures:
o Ammonia (mg/L)—commonly varies between 50 and 400
o Alpha Amino Nitrogen (mg/L)—commonly varies between 20 and 400
The YAN numbers are chiefly useful to indicate how much nutrient you
are going to have to add to the fermentation. However, the sugar and
acid numbers can serve as a guide to picking the grapes.
As grapes ripen, sugar levels rise and acid levels fall. Both sugar
and acid are important. Higher sugar levels are associated with grape
maturity and the higher levels of alcohol expected of superior
vintages. Higher acid levels (especially a low pH) provide better
wine-making circumstances. Thus, the trick in picking by the numbers
is to get to the optimum cross-over point, where sugars are high
enough and acid numbers still allow for good wine-making. The current
wisdom suggests you should be picking your grapes when the following
specs are realized.
The problem with the numbers in the above tables is that we seldom
see them in the grapes we get. Our 2003 Black Sage grapes illustrate
the issue. The following table is based on typical (median) readings
by Club members.
These parameters for 2003 Black Sage grapes suggest at least three
observations:
Only Chardonnay, Gewürztraminer, and Riesling meet most of the suggested specs, but
even Chardonnay and Gewürztraminer have relatively high pH levels and this puts then
at the high end of the acceptable index range, where reds are more to be expected.
Semillon also comes close to meeting the suggested specs. Most people would consider
pH more important than TA, and the Index value for Semillon seems to capture the
quality of the Semillon better than the simple Brix/TA Ratio does.
The general "numbers" problem with these Black Sage grapes is low acidity—a low TA
and a high pH. Only the Cab Sauv and perhaps the Syrah are clearly overripe in terms of
sugar. The phenomenon of low acidity without excessive sugar is a well-known
consequence of overripe grapes or a warm climate because of the respiration of malic
and other organic acids. This past summer in the Okanagan, high heat caused vines to
"shut down," stopping sugar production but increasing acid loss.
The Index and Ratio numbers are not that bad for Pinot Gris and Sauvignon Blanc, but
these were the white grapes that arrived in poor condition with obvious VA problems.
Appearance does matter!
Pro: Great, long-lived wines almost always come from grapes with
"good numbers" to start with. pH is particularly important for good
winemaking, and it is something that is very difficult to correct
even by adding acid later. No one wants to add water to an overly
sweet must and thereby dilute the intensity and flavor of the wine.
Con: The right numbers may occur fairly regularly in classic wine
areas like Bordeaux, but you will not get them in hotter, drier wine
areas like the Okanagan (or South Africa or Australia). In such
conditions, sugar development usually outpaces flavor development.
Picking on Taste
Taste and its development in grapes and wines is not well understood
scientifically, but there are some basic things we do know.
There are hundreds of different molecules that contribute to the flavor and aroma of
grapes and wine. Some of these have a a sensory impact at concentrations of only a few
parts per billion.
Flavors develop over the course of grape ripening, but much of the typical varietal flavor
and intensity comes very late. In one Oregon test (Watson), Pinot Noir was made from
the same grapes, but one picking was one to two weeks later than the other. Later,
when the finished wine was tested, more than twice as many flavor compounds were
isolated from the grapes picked later—a rather astounding differences. Of course, more
flavor elements do not necessarily mean a better flavor, but in this case the later picking
was definitely judged to be better in overall flavor and aroma.
Most flavor compounds are synthesized in the grape itself and this synthesis can
continue after grapes are biologically ripe and phloem flow has stopped.
Identifiable tastes change as grapes develop. The exact tastes differ according to variety,
but the usual sequence is more vegetative tastes early and more fruit tastes later. The
sequence for the appearance of flavors in Cabernet Sauvignon goes something like this:
1. Vegetation (plant matter)
2. Herbaceousness (straw, herb, vegetal, tobacco)
3. Unripe fruit (green apple, citrus rind)
4. Red fruit (cherry, strawberry, raspberry, cranberry)
5. Black fruit (plum, blackberry, black cherry)
6. Jam (prune, date, raisin)
This is not necessarily a sequence of tastes. Tastes deteriorate at different rates than new
ones appear. It is possible to have herbaceous and jammy tastes at the same time. This
leads Bisson (2001) to suggest that it may be better to taste for the absence of the
"unripe" tastes you want to avoid rather than to taste for mature complexity.
Many of the flavors that are most desired in a mature wine are not taste-able in the
grape. They are locked-up with glucose in complex molecules that are technically known
as glycosides and more popularly known as flavor precursors. Some of these flavor
precursors can be unlocked with pectic enzyme, but some emerge only with
fermentation or aging. What can be tasted most easily in the grape are the fruit flavors
associated with a young wine.
o NOTE: There is a test called the Glycosyl-Glucose Assay (or G-G Assay) that is
used to establish the total level of glycosides in juice or wine. At one point in the
1990s there was hope that this test would provide a more objective basis for
establishing the flavor potential of grapes. It has not so far proved to be a magic
bullet.
