Control of alcoholic fermentation in winemaking : some new prospects. 15th Australian Wine Industry Technical Conference,

(1)

HAL Id: hal-01837717

https://hal.archives-ouvertes.fr/hal-01837717

Submitted on 6 Jun 2020

HAL is a multi-disciplinary open access archive for the deposit and dissemination of sci- entific research documents, whether they are pub- lished or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers.

L’archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d’enseignement et de recherche français ou étrangers, des laboratoires publics ou privés.

Control of alcoholic fermentation in winemaking : some new prospects. 15th Australian Wine Industry Technical

Conference,

Jean-Roch Mouret, Sumallika Morakul, Stéphanie Rollero, Jean-Marie Sablayrolles

To cite this version:

Jean-Roch Mouret, Sumallika Morakul, Stéphanie Rollero, Jean-Marie Sablayrolles. Control of alcoholic fermentation in winemaking : some new prospects. 15th Australian Wine Industry Technical Conference,. 15. Australian Wine Industry Technical Conference, Jul 2013, Sydney, Australia. �hal- 01837717�

(2)

Control of alcoholic fermentation in winemaking : some new prospects

J.R. Mouret, S. Morakul, S. Rollero, J.M. Sablayrolles UMR Sciences pour l’Œnologie, Montpellier, France

(3)

Alcoholic fermentation

Yeast

Grape - must

Control

Sydney - July 15 , 2013

(4)

Wine yeast characterization

(5)

Impact on :

. Fermentation kinetics Needs for nutrients Resistance to ethanol . Wine characteristics Aroma

Colour, acidity …

How to standardize ?

In which conditions ?

Significance of the tests?

Interest of dedicated tools . Some examples

(6)

About kinetics

1 1050 fermentations

Interest of on line monitoring

Small … but significant differences

Aguera et al., 2005

(7)

Nitrogen needs

dCO 2/dt (g/L.h) Added nitrogen (mg/L)

Nitrogen deficiency slow fermentations

Nitrogen addition

Regulation of the CO₂ production rate by addition of assimilable

nitrogen

Comparison in the same conditions (same fermentation kinetics)

Added nitrogen

Julien et al., 2000

(8)

Oxygen needs

How to measure oxygen needs ?

Oxygen deficiency sluggish ou stuck fermentations

CER max

Anaerobic conditions : ability to grow and ferment without oxygen

S2

De-aerated Chardonnay must

S11

Julien et al., 2000

(9)

Sydney - July 15 , 2013 8

In the future?

. A major challenge :

. to analyse complex criteria : typicity, aroma production … . using tests:

. mimicking industrial conditions, including variability . in standardized conditions

. New approaches :

. Genomic – post genomic – system biology approaches .

More and more affordable

. Not yet feasible for a large number of strains

. High throughput approaches

. M

ethods for phenotyping?

(10)

Sydney - July 15 , 2013 9

A new tool : continous multi stage bioreactor

- Mimics 4 stages of a batch fermentation

- Yeasts in a stable physiological state

(11)

10

On line monitoring of fermentation

(12)

11

Feasibility and interest

Precision – fermentation rate

Microbiological interest Technological interest

On line monitoring

CO₂, density, refractometry First industrial sensors

Database Remote control

(13)

12

Red winemaking

0 0,2 0,4 0,6 0,8 1 1,2 1,4 1,6 1,8 2

0 20 40 60 80 100 120 140 160

Time (h)

dCO2/dt (g/l.h)

Aguera et al. (2005)

Precision

(14)

13

On line monitoring of fermentation rate

New control strategies

Control tank by tank

taking into account the must variability

New control strategies

Detection of nitrogen deficiency

Best timing for nutrient additions

Control of the fermentation rate

(temperature)

(15)

14

Kinetic model

. Physiology . Validated

. Not suitable for sluggish fermentations

. Must composition . Temperature profile

. Fermentation kinetics

. Energy requirement

Malherbe et al., 2004; Colombie et al., 2005, Colombie et al., 2007, Goelzer et al., 2009

Thermal model

Modeling. Prediction of fermentation kinetics

(16)

Nass = 170mg/l

Which effect of increasing temperature (16 vs 16-22)?

27%

36%

(17)

Which effect of adding nitrogen (400 mg/l DAP) ?

Nass = 100mg/l Température: 20°C

38%

(18)

Monitoring and control of marker molecules synthesis

Example of esters and higher alcohols

(19)

18

Higher alcohol and esters

Markers of :

. aroma compounds : volatility

. metabolism : key pathways of carbon and nitrogen metabolism

Metabolic pathways well known but : . complex regulations

. example : no direct relationship between must nitrogen composition and higher alcohol synthesis

Needs for new approaches to better describe, understand and control

. Improvement of monitoring on-line monitoring

. Gaz - liquid transfer production, accumulation and losses

(20)

Cold trap - GC Heated lines

Valve selector

16 compounds

Maximum frequency analysis : 1 h

On line monitoring

(21)

Compound name Metabolic or flavor interest Aldehyde Acetaldehyde non flavor compound but key

metabolic intermediary Propanol

Isobutanol Isoamyl alcohol 2-phenyl ethanol

Isobutyl acetate Isoamyl acetate Phenyl ethyl acetate

Ethyl acetate non flavor compound but the most abundant ester in wine

Ethyl butyrate Ethyl hexanoate

Ethyl octanoate Ethyl decanoate Ethyl dodecanoate

Hexyl acetate Diethyl succinate Higher alcohols

Esters

esters from the metabolism of the keto-acids

esters from the metabolism of the fatty acids

other esters

higher alcohols from the metabolism of the keto-acids

(22)

