Professional Documents
Culture Documents
Introduction
Wine has been with us since the dawn of civilization and has followed humans and
agriculture along diverse migration paths .Serendipity presumably played a part in
its genesis more than 7,000 years ago: damaged grapes spontaneously fermented
in harvesting vessels; curious farmers tasted the resultant alcoholic beverage; the
curious farmers liked what they tasted and enjoyed its effects; said farmers
preferred fermented grape juice to the unfermented fruit. The fate of the grape was
sealed.
The natural occurrence of fermentation means it was probably first observed long
ago by humans. The earliest uses of the word "fermentation" in relation to
winemaking was in reference to the apparent "boiling" within the must that came
from the anaerobic reaction of the yeast to the sugars in the grape juice and the
release of carbon dioxide. The Latin fervere means, literally, to boil. In the mid-19th
century, Louis Pasteur noted the connection between yeast and the process of the
fermentation in which the yeast act as catalyst and mediator through a series of a
reaction that convert sugar into alcohol. The discovery of the EmbdenMeyerhof
Parnas pathway by Gustav Embden, Otto Fritz Meyerhof and Jakub Karol Parnas in
the early 20th century contributed more to the understanding of the complex
chemical processes involved in the conversion of sugar to alcohol. [
One might argue that the most important test tube in the birth and growth of the
modern life sciences is the fermenter
Figure 1
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A generalized scheme of the spread of Vitis vinifera noble varieties of grapevine and
winemaking from their centre of origin in Asia Minor to other parts of the world.
One might argue that the seeds of science and technology, particularly
biotechnology, were also sown at this time. Empirical observations of natural events
and processes were harnessed in repeat experiments'which is to say, vintages
and improvements were made by trialling modifications to practices, retaining those
that were beneficial and discarding failures, with the results communicated down
through the generations. At that time, there was no EMBO reports or alternative
means by which to facilitate horizontal dissemination of information, but the
principle of developmentsans peer reviewis clear: experimentation and
invention lead to progresstechnological and otherwiseand new knowledge is
shared and built upon.
Of course, early inventions and innovations in grape and wine production were
based on little or no knowledge of the biology of grapevines or the microbes that
drive fermentation. In fact, it would be several thousand years before it was even
known that microscopic organisms exist: using a primitive microscope, Antonie van
Leeuwenhoek observed cells for the first time in 1680 (Fig 2).
Figure 2
Selected milestones that mark the path of research in microbiology and yeast
biology that have affected, directly or indirectly, wine science and winemaking.
Scientific knowledge grows at an exponential rate, and nowhere is this more evident
than in the historical milestones of chemistry and biology that have shaped our
understanding of the biology of the microorganisms that drive fermentation (Fig 2).
This progress has been adorned with some of the most significant names in the
chemical and biological sciences, including van Leeuwenhoek, Lavoisier, Gay-
Lussac, Pasteur, Buchner and Koch. One might argue that the most important test
tube in the birth and growth of the modern life sciences is the fermenter, and the
most important model organism has been the yeast Saccharomyces cerevisiae
commonly known as baking, brewing or wine yeast. As readers might know, this is
exemplified in the origin of the word enzymeen' meaning within and zyme'
meaning leaven. Yeast has been integral to pioneering work in microbiology and
biochemistry, particularly in the fields of metabolism and enzymology (Barnett,
1998, 2000; Barnett & Lichtenthaler, 2001).
Throughout the early decades of the twentienth century the place for S. cerevisiae
in fundamental research was affirmed, and there are several good reasons for this.
Our close relationship with this yeast in food and beverage production over
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The 1970s set the stage for another explosion of knowledge, sparked by the advent
of gene technology and driven by a convergence of genetics, biochemistry, cell
biology, microbiology, physical and analytical chemistry, as well as computing
brought together under the banner of molecular biology (Fig 3). Yeast molecular
biology was established when Gerald Fink's group in the USA demonstrated that
yeast could be transformed with foreign DNA (Hinnen et al, 1978). In the same year,
Jean Beggs in the UK developed a shuttle vector between Escherichia coli and S.
cerevisiae that enabled cloning in yeast (Beggs, 1978). The research community
now had a eukaryotic host that was amenable to genetic engineering, benefiting
both fundamental research and offering the potential of precise engineering of
novel strains for industrial applications. It was the first host cell for industrial-scale
production of a recombinant vaccine against hepatitis B and a recombinant food-
grade enzyme, chymosin, which is used in cheese processing (Pretorius et al, 2003).
Figure 3
Selected milestones that mark the path of research in genetics and molecular
biology that have affected, directly or indirectly, wine science and winemaking.
Ever since, S. cerevisiae has been one of the most important model organisms in
molecular biology and emerging fields; breakthroughs and technological advances
in molecular, systems, and now synthetic biology rarely happen without S.
cerevisiae figuring somewhere prominently in the story (Fig 3). The international
yeast science community has been particularly progressive and proactive in
establishing large collaborative projects and building resources that are available to
the scientific community. S. cerevisiae was the first eukaryote to have its genome
sequenced (Goffeau et al, 1996), a feat that was achieved through an international
effort that involved 600 scientists, which paved the way for the first chip-based
gene array experiments (Schena et al, 1995). It was the first organism to be used to
build a systematic collection of bar-coded gene deletion mutants (Winzeler et al,
1999; Giaever et al, 2002), in which there are deletion strains for most of the open-
reading frames in the S. cerevisiae genome. This has enabled high-throughput
functional-genomic experiments, and anyone seeking information on just about any
aspect of S. cerevisiae biology has access to the amazing community resource: the
Saccharomyces Genome Database (SGD; http://www.yeastgenome.org/).
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All of this is important to wine research; our winemaking workhorse is centre stage
in thousands of research projects worldwide, so we know more about this humble
eukaryote than any other organism on the planet. It is therefore unsurprising that
wine research has benefited enormously from the privileged place that S. cerevisiae
occupies in life sciences research. This is particularly evident in the impact that
advances in molecular biology and related fields have had on winemaking.
One of the reasons for the yeast-induced variation in wine quality is that, during
fermentation, S. cerevisiae produces an abundance of aroma-active secondary
metabolites and releases many aroma compounds from inactive precursors in grape
juice, which greatly affect the sensory properties of the wine (Swiegers & Pretorius,
2007). Thus, any genetic variation in wine yeast that affects the production or
release of sensorially important molecules will affect wine quality. In this context it
has been demonstrated, for example, that different commercial yeast strains
generate wines with very different profiles of volatile thiols (Swiegers et al, 2009).