Sources
ETS Laboratories (2001). "Tools for Grape and Must Analysis." Useful
explanation of grape testing terminology. the 2003 version of this
paper is available at: http://www.etslabs.com/pagetemplate/blank.asp?
pageid=195. There are other useful technical papers available through
the ETS website.
Master of Faults
Sam Harrop, a co-chair at the
International Wine Challenge in
London, spends half of his time
at the faults table. A New
Zealander by birth, a London-
based international winemaking
consultant by trade, Harrop
loves complex sulfides as much as minerality...
which, for him, are often one in the same.
For the ten years before Harrop started with
the Challenge, the organizers had been tracking
corked wines, flagged by a range of different
judges. In those years, Harrop says, wines
presumed to have cork taint ranged between six
and seven percent of the total. Four years ago,
when Harrop joined the competition, he saw a
range of faults beyond cork taint coming through
from the panels. "On the hoof, I created a little
program on the computer and started logging
them," he recalls. Now he tracks faults through
14,000 to 15,000 bottles over the course of the
annual two weeks of judging.
"I found in my first year that the figure for cork
taint was closer to three percent. Total faults
were around seven percent–a similar figure to
previous years; so the balance must be other
types of faults. The cork industry has been going
on for decades that seven percent is a load of
rubbish. And though I'm no apologist for the cork
producers, sure enough, tasters were flagging
cork taint when it was actually Brettanomyces."
Harrop's interest in faults began when he was
winemaker at Marks & Spencer, the up-market
British retail chain. "I didn't actually make wine,"
he says. "But when you're a retailer selling five
million cases a year of own-label wine, you own
the product and have to make sure the faults are
minimized.
"I started studying for the MW in 2001 and
realized that faults can actually be the making of
a wine," he recalls. Soon, for his MW dissertation,
he began to research Brettanomyces and its role
in high-quality syrah. "I'd been told that Brett was
the pox–its nickname in Australasia, where it's
treated as some kind of plague. Here I was in the
Rhône hearing winemakers wax lyrical about
wines I was taught to hate. I collected syrahs from
some of the most well-known producers around
the world, tasted them with MWs and asked them
to rate and rank them, and whether they
perceived Brett. Then I analyzed the wines for
volatile phenols. It was clear that just a little bit–
just over threshold–is fantastic. In the highest-
scoring wines, it brings out the varietal character
and complexity. Too little and the wine is simple.
A little Brett is good; too little is less good. On
the other end, the worst wines in the ranking
were full of volatile phenols."
Harrop believes the New World winemakers who
insist that any Brett is bad Brett are just as
misguided as those who let it run rampant. "It's
easily controlled if you monitor it," he says, "and
easily stabilized through filtration. I don't know
who spread the word that unfiltered wines were
better. I've done trials and the filtered wines
come out best, both in texture and expression."
Harrop surveys the damage from the Apple iMac
surrounded by white-sheathed bottles at his
faults table. "A lot of people think I'm some kind
of masochist. But I have the last laugh." For
Harrop, faults are the way not only to
understanding great wine, but to making great
wine. At below, he describes the marker for each
of the major faults, and some of the trends he has
found over time.
Lactic acid bacteria (LAB) are responsible for many fermented foods such as sauerkraut, pickles and
yogurt. They have also been isolated from wines at various states of vinification. In wines they are
responsible for malolactic fermentation (MLF) which can be beneficial in some cases and undesirable in
others. Besides conducting MLF, these bacteria under certain conditions can also cause undesirable
changes in wine flavor which renders the wine undrinkable. Many species of LAB do not conduct MLF
and their growth in wine can cause some serious wine spoilage.
These organisms are gram positive, catalase negative, nonsporing cocci, coccobacilli or rods. They are
microaerophilic that means they grow well under conditions of low oxygen content. Since they can grow
under low oxygen conditions, they can grow throughout the wine (as opposed to on the surface of the
wine) even though the container is kept full. The bacteria can metabolize sugars, acids and other
constituents in wine and produce several compounds. Some of these are undesirable and constitute
spoilage.
After MLF the fate of lactic acid bacteria depends on wine composition and how the wine is handled. If
the wine pH is high (>3.5), and the SO 2 level is inadequate, then the spoilage causing species of LAB
can grow and spoil the wine. For this reason special attention should paid to the wines during storage
after MLF.
1. Fermentation of Sugars
LAB, including those involved in MLF, metabolize sugars such as glucose and fructose, and produce
lactic acid and acetic acid. The resulting wine acquires a sour vinegar-like aroma due to high VA levels.
