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nonnon--SaccharomycesSaccharomyces winewine yeastyeast
Annicka Bunte 16 June 2009
History of yeast starters
First starter culture: Müller-Thurgau 1890First commercial wine yeast starter: California1964-65Today: 90% starter fermentations in California[Mortimer(2000) Genome Res. 10 (4): 403-409 ]
Process consistency gainedat the expense of some positive wine qualities
Lost qualities associated with ‘‘wild’’ yeast
FRUIT.
COMPLEXITY.
MOUTHFEEL.
Reintroducing the good “wild” characters
Aiming to reintroduce fruit, complexity and mouthfeel likespontaneous fermentations
Keeping process control
Screening of non-Saccharomyces isolates
Laboratory and field trials ensuring good vinification
Safety aspects tested (low urea/ethyl carbamate)
Sensoric impact evaluated in field trials
Chr Hansen screening of wine related ‘‘wild’’ yeasts started in the 1990’s
Brettanomyces / Anamorph DekkeraCandidaCryptococcusDebaromycesHanseniaspora / Anamorph KloeckeraHansenulaKluyveromycesTorulasporaMetschnikowiaPichiaRhodotorulaSaccharomycesSaccharomycodesSchizosaccharomycesZygosaccharomyces
Saccharomyces drives the alcoholic fermentation
0102030405060708090
100
0 5 10 15 20
Time (days)
Glu
cose
/fruc
tose
(g
/L)
1,E+03
1,E+04
1,E+05
1,E+06
1,E+07
1,E+08
Log
CFU
(/m
l)
Glucose Fructose CFU
0102030405060708090
100
0 5 10 15 20
Time (days)
Glu
cose
/fruc
tose
(g/L
)1,E+03
1,E+04
1,E+05
1,E+06
1,E+07
1,E+08
Log
CFU
(/m
l)
Glucose Fructose CFU
S. cerevisiae T. delbrueckii
sugarconsumed
residualsugar
non-Saccharomyces mixed starters
Saccharomyces yeast needed to finish fermentationnon-Saccharomyces yeasts and Saccharomyces yeast shouldferment under similar conditions2003 big scale field trials2004 first non-Saccharomyces containing starter culturesintroduced by Chr Hansen
Temperature optimum similar, temperature tolerance differs slightly.
Torulaspora delbrueckiiOptical density (620 nm)
00,020,040,060,080,1
0,12
10 15 20 25 30 35 40
Temperature (°C )
Max
. sp
ecif
ic g
row
th
rate
(h
-1 )
Saccharomyces cerevisiae
Optical density (600 nm)
0
0,05
0,1
0,15
0,2
0,25
5 10 15 20 25 30 35 40
Temperature (°C)
Max
. sp
ecif
ic g
row
th r
ate
(h-1
)
HARMONY.nsac & MELODY.nsac
2 Blends of:
Torulaspora delbrueckii (Td)Kluyveromycesthermotolerans (Kt)Saccharomyces cerevisiae (Sc)
20%mild effect
40%strong effect
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
HARMONY MELODY
HARMONYHARMONY MELODYMELODY
SYMPHONY.nsac & RHYTHM.nsac
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
SYMPHONY RYTHM
2 Blends of:
Kluyveromycesthermotolerans (Kt)Saccharomyces cerevisiae(Sc)
20%mild effect
40%strong effect
SYMPHONYSYMPHONY RHYTHMRHYTHM
Example fermentation Cabernet sauvignon, HARMONY.nsac
D80%MERIT/10%T.delb/10%K.th
1,E+00
1,E+02
1,E+04
1,E+06
1,E+08
0 2 4 6 8 10Days
Log
CF
U/m
l
0
5
10
15
g/L
ØA ØA 37°C LysinGlucose/10 Fructose/10 EthanolGlycerol Figure 4
non-Saccharomycesnumbers declineover time, like in spontaneousfermentation
Saccharomycesdrives thefermentation
Mixed starters vs pure Saccharomyces cultures
0%
40%
80%
120%
160%
200%Acidity
Fruitiness
Body
Bitterness
Lemon
PineapplePeach
Appel
Honey
Mango
Green beans
3001 3004 3005 3006 3007
½
ff
Data kindly provided by University of Neustadt, Germany and E: Begerow GmbH & Co, Germany
S. cerevisiae/
T. delbrueckii/
K. thermotoleransh
Experience with starters
Improves mouthfeel and smoothness / roundness of winesIncreases longevity of flavours and aromas
