Yeast fermenta+on management

Stuck fermenta+on
YEAST GROWTH
 Yeast growth cycle : three main
Cellules/mL
phases

1 2 3 4 5

temps
1 : Lag phase : adapta+on of cells to the medium
2 : Start Phase : the growth starts
3 : Exponen+al phase : constant mul+plica+on kine+c
4 : Sta+onary phase : compounds necessary for growth disappeared
5 : Decline phase : mortality rate increased, autolysis
Yeast growth cycle during alcoholic fermenta+on

Fermented sugar

Total yeast popula+on

Viable yeast popula+on

 Yeast growth cycle during alcoholic fermenta+on

•Growth is limited to 4 to 5 generations

•The growth stop is not the result of a disappearance of energy
nutrients
•The duration of the different phases is not equal (death phase >>>
growth phase)
•Stationary phase : low activity of the yeast but high viability
•Fermentation kinetic is directly correlated with the growth cycle

Ø AF kinetic f (% nitrogen, yeast strain, AF temperature)

 Grape must is a highly fermentable
medium
-Carbon source (glucose and fructose)
-Nitrogen source
Total nitrogen:
-3-10%=amonium YAN = Yeast Assimilable Nitrogen
-25-30% = amino acids Deficient when <130-140 mgN/L
-25-40% = polypep+des
-5-10% = proteins
 Grape must is a highly fermentable
medium
-Carbon source (glucose and fructose)
-Nitrogen source
-Vitamins (thiamine, bio+ne, panthotenic acid, nico+nic
acid, …
-Mineral salts (phosphate, sulfate, potassium, calcium,
magnesium)
one strategy for introducing micronutrients and vitamins is by directly making
them available to the yeast in the rehydra+on water
Fermenta(on ac(vators
Growth factors
Yeast Mul+plica+on and cellular
ac+vity
metabolism

Vitamines
Mineral salts

Ammonium salts, organic nitrogen

from inac2vated yeasts, vitamins

Nitrogen addi+on : 1. beginning of yeast cell growth

2. end of yeast cell growth
 Fermenta(on ac(vators

Survival factors
End of AF
Synthezised by yeast in presence of
oxygen

Sterols
Long chain fa_y acids

Yeast cell walls, hydrolyzed

Op+mal +me to aerate or add
oxygen (4-6 mg/L): beginning of cell
yeast extract
growth, in prac+ce consump+on of
15-30 g/L sugar
Fermenta(on ac(vators
 Main inhibitors of the
alcoholic fermentation
High level of sugars/ ethanol toxicity
n The kine+cs of sugar u+liza+on by S. cerevisiae is
largely driven by sugar transport
n Glucose is typically consumed as a faster rate than
fructose
n A high concentra+on of alcohols : fructose use is
compromised, thus resul+ng in stuck fermenta+on

èaerobic survival factors to maintain the yeast

membrane fluidity = aera+on or oxygen addi+ons
 Main inhibitors of the
alcoholic fermentation
n Under or over clarified juice at low
temperatures
n Juice clarification removes materials that may be a
source of grape sterols and long-chain fatty acids =
nutritional value for yeast
n Juice turbidity should be adjusted at 150-200 NTU
 Main inhibitors of the
alcoholic fermentation
n Critical temperature during AF; heat shock
n Lack of oxygen
n Lack of nutrients (nitrogen, vitamines, …)
n Inhibition phenomenon (short chain fatty
acids, pesticides residues, …)
n Yeast/yeast and yeast/bacteria interactions
 Difficult fermenta+on condi+ons
Tall and narrow tanks
– CO2 helps to keep the yeast cells in suspension
– However, some cells tend to se_le (end of AF) ;
leaving fewer cells in contact with sugars
• In prac+ce : it is recommended to re-suspend
the yeast frequently
– S+rring by hand in barrel
– Deep punching down in open tanks (delestage)
– Nitrogen or air addi+on, …
 How to prevent stuck fermenta+on
• Daily monitoring tank density and temperature (to avoid
sudden shifs as well as temperature extremes)
• Nitrogen supplementa+on if necessary (190-200 mg/L) (with
thiamine)
• Adding oxygen at the start of fermenta+on during the growth
phase of yeast fermenta+on
• The use of yeast hulls : fixing by-products inhibitors (toxic
fa_y acids

