BACTERIA
Malolactic Bacteria Selected from Nature
Lallemand offers two popular formats of freeze-dried bacteria cultures.
MBR®
The MBR® form of malolactic bacteria represents a Lallemand acclimatization process that subjects the
bacteria cells to various biophysical stresses, making them better able to withstand the rigors of direct addition to wine. The conditioned MBR® bacteria that survive are robust and possess the ability to conduct
reliable malolactic fermentation (MLF), even under difficult wine conditions.
Easy rehydration and inoculation protocol for MBR® bacteria cultures
2
1-Step®
The 1-Step® starter kit contains one of our popular MBR® bacteria and an activator mix that require a
short acclimatization protocol to “wake-up” the bacteria and activate their metabolism. Unlike the MBR®
cultures, the 1-Step® cultures are not preconditioned at our Lallemand bacteria facility and therefore
require a simple rehydration/activation step 24 hours before they can be inoculated into the wine.
Three popular MBR® bacteria – Enoferm ALPHA™, Lalvin VP41® and PN4™ – are now available as 1-Step®
starter kit cultures. The 1-Step® cultures perform as well as or better than MBR® cultures, especially under
more challenging MLF conditions, and they are recommended for restarting stuck MLF. Refer to pages XX
and XX for more information.
Easy rehydration and inoculation protocol for 1-Step bacteria culture
Mix and dissolve content of activator sachet in 100 L of
drinking water (temperature between 18° and 25°C).
Add content of the bacteria sachet and dissolve carefully
by gentle stirring.
Wait 20 minutes.
Mix the 1-Step® preparation (activator and
bacteria dissolved in 100 L of drinking water)
with 100 L of wine, pH > 3.5 (temperature
between 18° and 25°C). Wait 18 to 24 hours.
If malic content is below 1-2 g/L, wait only
8-12 hours.
Transfer the preculture to 1000 hL of wine.
Maintain temperature between 18° and 22°C.
Check malolactic fermentation activity (malic
acid degradation) every 2 to 4 days.
BACTERIA MBR® AND 1-STEP OENOCOCCUS OENI CULTURES
Enoferm ALPHA™
Structure and fruit, adapted to high alcohol wines
Enoferm ALPHA™ was selected by the Institut Francais de la Vigne et du Vin (IFV) from spontaneous malolactic fermentations
showing good fermentation activity and sensory contribution. ALPHA™ is a dominant strain and has the capacity to achieve
reliable MLF, even showing good resistance to botrycides. The contribution of ALPHA™ to wine is usually described as enhancing
the mouthfeel, while respecting the wine’s varietal character. The lower perception of green and vegetative flavors is the result
of the very positive impact of ALPHA™ on wine complexity.
Enoferm BETA™
For co-inoculation
Enoferm BETA™ was isolated in Italy and is best used to enhance tannin structure and red berry varietal character in reds. The
name “Beta” comes from its capacity to increase levels of beta-damascenone and beta-ionone, which contribute floral notes,
especially in Merlot. BETA™ benefits from the addition of a malolactic nutrient, such as Acti-ML™. BETA™ is available in MBR®
and BETA CO-INOC form. Refer to page XX for co-inoculation in whites and reds.
Lalvin 31®
For cool climate, adapted to low pH and low temperature wines
Lalvin 31® (MBR®) was selected by the Institut Francais de la Vigne et du Vin (IFV), and performs well under stressful low pH
or low temperature conditions. Lalvin 31® is noted for its good sensory balance in Pinot noir and white wines, and for low
production of biogenic amines; Lalvin 31® benefits from the addition of a malolactic nutrient such as Opti’ML Blanc. Available
only in MBR® form.
Lalvin Elios 1™
For spice contribution and integration of highly ripened fruit
Lalvin Elios 1™ was isolated by the Institut Coopératif du Vin (ICV) in Montpellier, France, from spontaneous malolactic fermentations showing very good fermentation performance, as well as positive sensory profiles. Lalvin Elios 1® consistently
demonstrated good fermentation kinetics under such difficult MLF conditions as high alcohol. This malolactic bacteria culture
enhances the perception of overall tannin intensity, while avoiding green and vegetative character development. Available only
in MBR® form.
Lalvin VP41®
Supports red berry fruit and very low diacetyl production
Lalvin VP41® was isolated in Italy during an extensive European Union collaboration to research natural Oenococcus oeni
strains. The positive mouthfeel contribution of Lalvin VP41® stood out in tastings when compared to other ML bacteria strains.
The very good implantation, high alcohol and SO2 tolerance, plus the steady fermentation kinetics of Lalvin VP41®, make it a
very reliable malolactic fermentation culture to use when a significant impact on wine structure is desired.
O-MEGA™
For fruit-driven wines
O-MEGA™ was isolated and selected in the south of France by the Institut Français de la Vigne et du Vin (IFV) in Burgundy for its
capacity to quickly achieve malolactic fermentation (MLF) in a wide range of applications. O-MEGA™ is a secure and effective
strain that tolerates low pH or high alcohol conditions, and is easy to use. O-MEGA™ complements fresh and fruit-driven wines,
and helps stabilize red wine color by degrading acetaldehyde more slowly. Available only in MBR® form.
PN4™
The Rocket, supports tannin structure
The PN4™ bacteria was isolated from a spontaneous malolactic fermentation in a Pinot noir by the Institute of San Michele in
Trentino, Italy. This bacteria demonstrates its capacity to achieve malolactic fermentation for red and white wines in difficult
conditions of pH, alcohol and S02. The PN4™ bacteria is well suited for spicy and structured Pinot noir wines, and may also be
used to carry out malolactic fermentation in Chardonnay.
3
MALOLACTIC BACTERIA NUTRIENTS
Acti-ML™
The nutrient for malolactic bacteria rehydration
Acti-ML™ was developed by the Lallemand bacteria R&D team led by Dr. Sibylle Krieger. For MLF
in difficult wines, add Acti-ML™ to the bacteria culture’s rehydration water. Acti-ML™ is a specific
blend of inactive yeasts rich in amino acids, mineral cofactors and vitamins. These inactive yeasts
are mixed with cellulose to provide more surface area to help keep bacteria in suspension.
Dosage recommendation: Dissolve 20 g of Acti-ML™ in 200 mL of 25°C (77°F) clean water then add
1 g of bacteria. Wait 15 minutes and add the suspension to 100 L of wine.
ML RED BOOST™
The nutrient for high maturity red wines
4
ML RED BOOST™ is a malolactic fermentation nutrient specifically formulated from inNEW activated yeast fractions for the application of conducting MLF in challenging red wines
with high maturity (Malbec, Merlot, Syrah or Zinfandel). ML RED BOOST™was developed by
Lallemand after investigating lactic acid bacteria’s specific nutritional requirements, and the role
of specific yeasts fractions that improved the resistance of wine bacteria against the inhibitory
effects of high polyphenolic content in red wines. The bioavailability of certain peptides strongly
favor the growth of Lallemand selected wine bacteria, and the quality of specific polysaccharides
included in ML RED BOOST™ are particularly effective in reducing the MLF duration especially in
challenging red wine environments.
Dosage recommendation: Add 20 g/hL (1.6 lb/1000 gal) calculated on final wine volume to a small
amount of water or wine then add directly to the wine, 24 hours before the addition of bacteria.
Opti’Malo Blanc™
The nutrient for white wines
Opti’Malo Blanc™ is a malolactic fermentation nutrient specifically formulated from inNEW activated yeast fractions for the application of conducting MLF in challenging white
wines. Opti’Malo Blanc™ was developed by Lallemand after intensive investigations into specific
nutritional requirements of lactic acid bacteria. The bioavailability of certain peptides strongly favor the growth of Lallemand selected wine bacteria and are particularly effective in reducing the
MLF duration especially under more difficult white winemaking conditions.
Dosage recommendation: Add 20 g/hL (1.6 lb/1000 gal) calculated on final wine volume to a small
amount of water or wine then add directly to the wine.
ENZYMES
Lallzyme LYSO-EASY™
Ready-to-use solution of lysozyme
IMAGES COURTESY OF
INOVATECH
Lallzyme LYSO-EASY™ is a purified natural enzyme preparation extracted from egg whites and is specific for inhibiting
Gram-positive bacteria. Lallzyme LYSO-EASY™ is a very efficient lysozyme solution (22% solution, no preservatives, stable
at room temperature for 18 months or longer when refrigerated) Figure 1. Effect of lysozyme on Lactobacillus sp.
that can be added directly to the must, juice or wine. It works by Before lysozyme treatment After lysozyme treatment
lysing the protective outer membrane of Gram-positive bacteria
(see figure 1). It does not inhibit yeast or Gram-negative bacteria,
such as Acetobacter.
To reduce the risk of spoilage lactic acid bacteria, use Lallzyme
LYSO-EASY™ during cold soak, spontaneous fermentations,
­
­extended maceration and micro-oxygenation.
5
SELECTING THE APPROPRIATE BACTERIA CULTURE
Guidelines for Selecting the Appropriate
Lallemand MBR® Culture
There are two basic considerations when selecting an MBR® culture: security or the culture’s
compatibility to the wine environment and the culture’s sensory attributes.
