Assessments of Microbial Activity in Wine Linda F. Bisson Department of Viticulture and Enology, UCD December 13, 2013
Dec 17, 2015
Assessments of Microbial Activity in Wine
Linda F. BissonDepartment of Viticulture and Enology,
UCDDecember 13, 2013
Post-Fermentation Microbial Activity• Turbidity• CO2 Production
• Film Production• Polysaccharide Production• Positive Character Production• Negative Character Production
– Odor– Taste– Color– Sediment– Haze
Mechanisms to Assess Post-Fermentation Microbial Presence
• Microscopically• Appearance of Wine
– Turbidity– Effervescence– Films
• Analysis of Metabolic Activity
The Types of Impacts of Microbial Metabolites
• Positive character synthesis• Negative character synthesis• Modify existing character• Enhance or amplify perception of existing characters• Diminish or buffer perception of existing characters• Matrix effects: change in chemical properties
Positive Contributions to Aroma
• Direct synthesis• Release of bound molecules• Autolysis factors• Indirect (chemical) effects• Matrix effects
Types of Microbial Transformations of Flavor Compounds
• Primary roles– Production of flavor compounds de novo from
nutrients– Liberation of grape flavor components from
precursors• Secondary roles
– Provide chemical reactants – Enzymatic modification of grape/oak flavors– Impact Redox status and buffering capacity
Microbial Components Impacting Wine Flavor
• Metabolites• Enzymes• Catalysts• Mannoproteins and Polysaccharides
Release of Bound Molecules
• Thiols in Sauvignon blanc– Cysteine conjugates– Carbon sulfur lyase activity
• Terpenes– Β-Glucosidase activity
• Norisoprenoids– Β-Glucosidase activity
Autolysis Factors
• Release of active enzymes: hydrolases• Release of cellular macromolecular
components• Breakdown of cellular macromolecular
components
Indirect (Chemical) Effects
• Creation of reactive molecules– S-containing compounds– Diacetyl– Aldehydes
• Alteration of pH• Alteration of Redox status of juice
Matrix Effects
• Alteration of solution chemistry• Production of flavor enhancers
– Dimethyl sulfide
• Production of masking factors– Fusel family– Hydrogen sulfide
Post-Fermentation Microbiota
• ML Fermentation• Surface films• Brettanomyces spoilage• Yeast spoilage
Monitoring Lactic Acid Bacterial Activity in Wine
• Measure decrease in malate levels• Assess change in pH• Note presence of effervescence• Follow increase in wine turbidity• Evaluate aroma profile
Monitoring Lactic Acid Bacterial Activity in Wine
• Measure decrease in malate levels– Will tell you when it is done
• Assess change in pH– Something is happening, will not tell you it is complete
• Note presence of effervescence– Something is producing CO2
• Follow increase in suspended wine turbidity– Something is growing in the wine
• Evaluate aroma profile– Characteristic compounds of lactic acid bacteria
• Diacetyl• Acetic acid• Off-characters
ML Fermentation• Monitor disappearance of malate
– Paper chromatography– Enzymatic
• By hand• Automated
– Spectroscopy• UV/VIS• FTIR (Fourier Transform Infrared)
– Photometric analysis
Note on Measurements
• Some methods are direct– Interference is minimal– Misidentification of compound rare
• Some methods are indirect– Need to make sure peak is correctly being
identified– Interference can be a problem– Misidentification can occur if unexpected
components are present
Lactic Acid Bacteria Off-Odors
• Mousiness• “Poo” off- notes• Animal characteristics: fur, meaty• Floral taints: geranium• Metallic• Vomit• Rancidity
Mousiness
Several compounds(oxidation products of lysine) have been implicated in this off-character:
2,4,6-trimethyl-1,3,5-triazine
2-ethyl-3,4,5,6-tetrahydropyridine
2-acetyl-3,4,5,6-tetrahydropyridine
Monitoring Lactic Acid Bacterial Activity in Wine: Best Practices
• Microscopic analysis• Aroma analysis• Taste analysis• Monitor malate levels
Surface Films
• Bacteria or Yeast • Most Often Both• Off-Character Production• Film Microbiota Can Produce Copious
Amounts of Off-Characters Due to Availability of Oxygen
Brettanomyces
• Problem in barrels• Can monitor 4-ethyl phenol or 4 ethyl guaiacol
formation by nose or analytically• Some wines suppress odors from these
compounds; the mechanism is not understood• Monitor via microscopy; sometimes
Saccharomyces looks like Brettanomyces• Genomic/molecular monitoring of presence
Spoilage Yeasts
• Use residual sugar• Turbidity and effervescence in bottle• Zygosaccharomyces• Saccharomyces