For white wine vinifica+on :

– adjust the turbidity level (100-200 NTU), temperature
For red wine vinifica+on:
– temperature (to start alcoholic fermenta+on)
– in the cap
 How to prevent stuck fermenta(on

Inocula+on with Ac+ve Dry Yeast (ADY)

– Rehydra+on = cri+cal in order to avoid fermenta+ons
problem
• 500 g of ADY into 5 L of warm water or water with
sugars
• Afer 5 min : s+r the suspension
• Temperature: 35-40°C, dura+on: 20-45
• Inocula+on rate : 5.106 cfu/mL or 15-20 g/hL
– Hygiene : inoculate the must with ADY as soon as
possible
-White wine vinifica+on : avoid cold-shocking the yeast by
adjus+ng the yeast suspension temperature
-Red wine vinifica+on : through
pumping over, in the cap (lipids)
Stuck fermenta(ons

èHigh risk of bacterial spoilage

 Bacterial spoilage

• Lactic acid bacteria spoilage

– Volatile acidity : after AF and MLF: normal level : 0.36 à 0.48 g/L expressed
in acetic acid
– EEC limits: white and roses wines: 1.07 g/L expressed in acetic acid ; red
wines: 1.20 g/L expressed in acetic acid
– If VA<0.72 g/L expressed in acetic acid, not easily detectable

Alcoholic fermentation
Yeast

Malolactic fermentation
Bacteria
 Bacterial spoilage

• Acetic acid bacteria

– Due to a large surface contact between air and wine : major increase of volatile
acidity :
Ethanol è acetic acid è ethyl acetate = acescence
– Ethyl acetate: perception threshold around 150 mg/L ; no impact on flavour at
concentration below 120 mg/L but if > 160 -180 mg/L : affect the after taste and
accentuates an impression of harshness
• Related to wine storage conditions
– Containers must be kept full and perfectly sealed with airtight bungs
– Temperature (spoilage is twice rapid at 23°C (73.4°F) than at 18°C (64.4°F)) ; optimum
storage temperature is at 15°C (59°F)
– Cleanliness and proper maintenance of all containers
Acescence : easy to avoid by taking basic precautions
 What to do in case of stuck
fermenta+on?
• When density is stable during 48 to 72 hours…
• For red wines:
– Wines should be drained rapidly = separate the juice
from the cap (rich in bacteria) even if the skin and
seed macera+on is not complete
– Elimina+on of bacterial contamina+on source, oxygen
is adding, temperature decrease
– In some case fermenta+on restarts spontaneously
• For white wines:
– Lightly sulfited, use of yeast starter
Restarting a fermentation…
Pied de cuve implementation

5) Starter is ready to use

tanck
100 hL 4) Monotoring
sugars
when < 2 g.L-1 20°C

1) 5hL stuck fermenta(on wine

2) 10-11% alcohol
Ø15-25 g.L-1sugars 3) + 5 hL stuck fermenta(on wine
ØSO2 addi(on
Ø5 g.hL-1 NH4+ salts
Ø10-20 g.hL-1 yeast cell wall
2) Monitoring sugars
when < 5 g.L-1
LSA

20°C
15-25 g.hL-1
Ethanol tolerant strain
 Restarting a fermentation…
• Should be closely monitored
– By measuring vola+le acidity
– Judicious sulfi+ng to prevent contamina+on
• Tanks with stuck fermenta+on must be
restarted as soon as possible
• It is preferable to wait un+l the following
spring