This is a serious spoilage and occurs in must with stuck fermentation or wines with higher residual
sugars (sweet wines).
A less serious form of lactic spoilage can occur in dry wines. In these wines the LAB utilizes pentose
sugars, trace amounts of glucose and fructose, and produces lactic and acetic acid as a by-product.
When sugars are attacked by LAB, lactic and acetic acids are produced. Formation of these acids
increases the titratable acidity and lowers the pH. The decrease in pH restricts the growth of those
organisms.
2. Degradation of Glycerol
Breakdown of glycerol by LAB results in the formation of lactic acid, acetic acid and acrolein. The wine
smells acetic, butyric and acquires a bitter taste due to acrolein.
The mousy aroma has been attributed to the formation of a compound called acetyltetrahydropyridine.
Two species of lactobacillus have been shown to produce these mousy odor compounds.
Sometimes a wine can develop a geranium-like odor which makes it undrinkable. This odor is caused by
a compound known as 2-ethoxyhexa-3, 5-diene. This compound is produced from the decomposition of
sorbic acid by the LAB. In sweet wines sorbic acid is often added to prevent the growth of unwanted
yeast (yeast growth can cause refermentation). When the sorbic acid is attacked by LAB, 2-ethoxyhexa-
3,5-diene is formed which imparts the geranium-like odor to the wine. To prevent this odor the growth of
LAB in sweet wines containing sorbic acid should be controlled.
Must/Wine Composition
Wine pH - Wine pH is one of the most important factors influencing the growth of LAB. It affects the
initiation and duration of malolactic fermentation MLF, it influences the type of species of bacteria that
may develop in wine and it also affects the metabolic behavior of the organism and thereby determines
the kind of by-products formed as a result of bacterial activity.
In the wine pH range of 3.0 to 4.0, the time needed for the completion of MLF decreases with an
increase in pH. Bousbouras and Kunkee (1971) reported that at pH 3.15 it took 23.4 weeks to complete
MLF; whereas at pH 3.83, it was completed in just two weeks.
Many researchers have noted the effect of pH on the species of bacteria that can grow in wine.
Generally at pH below 3.5, the MLF is often dominated by Leuconostoc, whereas; above pH 3.5,
species of Pediococcus and Lactobacillus seem to flourish. It should be noted here that many strains of
Lactobacillus are involved in wine spoilage.
Another important pH effect not commonly realized is the effect of pH on the metabolic behavior of the
organisms. For example at pH 3.5 and above, LAB are more likely to decompose sugars, tartaric acid
and citric acid. As mentioned earlier, fermentation of sugar leads to higher volatile acidity (VA) levels in
wine. From the foregoing discussion it should be obvious that controlling wine pH is one of the keys to
controlling wine spoilage by LAB.
Sulfur dioxide (S02) Sulfur dioxide is an effective germicide commonly used by the winemakers to
control the growth of harmful bacteria. The SO2 in wine exists in free and bound forms. All these forms
remain in an equilibrium which is influenced by pH. Concentration of the molecular SO2 form of free SO2
which is also the most toxic form increases with a decrease in wine pH. Therefore, maintaining low pH is
helpful in making SO2 the most effective tool to control LAB. The bound form of SO 2 has also been
reported to have a detrimental effect on LAB. In wine, SO 2 is bound to certain carbonyl compounds such
as acetaldehyde. When LAB attacks the carbonyl compound, the bound SO 2 is released. It is this
liberated free SO2 that prevents further growth of the bacteria.
SO2 is an effective germicide and concentrations of 0.8 ppm molecular SO 2 will be adequate to control
the growth of LAB in wine.
Alcohol - Generally LAB can survive and grow in table wines. There is some variation between various
species regarding alcohol tolerance. For example; Lactobacillus trichods has been found in wine
containing 20% alcohol. The alcohol tolerance is influenced by pH and storage temperature.
Oxygen and Carbon Dioxide - Although microaerophilic conditions are desirable for the growth of LAB,
the evidence suggests that a small amount of 02 may be necessary. It is however, widely recognized
that the presence of CO2 stimulates the growth of LAB. This may be a factor stimulating MLF in wines
left on the lees which would contain a fair amount of dissolved CO 2. Kelly, Asmudson, and Hopcroft
(1989) concluded that this was likely due to low levels of O2 as they observed the same effect using N2.
Nutrients - LAB require a source of energy such as carbohydrates and inorganic salts. In addition they
also need other growth factors such as vitamins and amino acids. Yeast autolysis (which occur during
prolonged lees contact) resulting in increased nutrient content can render a young wine prone to attack
by LAB.