Note: The four main Environmental Limits have a cumulative effect on the MBR® cultures and
are the limits the MBR® cultures can normally tolerate.
Table 1. Environmental limits and sensory impact
ENVIRONMENTAL LIMITS
MBR®
BACTERIA
6
Alcohol
(% v/v)
pH
< 15.5
> 3.2
< 15.0
> 3.2
< 14.0
> 3.1
< 15.5
> 3.4
O-Mega®
< 16.0
> 3.1
Lalvin
VP41
< 16.0
> 3.1
PN4
< 16.0
> 3.0
Enoferm
ALPHA
Enoferm
BETA
Lalvin
31
Lalvin
Elios 1
Typical
Impact
Total Temperature Impact
MLF
on
on
SO2
kinetics
(°C)
mouth- fruiti(mg/L)
Temp.
feel
ness dependent
Quick start,
< 50
> 14
****
***
slow finish
Slow start,
< 60
> 14
**
***
fast finish
Slow start,
< 45
> 13
*
***
fast finish
Reliable
< 50
> 18
***
**
in reds
start
< 60
> 14
**
**** Quick
and finish
Moderate
start and
< 60
> 16
***
****
finish
Moderate
< 60
> 14
**
**
start and
fast finish
Nutrient
demand
Low
High
High
Medium
Low
Low
Medium
VINIFICATION GUIDELINES
GORDON SPECHT
The following vinification guidelines for malolactic fermentation (MLF) management were developed to help winemakers minimize risks and achieve their winemaking goals. These guidelines
highlight the reasoning, the key points to consider, and the winemaking strategies used for managing MLF. Whether deciding when to inoculate, predicting a wines’ potential difficulty to get through
MLF or handling selected wine lactic acid bacteria (LAB) starter cultures, this information will help
the winemaker optimize MLF management. These guidelines are based on more than 25 years of
experience at Lallemand in LAB production, research, and assistance to winemakers in the optimization of the use of wine LAB and management of MLF.
PRACTICAL GUIDE TO PREDICTING MALOLACTIC FERMENTATION
Why?
• Numerous parameters influence the development and activity of malolactic bacteria in wine.
•M
LF that is spontaneously triggered too early or too late can result in loss of wine quality.
KEY POINTS
pH/SO2
pH/SO2
pH/SO2
pH/SO2
pH/SO2
Interactions of pH/SO2
Free SO2 has a powerful toxic effect on wine LAB, which is enhanced when the pH is
low. However, total SO2 must also be taken into account as the bacteria can convert
the total SO2 into the active form. Attention should also be paid to high pH levels,
which can lead to microbial growth that is often detrimental to the quality of the wine.
Alcohol/temperature interactions
Bacteria are sensitive to ethanol. High temperatures (>25°C/77°F) increase the toxicity
of alcohol to the bacteria. Low temperatures (<16°C/61°F) limit the multiplication of
the bacteria and their activity.
Initial L-malic acid content
At very low levels of L-malic acid, the bacteria will have much more difficulty starting
initiating the MLF. Excessively high levels can also be harmful and cause delays.
Initial L-lactic acid content
The initial presence of L-lactic acid inhibits MLF. At concentrations of 1.5 g/l, the inoculated wine LAB population does not grow and the malic acid degradation is slow. At
L-lactic acid levels of 3.0 g/l, the wine lactic acid bacteria population dies-off.
Inhibition by alcoholic fermentation
The yeast can consume amino acids in the must/juice that are required by the LAB,
especially if the yeast has high nitrogen requirements. If the alcoholic fermentation
(AF) is either very fast or sluggish, toxins that may inhibit MLF are generated.
Timing of inoculation
The point at which malolactic bacteria are added can impact the successful start of
MLF.
7
VINIFICATION GUIDELINES
MLF Scorecard
To take some of the guesswork out of winemaking, Lallemand has developed this scoring
system to assess the malolactic fermentation potential of a wine. Each relevant condition is
assigned a score, and the total score indicates whether MLF is likely to be easy or difficult.
Scorecard for determining the ease of malolactic fermentation
CONDITION
Alcohol (% vol)
pH
Free SO2 (mg/L)
Total SO2 (mg/L)
Temperature
(°C)
Yeast’s
nutritional
needs
8
Ease of alcoholic fermentation
1 point
each
<13
>3.4
<8
<30
18 - 22
2 points
each
13 - 15
3.1 - 3.4
8 - 12
30 - 40
14 - 18 or
22 - 24
8 points
each
15 - 17
2.9 - 3.1
12 - 15
40 - 60
10 - 14 or
24 - 29
10 points
each
>17
<2.9
>15
>60
<10 or >29
Low
Medium
High
Very high
No
problems
Transient
yeast
stress
4 - 5 or
1-2
Sluggish /
stuck AF
Prolonged
yeast
contact
Score
5 - 7 or
Initial level of
>7 or <0.5
2-4
0.5 - 1
malic acid (g/L)
Maximum AF
rate (maximum
<2
2-4
4-6
>6
loss
of brix/day)
Note: Other, currently less well-known factors that are not considered in this scorecard may include the level of dissolved oxygen, polyphenolic content, lees compacting,
pesticide residues, etc.
Total score for the ease of malolactic fermentation:
RESULTS
Favourable: <13 points – Pay attention to development of indigenous flora
(Brettanomyces, contaminating bacteria, etc.) Inoculate rapidly with selected bacteria.
Not so favourable: 13-22 points – Choose the bacteria adapted to your wine. A
specific bacteria nutrient may be necessary.
Difficult: 23-40 points – Give preference to a 1-STEP® bacteria adapted to your wine.
Adjust conditions to optimize MLF: temperature, bacterial nutrition, etc.
Extreme: >40 points – Run quick test or consult your Lallemand representative.
Decrease the obstacles before inoculating: blending, deacidification, temperature,
bacterial nutrition, etc.
VINIFICATION GUIDELINES
PRACTICAL GUIDE TO GOOD PRACTICES
FOR SELECETD WINE LACTIC ACID BACTERIA
Why?
Proper rehydration and inoculation of wine lactic acid bacteria preparations is a crucial step for bacterial survival and malolactic activity.
KEY POINTS
Impact of chlorine
Selected wine LAB are highly sensitive to the chlorine in tap water. Consequently,
spring water or mineral water must be used for rehydration of direct inoculation bacteria (MBR®). With 1-Step® kits, the impact of the chlorine is neutralized by the presence
of the specific rehydration activator, therefore rehydration can be carried out with
normal drinking water.
Adhere to the recommended temperatures
Studies carried out by Lallemand have demonstrated the importance of rehydration
temperature on the survival of the bacteria and subsequent malolactic activity.
Complying with the recommended temperature for each step is vital.
Adhere to time limits
The wine LAB must not lack for L-malic acid for too long, whether during rehydration or
acclimatization, or their population will rapidly decrease.
Use a nutrient or a wine LAB activator if necessary
In cases of nutritional deficiency, or of wines in which the MLF is difficult to start, nutrients or activators may be required.
9
VINIFICATION GUIDELINES
Selected Wine Lactic Acid Bacteria Inoculation Step by Step
Direct Inoculation
Bacteria
1-STEP
Acclimatization Kits
Remove bacteria packets from freezer 30 minutes before use.
1
1A
Add MBR® lactic bacteria
(1 g/hL of wine or must)
and dilute carefully.
1B
Stir specific 1-Step® activator (dosage for
100 hL) into water.
Carefully stir in 1-Step® bacteria (dosage for
100 hL).
®
Non-chlorinated water
at 20°C
10
MBR
ria
bacte
MINERAL WATER
Volume =
20 mL/g of MBR
15 minutes
maximum
2
1B ®
p
1-Ste
ria
bacte
WATER
Volume
= 10 L
Water temperature
must remain constant, between
17° and 25°C.
Avoiding excessive air, stir this bacteria
starter into the wine/must until evenly
mixed.
Wine
temperature
must remain
constant,
ideally between
18° and 22°C.
1A
1-Ste
p
activa ®
tor
2
Double the volume with the wine/must
to be inoculated.
Water + wine
temperature must
remain constant,
between
17° and 25°C.
3
20 minutes
WINE
+
WATER
Volume
= 20 L
pH > 3.5
18 to 24
hours
Avoiding excessive air, stir this bacteria
starter into the wine/must until evenly
mixed.
Wine temperature
must remain constant, ideally
between
18° and 22°C.
Volume
= 100 hL
BACTERIA
STARTER
Regularly monitor MLF (using malic acid analysis every 2 to 4 days).
VINIFICATION GUIDELINES
PRACTICAL GUIDE TO TIMING THE INOCULATION OF SELECTED WINE
LACTIC ACID BACTERIA
Why?
Numerous parameters are involved in selecting bacteria for MLF. Choosing the right moment for
inoculating wine with selected wine LAB is a factor in successful MLF. In addition, choosing the proper time for wine LAB inoculation controls the MLF according to the wine style desired, and allows for
adaptation to technical constraints.
KEY POINTS
Knowledge of resources
The technical resources available (facilities, temperature controls, etc.) and the human resources available (skill, number of personnel, etc.) are critical to properly
choosing the point in time at which the selected wine LAB should be added. Certain
processes, such as co-inoculation, require good technical skills and a higher level of
attention.