These thiolswhich are present in grape juice as non-volatile cysteinylated
precursors (Tominaga et al, 1998)are often described as passionfruit', tropical
fruits' and citrus' by tasters, flavours that are particularly important in wine
varieties such as Sauvignon Blanc (Dubourdieu et al, 2006).
Molecular biology and its tools are crucial to our understanding of the genetic and
molecular bases of yeast-driven volatile thiol release from non-volatile precursors in
grape juice. Howell et al (2005) have used bioinformatic tools and the SGD to
identify candidate S. cerevisiae carbonsulphur lyase genes that might be involved
in the release of volatile thiols from cysteinylated precursors during fermentation.
The researchers used targeted gene deletion to remove these candidate carbon
sulphur lyases from the wine and laboratory yeast strains, and they identified four
genes that potentially contribute to the release of these important aroma
molecules.
A similar approach has been used to engineer yeasts for the enhanced production of
fruity esters (Lilly et al, 2006a) and to increase the production of higher, fusel
alcohols (Lilly et al, 2006b)all of which contribute to the flavour profiles of wines.
Although this work is in the early stages of development, it shows the value of yeast
molecular biology, and the amazing resources that come with it.
Wine alcohol content is of growing importance to the wine industry. In some wine
regions, it has been increasing during recent decades (Godden & Muhlack, 2010).
The main reason for this increase is that grapegrowers tend to leave fruit on the
vine as long as possible to increase fruity characterswhich develop as berries
matureand reduce undesirable green' characters. This practice, however,
produces fruit with a higher sugar content, which translates to higher ethanol
concentrations in the wine.
timing of flocculation is crucial; it must not happen too early, as yeast in large flocs
are inefficient at sugar utilization and can generate suboptimalstuck or sluggish
fermentations (Pretorius, 2000).
Generally, wine yeasts are not good at flocculation; they do not form large clumps
that settle out of suspension. Many years of research using laboratory strains of S.
cerevisiae led to the identification and characterization of genes that encode cell-
surface glycoproteinsincluding lectin-like flocculinsthat cause, among other
things, flocculation and subsequent settling to the bottom of the fermentation
vessel (Pretorius, 2000).
The results of this work were promising, but, when they were carried over to wine
yeast, the findings were rather different. There were even substantial differences
between wine yeast strains, leading the authors to caution that optimisation of the
flocculation pattern of individual commercial strains will have to be based on a
strain-by-strain approach (Govender et al, 2010). Nonetheless, controlled
expression of FLO genes at the end of fermentation remains a plausible technique
for improving the performance of wine yeast, but the strategies required to achieve
a desirable outcome might be more complex than was originally thought.
While the complexity of biological systems is a cause for excitement and wonder to
most biologists, it can make engineering novel strains for industrial applications
trickier than molecular biology and biotechnology textbooks might suggest. For
those of us working on industrial yeast strains, it might be pertinent to directly
tackle the issue of complexity and use systems biology approaches to better
understand the workings of yeast metabolism. This should lead to more accurate
modelling of metabolic processes for better-informed manipulations, to achieve
targeted, predictable outcomes.
However, molecular biologists face one important obstacle to this progress: near
worldwide refusal to permit the use of GMOs in the production of foods and
beverages
S. cerevisiae has been at the forefront of -omics' research. This provides us with
enormous opportunities to improve understanding of wine yeast complexity, which,
in turn, will inform the design of new strains for industrial applications. Increased
and improved knowledge from a huge number of studies investigating strains of S.
cerevisiae at the various -omic levels gives wine yeast scientists a head start in this
field (Borneman et al, 2007; Petranovic & Vemuri, 2009).
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One of the most interesting developments has come from the sequencing of a wine
yeast genome, and its comparison with the genomes of a laboratory strain and an
opportunistic pathogenic S. cerevisiae (Borneman et al, 2008). The authors found a
difference of about 0.6% in sequence information between the wine yeast and the
other strains. They also found, perhaps more importantly, 100 kb of additional
genome sequence in the former; enough to carry at least 27 genes. Open reading
frames (ORFs) in the additional sequences do not resemble anything found in other
strains of S. cerevisiae, but seem to be similar to genes found in distant fungal
relatives. BLAST searches have indicated that some of the genes that are specific to
wine yeast are similar to those encoding cell-wall proteins. This might contribute to
the greater robustness of wine yeast, compared with laboratory strains. Other genes
might encode proteins associated with amino acid uptake, which is significant in the
context of wine sensory attributes; amino acid metabolism is central to the
production of many sensorially important volatile aroma compounds.
Novo et al (2009) published similar findings from a different wine yeast strain
(EC1118) and suggested that the extra sequence was probably the result of
horiziontal gene transfer. Further work using functional geneticsto determine the
effects of knocking out and overexpressing the ORFsshould enable
characterization of the phenotypes of these ORFs, determine their relevance in the
context of winemaking and might also reveal their origins.
There have also been numerous studies describing transcriptomic, proteomic and
metabolomic analyses of wine-yeast fermentations. This work is beginning to
provide insights into wine-yeast fermentations, but it is still early days. It should
also be noted that much of the -omics work on wine yeast has used resources and
databases that are based on laboratory strains. It is now clear that there are
genomic differences between wine and lab strains of S. cerevisiae, and these might
affect -omics data acquisition and analysis. For example, gene-array chips based on
the reference laboratory strain S288c will not include the additional ORFs found in
wine strains. This does not suggest that earlier work is invalid, but that there are
likely to be gaps in it.
The engineered wine yeast strains described in this paper show the potential of
novel yeast strain development to improve wine quality. But molecular biologists
face a major obstacle to this progress: near world-wide refusal to permit the use of
GMOs in the production of foods and beverages, at least in developed' countries
(Gross, 2009; Pretorius & Hj, 2005). Wine industries in most parts of the world have
eschewed the use of GMOs in commercial winemaking, leaving most new-
Page 8 of 36
generation wine yeasts on the laboratory shelf, where they await more enlightened
times.
Two genetically modified wine yeast strains have been released to market in a
limited number of countries including the USA, Canada and Moldova: ML01 and
522EC. ML01, a transgenic wine yeast, has genes that enable it to perform
malolactic fermentation (MLF), a deacidifying secondary fermentation in which malic
acidpresent in grape juiceis decarboxylated to lactic acid. MLF is usually
performed by the lactic acid bacterium Oenococcus oeni after alcoholic
fermentation. However, this bacterium is rather fastidious, being inhibited by a
range of conditions that are typical of fermented grape juicelow pH, high alcohol
content, poor nutrient availability and the presence of sulphur dioxideand can
become stuck' or take considerable time to complete fermentation (Davis et al,
1985). In addition, lacitic acid bacteria can produce a range of biogenic amines,
which are associated with health risks (Lonvaud-Funel, 2001).
