TU3 - Microorganisms
and fermentation
managment
Main factors that impact the
organoleptic characteristic of the
wine
Vinification
process
Terroir:
soil,
climatic
conditions, Wine
human ageeing
factors
Microorganisms
Fermentations managment
Current context of enological
practices
n Spontaneous fermentations are very popular ; organic wines
n Low level of SO2 wine without added sulfites
n Climate change
¨ Grape juice with high sugar content, high ethanol level = higher risk of
sluggish and stuck fermentations
¨ higher pH : major impacts on wine microbial stability (microbial diversity,
active SO2 )
Knowledge of the microbial diversity and dynamic is a key
issue to manage alcoholic fermentation and to prevent
microbial spoilage
Main microorganism’s families
PHYLLOSPHERE
AF MLF
Yeasts Lactic acid bacteria
GRAPE Oenococcus oeni
Saccharomyces
Yeasts 106 to107 UFC/mL
cerevisiae
Bacteria Brettanomyces
107 to 108 UFC/mL
Fungi Acetic acid bacteria
Non-Saccharomyces
104 to 106 UFC/g grapes 101to103 UFC/mL
102 to 106 UFC/mL
Lactic and acetic acid
bacteria
102 to 103 UFC/mL
The Vinification process
Winemakers:
managment of
Key role of microorganisms alcoholic fermentation:
SO2 addition,
temperature control,
inoculation, …
1) Alcoholic fermentation; yeasts
¨ Saccharomyces cerevisiae
¨ C6H12O6 è 2C2H5OH + 2CO2 + heat
2) Malolactic fermentation; lactic acid bacteria
¨ Oenococcus oeni
¨ Ac L-malique è Ac L-lactique + CO2
Two main strategies to manage
fermentations
n Spontaneous
¨ Inoculation of the grape must by the grapes and cellars
microorganism (Yeast)
¨ Œnococcus œni population growth at the end of alcoholic
fermentation and start of malolactic fermentation
n Use of selected starters
¨ Selected yeast (could be pure culture of Saccharomyces
cerevisiae, but also non-Saccharomyces / S. cerevisiae mixed
culture) : to better manage alcoholic fermentation, to prevent
wine spoilage, parameter of the final wine quality)
¨ Œnococcus œni malolactic starters : to prevent inadequate
indigenous population ; success not always ensured due to
the inoculation in wine
The Vinification process
Dramatic change of the Gradual change of the
medium composition medium composition
Semi-anaerobiosis, SO2 addition,
Semi-anaerobiosis, alcohol 13-
220-250 g/L of sugars
15%, î nitrogen, vitamins
Microbial diversity
« Microbial waves »
(Renouf 2006)
Population
level MLF
Cold soak AF
Non- Saccharomyces
F Oenococcus oeni
Saccharomyces Brettanomyces
Brettanomyces
Time
üDiversity microbial is decreasing
üSuccession of microorganism over time
Yeasts/bacteria – Species - Strains
Link with wine quality
n Alcoholic and malolactic fermentation
n Production of fermentative aromas
n Aromatic potential liberation : from precursor form to
volatile compounds
SH
Grape aroma OH
Odoriferous thiol
precursor S. cerevisiae
released in the wines of
numerous varieties
Murat et al. 2001 Am J Enol Vitic.
n Dead yeast= lees
n Autolysis = positive contribution : aroma stabilization,
wine limpidity, enhanced sweet taste
Link with wine spoilage
n Brettanomyces bruxellensis and the volatile phenols
n Could increased during post maceration and wine aging
¨ B : ETHYL-4-PHENOL : stables and horse’s sweaty
¨ A : ETHYL-4-GAÏACOL : smoky, spicy
¨ E-4-P/E-4-g ratio is on the order of 8:1
¨ Preference threshold: mixture 10/1 : 380 µg/L
¨ Consequences : mask the fruity character of the red wines (from the
concentration of 9µg/l)
n 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: solvent-like and nail polish remover aroma Sulfur dioxyde
¨ Related to wine storage conditions
easy to avoid by taking basic precautions
The outline
n Introduction to the wine microbiology
n Yeasts and AF
¨ Ecology, Taxonomy, life cycle, cell wall, killer phenomenon, …
¨ Metabolisms: alcoholic fermentation, sugar pathways and regulation, nitrogen
metabolism
¨ Aromas
Evaluation methods
n Stuck fermentation ✓Class participation
✓Take home exam
¨ Yeast growth and fermentation kinetics ✓Sensory analysis exam
n Starters
¨ Implementation
¨ Wine microorganisms selection
n Lactic acid bacteria and MLF
¨ Taxonomy, bacteria cell
¨ Metabolisms: malolactic fermentation, sugars, organic acids
n Wine defect and Spoilage Microorganisms