Understand the critical parameters of must and wine
As numerous parameters play a role in the feasibility of MLF, it is important to have
a thorough knowledge of them in order to choose the best form of bacterial preparation to use, the appropriate wine LAB strain to use, and the timing of
inoculation.
Yeast and wine lactic acid bacteria interactions
Lallemand has studied the interactions between various yeast and wine LAB strains.
Interactions between the yeast used in the primary fermentation and the selected
wine LAB used to conduct the MLF may positively or negatively affect the MLF itself.
In addition, such interactions may impart specific sensory advantages to the resulting wine. The fact that wine lactic acid bacteria require amino acids and peptides
may necessitate their addition to facilitate bacterial growth and the ensuing MLF.
Make informed choices
The type of wine to be produced as well as resource availability will determine the
type of vinification procedure to be used, and when the selected lactic acid bacterial inoculation is to be added. If the aim is to produce a structured, complex wine
with a long macerating time, the bacterial inoculum should be added after AF.
However, if there is a risk of microbiological contamination (due to high pH,
Brettanomyces, etc.), the co-inoculation technique should be used.
11
VINIFICATION GUIDELINES
Timing MLF according to PRODUCT OBJECTIVE
and/or TECHNICAL CONSTRAINTS
Co-Inoculation
24 to 48 hours after adding yeast
Fast turnover wine
Varietal, fruity white wine
Fresh, fruity red wine
Delayed
Inoculation
maximum 1 month after AF
PRODUCT
OBJECTIVE
Early
Inoculation
2/3 of the way through AF
Ripe berries, red wine
12
Complex, structured
red wine
Sequential
Inoculation
at the end of AF
Co-Inoculation
24 to 48 hours after adding yeast
Complex white wine
Short post-fermentation
maceration
Risk of spoilage contamination
(Brettanomyces)
Red musts
with high pH
Delayed
Inoculation
maximum 1 month after AF
TECHNICAL
CONSTRAINTS
2/3 of the way through AF
High potential
alcohol level
Willingness to work with
micro-oxygenation
Low colour stability
and/or extractability
Early
Inoculation
High-risk AF
Sequential
Inoculation
at the end of AF
Poor temperature control
VINIFICATION GUIDELINES
PRACTICAL GUIDE TO PROPER CO-INOCULATION OF YEAST AND SELECTED WINE LACTIC ACID BACTERIA
Why?
•Co-inoculating a must with selected yeast and wine lactic acid bacteria not only accelerates
the onset of MLF, it makes MLF possible under difficult conditions. This time savings can be of
benefit for finishing fruit-driven and/or quick turnover wines.
•It allows MLF to occur in a less oxidative medium, which reduces the production of buttery
aromas.
• It minimizes the risk of spoilage organism development.
KEY POINTS
??
?
?
On which musts?
• Musts intended for fruit-driven wines or those that will be on the market quickly.
• Musts with a low pH (3.1 to 3.4). If working with > 3.5 pH must VA can increase when
AF is sluggish
• When the cellar and wine temperatures are not conducive to MLF.
• Musts with high risk factors for microbial spoilage by such organisms as Brettanomyces
or indigenous wine LAB producing biogenic amines or off-flavours.
Good management of yeast development
Choice of a reliable selected wine yeast strain. Nutritional supplements must be used
to avoid a stuck AF and aid in the completion of MLF. Co-inoculation is not recommended above 3.5 pH when there is a risk of VA formation due to sluggish and stuck AF.
Temperature control
%
%
%
%
Excessively high fermentation temperatures are detrimental to both yeast and malolactic bacteria. During co-inoculation maintain temperatures < 25oC (77 oF) when
alcohol level is >12.5%vol.
Avoid excessively high degrees of potential alcohol
Co-inoculated wines with a potential alcohol over 14% are more prone to stuck AF.
Avoid excessive use of sulphur dioxide
SO2 rapidly kills wine LAB bacteria. If the harvest is highly contaminated with undesired
micro-organisms and higher additions of SO2 are required, yeast/wine LAB co-inoculation is not recommended..
13
VINIFICATION GUIDELINES
USING BETA CO-INOC FOR PROPER CO-INOCULATION OF YEAST AND BACTERIA
Co-inoculation for White Wines
1
pH >3.1
2
14
Must >16°C
>60°F
3
Optimum
between
16° and 18°C
60° and 65°F
Alcoholic
Ferm.
(AF)
Co-inoculation for Red Wines
4
Towards the
end of AF,
between
16° and 20°C
60° and 68°F
• Selected, properly rehydrated and protected wine yeast.
• Choose a selected wine yeast with low nitrogen requirements
adapted to the style of wine desired.
• Bacteria addition timing depends on SO2 added:
<50 ppm of SO2 added: wait 24 hours
50 to 80 ppm of SO2 added: wait 48 hours
>80 ppm of SO2 added: wait 72 hours
Note: If measuring free SO2 24 hours after addition, at pH >3.3,
the free
SO2 should be <25 ppm and <10 ppm when pH is below 3.3 pH.
• Open BETA CO-INOC™ malolactic bacteria sachet and add
directly
to juice/must at a dosage of 0.5 g/hL (~25 gallons)
• For better dispersion, quickly rehydrate the sachet contents into
clean, chlorine-free drinking water at 20°C (68°F).
• Avoiding excessive air, stir bacteria into must until evenly mixed,
based on the SO2/bacteria addition timing above.
• Complex yeast nutrition one third of the way through alcoholic
fermentation.
• Monitor temperature, malic acid and volatile acidity.
• Top off tank after AF.
• When malolactic fermentation finishes during AF, monitor
volatile
acidity. If there is a 0.1 g/L increase per day, add 20 ppm SO2 or
use lysozyme.
• When MLF finishes after AF, rack and stabilize the wine after
MLF.
Malolactic
Ferm.
(MLF)
Same procedure as above except different temperatures in Steps
3 and 4. Step 3 nor-mal red must
starting temperature at the beginning of AF is 16° to 25°C (60° to
77°F), and once the alcohol level reaches 12.5%
keep it below 25°C. Step 4 optimum temperature to finish MLF
post-AF is 18° to 22°C (65° to 72°F).
RESTARTING STUCK MALOLACTIC FERMENTATION
Recommendations from Lallemand North America
Winemakers are aware that Oenococcus oeni bacteria, traditionally responsible for malolactic fermentation (MLF), are
successful only when the bacteria can adapt to the harsh environment of a fermenting must or a finished wine. The usual
factors influencing the success of MLF include pH, temperature, alcohol and SO2 (both free and total). Problems can arise
when pH levels are low (under 3.4), alcohols are high (greater than 14%), the temperature of the wine is low (less than
18°C [65°F]), or the molecular SO2 is high (greater than 0.3 mg/L). Other inhibitory factors, such as nutrient availability,
high volatile acidity (greater than 0.4 g/L), high levels of condensed tannins and/or agrochemical residues, may also cause
stuck MLF. Under cool climate conditions, where the initial L-malic content is in excess of 5 g/L, LAB can be inhibited by
L-lactic acid contents in excess of 1.5 g/L. Under these conditions, MLF will initiate, but the resulting high L-lactic acid
content can inhibit the malolactic bacteria (MLB), thus preventing MLF from proceeding to completion. Limiting conditions
have a combined cumulative effect, making life truly difficult for the MLB. If several stressful conditions coincide, often MLF
will not go to completion, resulting in a stuck MLF.
To restart and complete a stuck MLF, sometimes it is enough to simply add and gently mix in 20 g/hL (1.7 lb/1000 US gal) of
a nutrient such as ML Red Boost™ for red wines or Opti’ML Blanc™ for white wines. If that fails to restart the MLF, a more extensive protocol using a 1-Step® starter kit is necessary. The Lallemand MLF R&D team, headed by Dr. Sibylle Krieger-Weber,
has developed a wine MLB acclimatization strategy for use with the 1-Step® starter kit, which allows wines undergoing a
stuck MLF to proceed to completion. Lallemand offers four different sizes of 1-Step® starter kits, each containing a sachet
of a selected MLB and another sachet containing an activator. The activation and adaptation of the MLB can be critical in
reducing the effect of an unfavorable wine matrix on the bacteria, thus favoring successful completion of MLF.
If an MLF is encountered that will not proceed to completion, Lallemand has devised a fermentability test to determine
whether a wine is capable of completing MLF when re-inoculated with the 1-Step® starter kit. The test can be performed
by utilizing the following procedure.
1. Immediately before undertaking this test, accurately determine the L-malic acid content (mg/L) of the stuck wine and
record this value as MA 1.
2. Rehydrate the contents of a 2.5 hL sachet of VP41 MBR® or ALPHA MBR® in 50 mL of clean chlorine-free water at 20°C
[68°F]. Gently stir and let rehydrate for 10 minutes.
3. Inoculate 1000 mL of the stuck wine with 2 mL of the bacterial suspension from step 2. This gives a 10x higher inoculation
rate.
4. Maintain the temperature of this 1000 mL of stuck wine at 20°C (68°F) for 72 hours.
5. After the 72 hours, accurately determine the L-malic acid (g/L) content of this 1000 mL stuck wine and record this value
as MA 2.
6. Using the L-malic acid content values measured before inoculation and again after 72 hours, calculate the percentage of
malic acid decrease by using the following formula: (MA 2 / MA 1) x 100 = % of malic acid decrease.