A wine yeast that completes both primary and secondary fermentations should
therefore have great potential in the wine industry. The genetically modified wine
yeast ML01 carries two foreign genesthe Schizosaccharomyces pombe malate
transporter gene (mae1) and the O. oeni malolactic enzyme gene (mleA)which are
both chromosomally integrated and regulated by the S. cerevisiae PGK1 promoter
and terminator (Husnik et al, 2006). This enables the host wine yeast to perform
MLF, in parallel with alcoholic fermentation.
The researchers went to great lengths to ensure the safety of ML01. The transgenes
came from microorganisms found in wine, there were no antibiotic resistance genes
or vector sequences carried by the yeast and transcriptome and proteome analysis
showed no important differences in gene expression profiles between the
genetically modified strain and its parent. The FDA granted Generally Regarded As
Safe' status to ML01, but it has not been widely adopted, even in countries where it
is approved for use. This is largely owing to concerns about export markets that do
not tolerate GMOs. In fact, wine industries in many countries have banned the use
of GMOs in wine production, in order to avoid jeopardizing their exports.exports.
The genetically modified wine yeast 522EC was engineered to reduce the risk of
ethyl carbamate production during fermentation. Ethyl carbamate, a potential
carcinogen, is the product of yeast-derived urea reacting with ethanol. It is usually
produced at such low levelsif at allthat it is not a cause for concern, but it
sometimes can make an appearance in some wine-producing regions.
by using techniques that involved the manipulation of DNA in vitro, the regulations
of many countries classify it as a GMO. Again, to the best of our knowledge, this
yeast is not being used in the industry. This might be because ethyl carbamate
production is not a widespread problem, but it probably also reflects the influence of
GMO bans and the reluctance of winemakers to risk losing market share in countries
that harbour strong anti-GMO sentiment.
Who knows what bottled masterpieces await us as we sculpt novel yeast strains in
the laboratory using molecular, systems and synthetic biology
Winemaking, science and technology have interwoven histories and have grown
together over the millennia, benefiting from each other. Although science is an
important part of an oenologist's training and scientific methods and equipment are
routinely employed in the winery, winemakers are not scientists per se. They are,
perhaps more appropriately regarded as artisans, with the emphasis on the art'. As
for many human endeavours, the Arts progress with developments in technology;
think of the use of acrylic paint in the fine arts since its introduction in the 1950s, or
David Hockney's use of a Polaroid camera to create photocollages. In the way that
acrylic paint and photography have provided more options to artists, enabling them
to broaden their horizons, yeast science and technology is adding to the
winemaker's palette. Who knows what bottled masterpieces await us as we sculpt
novel yeast strains in the laboratory using molecular, systems and synthetic biology.
The only real obstacle that we face is consumer acceptance of GMOs; we can only
hope that rationality will eventually prevail.
Yeasts are what ferment the sugar in grape juice into alcohol, transforming it into
wine. Yeasts exist everywhere in our environment, and certain strains are
indigenous to certain places. This is why San Francisco sourdough is so unique we
have our own strain of wild yeast that lives in the foggy San Francisco air, giving our
bread its own unique flavor.
Many wineries use commercially-sold yeasts to ferment their wines. These yeasts
are selectively bred or genetically altered by laboratories to enhance favorable
flavor profiles in a wine (such as spiciness or fruit flavor), to tolerate heat extremes
(cold and hot), to limit malodor in fermentations, and to produce wines with
consistent flavor profiles year after year. These yeasts are also resistant to SO2, the
chemical additive used in wine to prevent unfavorable microbal growth and
oxidation. There are many labs out there making hundreds of yeast strains available
in freeze-dried or liquid form for wineries, and each strain comes with its own name
and marketing materials explaining its virtues. These commercially-sold yeasts are
strong and hearty; pitch some into your grape must and they will quickly overpower
any wild yeasts living in it.
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Whos to say which yeast fermented a wine? You would have to be a scientist to
figure it out. My belief is that less is more, and I would rather sell a wine
fermented spontaneously, even if that means the fermentation started with a
commercial yeast that just happens to be part of the landscape. So many new
winemakers are making wine in shared custom crush facilities and co-ops, its
hard to say anymore. Unless your wine came from a centuries-old chateau in France
that has never seen a commercial yeast, its hard to say for sure whether or not
your wine is truly indigenous or native yeast fermented.
It would be nave for us to be dogmatic about this topic, insisting that we only sell
native yeast wine, so instead we only offer wines that are fermented
spontaneously, and never through direct inoculation with commercial yeasts. We
simply feel that these wines are better they are more interesting, delicious, and
unique.
The fermenting of grapes and grape juice into wine, is a natural process that has
been enjoyed by mankind for thousands of years. The choices made about soil and
particular grapes in the vineyard and winery, are of relevance to the wine
enthusiast as they directly effect the final product. Wine derives its alcohol by the
Page 11 of 36
process of fermentation. Grapes on the vine are covered with yeast, mold and
bacteria. By putting grape juice into a container at the right temperature, yeast will
turn the sugar in the juice into alcohol and carbon dioxide. The grape juice will have
fermented and become wine. But fermentation is not the only step in the process of
making wine. The next several pages will discuss in basic terms the total process of
wine making. Links to individual steps in the process are listed to the left if you
prefer to skip around. To start at the beginning of the delicate process of making
one of the world's oldest drinks click on harvesting, where the grapes take their first
step to reaching the goblet.
For example, if you had five gallons of juice that had 10 pounds worth of sugar
in it, and you fermented all of that sugar with yeast, you would end up with 5
gallons of juice that has roughly 5 pounds of alcohol in it.
The other five pounds of sugar would dissipate into the air as CO2 (carbonic)
gas. So in fact the five gallon batch would become five pounds lighter than it
was before the fermentation started.
Realize that the breakdown of alcohol verses gas would not be exactly half and
half, but usually it would be very close. Some variances do occur depending on
external factors such as the amount of available air, nutrients as well as the
type of yeast used. But, rest assured that it would be within 46% one way or
another.
It is important to note here that the 10 pounds of sugar that was in the five
gallon batch may not have come all from sugar you added, but partially from
the fruit as well. And in some cases, such as when making a wine from grapes,
there may be no sugar required at all. In these cases enough sugar is already in
the fruit itself to produce a wine with 11 or 12 percent alcohol.
