malolactic fermentation management - home : …€¦ · · 2014-04-07bacteria in juice and/or in...
TRANSCRIPT
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MALOLACTIC FERMENTATION
Management
Gordon Specht February 25, 2010
Grand Rapids
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Lallemand produces both wine yeast & wine bacteria products
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St. Simon, France
Bacteria Production Facility Virtual Tour:
http://stsimon.lallemand.com/tour_coeno.htm
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Malolactic Fermentation Topics:
• Background
• Monitoring
• Selecting MLB Starter Cultures
• Yeast & Bacteria Interaction
• Timing of MLB Inoculation
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Bacteria in juice and/or in wineLACTIC BACTERIA – gram positive rods and cocci• Oenococcus oeni (ex Leuc. oenos)• Leuconostoc mesenteroides• Lactobacillus plantarum • Lactobacillus casei• Lactobacillus brevis• Pediococcus damnosus• Pediococcus pentosaceus• Lactobacillus hilgardi
ACETIC BACTERIA – gram negative cocci• Gluconobacter oxydans• Acetobacter aceti• Acetobacter pasteurianus
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Oenococcus oeni(formerly Leuconostoc oenos) cells are coccoid to rod-shaped, dividing into long chains.
Oenococcus oeni cells are among the smallest that grow in wine, with 1 µm diameters; lengths typically run 1-3 µm, but may be much longer.
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Malolactic Fermentation (MLF)
Lactic bacteria
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wine type
inoculation rate
fermentation time
O. oeni strain
SO2
aeration
temperature
contact withyeast lees
Diacetyl concn
variable
white - lowerred - higher
104 - higher
106 - lower
longer MLF - higher
Diacetyl concn
18°C - higher25°C - lower
binds to diacetyl- sensorially inactive
air - higheranaerobic - lower
long contact- lower
lower pH may favourpH
Diacetyl - management during winemaking
From: Dr. Eveline Bartowski (AWRI) Trier (D) April 2008
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Environmental Factors that Impact Malolactic Fermentation
Good Moderate Difficult Extreme
Alcohol (% vol) <13 13 - 15 15 - 17 >17
pH >3.4 3.1 - 3.4 2.9 - 3.1 <2.9
Free SO2(mg/L) <8 8 - 12 12 - 15 >15
Total SO2(mg/L) <30 30 - 40 40 - 60 >60
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More Environmental Factors that Impact Malolactic Fermentation
Good Moderate Difficult Extreme
Temperature (°C) 18 - 22
14 - 18 or
18 - 24
10 -or
24 -
14
29or
>29
<10
Yeast's nutritional
needsLow Medium High Very high
Ease of Alcoholic
Fermentation
No problems
Transient yeast stress
Sluggish / stuck
AF
Prolonged yeast
contact
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More Environmental Factors that Impact Malolactic Fermentation
Good Moderate Difficult Extreme
Initial level of malic
acid (g/L)2 - 4
4 - 5 or
1 - 2
5 -or
0.5 -
7 1
>7 or
<0.5
Maximum AF rate
(maximum loss of
brix/day)
<2 2 - 4 4 - 6 >6
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Other, currently less well-known factors that may impact Malolactic
Fermentation and are not considered in this scorecard may include the level of
dissolved oxygen, polyphenoliccontent, lees compacting, pesticide
residues, etc.
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Malolactic Fermentation Topics:
• Background
• Monitoring
• Selecting MLB Starter Cultures
• Yeast & Bacteria Interaction
• Timing of MLB Inoculation
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Monitoring MLF• Most accepted methods for
following malic acid degradation used by Winemakers…– Paper Chromatography– Enzymatic
• Most accepted method for following MLB used by winemakers…
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–MLF has not started yet if you see double cells or very short chains rather than long chains. –You should see about 5 short chains per view and the cells should give contrast. If they look shiny they are most probably dead.
The Microscope…
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The Microscope…
Pure cultures of Oenococcus such as in these ML starter cultures ready for buildup often have long chains of cells; on the other hand, some cultures don't make very long chains under winery conditions.