Contact your local Lallemand representative for advice if the result is less than 20% malic acid degradation within 72 hours,
as there is a high level of MLF inhibition in the wine.
However, if the result of the fermentability test shows a greater than 20% malic acid decrease within 72 hours, the 1-Step®
restart protocol may be used to restart the stuck MLF. The procedure is described on the next page.
15
RESTARTING STUCK MALOLACTIC FERMENTATION
Adaptation protocol for handling stuck malolactic fermentations
Stage 1. Pre-treat wine and adjust temperature
Prepare the wine undergoing stuck MLF to receive the activator and selected MLB contained in the 1-Step® starter kit.
The wine should be separated from any lees either by racking or by normal lees removal filtration. This step will help to
remove any potential inhibitory factors or unwanted spoilage organisms. In addition, cross-flow or 0.45 micron membrane
filtration may be necessary to control undesirable bacteria such as Lactobacillus or Pediococcus.
In some cases of stuck MLF, the wine may be suspected of containing substances which may be toxic to malolactic bacteria.
If that is the case, Lallemand recommends pre-treatment of the wine with Nutrient Vit End™ at 30 g/hL (2.4 lb/1000 US
gallons). The Nutrient Vit End™ suspension is prepared in either water or wine then added to the stuck wine with sufficient
mixing. Rack the wine off the Nutrient Vit End™ lees after 48 hours.
Finally, adjust the temperature of the stuck-MLF wine to 18° to 22°C (65° to 72°F).
Stage 2. Prepare the 1-Step® starter kit
Refer to the LIQUID VOLUME TABLE below to determine the size of 1-Step® starter to be used, keeping in mind the 1-Step®
starter kit for restarting a stuck MLF requires twice the normal dosage to compensate for the more difficult MLF environment encountered in a stuck MLF.
16
Rehydration Phase
1. Mix and dissolve contents of the activator sachet in drinking water.
2. Adjust temperature between 18° and 22°C (64° and 72°F)
3. Add contents of the bacteria sachet and dissolve carefully with gentle stirring.
4. Wait 20 minutes.
Acclimatization Phase
1. Mix the 1-Step® preparation (activator and bacteria dissolved in drinking water) with wine, ensuring a pH greater than 3.5.
2. Verify that the temperature is between 18° and 22°C (64° and 72°F).
NOTE: If the stuck wine has less than 1.0 g/L of L-malic acid, wait 6 to 8 hours before inoculation. If the stuck wine has
greater than 1.0 g/L of L-malic acid, wait 12 to 18 hours before inoculation.
Inoculation
1. Inoculate the wine with the acclimated culture.
2. Maintain the temperature between 18° and 22°C (64° and 72°F)
3. Check for MLF activity by analyzing for L-malic acid degradation every 2 to 4 days.
LIQUID VOLUME TABLE
Rehydration Phase
Acclimatization
Phase
Inoculation
ALPHA™ for 25 hL
Dissolve ALPHA™
Mix ALPHA™ (2.5 L)
Inoculate 12.5 hL (330 gals) of wine
(660 gals)
in 2.5 L of water
with 2.5 L of wine
with ALPHA™ culture (5 L)
VP41™ for 100 hL
DissolveVP41™
Mix VP41™ (10 L)
Inoculate 50 hL (1,320 gals) of wine
(2,640 gals)
in 10 L of water
with 10 L of wine
with VP41™ culture (20 L)
VP41™ for 500 hL
Dissolve VP41™
Mix VP41™ (50 L)
Inoculate 250 hL (6,600 gals) of wine
(13,200 gals)
in 50 L of water
with 50 L of wine
with VP41™ culture (100 L)
VP41™ for 1000 hL
Dissolve VP41™
Mix VP41™ (100 L)
Inoculate 500 hL (13,200 gals) of wine
(26,400 gals)
in 100 L of water
with 100 L of wine
with VP41™ culture (200 L)
Berry, leaf and stem sort then
crush
Cluster sort at destemmer
Hand harvest early AM
No adjustment made to must
or wines
Add 40-80 ppm of SO2 at crush
Harvest data: 24-29° Brix; 3.43.9 pH; 0.4-0.8 g/100 mL TA
Cold soak at 10°C (50°F) for
2-10 days
Juice or Must Treatment
Grape Handling
Other:
Opti-Red®
Nutrients:
FERMAID® 2133,
DAP (if needed)
Yeast:
LALVIN ICV D254™
Recommended selections for
primary fermentation:
Caution:
Avoid temperature shock to
the yeast!
Add 2.5 lb/1000 gal (31 g/
hL) Opti-Red® ~1/4 sugar
depletion add an additional
2 lb/1000 gal (25 g/hL)
FERMAID® 2133
Add DAP as needed to adjust
nitrogen
Add 2 lb/1000 gal (25 g/hL)
FERMAID® 2133 at start of
ferment
Add properly rehydrated
LALVIN ICV D254™
Yeast Inoculation
Post AF extended maceration,
drain and press directly to barrels
Practise extended maceration for
3-8 weeks depending on variety
Start fermentation at 32° (90°F)
and ramp up to 35°C (95°F)
Practise pumpovers and
délestage (rack and return) –
frequency depends on grape
variety
Yeast Fermentation
Management
BORDEAUX- STYLE RE D
Rutherford Cabernet sauvignon, Merlot,
Cabernet franc, Petit verdot and Carmenère
GOAL: Bordeaux-Style Red
Wine Style… Techniques for Success
17
Bacteria:
LALVIN® ELIOS 1
Recommended selections for
secondary fermentation:
No filtration or clarification prior to bottling
All clarification is done by settling in barrels or
after blending in tank
Rack to clarify every 3-4 months
Rack to tank, clean barrels, adjust SO2 and
return wine to barrel
Stir lees once per week
Inoculate in barrel with
­LALVIN® ELIOS 1
MLF is complete mid-late
December
Malolactic Fermentation
Wine Stabilization
Management
Malolactic Bacteria
Inoculation
Add acid (if needed) to keep
pre-ferment pH between 3.4-3.5
Thin shoots early to expose
developing berries to sunlight,
accelerating phenolic and
flavour ripeness
Do not crush! Whole berries
for fruit intensity, definition and
gentle tannin extraction
30 ppm of SO2 added to tank
before and after destemming
Hand sort before destemming
Potential alcohol between
13-14%
Yield: 1.5 tons/acre
Other:
LALLZYME® EX
Nutrients:
FERMAID® K, DAP
Nutrients:
ACTI-ML
Bacteria:
Enoferm® Beta
Yeast:
LALVIN ICV D80™, LALVIN ICV
D254™
Rack wine to barrels to complete MLF
Add ACTI-ML to wine
Batch 2: Post AF, rack to
tank for inoculation with
­ENOFERM® BETA
Recommended selections for
secondary fermentation:
If sulphides develop, plunge excessively more than once per day
and délestage (rack and return)
at mid-fermentation
Caution:
Monitor VA and microbes
Split fermentations into 2
volumes:
Batch 1 (25% vol): Drain off at
10 g/L residual sugar and rack to
500 L new French oak barrel for
MLF – Keep pressings separate
to avoid excessive bitterness and
green flavours
Batch 2 (75% vol): Drain off
after 5-10 days extended maceration (70°F)
Small batch fermentations are
easier to handle, but watch for
sulphides and take corrective
action if needed
Gently hand plunge in open top
vessel twice a day
Batch 1: Post AF, inoculate
with ENOFERM® BETA directly
in 500 L oak barrel
If fruit has good phenolic and
flavour ripeness, ferment cool
at ~22-25°C (72-77°F) to retain
fruit intensity and definition
Add ACTI-ML to wine
Malolactic Bacteria
Inoculation
Yeast Fermentation
Management
Recommended selections for
primary fermentation:
Caution:
Do not add DAP during yeast
rehydration
Add 10 g/hL (0.8 lb/1000 gal)
DAP at ~14° Brix
Cold soak at temperature of
incoming fruit, at ~7-10°C
(45-50°F) for 24 hours
Caution:
Practise good hygiene
After yeast inoculation, add
25 g/hL (2 lb/1000 gal)
FERMAID® K
After cold soak, add 25 g/hL
(2 lb/1000 gal) properly
hydrated LALVIN ICV D254™ or
LALVIN ICV D80™
Yeast Inoculation
Gently plunge to mix enzymes
and acid
Add LALLZYME® EX macerating
enzyme
Juice or Must Treatment
Hand pick grapes at flavour and
phenolic ripeness
JE N’AVAIS PAS EXACTEMENT LA MÊME ENTÊTE QUE SUR LE PDF
LE SIXIÈME WINESTYLE EST AUSSI UN ULTRA PREMIUM
CABERNET SAUVIGNON MAIS PAS AVEC LA MÊME DESCRIPTION
VÉRIFIER CELLE-CI AU COMPLET
Watch out for:
• Formation of excessive volatile aromas before, during
and after fermentation – work towards absolute purity
and enhanced flavour definition
• E xcessive dryness – work towards a supple, balanced
texture
Caution:
Monitor VA levels and unwanted microbes, especially if
off-flavours develop
Maintain aging temperature
below 15°C (59°F)
No lees stirring after MLF
When MLF is complete, add
30 ppm of SO2 (ideal target