Alcoholic Fermentation
The important bit converting the sugars to alcohol. Fermentation requires the
action of yeasts to convert the sugars to alcohol. These yeasts can be the natural
yeasts from the vineyard, or specially selected, cultured yeasts. Cultured yeasts are
much easier to control and ensure a more consistent fermentation. Natural yeasts,
on the other hard, ensure a truer manifestation of the vineyards terroir, but are less
Page 12 of 36
Fermentation vessel is a big decision oak, stainless steel or other inert vessel.
Because of its affinity with oak, Chardonnay is often fermented in small oak barrels.
In contrast aromatic grapes such as Riesling or Sauvignon Blanc are typically
fermented in stainless steel or other inert vessels to preserve their vibrant aromas
and flavors.
Fermentation temperature also impacts the wine. Overall, white wines are typically
fermented at cooler temperatures than red. The cooler the temperature the more
well-preserved the primary fruit aromas and flavors. Warmer temperatures make for
a more structured wine.
Typically when the yeasts have converted all the sugars to alcohol the fermentation
is over and you have a dry wine. However, if the intended style off dry or medium
sweet, the winemaker will stop the fermentation before all the sugars have been
converted, leaving the desired amount of residual sugar.
* The Primary Fermentation will typically last for the first three to five days. On
average, 70 percent of the fermentation activity will occur during these first few
days. And in most cases, you will notice considerable foaming during this time
of rapid fermentation.
Here's how important. The little packets of yeast that is generally called for in a
five gallon wine recipe will typically be multiplied up to 100 to 200 times during
the few days of primary/aerobic fermentation. Without air this multiplying stage
is hindered. That is why it is important that you do not use an air-lock during
the first few days of fermentation and allow the fermentation to be open to air.
So as you can start to see, the secondary fermentation is much slower with less
activity at any given time. You will also notice the activity becoming slower and
slower with each passing day.
If the fermentation temperature is too warm, the yeast may ferment fine, but
the flavor of the wine will usually suffer. This is because of the increased
production of unwanted enzymes by the yeast and the possible growth of
micro-organisms that thrive in warmer temperatures.
* Throughout the fermentation process you will need to transfer the wine off the
sediment into a clean container. This is a process that is referred to as "racking"
in most wine making books.
This should be done at the end of the primary fermentation or when the
Specific Gravity reading on your hydrometer reaches approximately 1.030. It
should also be racked after the secondary fermentation as well as right before
bottling the wine.
Once you feel comfortable with the wine making process, your next step is to
purchase the necessary equipment that goes along with each step. You can find
everything you need to start your wine making career at EC Kraus, including a
Wine Making Kit.
Going beyond ideal fermentation temperatures can cause problems. Ferment too
hot or too cold and your wine will suffer.
Fermentations that get too hot not only ferment too fast but it could lead to
cooked flavors. Your wine will taste like it was boiled on the stove. Additionally,
yeast can only tolerate fermentation temperatures that are so high. Go beyond their
maximum temperature tolerance and theyll die.
Keep in mind that the fermentation process is exothermic which means that heat is
produced as the yeast are doing their work. So even if your wine is stored where the
room temperature is within the ideal temperature range your wine could still over
get over heated.
At the other extreme if your wine gets too cold your yeast will go dormant. The good
news here is that when your fermentation temperature rises again the yeast will
likely wake up again and continue fermenting. Even if they dont come back you
can pitch more yeast and continue where you left off. Your wine wont get damaged
by excessively cold temperatures like it will with excessively warm temperatures.
Another option is to get a self adhesive temperature strip. It sticks right to the side
of your carboy or fermenter and displays the internal temperature. While these
Page 16 of 36
arent accurate to the tenth of a degree or anything they will at least give you a
good idea of where you are in the temperature range.
There are several inexpensive products to help you warm up your wine when
fermenting in a cool environment. The least expensive option is using an insulative
wrap on your carboy. These are passive and only trap heat produced by
fermentation.
Another option is to use a heating belt or a ferm wrap. These are electric heaters
that wrap around your carboy or fermenter. Once plugged in they will raise the
temperature of your wine by approximately 10 degrees F.
To get finer control of how warm your wine is you can pick up a thermostat which
allows you to dial in the temperature to within a degree of what you set it
at. Heating bands usually run around $20 and the thermostat is another $80.
Another similar device is a heating panel. These products are just a flat pad that you
place your carboy on top of. When plugged in they heat up to a pre-determined
temperature. Again, you can pick up thermostats for these too.
The easiest way to bring down a high fermentation temperature is to place the
fermenter or carboy in an ice bath. Simply place your container in a plastic tub, add
cool water, and then add ice. Be very careful when moving your carboys especially
if theyre wet. Remember, if you go too far your fermentation can stop.
Wine cooling equipment can be quite expensive. There are a few products on the
market to help you with this task, however, most solutions cost well over $1000.
Beyond using ice water you could opt for a glycol chiller to use on the wine itself or
you can cool the entire room. The latter may not be practical if you dont have a
thermally isolated room to ferment your wine in. Glycol chillers start at around
$1600 and go up from there. Theyre designed to work on larger fermentation tanks.
One intermediate option is to pick up a stainless steel tubing coil or heat exchanger
used in glycol chiller system and run cold water through it. This works a little more
slowly and youll have to figure out your own system of recirculating the cold water
or find a garden to pump it into. Coils start at around $300.
Desirable fermentation temperatures vary for red and white wines. Red wine
fermentation temperatures are optimally between 68-86F (20-30C), while white
wine fermentation temperatures are recommended at or below 59F (15C)
(Reynolds et al. 2001). Higher temperatures are favorable in red winemaking to
enhance extraction of color, phenolics, and tannins from skins (Reynolds et al.
2001). The goal of white wine fermentation temperature control is to preserve
compounds that contribute to aroma and flavor (Molina et al. 2007).
versus 28C. These findings exemplify that temperature creates differences in
flavor and aroma.
Temperature control has also been shown to prevent the development of off-
flavors, specifically volatile
sulfur-containing compounds like hydrogen sulfide. Higher (above 30C/86F)
fermentation temperatures increase the rate of fermentation, which may enhance
production of undesirable byproducts produced by the yeast (Reynolds et al.