0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

100%

0 50 100 150 200

Time (h)

Arbitrary Units

Ethyl acetate

High reproducibility

Average error < 7% for 15 compounds

(23)

. High frequency analysis

Ethyl acetate

0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

100%

0 20 40 60 80 100 120 140 160

time (h)

Gasphase (arbitrary units)

production global rate exponential specific rate

Exponential phase

Ethyl acetate Ethyl acetate

Ethyl acetate

0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

100%

0 20 40 60 80 100 120 140 160

time (h)

Ethyl acetate

0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

100%

0 20 40 60 80 100 120 140 160

time (h)

Exponential phase

Ethyl acetate Ethyl acetate

. Rate of production . Metabolic flux

Rate Specific rate

(24)

Relation between an ester and its precursor higher alcohol

Isobutyl acetate = f(isobutanol)

0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

100%

0% 10% 20% 30% 40% 50% 60% 70% 80% 90% 100%

Isobutanol

Isobutyl acetate

(25)

Concentrations in the liquid phase

Interest of measurements in the gas phase

C° liquide C° gaz

+ Yeast metabolism

Losses : importance for control

Gaz-liquid partitioning

Organoleptic interest

Gas – liquid partitioning

(26)

25

Gas – liquid balance : production, accumulation, losses

Gas – liquid balance

Ethanol

On line monitoring

C ° gas

dCO

₂

/dt

Temperature

Model C ° liquid

(27)

26

Relative losses (% of total production)

Very important losses for ethyl hexanoate

Losses

(28)

27

Ethyl hexanoate

Isobutanol

Anisothermal fermentation 15 ° C – 30 ° C

Temps (h)

Balance of production, accumulation and losses

(29)

28

Anisothermal fermentation 15 – 30 ° C

End of accumulation (liquid)

Increase of losses throughout fermentation (> 90% at the end) Important to optimize temperature profiles

Ethyl hexanoate

Time (h)

Loss rate/ production rate

(30)

Effect of temperature and environmental parameters

(31)

Effect of temperature:

 Higher alcohols: increase at high temperature (not systematic)

 Esters: decrease at high temperature (especially for ethyl esters)

Effect of nitrogen

 Higher alcohols: maximal production between 200 and 300 mg/L of N_ass

 Esters: increase at high N_ass(acetate and ethyl esters)

Effect of lipids (solid particles) and/or oxygen

 Higher alcohols: increase when addition of oxygen and/or lipids

 Esters: complex, effect on the ratio acetate/ethyl esters)

Effect of temperature and nutrients

Beltran 2005, 2008; Jimenez-Marti 2006; Mauricio 1997; Miller 2007; Molina 2007 ; Swiegers 2005;

Torrea 2011; Vilanova 2007; Varela 2012

+ effect of yeast strain and interactions

(32)

Box-Behnken

Combined effects

(2 strains)

(33)

Température fixée Azote fixé

Propanol

Only effect of nitrogen

Same response for the 2 strains

(34)

Phenyl ethanol

Optimum when 24°C, 180 mg /l nitrogen , 5 mg/l lipids

Difference between strains = f(conditions)

(35)

Isoamyl acetate

Major effect of nitrogen

Low effect of temperature and lipids

Difference between strains = f(conditions)

(36)

Température fixée

Azote fixé

Ethyl hexanoate

Major effect of nitrogen

Intersection between the 2 strains

(37)

36

About temperature

Is the effect of temperature on the final

concentrations of esters in wine due to microbial or physico-chemical phenomena?

Is the higher concentration at low temperature due to a higher production or is it due to lower

losses?

(38)

Risk of important overestimation of the temperature effect.

x 2.5 x 1.3

37

Ethyl hexanoate

(39)

Partly the same effect than lipids

Control of oxygenation – difficulties :

Small amounts (several mg/l) During fermentation

‘Useful oxygen’ = oxygen transferred into the medium

Oxygen additions : firstly to limit risks of stuck fermentations …

About 10 mg/l

Best moment : end of celle growth (about 20% of fermentation)

… but also to modify wine aroma

needs for additional studies needs for new tools , e.g.,

possibility of very slow addition (transfer)

addition = f(time) or f(fermentation progress), e.g. 1 mg/l/% ethanol

About oxygen

Sydney - July 15 , 2013 38

(40)

Sydney - July 15 , 2013 39 dCO2/ dt (g.L-1.h- 1 )

Effect of adding 10 mg/l O₂ - in 1 h

- in 40 h

Different effects on kinetics and ester production

Farines et al., 2013

(41)

Conclusion

(42)

41

On line monitoring of fermentation

Understanding of metabolism

On line monitoring of quality markers

Effect of environmental parameters

New control

strategies

^Kinetics

Characteristics

Wine yeast characterizatoin

(43)

Sydney - July 15 , 2013 42

(44)

43

Many thanks to

^:

Evelyne AGUERA Magaly ANGENIEUX Violaine ATHES

Vincent FARINES Pamela NICOLLE Marc PEREZ Christian PICOU Christian TRELEA

The research leading to these results has received funding from the European Community's Seventh Framework Programme (FP7/2007- 2013) under grant agreement CAFÉ n° KBBE-212754