for microbe control is 0.5
molecular SO2)
Regularly monitor MLF
progress
Stir wines every 2 weeks
Maintain temperature between
18-20°C (64-68°F)
Caution:
Avoid Brettanomyces by maintaining good
SO2 levels especially over holidays and during
warm months
Prior to bottling, filter lightly and settle for
30 days at 10°C (50°F)
After aging, rack and adjust SO2 then, if needed, gently fine wine with egg whites
Regularly top barrels, monitor VA, volatile
phenols and sulphides
Age for 12-14 months in barrel with only 2
rackings. At first racking, blend the batches to
homogenize flavour, texture and microbial
stability
Monitor SO2 every 4 weeks and maintain 0.5
molecular SO2 to control Brettanomyces
Total barrel mix for both batches is 35% new
French oak, 35% 1-year French oak, and 35%
2-year French oak
Batch 1: Rack off gross lees, clean barrels and
return wine to barrels
Batch 2: Rack off gross lees and send this
batch to older barrels
Malolactic Fermentation
Wine Stabilization
Management
U LT R A P R E M I U M C A B E R N E T S A U V I G N O N
Grape Handling
Languedoc, France
GOAL: U
ltra Premium
Cabernet Sauvignon
Wine Style… Techniques for Success
18
Juice or Must Treatment
Adjust SO2 based on adjusted
volume
Measure for potential alcohol
and hydrate as needed
Crush and destem
Caution:
Recheck °Brix after overnight
soak; add more H2O and adjust
SO2 for volume increase
Grape maturity parameters for Cold soak for ~2 days
harvest:
Sample for Yeast Available
(1) By analysis: °Brix, pH, TA
Nitrogen (YAN)
(2) By experience: the physical
softening and flavour of berries,
seed colour and texture
Grape Handling
Nutrients:
GO-FERM®, FERMAID® K and
DAP (if needed)
Yeast:
LALVIN® L2226
Recommended selections for
primary fermentation:
Caution:
Warm must to 18-21°C
(65-70°F) prior to inoculation
Add 1 lb/1000 gal (12.5 g/hL)
FERMAID® K and (if necessary) additional DAP at a third
sugar depletion
Add 1 lb/1000 gal (12.5 g/hL)
FERMAID® K and 2 lb/1000 gal
(25 g/hL) DAP (if necessary)
prior to inoculation
Hydrate 2 lb/1000 gal (25 g/
hL) LALVIN® L2226 with
2.4 lb/1000 gal (30 g/hL) GOFERM®
Yeast Inoculation
Caution:
Watch for H2S formation
Press when AF is complete
Perform aerative pumpovers
during the first 2 days of active
fermentation
Set the tank jackets to 29°C
(85°F)
Yeast Fermentation
Management
Nutrients:
ACTI-ML
Bacteria:
ENOFERM® ALPHA
Recommended selections for
secondary fermentation:
Caution:
Caution:
Monitor for spoilage organisms; Watch for spoilage organisms
check VA weekly
Perform MLF in barrel with
cave temperature of 14°C
(58°F) or ferment in tank
(as space allows) for better
temperature control
Adjust temperature to ~21°C
(70°F) at ML inoculation
Monitor malic acid degradation
and VA
Inoculate with ENOFERM®
ALPHA at the end of AF during
or after pressing
Separate and retain lees for volume recovery
Upon confirmation of MLF completion, add
between 0.4-0.5 molecular SO2 and return
wine to barrels for aging
Rack-off lees to tank and retest for L-malic acid
Check for completion of MLF (barrel composite or tank sample) by enzymatic testing for
L-malic acid
Malolactic Fermentation
Wine Stabilization
Management
Watch out for:
• H2S production
• Nutrient deficiencies
• High pH
• Potential alcohol
Malolactic Bacteria
Inoculation
U LT R A P R E M I U M B A R R E L - A G E D R E D
Napa and Sonoma Counties, California
GOAL: Ultra Premium Barrel-Aged Red
Wine Style… Techniques for Success
19
Destem and crush
Bacteria:
LALVIN 31™
Yeast:
LALVIN® BM45
Other:
Oenological tannins,
macerating enzymes
Nutrients:
GO-FERM®,
FERMAID® K,
DAP
Recommended selections for
secondary fermentation:
Recommended selections for
primary fermentation:
Add LALVIN 31™ to wine in
stainless steel tank
Malolactic Bacteria
Inoculation
Caution:
Wait until AF is complete
before adding bacteria!
Drain and press when dry
(~6-7 days)
No seed removal
Pumpover twice daily for
~20 minutes
Add oenological tannins 24 hours
after yeast
Ferment in 1000 gal (3,785 L)
stainless steel tanks – 2 tanks set
at 29°C (85°F) and one at 32°C
(90°F) to give different flavours
Yeast Fermentation
Management
Caution:
Rehydrate and acclimatize yeast
properly
At a third sugar depletion, add
FERMAID® K
Add macerating enzymes while
filling tank
No cold soak
Add DAP on second day
Add 50 ppm of total SO2
Mechanically harvest very
early AM
Properly rehydrate LALVIN®
BM45 with GO-FERM®
Goal is 20% whole cluster, but
limited by equipment
Syrah harvest parameters:
4.5-5.8 tons/acre; average
23° Brix; 3.6 pH; 0.66 g/100 mL
TA
Yeast Inoculation
Juice or Must Treatment
Grape Handling
RHÔN E- STYLE RE D
Soft, light, fruit-driven, medium-bodied dry red wine
GOAL: Rhône-Style Red
Wine Style… Techniques for Success
20
Confirm ML completion
using paper chromatography
(~4 weeks)
Ambient cellar temperature
~18°C (65°F)
Caution:
Clean barrels between rackings
Prior to bottling, wine is settled and filtered
several times through pads
No cold stabilization or fining
Oenological tannins are added by taste to
barrels
Top regularly and check SO2 and VA
Clean the barrels and rack the other 50% of
Syrah to barrels
Rack half of Syrah to new American oak
barrels and 6 months later, remove this wine
to tank
Rack to tank 2 or 3 times until almost clear
After MLF is complete, add 30 ppm of free
SO2
Malolactic Fermentation
Wine Stabilization
Management
Caution:
Limit yield!
Destem and crush
Usually harvest in early
November
Harvest parameters:
Average 24.5° Brix; 3.3-3.4 pH;
0.8 g/100 mL TA
Thin crop from 4 tons/acre to
1.8 tons/acre
Juice temperature 4-7°C
(40-45°F)
No settling
Tank press directly to barrels
Other:
Clarification enzymes
Nutrients:
FERMAID® K
Yeast:
LALVIN® CY3079 (80% of
juice)
LALVIN® EC-1118 (~20% of
juice)
Some LALVIN® BA11
Recommended selections for
primary fermentation:
Caution:
Acclimatize yeast to COLD juice
temperature before inoculation!
Ferment remaining 20% juice
with properly rehydrated
LALVIN® EC-1118 (sometimes
LALVIN® BA11)
Ferment 80% of juice with
properly rehydrated LALVIN®
CY3079
Add clarification enzymes
Proprietor’s Reserve is from
older (1986) vineyards and
only in very good quality years
Add 30 ppm of SO2 (more if rot
is present)
Yeast Inoculation
Juice or Must Treatment
Grape Handling
AF usually completes in 8-9 days
Barrel ferment temperature
~12.8-18°C (55-65°F)
Bacteria:
LALVIN 31™
Recommended selections for
secondary fermentation:
Caution:
Only add bacteria after AF is
complete!
Add LALVIN 31™ to barrels
2-3 days after yeast inoculation,
add 2 lb/1000 gal (25 g/hL)
FERMAID® K
100% barrel ferment in new
French oak
Malolactic Bacteria
Inoculation
Yeast Fermentation
Management
C O O L R E G I O N C H A R D O N N AY
Light, delicate fruit without buttery taste
GOAL: Cool Region Chardonnay
Wine Style… Techniques for Success
21
MLF completion verified by
paper chromatography
Cellar temperature increased to
ensure MLF completion by mid
to late December
Prior to bottling, sheet and sterile filter
Heat and cold stabilize
Create Proprietor’s Reserve blend and adjust
SO2 to 0.8 molecular
Age in barrel 7-8 months
Rack in February
Return wine to clean barrels
After MLF is complete, rack and adjust to 0.8
molecular SO2
Malolactic Fermentation
Wine Stabilization
Management
• Cold cellar temperatures during MLF
Watch out for:
• Temperature shock to the yeast!
Sort berries and remove leaves
and stems
Sort clusters and destem
Harvest parameters:
24.5-26° Brix; 3.6-3.8 pH
(adjust to 3.65 pH);
0.60-0.65 g/100 mL TA in must
Incoming fruit temperature
~18°C (65°F)
Vineyard is located
~10 minutes from winery
Ferment in 8-ton open-top
fermenters
Cold soak at 16-18°C (60-65°F)
for 24-48 hours
Recommended selections for
secondary fermentation:
Bacteria:
LALVIN VP 41™
Nutrient: OPTI’MALO® PLUS
Recommended selections for
primary fermentation:
Yeast:
LALVIN ICV D21™
Nutrients:
FERMAID® K,
DAP (if needed)
Other:
Macerating enzymes,
Opti-Red®
Caution:
Wait until AF is complete
before adding bacteria!