2001). The most common off-flavors associated with high temperature and rapid
fermentation are sulfur-derived compounds including hydrogen sulfide,
mercaptans (thiols) and disulphides (Gladish 1999). These compounds have been
associated with aromas or flavors reminiscent of rotten eggs, cooked cabbage,
skunk, and landfill, amongst other descriptors. While some of these off-aromas
may be remediated with copper additions, use of clean lees, or other product
additions, these techniques can be challenging to achieve a clean wine and time
consuming on cellar personnel. Furthermore, additional flavor stripping of
desirable aromatics may occur (Pickering et al. 2005).
Cooler fermentations have been shown to improve the clarity of wine (Comfort
2012). Clarity is typically achieved through racking, cold settling, cold
stabilization, and fining, but low temperature fermentation may be an alternative
option for enhanced clarity. At lower temperatures, yeast cells are less likely to
give off colloids, thus improving clearness (Jackson 2014). Colloids derive from
yeast and pectin, which form polysaccharides and aggregates after a
physiochemical change (i.e. temperature), diminishing clarity (Drioli et al. 2010).
Therefore, the use of lower temperature fermentation may minimize potential
for cloudy or hazy wines by the end of primary fermentation.
Lowering fermentation temperature can also better control microbial growth and
Page 18 of 36
The harvest season marks a monumental time where it all comes together for grape
growers and winemakers. The lack of rainfall during the last few weeks of the
growing season usually transpire into a relatively ideal harvest. Warm, dry days and
cool, dry nights allow grapes to mature optimally.
In the eastern U.S., however, each growing season is unpredictable. Preparation for
incoming, diseased fruit can help winemakers make well-informed decisions during
processing to minimize the influence of disease on wine quality. Botrytis, which
typically flourishes in more humid or higher precipitated years, is one disease that
winemakers should be prepared to handle with incoming fruit. While management
and prevention in the vineyard is obviously the best strategy for dealing with
Botrytis, some years it's presence is unpreventable.
In Loinger, C. et al. (1977), Semillon grapes were fermented to assess the effects of
Botrytis rot on chemical composition of grape must, the wine qualitys sensory
attributes, and the chemical composition of wine. This research showed several
changes in the must and wine chemical composition chemically, associated several
sensory generalizations (i.e., color, aroma, flavor) in wines produced with rot.
Additionally, Bruce Zoecklein has noted in his Enology Notes that Botrytis routinely
causes a loss of fruitiness in wine, and may produce off-flavors that are phenolic
and iodine-like in their descriptions. Loinger et al.s guidelines on cluster rot
infestation can be summed up as follows:
The paper concluded, although not tested, that greater than 40% rot on clusters
would create a wine in the range of low quality to commercially unacceptable. It is
reasonable for winemakers and cellar assistants to evaluate incoming fruit,
regardless of disease or rot infestation, and decide if it is worth the time and
finances to ferment the received crop.
If you are processing more than 1 variety in 1 day, process the cleanest fruit
first. You should end with the dirtiest fruit.
Wash and sanitize all equipment prior to use. Theres no point in allowing
dead bugs, old mildews, and dirt in your product. This practice decreases the
potential risk that can be detrimental to wine quality as soon as grapes get to
the processing area.
Physically wash off equipment in between lots of grapes. This is just good
practice. Use water to initially clean leftover fruit and debris from the
processing equipment, and manually scrub off any debris before properly
sanitizing equipment. In our research winery, we use a citric acid/SO 2 rinse,
but you can also use commercial cleaners and sanitizers. If the sanitizer
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requires a neutralizing agent or water rinse, make sure you do this before
putting in the next lot of fruit. Always follow supplier safety instructions.
Make sure you take the time to properly wash and sanitize processing
equipment at the end of the day. This keeps your equipment in good
condition, reduces biofilm formation, and minimizes chances for any external
contamination.
With whites and reds, limit contact time with the skins
Botrytis resides on the skins with the grapes, and minimizing skin contact with the
pulp will inhibit extraction of Botrytis-related components into your wine. Remember
that red wines require skin contact in order to extract the red pigments
(anthocyanins) from the skins. However, there are several available options for
winemakers. Pre-fermentation flash pasteurization techniques that will quickly
increase red pigmentation and inhibit Botrytis. It should be noted that this
technique does impart its own flavor characteristics on the wine, but it will likely be
more beneficial than the disease-related flavors and aromas. If flash pasteurization
is not an option, consider removing
Press lightly this is not the time where you want to over-extract
With high Botrytis-infected fruit, pressing for increased yield is not recommended.
The increased pressure will not only over-extract tannins, but also extract more rot-
associated compounds and flavors that are detrimental to wine quality.
Limit your oxygen exposure to crushed fruit: use Nitrogen and Argon gas blanketing
Although this will slightly inhibit your commercialized yeast strain, the lack of
oxygen will completely knock out invasive yeasts and microorganisms, including
Acetobacter, which are usually more prevalent in rotted grapes. Ask your supplier if
the commercial yeast can handle this treatment.
If you have more than 10% rot on clusters, consider getting a Laccase Test from a
wine lab
Laccase is an oxidative enzyme that is produced in higher concentrations with
Botrytis infection. Laccase generally progresses the rate of oxidation (i.e., turning
wines brown or making them taste like sherry) even in very young wines. The
results of the Laccase Test will tell you whether or not you need to go extreme
treatments (e.g., heat treatment, early bentonite fining) to avoid rapid oxidation.
Binds with some of the active enzymes that may be destructive towards the
fermentation (e.g., laccase).
Will offer binding materials for anthocyanins to enhance for better color
stability throughout and after fermentation.
Enhance the mouthfeel of the red wine. Botrytis (and disease in general)
tends to thin the mouthfeel of many wines, which extenuates the off-flavors
associated with Botrytis. Building mouthfeel will off-set this phenomena.
Use a gluconase enzyme product, especially in white wines during cold settling
and before fermentation
This enzyme will help break down some of the solids associated with Botrytis
infections. Additionally, this enzyme should help make clarification and filtration
easier on the wine after fermentation. Both processes are often challenging
for Botrysized white wines.
Manage your free SO2 after primary fermentation and malolactic fermentation (MLF)
Checking the free SO2 concentration every other week or once every month can
help inhibit spoilage microorganisms from proliferating during times when the wine
is not actively worked on. Remember that the molecular SO 2 concentration is always
changing and heavily associated with pH. Every time the wine is moved, the
free SO2 should be confirmed to ensure that the proper molecular level is being
obtained in the wine.
your fermentation is dry to avoid any microorganisms from utilizing left over sugar),
volatile acidity (VA) to ensure that it is not high and to know the starting level post-
primary fermentation, and malic acid concentration. Make sure that your MLF is
complete by getting the malic acid concentration checked enzymatically after a
paper chromatography result shows that it has completed. Regularly
monitoring SO2 is also an important step.