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Malolactic Fermentation Topics:
• Background
• Monitoring
• Selecting MLB Starter Cultures
• Yeast & Bacteria Interaction
• Timing of MLB Inoculation
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MBR® Bacteria PROCESS: PLANT SCALE FERMENTATION - PHYSIOLOGICAL ADAPTATION
STRESSES
Multiplication stage Adaptation stage
5 to 7 days
STRESS CSTRESS A
STRESS B
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COMPARISON SUMMARY &
Controlled MLF 1-Step
Inoculation Direct After acclimatization
MLF Kinetics Medium-fast Medium-fast
Acclimatization/Handling
Lallemand does the
acclimatization
Winemaker does the acclimatization
COST $$$ $$
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Winemaker selection criteria for ML Bacteria
1. MLF Reliability
Alc.(%) pH
Total SO2
(mg/L)Temp(oC)
Typical
fermentation Kinetics
Nutrient Demand
Lalvin 31 < 14 > 3.1 < 45 > 13 Slow start,
fast finish Medium
PN4< 15.5 > 3.0 < 60 > 16
Moderate start & fast
finishMedium
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1. - MLF Reliability…
How to use 1-STEP…
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Winemaker selection criteria for ML Bacteria
MLF Reliability for extreme Low pH
Champagne base wine
CIVC protocol
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Extreme Low pH Champagne base wine
CIVC protocol Inobacter STANDARD freeze-dried culture
• Acclimate in Juice• No pH adjustment needed• Shorter reactivation step (3 days
instead of 7), duration of Pied de Cuve is maximum 12 days
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Winemaker selection criteria for ML Bacteria
• MLB Sensory Effect
Impact on Mouthfeel
Impact on Fruitiness
Lalvin 31 * ****PN4 ** ***
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AWRI / Lallemand Research Project Impact of ML Bacteria on Aroma
(Bartowski/Costello) 2006-2009
To demonstrate the role of MLF in affecting the varietal aroma (red berry fruit)
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Parameters and details of the Cabernet Sauvignon fruit and wine for the 2008 MLF trial
Batch No. I II III IV Fruit Source Clare
Valley Langhor
ne Creek
Padthaway
Adelaide Hills
FINISHED WINE Alcohol† (% v/v) 13.9 14.7 14.4 14.0 pH* 3.42 3.44 3.44 3.46 TA* (pH
8.2;g/L) 8.3 8.0 8.0 7.9
Specific gravity 0.9949 0.9936 0.9937 0.9938 Glu+Fru† (g/L) 0.4 0.3 0.5 0.6 Free SO2
†† (mg/L) nd** nd nd nd
Total SO2†† (mg/L) 11 8 10 10
L-Malic acid††† (g/L) 1.60 1.76 1.94 2.18 VA†‡ (g/L) 0.2 0.37 0.2 0.25 † analysis performed by NIR (Winescan) †† analysis performed by FIA ††† analysis performed by enzymatic procedure
AUSTRLIA: AWRI
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Malic acid metabolism during MLF of 2008 Cab. Sauvignon wines I, II, III and IV. Oenococcus oeni strains R1106 (BETA)
Un-inoculated wines treated with 60mg/L total SO2 served as non-MLF controls. - AWRI
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1105
1106
No MLF
Dark berries
Red berries
Jammy/Cooked fruit
Mint/Eucalyptus
Chocolate
SpiceMusty
Boiled Egg
Overall fruit flavour
Dark berries flavour
Red berries flavour
Mint/Eucalyptus flavour
Fresh green flavour
Chocolate flavour
Astringent
Bitter
Fruit after taste
PC1 (68%)
PC2 (32%)
1105
1106
1118
No MLF
Dark berries
Red berries
Jammy/Cooked fruit
Mint/Eucalyptus
Chocolate
Spice
MustyBoiled Egg
Overall fruit flavour
Dark berries flavour
Red berries flavour
Mint/Eucalyptus flavour
Fresh green
flavour
Chocolate flavour
Astringent
Bitter
Fruit after taste
PC1 (48%)
PC2 (32%)
1105
1106
1118
No MLFDark berries
Red berries
Jammy/Cooked fruit
Mint/Eucalyptus
Chocolate
Spice
Musty
Boiled Egg
Overall fruit flavour
Dark berries flavour
Red berries flavour
Mint/Eucalyptus flavour
Fresh green
flavour
Chocolate flavour
Astringent
Bitter
Fruit after taste
PC1 (63%)
PC2 (21%)PadthawayLanghorne Creek
Adelaide Hills Clare Valley