Vinification Practices
Many vinification practices can influence growth of LAB in a winery. Some of the important practices
include: fruit condition, must treatment (adjustment), clarification, fermentation conditions, skin contact
time (in case of red wine), lees contact, wine clarification, storage and winery sanitation.
Sound fruit has a low population of LAB on the surface, therefore; using clean and healthy fruit is
important in reducing the number of microbes that would enter the winery at harvest. Sulfur dioxide is
often added at the crush. It is one of the most effective measures in controlling the growth of LAB.
Winemakers not sulfiting the must at crush in order to reduce sulfite's in wine are taking a bigger risk in
exposing their wines to bacterial spoilage. High pH musts usually contain low acid levels. The acidity
and pH of such a must should be adjusted with tartaric acid additions before fermentation. This will
enable fermentation to occur at low pH, and thus reduce the chances of spoilage by LAB. Clarifying
white must by settling or other means reduces the suspended solids in the must. This practice is
suggested for discouraging MLF in white wine.
Fermentation conditions affect the growth of LAB. For example, in case of a stuck fermentation, LAB
can attack sugar and increase V A levels in wine. Controlling the fermentation so that it proceeds rapidly,
evenly, and reaches dryness, is a sound enological practice to prevent any damage from LAB. A young
wine left on the lees for a long time will be prone to MLF. This is due to the availability of nutrients
released by yeast autolysis and a reduced CO2 environment. For controlling LAB, early racking is
recommended. Wine clarification, especially using tight filter pads or a .45 micron membrane filter will
reduce the bacterial population and consequently the chance of spoilage.
Of all the winery practices, cleaning and sanitization of equipment and containers is one of the most
important practices that a winemaker must employ to control the wine spoilage.
Contrary to the antagonistic effects of the yeast, there are however, some reports that suggest that
yeast may have stimulatory influences on the growth of LAB. For example, prolonged contact with the
lees can result in enrichment of young wine by yeast autolysis. This in turn can stimulate
the growth of LAB.
Other microorganisms such as Botrytis cineria and acetic acid bacteria have been reported to have a
stimulating effect on LAB. LAB are often found in association with acetic acid bacteria and there is some
evidence indicating a symbiotic relationship between these organisms.
Bacteriophages are known to destroy the LAB. These phages have been isolated from wine. Not much
is known about the inhibitory impact of these phages on LAB in wine and its influence on wine quality.
Recommendations to Winemakers
Since LAB are involved in MLF as well as wine spoilage, a winemaker needs to decide up front whether
to encourage MLF.
If the choice is to encourage MLF (and avoid spoilage), then the following recommendations should be
followed and MLF must be conducted under controlled conditions.
If the winemaker's choice is not to encourage MLF then the following recommendations should be
followed as a guide to prevent MLF as well as spoilage due to LAB:
1. Use sound fruit for making wine.
2. Add SO2 at crush, about 50 to 75 ppm based on must pH.
3. In a low acid and high pH must, add tartaric acid to bring the pH to 3.3 or lower.
4. In the case of white wine, clarify the must (reduce suspended solids) before fermentation.
5. Control fermentation temperature. Use well prepared, pure culture yeast starter. Use yeast nutrient
if needed.
6. In the case of red wine, prevent must temperature from exceeding 85° F. Punching the cap and
keeping the cap moist is important.
7. After the must is fermented dry, promptly rack the wine off the lees and add enough SO 2 to attain
0.8 ppm molecular SO2 level.
8. Clarify and stabilize the wine and store in clean containers.
9. Clean and sanitize equipment and containers before processing the wine.
10. Sterile filter and store wine at cool cellar temperatures.
SUMMARY
Lactic acid bacteria are present on grapes, contaminated winery equipment and storage vessels. Some
of the LAB primarily decompose malic acid and under certain conditions, attack sugar and malic acid.
These are often involved in MLF and rarely in wine spoilage. Certain other LAB grows in low acid
conditions; metabolize sugars (pentose), tartaric acid and glycerol. These are more dangerous
organisms and cause serious spoilage. Conditions such as moldy fruit, low alcohol, low SO 2, high pH
(3.5.and above), low acidity, presence of fermentable sugars and warm temperatures such as 25° C
(78° F), favor the growth of LAB and can cause wine spoilage. Maintaining an adequate SO 2 level, low
pH, and sanitary conditions during processing can prevent the spoilage.
Literature cited.
Bousbouras, George E. and Ralph E. Kunkee.
1971. Effect of pH in malolactic fermentation in wine. Am. J. Enol. Vitic. 22:121-6.
Kelly, W.J., R.V. Asmundson, and D.H. Hopcraft. 1989. Growth of Leuconostoc olnos under anaerobic
conditions. Am. J. Enol. Vitic. 40:277-282.