These are all suggestions that will depend on the extent of Botrytis infection on the
incoming fruit, and what you are capable of accomplishing at your facility. If you can
estimate that less than 10% of the fruit is infected, minor treatments are usually
needed to ensure a successful fermentation and quality product. Above 15%
infected fruit is where processing operations can get a bit tricky to maintain wine
quality.
The knowledge and the understanding of the microbial terroir how the microbiome
contributes to the natural environment of grapes and to the identity of wine, is a
process that starts at the vineyards, at the harvest of grapes, and then evolves
along the different stages of fermentation (Van Leeuwen and Seguin, 2006; Bokulich
et al., 2013). Indeed, it is known that grapes harbor a complex microbiome,
including a high range of filamentous fungi, yeasts and bacteria with different
physiological and metabolic characteristics (Pretorius, 2000; Fleet, 2003; Barata et
al., 2012). The microflora of the grapes is highly variable, mostly due to the
influence of external factors as environmental parameters, geographical location,
grape cultivars and application of phytochemicals on the vineyards (Pretorius, 2000;
Cadez et al., 2010; Pinto et al., 2014). These microbial communities play an
Page 24 of 36
important role during the winemaking process, as they metabolize the sugars from
the grapes and produce a whole set of secondary metabolites that influence the
wine aromatic quality (Fleet, 2003). In fact, the natural diversity of those metabolic
pathways, and the contribution of the different microorganisms involved on the
fermentation process, is well documented (Setati et al., 2012). Therefore, unveiling
the microbial biodiversity of grapes and during their fermentation will expand our
understanding on fermentation dynamics, on its control (Bisson, 1999; Bisson and
Butzke, 2000) and may also contribute to the identification of novel starter cultures
(Fleet, 2008; Ciani et al., 2010).
The spontaneous wine fermentation is carried out by indigenous microbiota (Heard,
1999; Pretorius, 2000; Ciani et al., 2006; Renouf et al., 2007). Species of
Metschnikowia, Candida, Hanseniaspora, Pichia, Lachancea (Kluyveromyces), and
Saccharomyces are often present at the initial stages of wine fermentations and
form the dominant consortium (Cocolin et al., 2000; Mills et al., 2002; Fleet, 2008).
However, during the wine fermentation, the ethanol content increases and
Saccharomyces cerevisiae strains dominate the alcoholic fermentation (AF; Fleet,
2008). Additionally, a deacidification may occur, by conversion of malic acid into
lactic acid. This process is known as malolactic fermentation (MLF) and is due to the
activity of lactic acid bacteria (LAB; Lonvaud-Funel, 1999; Lerm et al., 2011). The
LAB species associated with MLF generally belong to the Oenococcus, Pediococcus,
Lactobacillus, and Leuconostoc genera (Lonvaud-Funel, 1999). Indeed, MLF mainly
influences the organoleptic characteristics and the aging of wines (Lonvaud-Funel,
1999). On the other hand, acetic acid bacteria (AAB) may cause a negative impact
on the winemaking process, due to the production of undesirable metabolites, as
acetic acid, thus affect negatively the quality of wine and so are considered spoilage
microorganisms (Zoecklein et al., 2000).
The majority of the wine microbiology studies focus on the characterization of S.
cerevisiae strains (Pretorius, 2000; Fleet, 2008; Nisiotou et al., 2011). Nevertheless,
recent studies based on culture-independent methods, started to explore the
microbial communities associated with wine grapes (Bokulich et al., 2013; Taylor et
al., 2014). It is widely accepted that unveiling the indigenous microbial community
associated with particular grape varieties, from specific locations, could represent
an important source of distinctive metabolites and introduce an authenticity terroir
to the region (Heard, 1999; Jolly et al., 2006; Fleet, 2008). The biogeographical
distribution of the wine associated microorganisms has been recently investigated
in vineyards from different regions of California (Bokulich et al., 2013), New Zealand
Page 25 of 36
Many notable wine regions have been recognized with an integrated use of
temperature control, as this processing aid has been shown to improve wine
quality dramatically.
Temperature control is an essential tool when crafting
high-quality wines, but winemakers are encouraged to monitor its use. White,
ros, fruit, and aromatically delicate wines have a greater need for temperature
control during fermentation to enhance aroma and flavor retention. Red wines
require higher temperature fermentations for color and tannin extraction, but can
still benefit from temperature control throughout wine production.
There are a number of solutions for temperature control, based upon winery size
and financial flexibility. These options provide incremental improvement steps
directly related to wine quality, overall. Over time, wineries can slowly enhance
temperature control steps to impact wine quality as economic means are
retained.
B.Discussion
During the primary fermentation of wine, the two grape sugars, glucose and
fructose are converted to alcohol (ethanol) by the action yeast. The by-products of
primary fermentation are aromas and flavours, the gas carbon dioxide, and heat.
The production of heat during fermentation (i.e. it is an exothermic process) means
that during fermentation the temperature of the fermentation vessel will rise, and
will require action on the part of the winemaker to cool it down. White fermentation
is usually conducted in the range of 8-19OC, and red wine fermentions typically are
allowed to run at between 25 and 32oC. At temperatures higher than this, there can
be a loss of desirable aroma and flavour compounds, and unattractive aroma
characters in the spectrum of caramel, burnt or cooked characters can be produced.
There are many types of yeast, but two closely related types known as
Saccharomyces cerevisiae and Saccharomyces bayanus are the ones that are
responsible for fermentation.
These species of yeast are encouraged to conduct the fermentation because they:
Page 26 of 36
Are alcohol tolerant. That is, they can continue to ferment sugars to alcohol
even during the latter stages of fermentation when the sugar is low but the
alcohol content is high.
They ferment quickly and only stop when all the grape sugars have been
utilised. Otherwise we would be buying sweet low alcohol wines.
Are more tolerant to sulfur dioxide than other yeasts and bacteria.
They produce wine like aroma and flavour characters. A free list of wine
fermentation characters can be found here, or can be purchased as a beaut
full colour laminated wine tasting wheel here.
In white winemaking the juice is usually inoculated with yeast following its
clarification. The most common type of inoculation is that using an active dried
yeast culture. Yeasts are freeze dried and stored in vacuum packed tins by yeast
supply companies ready for use. The winemaker then re-hydrates the yeast in warm
water bringing them out of their state of suspended animation. The hydrated yeast
is then added to the juice.
