The potential of Saccharomyces

paradoxus as a commercial
wine starter yeast culture.

Angela Kristel A. Dalao

“While drinking wine to ‘gladden the

heart’ was acceptable, over-indulgence
most definitely was not”
- Proverbs, 20:1
 Wine
 Alcoholic beverage made
from the fermented juice
of fruits rich in sugar;
more commonly grapes.
 Wine
 Should not be confused with distilled liquors or
spirits.
 Wine Selection

1. Recommendations
from other people.

2. Type or variety.

3. Matches the food or the

meal.
Wine Connoisseurs…
 Color
 Aroma
 Flavor
 Body
Wine-making Process

1. Aging
2.
3.
4.
5.
6. Mechanical
Fermentation
Pressing
Racking
Bottling.
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sugar
crushing
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the end product.
 Wine Yeast Starter Culture
 Fermentation by yeasts
› Traditional fermentation
 Spontaneous

› Modern fermentation
 Commercial Starter Cultures

S. cerevisiae
 Saccharomyces paradoxus
 Member of the taxon Saccharomyces sensu
stricto group.
› S. cerevisiae, S. bayanus, S. pastorianus
› Isolated in fluxes of oaks and other broad-leafed tree
 Isolated from Croatian vineyards
 Used in making a fermented Mexican drink from
the sap of Agave.
 Objectives
 To determine specific desirable enological
characteristics of Saccharomyces paradoxus;
 To compare the enological performances of S.
paradoxus and S. cerevisiae under limiting
conditions; and
 To establish the potential of S. paradoxus as a
commercial wine starter yeast culture.
 Fermentation
 Alcoholic fermentation
› Conversion of sugars to ethanol and other by-
products.

 Malolactic fermentation
› Malic acid degradation.
 Alcoholic Fermentation
 Early stage of fermentation.
› Presence of natural microflora of grape skin.

Kloeckera sp.

Hanseniaspora sp. Candida sp.

 Alcoholic Fermentation
 Latter stage of fermentation.
› Presence of more ethanol – tolerant species.

Saccharomyces sp.
 Malolactic fermentation
 Biological deacidification
› Breakdown of malic acid which is one of the most
dominant acids in grape must.
› Produce buttery and creamy aromas.

 Traditionally done by adding lactic acid bacteria

at the latter stage of fermentation.
› S. paradoxus is capable of effective malolactic
fermentation.
 Wine Bouquet
 Depends on a balance of volatile
constituents such as acid, alcohols, and
esters and their threshold values.

 Affected by temperature at
fermentation.
 Wine Bouquet
 Esters
› Most important group influencing the aroma of wine;
fruity odor.
 Higher Alcohols (400 mg/L)
› Contributes to aroma and taste; affects wine
negatively above its threshold value.
 Volatile Acids (1.1 g/L)
› Adds complexity to wine; rancid and cheesy odor
above its threshold value.
 Wine Flavor
 Resulted from the complex winemaking process.

 Combination of different components such as

glycerol, tannins, residual sugars, alcohol, and
many more.
 Glycerol
 Provides sweetness and fullness to the wine.

 Produced in response to osmotic pressure,

released upon cell lysis caused by death.
 Wine Flavor
 Now, there is an increased demand for wines
with high glycerol and reduced ethanol content.

 Studies done in Australia and Korea resulted to a

higher preference for “sweet” wines.
› “Sweetness” is associated with the aroma.
Table 1.1. Wine style preferences in Australia

Wine style Level of importance Standard score

Shiraz 181 0.372

Cabernet Sauvignon 131 0.270

Wine from a particular region 129 0.265

Wine from a well known brand 106 0.218

Sauvignon Blanc 96 0.198

Cabernet / Merlot 72 0.148

Chardonnay 21 0.043

Rosé -132 -0.272

Red House Wine -162 -0.333

White Sparkling Wine -174 -0.358

White House Wine -269 -0.553

 Methodology
 Saccharomyces paradoxus and Saccharomyces
cerevisiae were introduced to artificial musts.