Extended maceration on remainder of fruit for 25-28 days
Extended maceration on third to
half fruit for 8-10 days
Pumpover with aeration in early
fermentation, twice per day for
20-30 minutes each
Add OPTI’MALO® PLUS
Add LALVIN VP 41™ during
extended maceration when AF
is complete
Set temperature to 29°C (85°F)
and eventually let fermentation
increase to 31°C (88°F)
Add additional FERMAID® K at
third to half sugar depletion
Malolactic Bacteria
Inoculation
Yeast Fermentation
Management
Caution:
Avoid temperature shock to
the yeast!
Add Opti-Red® at
2.5 lb/1000 gal (31g/hL)
Adjust Yeast Available Nitrogen
(YAN) with DAP 1-2 days after
yeast addition
Add properly hydrated LALVIN
ICV D21™ and FERMAID® K to
the must
Add 40 ppm of SO2 at crusher
Handpick in half-ton bins in
early AM
Add macerating enzymes
Yeast Inoculation
Juice or Must Treatment
Caution:
Complete MLF before sending
to barrel
Enzymatically verify whether
MLF is complete before going
to barrel
Drain, press to stainless steel
tank
Ambient temperature 18-21°C
(65-70°F)
Caution:
Monitor VA, SO2, and taste frequently
Top barrels regularly
Rack once or twice in tank for clarification prior
to going to barrel
Adjust total SO2 to 40 ppm
Malolactic Fermentation
Wine Stabilization
Management
U LT R A P R E M I U M C A B E R N E T S A U V I G N O N
Grape Handling
Napa Valley
GOAL: U
ltra Premium
Cabernet Sauvignon
Wine Style… Techniques for Success
22
Destem and crush
Temperature of grapes at winery typically 24-27° (75-80°F)
Vineyard is ~5.5 hours from
winery
Mechanical harvest at night
Harvest parameters:
2.5-3 tons/acre; 23.5-24° Brix;
3.9 pH; 0.52-0.60 g/100 mL TA
Caution:
Look under microscope for
unwanted lactic acid bacteria
Observe juice under microscope
for lactic acid bacteria; if found,
use lysozyme
Cool must overnight in 10 ton
fermenters, target 24° (75°F)
Other:
Lysozyme (if needed),
macerating enzymes
Nutrients:
FERMAID® K
DAP (if needed)
Yeast:
LALVIN ICV D254™
Recommended selections for
primary fermentation:
Set fermenters to 24° (75°F)
Add additional FERMAID® K at
a third to a half sugar depletion
Caution:
Fruit is fragile – avoid over-extraction
Drain and press at dryness
Add some Viognier skins to Carignane fermentation at ~18° Brix
for co-pigmentation
Check pH daily, sudden increase
could indicate lactic contamination
Adjust acid to 0.8 g/100 mL after
first irrigation
Until third day, drain tank through
screen to remove seeds during
pumpover
Pumpover twice daily with the
goal of turning over tank volume
once per day
Malolactic nutrient such as
OPTI’MALO® PLUS
Bacteria:
LALVIN 31™
Recommended selections for
secondary fermentation:
Caution:
Routinely check free and total SO2, VA and
taste
Cold stabilize, DE and pad filter before bottling
Blend lots 2-3 weeks prior to bottling
Age in 2-to-3-year-old French oak barrels for
~6 months
Rack again if needed to achieve clarity before
sending to barrels
Rack tank-to-tank and adjust SO2 to 0.5
molecular
Set tank temperature to 13° (55°F)
Confirm MLF is complete by
paper chromatography
Add ML nutrient to the wine
At yeast inoculation, perform first
pumpover with head spray
Add FERMAID® K after yeast
addition and DAP as needed to
boost nitrogen
Add macerating enzymes on way
to fermenter
Day temperature in vineyard
can be over 38° (100°F) for
15 days, delaying ripening
After MLF is complete, add SO2 to 30 ppm of
free and readjust after 48 hours
Normally MLF is complete in
5-7 days
Add LALVIN 31™ when wine
is still warm from alcohol
fermentation
Start fermentation at 24° (75°F)
then increase to 29° (85°F) after
first pumpover
Add 2 lb/1000 gal (25 g/hL)
properly hydrated LALVIN ICV
D254™
SO2 not added unless poor fruit
or detectable VA
Harsh west Texas vineyard
conditions dictate harvest
Malolactic Fermentation
Wine Stabilization
Management
Yeast Inoculation
Juice or Must Treatment
Grape Handling
Malolactic Bacteria
Inoculation
Watch out for:
• Contamination from unwanted lactic acid bacteria
• Over-extraction of tannins
Yeast Fermentation
Management
RHÔN E- STYLE RE D BLE N D
A blend of mostly Carignane
with Syrah, Mourvèdre and Grenache
GOAL: Rhône-Style Red Blend
Wine Style… Techniques for Success
23
Add 50 ppm of SO2 and 2 g/L
tartaric acid at crusher
Other:
Macerating enzymes (fruit
dependant)
Nutrients:
Complex yeast nutrient such as
FERMAID® K, vitamin complex,
DAP
Nutrients: Malolactic bacteria nutrient such as OPTI’MALO® PLUS
Bacteria:
LALVIN 31™, LALVIN VP 41™
Yeast:
LALVIN ICV-D254™,
LALVIN ICV-D80™, LALVIN
ICV-D21™, LALVIN ICV-GRE™ or
LALVIN® L2056
Caution:
Wait until alcohol ferment is complete before inoculating MLF
Maximum temperature at inoculation is 23°C (74°F)
Add ML nutrients
After AF is complete, inoculate with
MLB in stainless steel tank
Malolactic Bacteria
Inoculation
Recommended selections for
secondary fermentation:
Caution:
For lower H2S formation, keep ferment temperature cool (~24°C/75°F)
and aerate during pumpovers
(especially during growth phase)
Keep free-run and press fractions in
separate tanks
Press based on sensory and target
phenolic numbers, not residual sugar
Analytically monitor phenolics daily
and reduce to one pumpover per day
if extraction is high
Add 2 lb/1000 gal (25 g/hL) complete
yeast nutrient and 2 lb/1000 gal
(25 g/hL) DAP at ~20° Brix
Pumpover using venturi to add O2
twice a day, full tank volume
Some lots are fermented to 27°C
(80°F) to increase phenolic extraction
Begin ferment at 18°C (65°F) and
increase temperature to 24°C (75°F)
Watch for H2S or sluggish fermentations and add additional nutrients if
needed and increase pumpovers with
aeration
Yeast Fermentation
Management
Recommended selections for
primary fermentation:
Caution:
Practise proper yeast rehydration
Remainder fermented with a
combination of LALVIN ICV D80™
(bright fruit), LALVIN ICV D21™
(richness), and LALVIN® L2056 or
LALVIN ICV GRE™ (soft wines with
bright fruit)
~50% fermented with LALVIN ICV
D254™ (“workhorse,” fruit expression, alcohol tolerance, mouthfeel)
Yeast selection is determined by
sensory goals, alcohol tolerance,
fermentation kinetics
Inoculate with
2 lb/1000 gal (25 g/hL) properly
rehydrated yeast
Add 2lb/1000gal (25 g/hL) complete
yeast nutrient, 2 lb/1000 gal
(25 g/hL) DAP and micronutrients
If fruit has harsh or astringent tannins,
cold soak is extended to 72 hours
Add macerating enzyme (light dose)
depending on fruit maturity, vineyard
history and tannin content
Adjust pH/TA with tartaric acid if
needed
Minimum 24-hour cold soak at 13°C
(55°F)
Harvest parameters are based on
flavour – looking for “ripe, jammy”
fruit, typically at ~27° Brix when
grape begins to shrivel
Crush and destem
Yeast Inoculation
Juice or Must Treatment
Grape Handling
PRE M I U M SYRAH
Bright fruit, round mouthfeel, not overly extracted
GOAL: Premium Syrah
Wine Style… Techniques for Success
24
Cold temperatures are a problem
later in the season
Caution:
Monitor temperature
Rack off lees after MLF is complete
SO2 additions are delayed if diacetyl
is detected, to allow for it to be
re-metabolized
As malate starts to decrease, wines
are often moved to barrel to finish
MLF
Analytically monitor malate every
other day
If tank gets too cold, heat exchange
to 21°C (70°F)
Caution:
Monitor VA and sulphites before all rackings
Crossflow and sterile filtration prior to bottling
Age in barrel ~18 months
Fine with gelatin if needed
Rack to tank quarterly
Additions and topping done monthly
Maintain SO2 at ~30 ppm
Rack wines before sending to barrel
Malolactic Fermentation
Wine Stabilization
Management
Watch out for:
• Stuck fermentations due to high alcohol
• H2S production
• Over-extraction of tannins