1105
1118
No MLF
Dark berries
Red berries
Jammy/Cooked fruit
Mint/Eucalyptus
Chocolate
Spice
Musty
Boiled Egg
Overall fruit flavour
Dark berries flavour
Red berries flavour
Mint/Eucalyptus flavour
Fresh greenflavour
Chocolate flavourAstringent
Bitter
Fruit after taste
PC1 (51%)
PC2 (33%)
1106
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Malolactic Fermentation Topics:
• Background
• Monitoring
• Selecting MLB Starter Cultures
• Yeast & Bacteria Interaction
• Timing of MLB Inoculation
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Competition or availability of nutrients
Negative effects of the yeast on the bacteria (inhibition) could be
caused by:
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Yeast Strain Nitrogen DemandJ.M. Sablayrolles, A. Julien INRA-IPV Montpellier, France
0
0.5
1
1.5
2
2.5
mg
N2/
g C
O2
rele
ased
71B
EC
1118
K-1
V11
16
L205
6
RC
212
S6U
CY3
079
BM
45 VL1
Relative Nitrogen Demand
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or... Production of inhibitory metabolites :
- SO2 (variable from one strain to another)
- Medium chain fatty acids- Anti-bacterial compounds
Negative effects of the yeast on the bacteria (inhibition) could be
caused by:
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0.0
2.0
4.0
6.0OSU
Bac
teria
(log
CFU
/mL)
end of alc.ferm.
0 10 20 30 40 Days
SO2
(mg/
L)
20
30
40
50
60
70
80
Wädenswil
71-B
EC-1118
K-1yeast strains:
Chardonnay juice:20°Brix, pH 3.3, SO2 17/35 mg/L at 20°C
Production of SO2 by yeast & effect on Oenoccocus
growth
Eno
logy
Gro
up
CornellUniversity
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YEAST / BACTERIA COMPATIBILITY MLF
compatibilityNitrogenNeeds yeast
POOR MEDIUM GOOD
LOWDV10
EC1118PM
ICV D21 / QA23 Rhone 4600
AMH / ICV D47VRB / 71B
MEDIUM SIMI WHITE*
RHSTBRL 97 / L2056
RC212 / R2 / S6UBDX*
ICV D254*CEG*
HIGH**BM45*
ICV K1*
T306*RA17*/ R2*
L2226*
CY3079*ICV D80*
* Rehydration in GOFERM / ** GOFERM + FERMAID after 1/3 of AF
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The result of many years of studying the interaction between yeast and bacteria …the perfect couple
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Malolactic Fermentation Topics:
• Background
• Monitoring
• Selecting MLB Starter Cultures
• Yeast & Bacteria Interaction
• Timing of MLB Inoculation
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107
105
104
103
102
106
10
Popu
latio
n (c
fu/m
l)
HARVESTTRANSPORT
AF MLF AGEING
108
SelectedML Bacteria
LYSOZYMESelectedYeast
Model Microbial Populations for Post AF inoculation of ML Bacteria…
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107
105
104
103
102
106
10
Popu
latio
n (c
fu/m
l)
HARVESTTRANSPORT
AF & MLF AGEING
108
SelectedML culture
LYSOZYMESelectedYeast
Co-inoculation
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When to Add Malolactic Bacteria (MLB)?
• Co-inoculation – Yeast addition followed 24-72 hours later by Malolactic bacteria addition
• Why co-inoculate?
– Want positive mouthfeel impact of MLF
– Want to avoid MLF aromas masking fruit character
– Limited tank space faster turn over of fermentors
– Helps the MLB adapt to more difficult wine conditions
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When to Add Malolactic Bacteria (MLB)?
• Co-inoculation – Yeast addition followed 24 hours later by Malolactic bacteria addition
– Safe to practice if…• pH < 3.5
• Reliable Alcohol Fermentation
• Post Alcohol MLB addition – Less risky
– Stylistic choice - more classical MLF sensory character impact
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Sensory descritors for a 2003 Riesling - MLF induced simultaneously, after alcoholic fermentation and no MLF
0
1
2
3
4
5buttery
lactic
acid
reducedbalanced
body
varietal fruitsimultaneousEC1118
post AFEC1118
no MLFEC 1118
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Lallemand MLF available resources…