 After fermentation, highest population reached,

ethanol yield, residual sugar concentration, as
well as glycerol and acetic acid production were
determined.
 Limiting Factors
 Sugar content of the must
› Ratio of glucose and fructose

 Assimilable nitrogen content

› Cell metabolism
› Formation of organoleptic properties
 Table 1.2. Fermentations included in the factorial experimental design
(two yeast strains ・ four musts) used in the present work

Treatment Yeast strains Must composition

code
Sp – NG Saccharomyces paradoxus 300 mg/L assimilable nitrogen
100 g/L glucose + 100 g/L fructose
Sp – NF S. paradoxus 300 mg/L assimilable nitrogen
80 g/L glucose + 120 g/L fructose
Sp – SG S. paradoxus 50 mg/L assimilable nitrogen
100 g/L glucose + 100 g/L fructose
Sp – SF S. paradoxusS. Paradoxus 50 mg/L assimilable nitrogen
80 g/L glucose + 120 g/L fructose
Sc – NG Saccharomyces cerevisiae 300 mg/L assimilable nitrogen
100 g/L glucose + 100 g/L fructose
Sc – NF S. cerevisiae 300 mg/L assimilable nitrogen
80 g/L glucose + 120 g/L fructose
Sc – SG S. cerevisiae 50 mg/L assimilable nitrogen
100 g/L glucose + 100 g/L fructose
Sc – SF S. cerevisiae 50 mg/L assimilable nitrogen
80 g/L glucose + 120 g/L fructose
 Microbiological analysis
 At variable time interval, must samples were
taken, diluted, and plated on yeast extract agar.

 The plates were incubated at 25o C for 48 hours.

 Chemical analysis
 Final ethanol content, volatile acidity production,
and residual sugar content in the must were
quantified according to the Official EU Methods
for wine analysis.
 Chemical analysis
 Ethanol content
› Fractional distillation
 Volatile acidity
› By titration of the volatile acids separated from the
wine by steam distillation and titration of the distillate
 Residual sugar content
› By determination of glucose and fructose content
using an enzymatic method
 Chemical analysis
 Glycerol concentration was determined with an
enzymatic ⁄ colorimetric commercial kit
especially designed for wines.
 Results …
Figure 1. Growth ⁄ decay
plate count data (red) and
glycerol production (blue)
of (a) Saccharomyces
paradoxus and (b)
Saccharomyces cerevisiae
in NG must (300 mg l)1 of
assimilable nitrogen; 100
g/L glucose + 100 g/L
fructose).
Glycerol production
Treatment Code Glycerol Production (g/L)

Sp – NG 6.846
Sp – NF 6.676

Sp – SG 3.763

Sp – SF 4.394

Sc – NG 4.785

Sc – NF 4.171

Sc – SG 4.850

Sc – SF 4.447
 Final production of ethanol (%), volatile acidity (g/L) and residual
sugars (g/L) for the different fermentations

Treatment Code Ethanol (%) Volatile Acidity (g/L) Residual Sugar (g/L)

Sp – NG 10.70 0.230 0.333

Sp – NF 10.82 0.140 0.433
Sp – SG 11.35 0.290 0.466
Sp – SF 11.60 0.176 0.366
Sc – NG 11.15 1.140 0.400
Sc – NF 11.60 0.766 0.400
Sc – SG 12.10 1.066 0.466
Sc – SF 11.70 1.072 0.433
 On average…
Parameter S. paradoxus S. cervisiae

Population Higher Lower

Glycerol production Higher Lower

Ethanol production Lower Higher

Volatile acid production Lower Higher

Residual sugar left Lower Higher

 Summary
 It was seen that S. paradoxus strain possesses
enological properties of interest for the wine
industry, such as significant higher synthesis of
glycerol and lower production of volatile acids
than S. cerevisiae.
 Conclusion
 The enological characteristics shown by
Saccharomyces paradoxus makes it a better
alternative to S. cerevisiae as wine starter culture
based on the current winemaking trends.
THANK YOU!