Wineries interested in experimenting with own ratios, inoculation schemes and favourite Saccharomyces cerevisiaecommercial strains
PRELUDE.nsac single strain Torulaspora delbrueckii
Saccharomyces drives alcoholic fermentation: growth
Prelude
Saccharomyces drives alcoholic fermentation: sugar
Prelude
Saccharomyces drives alcoholic fermentation: ethanol
Prelude
non-Saccharomyces contribute little to SO2
Day
0, F
SO2
Day
0, T
SO2
Day
9, F
SO2
Day
9, T
SO2
M ERIT.ferm
T. delbrueckii
K. thermotolerans02468
1012
SO2 (ppm)
Day and SO2
yeast culture
FSO2 & TSO2 produced by single strain cultures 9 days after inoculation
MERIT.ferm
T. delbrueckii
K. thermotolerans
Prelude
non-Saccharomyces made little difference on standard fermentation parametres (field trials 2003)
-0.8
-0.6
-0.4
-0.2
0
0.2
0.4
-0.6 -0.4 -0.2 0 0.2 0.4 0.6 0.8 1.0 1.2 RESULT1, X-expl: 8%,8% Y-expl: 37%,14%
A
BCDW1
W2
W3
W4
W5
W6
W9
W11
W12
W13
W14
W15
Cabernet s
Chardonnay
Pinot grisPinot noirRiesling
Syrah
Nr of days
EtOH
TA
VA Lactic
F SO2
T SO2
RS
Citric aciSuccinic a
Tartaric a
Color Inte
Glycerol
PC1
PC2 X-loading Weights and Y-loadings
S. cerevisiae drives the alcoholic fermentation: field trials 2003
Finished first Finished last Total No. reported trials
S. cerevisiae 4 1 16
80% NS 0 6 16
”PRELUDE” 0 1 15
”SYMPHONY” 3 1 14
Always use single strain Torulasporadelbrueckii (PRELUDE) with a commercial
Saccharomyces cerevisiae!
Torulaspora delbrueckii (PRELUDE) in Pinot blanc
Average 20%M1/80%T.delbrueckii
1,E+00
1,E+02
1,E+04
1,E+06
1,E+08
0 2 4 6 8 10 12 14 16Days
Lo
g C
FU
/ml
0
5
10
15
Subs
t/P
rod.
(g/
L
vol%
)
total cfu T.delbrueckii M1Glucose/10 Fructose/10 EthanolGlycerol
Fast MLF with non-Saccharomyces
non-Sacch
Select your yeast product
Saccharomycescerevisiae
Non-Saccharomycesspecies
HARMONYHARMONY
RHYTHMRHYTHM
mild ‘wild effect’
strong ‘wild effect’
Palate weightstrong effect
High sugarcontent
MERITMERIT PRELUDEPRELUDE
20% 40% 100%0%
80% 60% 0%100%
MELODYMELODY
SYMPHONYSYMPHONY
100% Torulaspora delbrueckii
To be used with your favorite Saccharomyces cerevisiae
Gives rounder, smoother mouthfeel
Extend palate weight and flavors
Used in white/ rosé and premium red wines to get additional complexity
ALWAYS use your favorite Saccharomyces cerevisiæ in addition to PRELUDE.nsac
PRELUDE.nsacNEW!NEW!
Experience with PRELUDE.nsacSouth African field trials 2009
Chardonnay: Inoculation of PRELUDE.nsac 5 days prior to Saccharomyces cerevisiae (commercial strain)
“Remarkable” increased mouthfeel, palate weight and creaminess.Impact on red wines predicted to be very beneficialEffect similar to addition of mannoproteins
Sauvignon blanc: Inoculation of PRELUDE.nsac 2 days prior to Saccharomyces cerevisiae (commercial strain)
Preferred the PRELUDE.nsac treatmentImproved mouthfeel and lenght of flavours in the palateReduced VA notesMaintain Sauvignon blanc aromas produced by Saccharomyces
Experience with PRELUDE.nsacSouth African field trials 2009
Chardonnay and Sauvignon blanc: PRELUDE.nsac pre- and co-inoculated with S. cerevisiae (commercial strain)
PRELUDE.nsac wines increased mouthfeel and roundnessAllows Saccharomyces cerevisiae to express flavour and fermentation characteristicsCo-inoculation preferred by most
Shiraz trials with non-Saccharomyces single strainEnhanced varietal character, enhanced colour and great mouthfeel, full bodiedEffect similar to addition of mannoproteins