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