Another less interventionist approach is to let nature take its course. The grapes
have a bloom that contains active cells of Saccharomyces cerevisiae. After juicing
the grapes these grape yeast cells and other yeast cells picked up from the winery
equipment then start to convert the grape sugars to alcohol. This type of
fermentation is variously called natural, indigenous or spontaneous fermentation.
The former approach is common in New World winemaking whilst the latter is more
often practiced in Europe. So what are the advantages and disadvantages of each
approach to inoculation?
Consistency. All strains are selected for their ability to meet the general
necessary criteria given above. This maximises the chances of clean and
complete fermentations occurring.
Better Mouthfeel. Many commentators' report that they feel that spontaneous
fermentation promotes better mouthfeel in wines. That is, the wines are
thought to be softer and creamier than those made using single strain starter
cultures. This suggestion however has not been conclusively demonstrated
scientifically. The rational for the suggestion is that the natural yeast flora on
the grape is genetically heterogeneous, i.e. consists of multiple strains and
that this is the reason for the improved mouthfeel.
Lag time. Yeast cells increase in number by division. One becomes two, two
become four, four become eight etc. The initial numbers of yeast cells in an
un-inoculated juice are by nature low. Dividing from a low initial base of cells
means that it takes longer for fermentation to become active. A wait of 3-4
days is typical. In large commercial wineries where the vintage is planned to
work like clockwork, with tanks becoming free just when purchased fruit is to
arrive at the weighbridge, the uncertainties of spontaneous fermentation
present excessive risk. When inoculating, the winemaker adds about 5 billion
cells per litre. Quite a head start over spontaneous fermentation.
The white juice may either be entirely fermented in barrel, or in tank, and when
partially completed, transferred to barrel. With red wines, practicalities demand that
they be fermented in tank for most of duration of the ferment (so as to ensure
appropriate extraction of colour and tannin from the skins), before the coloured
partly fermented juice is drained into barrel to complete the fermentation.
Temperature control in barrel ferments is pretty basic. The barrels are placed in a
cool room and the wine is left to ferment. The smaller volume and relatively high
surface area of the barrel aids in temperature control. Barrel fermentation is
generally carried out for premium full bodied styles and used with varieties such as
Chardonnay, Semillon and occasionally Sauvignon Blanc. Any full bodied red wine
can benefit from partial barrel fermentation.
White wines are often left in contact with the dead yeast cells (lees) that fall to the
bottom of the barrel following the completion of fermentation. This contact is
usually for a period of six to nine months, and the lees can either be left or stirred
(battonage) at regular intervals. Barrel fermentation and lees contact increases the
Page 28 of 36
aroma and flavour complexity, imparting smokey, toasty and cheesy flavours to
white wines. Some winemakers also feel that barrel ferment results in better
integration of fruit and oak, and that imparts a creamy texture to the mouth-feel.
However if it is overdone, the wine can take on overt doughy and vegemite
characters.
Fermentation is completed when all the fermentable sugars have been converted to
alcohol. This end point is measured chemically. Once the wine is deemed free of
fermentable sugar, i.e. dry it is cooled to 4oC and the dead yeast cells (known as
gross lees) are allowed to settle. The relatively clear wine is then racked to remove
the gross lees before it it is stabilised and filtered ready for bottling.
Even a very complex wine is only alcoholic grape juice. The alcohol is produced by a
process called fermentation.
A ripe organic grape is full of natural sugars and there are wild yeasts living on its
skin. As soon as the skin of the grape is broken, fermentation can begin.
To make wine, all the winemaker has to do is collect his grapes and gently crush
them, releasing the sugary juice and exposing it to the yeasts.
Fermentation will continue until all the sugar has been turned into alcohol or the
level of alcohol in the juice reaches around fifteen percent, whichever is sooner.
At around fifteen percent alcohol, the yeasts will die naturally and any left over
sugars will remain in the wine.
Yeasts
A natural wine is fermented only with the wild yeasts native to its terroir.
Yeast strains vary widely from place to place and contribute significantly to the
odour of the finished wine. The yeasts indigenous to a particular area are an
important part of what gives its wines their character.
Conventionally grown grapes have little or no wild yeast living on their skin.
The winemaker will kill whatever yeast remains with sulphur dioxide, and reseed the
grapes with a single strain of commercially produced yeast.
Page 29 of 36
Wines fermented in this way have less personality, all using the same few
commercial yeast strains, and are less an expression of their terroir. This is one
reason they taste so similar.
They are also less complex, as each of the many wild yeasts present on an organic
grape will contribute something to the finished wine.
Sugars
The level of alcohol in the finished wine is determined by the level of sugar in the
grapes from which it is made.
More sugar means there is more for the yeast to convert into alcohol.
Grapes grown further north see less sun and therefore contain less stored sugar
than those grown in the south. Traditionally, therefore, northern wines contain a
lower level of alcohol.
Malolactic fermentation
Malolactic fermentation is a secondary process of bacterial conversion, which may
follow or overlap with primary fermentation.
Harsher tasting malic acid is converted into softer, and less acidic, lactic acid.
Carbon dioxide is also produced.
In practical terms this means a reduction in the acidity of the wine and an increase
in its complexity. The level of alcohol is unaffected.
If a wine is bottled quickly, it may take place inside the bottle. One reason SO2 is
used at bottling is to prevent this.
A natural winemaker has to wait for the malo to finish naturally before he can bottle
his wine.
Page 30 of 36
In winemaking, there are distinctions made between ambient yeasts which are
naturally present in wine cellars, vineyards and on the grapes themselves
(sometimes known as a grape's "bloom" or "blush") and cultured yeast which are
specifically isolated and inoculated for use in winemaking. The most common
genera of wild yeasts found in winemaking include Candida,
Klckera/Hanseniaspora, Metschnikowiaceae, Pichia and Zygosaccharomyces. Wild
yeasts can produce high-quality, unique-flavored wines; however, they are often
unpredictable and may introduce less desirable traits to the wine, and can even
contribute to spoilage. It should be noted that few yeast, and lactic and acetic acid
bacterial colonies naturally live on the surface of grapes, but traditional wine
makers, particularly in Europe, advocate use of ambient yeast as a characteristic of
the region's terroir; nevertheless, many winemakers prefer to control fermentation
with predictable cultured yeast. The cultured yeasts most commonly used in
winemaking belong to the Saccharomyces cerevisiae (also known as "sugar yeast")
species. Within this species are several hundred different strains of yeast that can
be used during fermentation to affect the heat or vigor of the process and enhance
or suppress certain flavor characteristics of the varietal. The use of different strains
of yeasts is a major contributor to the diversity of wine, even among the same
grape variety. Alternative, non-Saccharomyces cerevisiae, yeasts are being used
more prevalently in the industry to add greater complexity to wine. After a winery
has been in operation for a number of years, few yeast strains are actively involved
in the fermentation process. The use of active dry yeasts reduces the variety of
strains that appear in spontaneous fermentation by outcompeting those strains that
are naturally present.