Caution:
Avoid bunch rot – no touching
clusters in the vineyard
Ferment in 5-7 ton lots
Macerating enzymes and
oenological tannins added at
crusher
Add 50 ppm of SO2 to crusher
Add additional oenological tannins during second pumpover
Adjust nitrogen to 300 ppm
during second pumpover
Destem
Retain 30-50% whole berries
Analyze must (pH/TA, NOPA) and
make necessary adjustments
Pumpover the following morning
and shut off chiller
Light sorting at receiving
hopper
Harvest as cool as possible
Typically harvested at 28-30°
Brix
Other:
Macerating enzymes,
oenological tannins
Nutrients: GO-FERM®,
Complex yeast nutrient such as
FERMAID® K
Yeast:
LALVIN® BRL97, LALVIN ICV
D254™
Recommended selections for
primary fermentation:
Rehydrate 2.5 lb/1000 gal
(31 g/hL) LALVIN® BRL97
or LALVIN ICV D254™ with
GO-FERM® and add during
pumpover
After a 2-3 day cold soak,
inoculate with yeast
Process as quickly as possible
Harvest parameters determined
by flavour, not sugar
Modified cold soak – chill while
filling
Yeast Inoculation
Juice or Must Treatment
Grape Handling
Press at ~3° Brix to stainless
steel tank
Target peak temperature 32-33°C
(90-92°F), heat if necessary
Add nutrients at 5-10° Brix drop
(~13° Brix) or if off-odours occur
First 2 days pumpover using
venturi to inject O2
Bacteria:
LALVIN® ELIOS 1
Recommended selections for
secondary fermentation:
Caution:
Monitor wine parameters
before inoculation
Next day, rack off heavy lees
and send to barrel
Do not move wine until MLF is
complete
Inoculate with LALVIN® ELIOS 1
directly into tank at 21°C (70°F)
Pumpover with irrigator and
flexible impeller pump 3 times a
day (as evenly spaced as possible) pulling from racking valve to
avoid seeds
Prior to bottling, filter through pad and
membrane
Acceptable to have as much as 0.5% residual
sugar
Use lysozyme only if needed
Maintain 30 ppm of free SO2 after MLF
Rack after ML is complete
Malolactic Fermentation
Wine Stabilization
Management
Malolactic Bacteria
Inoculation
Watch out for:
• Bunch rot
• Problems associated with high potential alcohol
Yeast Fermentation
Management
WA S H I N G TO N Z I N FA N D E L
GOAL: Washington Zinfandel
Wine Style… Techniques for Success
25
Caution:
Avoid bunch rot – no touching
clusters in the vineyard
Ferment in 5-7 ton lots
Macerating enzymes and
oenological tannins added at
crusher
Add 50 ppm of SO2 to crusher
Add additional oenological tannins during second pumpover
Adjust nitrogen to 300 ppm
during second pumpover
Destem
Retain 30-50% whole berries
Analyze must (pH/TA, NOPA) and
make necessary adjustments
Pumpover the following morning
and shut off chiller
Light sorting at receiving
hopper
Harvest as cool as possible
Typically harvested at 28-30°
Brix
Other:
Macerating enzymes,
oenological tannins
Nutrients: GO-FERM®,
Complex yeast nutrient such as
FERMAID® K
Bacteria:
LALVIN® ELIOS 1
Yeast:
LALVIN® BRL97, LALVIN ICV
D254™
Caution:
Monitor wine parameters
before inoculation
Next day, rack off heavy lees
and send to barrel
Do not move wine until MLF is
complete
Inoculate with LALVIN® ELIOS 1
directly into tank at 21°C (70°F)
Prior to bottling, filter through pad and
membrane
Acceptable to have as much as 0.5% residual
sugar
Use lysozyme only if needed
Maintain 30 ppm of free SO2 after MLF
Rack after ML is complete
Malolactic Fermentation
Wine Stabilization
Management
Watch out for:
• Bunch rot
• Problems associated with high potential alcohol
Malolactic Bacteria
Inoculation
Recommended selections for
secondary fermentation:
Press at ~3° Brix to stainless
steel tank
Target peak temperature 32-33°C
(90-92°F), heat if necessary
Add nutrients at 5-10° Brix drop
(~13° Brix) or if off-odours occur
First 2 days pumpover using
venturi to inject O2
Pumpover with irrigator and
flexible impeller pump 3 times a
day (as evenly spaced as possible) pulling from racking valve to
avoid seeds
Yeast Fermentation
Management
Recommended selections for
primary fermentation:
Rehydrate 2.5 lb/1000 gal
(31 g/hL) LALVIN® BRL97
or LALVIN ICV D254™ with
GO-FERM® and add during
pumpover
After a 2-3 day cold soak,
inoculate with yeast
Process as quickly as possible
Harvest parameters determined
by flavour, not sugar
Modified cold soak – chill while
filling
Yeast Inoculation
Juice or Must Treatment
Grape Handling
WA S H I N G TO N Z I N FA N D E L
GOAL: Washington Zinfandel
Wine Style… Techniques for Success
26
Target alcohol potential 13.5%14% máximum
Add 40ppm of SO2 at crusher
Analyze must (YAN)
Add Optired at 2.5 lb/1000 gal
(31g/hL)
Analyze must (pH/TA) and make
necessary adjustments Add
tartaric acid to achieve a pH of
3.4-3.5
Nutrients:
Fermaid E
Yeast:
Lalvin YSEO 71B
Recommended selections for
primary fermentation:
Caution:
Avoid temperature shock to
the yeast!
Inoculate with Beta Coinoc
24-48h after the yeast
Add Lalvin YSEO 71B properly
hydrated with Goferm Protect
Add macerating enzymes
LALLZYME® EX 2g/100Kg
Destem and crush
Harvest grapes at 23.5ºBrix
Yeast Inoculation
Juice or Must Treatment
Grape Handling
Caution:
If it is necessary more than
40g/hL of Fermaid E, complete it
with DAP
Monitor density, VA and malic
acid
Drain off when the density reach
~1010 (3ºBrix) Keep presing
separate
Use Nutrient Vit End if sluggish
Perform anaerative pumpovers
twice a day (a third of fermenter
volume) until end of AF
Adjust Yeast Available Nitrogen
(YAN) with complex yeast nutrient Fermaid E; add half at the
beginning of AF and half at the
midlle of AF Perform 1 aerative
pumpover at the first third or add
O2 at 6-8 mg/L with cliqueur
Set fermenter temperature
control to 24ºC (76ºF)
Yeast Fermentation
Management
Q U I C K-TO-MAR K E T R E D
GOAL: Q
uick - To - Market Red
Wine Style… Techniques for Success
27
Nutrients:
Bacteria:
Beta Coinoc
Recommended selections for
secondary fermentation:
Regularly monitor MLF
progress
Inoculate with Beta Coinoc
24-48h after the yeast
Rack off the wine 1 week after
Settle and rack off
No lees stirring after MLF
Bottling
Filtered
Cold stabilized
Adjust free SO2 to 30 ppm
When MLF is complete, add
30 ppm of SO2 (ideal target
for microbe control is 0.5
molecular SO2)
Maintain aging temperature below 15°C
(59°F)
Rack off gross lees 24h after MLF
Malolactic Fermentation
Wine Stabilization
Management
Malolactic Bacteria
Inoculation
Watch out for:
Potential alcohol, Temperature, Initial SO2
Juice or Must Treatment
Target alcohol potential 13.5%14.5%
Add 40ppm of SO2 at crusher
Destem and crush
Harvest grapes at 23.5-24.5ºBrix Add macerating enzymes LALLZYME® EXV 2g/100Kg
Grape maturity parameters:
even ripening within cluster,
Analyze must (pH/TA) and make
sensory evaluation, colour
necessary adjustments Add
tartaric acid to achieve a pH of
Harvest at night when tempera3.4-3.5
tures are cooler
Analyze must (YAN)
Assess incoming fruit integrity.
Grape Handling
Nutrients:
Fermaid E
Nutrients:
Bacteria:
VP41 1-step
Yeast:
Lalvin YSEO ICV D80
Lalvin YSEO ICV D254
Lalvin YSEO CLOS
Inoculate VP41 1-step after 1-2
weeks
Caution:
Monitor malic acid and VA
No lees stirring after MLF
When MLF is complete, add
30 ppm of SO2 (ideal target
for microbe control is 0.5
molecular SO2)
Bottling
Filtered
Cold stabilized
Settle and rack off 48h-1 week after
Adjust free SO2 to 30 ppm Add Reduless at
1g/hL and Noblesse at 10g/hL
Monitor for Brettanomyces during aging
Rack off the wine once each 4 months
Age in 2-to-4-year-old American oak barrels of
225L for ~12 months
Adjust pH and TA
Maintain temperature at
~20°C (68°F)
Microoxigenate the wine with
15mg/L/months at the beginning and follow the evolution
of the acetaldehide by tasting.
Increase or decrease the dosage depending on the case.
