On-line monitoring and control of fed-batch fermentations in winemaking. Michal Dabros & Olivier Vorlet

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1 On-line monitoring and control of fed-batch fermentations in winemaking Michal Dabros & Olivier Vorlet Summer School on Advanced Biotechnology HES-SO//VS

2 Haute École Spécialisée de Suisse Occidentale 2

3 Winemaking Common method of making white wine HARVEST CRUCH PRESS CLARIFICATION BOTTELING FILTRATION AGING FERMENTATION source : saq.com 3

agitation From a few days to a month https://www.

4 Fermentation step Traditionaly a batch process Mainly tanks of to L Without agitation From a few days to a month 4

5 Sugar Ethanol Batch fermentation process Yeast eat sugar release CO 2 and ethanol and die 5

6 Sugar Ethanol Hyperosmotic stress High sugar concentration Low sugar concentration Little yeast Lots of sugar Stressful state for yeast Acetic Acid Acetaldehyde Other metabolic pathways Lots of yeast Little sugar 6

Effect of global warming - Higher concentration of sugar in must (and thus also of alcohol) - Higher environmental temperatures during fermentation High alcohol levels during fermentation require

7 Effect of global warming - Higher concentration of sugar in must (and thus also of alcohol) - Higher environmental temperatures during fermentation High alcohol levels during fermentation require more alcohol-tolerant yeast strains. High sugar concentrations in must cause a stress response in yeast, which leads to increased formation of fermentation co-products, such as acetic acid. Mira de Orduña, R., Climate change associated effects on grape and wine quality and production. 7

8 Drosophila Suzukii - Observed in Switzerland since Significant damage to vineyards in Probably responsible for the increase of acetic acid in grapes Fredrik von Erichsen 8

9 Impact on wine quality High sugar level + Stressed yeast = Poor Wine Quality Source: winefolly.com 9

10 Sugar Ethanol Constant low sugar concentration Low sugar concentration Feed Fermentation tank Must tank 10

11 Batch vs. fed-batch: preliminary studies C. Frohman, D. Widmer, R. Mira de Orduña, Revue Suisse Viticulture, Arboriculture, Horticulture, 2014, 46 (3),

12 Prototype for fed-batch fermentations 1. In-line measurement of sugar 2. Control of must feed 12

13 Prototype for fed-batch fermentations 2) Fermenter 3) Must tank 4) Thermostats 5) Feed pump 6) Temperature probe 7) IR-ATR probe for sugar measurement Cornelius Keg 20L 13

14 Mechanical aspect Must Tank Hole for IR-ATR probe ISMATEC rotary piston pump Cornelius Keg 20L Fermentation Tank Aquarium pump 14

15 Which measurement mode do we need? Off-line analysis Manual sampling Analysis in another laboratory by other operators At-line analysis Manual sampling Analysis in the same laboratory On-line analysis Automatic sampling Automated analysis and processing In-line analysis Without sampling In process analysis (Tank or pipe) 15

16 Overview of on-line and in-line methods Classical methods Viscosimeter Conductimeter ph-meter Physical properties Spectroscopic methods NIR, MIR, FT-IR, UV-VIS, Raman NMR MS, MALDI-TOF Composition, Structure Optical methods Fluorescence quenching Photoacoustic Composition Scattering methods Light Scattering X-ray Scattering Chromatographic GC HPLC, UPLC Structure, Morphology Composition, Structure 16

17 On-line measurement with flow cell Flow through cuvette with classical spectrometer Z-type X-type For absorption spectroscopy ( UV-VIS, IR,..) Sample IN Sample OUT W-type For emission spectroscopy (Fluorescence,..) ATR Crystal Detector IR Beam ATR Flow cell 17

In-line measurement with probe Transmission probe Measure of liquids Measure both transmitted and backscattered light Internal reflections may limit the rangeuv, VIS, NIR Reflectance probe Measure of

18 In-line measurement with probe Transmission probe Measure of liquids Measure both transmitted and backscattered light Internal reflections may limit the rangeuv, VIS, NIR Reflectance probe Measure of solids, powders, turbid liquids Color measurement, Fluorescence UV, VIS, NIR ATR probe Measure of liquids, - - high concentration, turbid Ideal for samples with high optical density Crystal : Sapphire or Diamond UV, NIR 18

19 midir instruments used Perkin Elmer Spectrum 100 FT-IR with ATR Flow cell Mettler Toledo ReactIR IC10 with ATR probe Sample IN Sample OUT Detector ATR Crystal IR Beam 19

20 Measurement of sugar & EtOH Superposing spectra chemometric (PLS) calibration model 20

21 Contol of must feed rate e(t): control error = C(t) sugar,setpoint - C(t) sugar F(t): must feed rate (manipulated variable) 21

22 Adaptive PI Controller t F t F0 exp t KP e KI e t dt 0 F 0 : initial flow rate: 1 F0 X 0V0 Y X / S C Feed μ : specific growth rate of yeast K P, K I : proportional and integral gains 22

23 Result with PI Controller and on-line monitoring On-line measurement of sugars with a Perkin Elmer Spectrum 100 FT-IR spectrophotometer and ATR Flow cell 23

24 Pump speed Sugar Concentration Model Predictive Controller (MPC) Δt = 3 min X X X X X Error Setpoint 24

25 Concentration (g/l) Result with MPC and in-line monitoring Sugar Sepoint Sugar Concentration Ethanol Concentration Time (days) In-line measurement of sugars with a Toledo ReactIR IC10 spectrometer with ATR probe 25

26 Perspectives Scale-up (in time and in volume) Monitor the concentration of the secondary metabolites Implementation with improved fermentation conditions Assess impact on the final product quality Source: rantnow.com 26

27 Acknowledgments Prof. Ramón Mira Prof. Jean-Pascal Bourgeois Giorgio Genasci Lucien Blanchard Samuel Unterhofer Ludivine Andrey Steven Viollet HES-SO programme P1 27

ch +41 26 429 67 09 http://olivier.vorlet.home.hefr.

28 Thank you for your attention myswitzerland.com hes-so.ch

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