The addition of cultured yeast normally occurs with the yeast first in a dried or
"inactive" state and is reactivated in warm water or diluted grape juice prior to
being added to the must. To thrive and be active in fermentation, the yeast needs
access to a continuous supply of carbon, nitrogen, sulfur, phosphorus as well as
access to various vitamins and minerals. These components are naturally present in
the grape must but their amount may be corrected by adding nutrients to the wine,
in order to foster a more encouraging environment for the yeast. Newly formulated
time-release nutrients, specifically manufactured for wine fermentations, offer the
most advantageous conditions for yeast. Oxygen is needed as well, but in wine
making, the risk of oxidation and the lack of alcohol production from oxygenated
yeast requires the exposure of oxygen to be kept at a minimum.
Page 31 of 36
Dry winemaking yeast (left) and yeast nutrients used in the rehydration process to
stimulate yeast cells.
Upon the introduction of active yeasts to the grape must, phosphates are attached
to the sugar and the six-carbon sugar molecules begin to be split into three-carbon
pieces and go through a series of rearrangement reactions. During this process, the
carboxylic carbon atom is released in the form of carbon dioxide with the remaining
components becoming acetaldehyde. The absence of oxygen in this anaerobic
process allows the acetaldehyde to be eventually converted, by reduction, to
ethanol. During the conversion of acetaldehyde, a small amount is converted, by
oxidation, to acetic acid which, in excess, can contribute to the wine fault known as
volatile acidity (vinegar taint). After the yeast has exhausted its life cycle, they fall
to the bottom of the fermentation tank as sediment known as lees. Yeast ceases its
activity whenever all of the sugar in must has been converted into other chemicals
or whenever the alcohol content has reached 15% alcohol per unit volume; a
concentration strong enough to halt the enzymatic activity of almost all strains of
yeast.
Some strains of yeasts can generate volatile thiols which contribute to the fruity
aromas in many wines such as the gooseberry scent commonly associated with
Sauvignon blanc.
Brettanomyces yeasts are responsible for the "barnyard aroma" characteristic in
some red wines like Burgundy and Pinot noir.[11]
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Carbon dioxide is visible during the fermentation process in the form of bubbles in
the must.
During fermentation, there are several factors that winemakers take into
consideration, with the most influential to ethanol production being sugar content in
the must, the yeast strain used, and the fermentation temperature. The biochemical
process of fermentation itself creates a lot of residual heat which can take the must
out of the ideal temperature range for the wine. Typically, white wine is fermented
between 18-20C (64-68F) though a wine maker may choose to use a higher
temperature to bring out some of the complexity of the wine. Red wine is typically
fermented at higher temperatures up to 29C (85F). Fermentation at higher
temperatures may have adverse effect on the wine in stunning the yeast to
inactivity and even "boiling off" some of the flavors of the wines. Some winemakers
may ferment their red wines at cooler temperatures, more typical of white wines, in
order to bring out more fruit flavors.
To control the heat generated during fermentation, the winemaker must choose a
suitable vessel size or else use a cooling device. Various kinds of cooling devices are
available, ranging from the ancient Bordeaux practice of placing the fermentation
vat atop blocks of ice to sophisticated fermentation tanks that have built-in cooling
rings.
A risk factor involved with fermentation is the development of chemical residue and
spoilage which can be corrected with the addition of sulfur dioxide (SO2), although
excess SO2 can lead to a wine fault. A winemaker who wishes to make a wine with
high levels of residual sugar (like a dessert wine) may stop fermentation early either
by dropping the temperature of the must to stun the yeast or by adding a high level
of alcohol (like brandy) to the must to kill off the yeast and create a fortified wine.
In winemaking, there are different processes that fall under the title of
"Fermentation" but might not follow the same procedure commonly associated with
wine fermentation.
Bottle fermentation
Bottle fermentation is a method of sparkling wine production, originating in the
Champagne region where after the cuvee has gone through a primary yeast
fermentation the wine is then bottled and goes through a secondary fermentation
where sugar and additional yeast known as liqueur de tirage is added to the wine.
This secondary fermentation is what creates the carbon dioxide bubbles that
sparkling wine is known for.
Carbonic maceration
The process of carbonic maceration is also known as whole grape fermentation
where instead of yeast being added, the grapes fermentation is encouraged to take
place inside the individual grape berries. This method is common in the creation of
Beaujolais wine and involves whole clusters of grapes being stored in a closed
container with the oxygen in the container being replaced with carbon dioxide. ]
Unlike normal fermentation where yeast converts sugar into alcohol, carbonic
maceration works by enzymes within the grape breaking down the cellular matter to
form ethanol and other chemical properties. The resulting wines are typically soft
and fruity.
Page 34 of 36
Malolactic fermentation
Instead of yeast, bacteria play a fundamental role in malolactic fermentation which
is essentially the conversion of malic acid into lactic acid. This has the benefit of
reducing some of the tartness and making the resulting wine taste softer.
Depending on the style of wine that the winemaker is trying to produce, malolactic
fermentation may take place at the very same time as the yeast fermentation.
Alternatively, some strains of yeast may be developed that can convert L-malate to
L-lactate during alcohol fermentation. For example, Saccharomyces cerevisiae strain
ML01 (S. cerevisiae strain ML01), which carries a gene encoding malolactic enzyme
from Oenococcus oeni and a gene encoding malate permease from
Schizosaccharomyces pombe. S. cerevisiae strain ML01 has received regulatory
approval in both Canada and the United States
References
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Bartowsky, E.J. and P.A. Henschke. 2004. The buttery attribute of wine
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Bisson, L. F. 1999. Stuck and Sluggish Fermentations. Am. J. Enol. Vitic.
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Blateyron, L. and J.M. Sablayrolles. 2000. Stuck and slow fermentations:
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Butzke, B. 2010. Wine Storage Guidelines. Purdue University Extension.
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Comfort, S. 2012. Controlling Temperature for a WhiteFermentation. More
Wine. 28 April 2015.
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Pretorius, I.S. and F.F. Bauer. 2002. Meeting the consumer challenge
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