Stir/mix twice weekly
Rack again if needed to achieve clarity before
sending to barrels
Regularly monitor MLF
progress
Delay MLF
Drop temperature to 15°C
(59°F)
Malolactic Fermentation
Wine Stabilization
Management
Watch out for:
Molecular SO2 in barrel. Clean barrel after rack off
Malolactic Bacteria
Inoculation
Recommended selections for
secondary fermentation:
Caution:
If it is necessary more than
40g/hL of Fermaid E, complete it
with DAP
Monitor density and temperature
Analyze residual sugar
Watch for H2S production
Drain off
Keep press fraction separate
Add Reduless at 1-2g/hL
Rack off gross lees 24h after
pressing
Perform a delestage once a day
Use Nutrient Vit End if sluggish
AF + Maceration = 12-15 days
Perform anaerative pumpovers
once a day (a third of fermenter
volume) until end of AF
Perform 1 aerative pumpover at
the first third or add O2 at 6-8
mg/L with cliqueur
Adjust Yeast Available Nitrogen
(YAN) with complex yeast
nutrient Fermaid E; add half at
the beginning of AF and half at
the midlle of AF
Set fermenter temperature
control to 26ºC (79ºF)
Yeast Fermentation
Management
Recommended selections for
primary fermentation:
Caution:
Avoid temperature shock to
the yeast!
Add Booster Rouge at
2.5 lb/1000 gal (31g/hL)
Add Lalvin YSEO ICV D80
properly hydrated with Goferm
Protect or Lalvin YSEO ICV
D254 or Lalvin YSEO CLOS
Yeast Inoculation
RIOJA CRIANZA
GOAL: R
ioja Crianza
Wine Style… Techniques for Success
28
CALCULATIONS AND CONVERSIONS
TEMPERATURE CONVERSIONS
C° = Degree Celsius
F° = Degree Fahrenheit
F° to  C° = (F° - 32) x (5/9)
C° to  F° = (C° x 9/5) + 32
C° -18 -15 -10 10 16 21 27 32 38 49
F°
0
5
14 50 60 70 80 90 100 120
MASS CONVERSIONS
VOLUME CONVERSIONS
1 kg
=
1000 g
1 mL
=
0.035 US fl oz
1 kg
=
2.205 lb
1 US fl oz
=
30 mL
1g
=
1000 mg
1L
=
1000 mL
1 lb
=
453.6 g
1L
=
0.2642 US gal
1 lb
= 0.4536 kg
1 US gal
=
3785 mL
1 metric tonne =
1000 kg
1 US gal
=
3.785 L
1 metric tonne =
2205 lb
1 hL
=
100 L
1 US ton
=
2000 lb
1 hL
=
26.4 US gal
1 US ton
=
907 kg
WEIGHT/VOLUME EQUIVALENTS
1 lb/1000 gal = 454 g/1000 gal = 0.45 g/gal = 0.12 g/L = 120 ppm = 12 g/hL
2 lb/1000 gal = 0.90 g/gal = 0.24 g/L = 240 ppm = 24 g/hL
1 g/hL = 1 g/26.42 gal = 0.038 g/gal = 0.084 lb/1000 gal
OTHER CONVERSIONS
1 kg/hL = 1000 g/hL = 10,000 mg/L = 10 g/L = 10 mg/mL
1 lb/1000 US gal = 454 g/1000 US gal = 0.454 kg/1000 US gal = 120 mg/L = 0.120 g/L
1 ppm = 1 mg/L
1 ppb = 1 mg/1000 L
1°Brix = 1% sugar (wt/vol)
1 Vol. % = 1 mL/100 mL
1 Gew. % = 1 g/100 g
29
CALCULATIONS AND CONVERSIONS
Free SO2
(mg/L)
MOLECULAR SO2 AT DIFFERENT ETHANOL LEVELS
Molecular SO2 (mg/L) – 18° – ethanol at 8% v/v
Molecular SO2 (mg/L) – 18° – ethanol at 13% v/v
pH 3.0
pH 3.2
pH 3.4
pH 3.6
pH 3.0
pH 3.2
pH 3.4
pH 3.6
5
0.23
0,14
0.09
0.06
0.41
0.26
0.16
0.10
8
0.37
0.23
0.15
0.09
0.66
0.42
0.26
0.17
10
0.46
0.29
0.18
0.11
0.83
0.52
0.33
0.21
15
0.69
0.43
0.27
0.17
1.24
0.78
0.49
0.31
The following table is meant as an aid to help the user to calculate density data in any of
the three worldwide units: Balling/Brix, Baumé and Oechsle.
COMPARISON OF DIFFERENT DENSITY DATA
30
Gew.
Verh.
20°/20°
1.00000
1.00078
1.00155
1.00233
1.00311
1.00389
1.00779
1.01172
1.01567
1.01965
1.02366
1.02770
1.03176
1.03586
1.03998
1.04413
1.04831
1.05252
1.05667
1.06104
1.06534
1.06968
1.07142
1.07404
1.07580
1.07844
1.07932
1.08021
1.08110
1.08198
1.08287
1.08376
1.08465
1.08554
1.08644
Degree
Balling
/Brix
0.0
0.2
0.4
0.6
0.8
1.0
2.0
3.0
4.0
5.0
6.0
7.0
8.0
9.0
10.0
11.0
12.0
13.0
14.0
15.0
16.0
17.0
17.4
18.0
18.4
19.0
19.2
19.4
19.6
19.8
20.0
20.2
20.4
20.6
20.8
Degree
Baumé
Degree
Oechsle
0
0.1
0.2
0.3
0.45
0.55
1.1
1.7
2.2
2.8
3.3
3.9
4.4
5.0
5.6
6.1
6.7
7.2
7.8
8.3
8.9
9.4
9.7
10.0
10.2
10.55
10.65
10.8
10.9
11.0
11.1
11.2
11.35
11.45
11.55
0
1
2
2
3
4
8
12
16
20
24
28
32
36
40
44
48
53
57
61
65
70
71
74
76
78
79
80
81
82
83
84
85
86
86
Gew.
Verh.
20°/20°
1.08733
1.08823
1.08913
1.09003
1.09093
1.09183
1.09273
1.09364
1.09454
1.09545
1.09636
1.09727
1.09818
1.09909
1.10000
1.10092
1.10193
1.10275
1.10367
1.10459
1.10551
1.10643
1.10736
1.10828
1.10921
1.11014
1.11106
1.11200
1.11293
1.11386
1.11480
1.11573
1.11667
1.11761
1.11855
Degree
Balling
/Brix
21.0
21.2
21.4
21.6
21.8
22.0
22.2
22.4
22.6
22.8
23.0
23.2
23.4
23.6
23.8
24.0
24.2
24.4
24.6
24.8
25.0
25.2
25.4
25.6
25.8
26.0
26.2
26.4
26.6
26.8
27.0
27.2
27.4
27.6
27.8
Degree
Baumé
Degree
Oechsle
11.7
11.8
11.9
12.0
12.1
12.2
12.3
12.45
12.55
12.7
12.8
12.9
13.0
13.1
13.2
13.3
13.45
13.55
13.7
13.8
13.9
14.0
14.1
14.2
14.3
14.45
14.55
14.65
14.8
14.9
15.0
15.1
15.2
15.3
15.45
87
88
89
90
91
92
93
94
95
95
96
97
98
99
100
101
102
103
104
104
106
106
107
108
109
110
111
112
113
114
115
116
117
118
119
CALCULATIONS AND CONVERSIONS
Evaluation of the titratable total acid
In such countries as Switzerland and Germany, the titratable acids of wine are calculated
as tartaric acid and are indicated as a decimal in grams per liter. In France and in other
Roman countries, the titratable total acid is calculated as sulfuric acid. For fruit and berry
juices the titratable total acid is often calculated and indicated as citric acid and malic acid.
In order to perform the appropriate conversions, multiply the titratable acid by the factors
given in the following table.
The total acid
was titrated and
calculated as
Wine acid
Wine
acid
—
The total acid is to be expressed as
Apple
Citric
Lactic
Sulfuric
acid
acid
acid
acid
0.893
—
Acetic
acid
0.853
1.2
0.653
0.8
0.955
1.343
0.731
8.896
1.406
0.766
0.938
0.544
0.667
Apple acid
1.119
Citric acid
1.172
1.047
Lactic acid
0.833
0.744
0.711
Sulfuric acid
1.531
1.367
1.306
1.837
Acetic acid
1.25
1.117
1.067
1.5
—
—
—
0.817
1.225
—
Enological Guidelines
1. Basic Conversions and Addition Rates
a. Brix to alcohol conversion factor use 0.60-0.64% EtOH per °Brix
b. 1 ton of grapes yields roughly 200 gallons must or 155-175 gallons of juice/wine
c. Calculate fermentation additions based on total volume (including skins, etc.)
d. Tartaric additions: 8 lb tartaric/1000 gal adds roughly +0.1 g/100 mL shift in Titratable
Acid
e. Dry ice additions use 7 lb dry ice/ton of grapes to lower 1°F
f. Water additions:
i. (initial Brix - target Brix)/initial Brix = % water to add
Example: Initial Brix = 27
Target Brix = 24
(27-24)/27 = 11% water to achieve target Brix level
ii. C1V1 = C2V2
C1= potential alcohol if all sugar fermented (use 0.60-0.64 conversion rate)
V1 = initial volume of wine
C2 = final alcohol desired
V2= final volume of wine
C1V1 = C2V2 Example:
Current alcohol = 15.53%
Current sugar = 7,260 mg/100 mL
Potential alcohol = 19.88% @ 0.60 conversion rate, 60 gallons wine
(19.88%)(60 gal) = (15.5%)(x gal)
x = 73.5 gal
73.5 gals of final wine - 60 gals of initial wine = 13.5 gal of water
required to bring total alcohol to 15.5%
31
32