Prevention of Wine Oxidation Potential Targets and Treatments

Varietal Flavor/Aroma Location Only a small number of the dozens of known grape compounds important for flavor are represented here. Potentially volatile compounds such as terpenes, norisoprenoids, and thiol precursors are stored as sugar or amino acid conjugates in vacuoles of grape skin cells. Lund and Bohlmann, Science 311, 804 (2006)

Flavor/Aroma Structures Norisoprenoids The oxidizable portion of flavor/aroma compounds are hydroxyl and keto groups Monoterpenes 4MMP 4 mercapto methylpentanol 3MMB 3 mercapto methylbutanol 3MH 3 mercaptohexanol 3MHA 3 mercaptohexan acetate Thiols

FRESHArom® Glutathione as an Antioxidant in Wine FRESHArom® is an OMRI Listed yeast derived product naturally rich in S- amino acids (NAC, Cystein, GGC, Glutathione) which create antioxidant power for aroma protection N-Acetyl Cysteine (NAC) Glutathione added directly to water or finished wine can be rapidly oxidized and with no Glutathione Reductase to recycle it loses antioxidant properties Cysteine ɣ - Glutamylcysteine (GGC) Glutathione FRESHArom® added during the first third of fermentation allows yeast to accumulate and slowly release glutathione during lees ageing - autolysis

FRESHArom Protection From Laffort Export Seminar 2014 – Palermo, Italy

FRESHArom Protection of Wine from Oxidation Winemaking Stages and Oxidation Risk Juice/Must – Glutathione Loss-GRP (Glutathione-Caftaric Acid) maintain reductive conditions, fermentation tannins, SO2, CO2 Fermentation – Low risk yeast oxygen demand, CO2 evolution, conjugated precursors Racking – Fining – Stabilization – Bottling- FRESHArom glutathione, SO2, best cellar practices Highest Risk

FRESHArom Protection FRESHArom is added during the first third of Alcoholic Fermentation Correct any nitrogen deficiency in the must during AF with proper nutritional supplements Add FRESHArom directly to the fermentation tank Saccharomyces can assimilate the Glutathione and precursor molecules efficiently at this stage of the fermentation Wine glutathione content is increased as yeast undergo autolysis FRESHArom aids in the protection of the aromatic potential of wines and delays the appearance of oxidized notes eg. Sotolon, phenylacetaldehyde FRESHArom also provides protection against the mechanisms of browning reactions in white and rose’ wines

Klosterneuburg - Austria Grüne Veltliner 2011 FRESHArom Protection Klosterneuburg - Austria Grüne Veltliner 2011 Parameters FRESHArom Control Alcohol (% Vol.) 12,2 12,1 GF (g/L) 0,9 0,8 VA (g/L H2SO4) 0,12 TA(g/L H2SO4) 4,12 4,04 pH 3,23 Malic acid (g/l) 1,75 1,74 FRESHArom protects the varietal and the fermentative aromas FRESHArom Control

FRESHArom FRESHArom FRESHArom FRESHArom

ARC Projects - 2015 Impact of FRESHArom on Redox Status in White Wines Foundation – Glutathione / Redox Research More reactive compounds protect less reactive compounds through kinetic preference

ARC Projects - 2015 Impact of FRESHArom on Redox Status in White Wines Feasibility – Glutathione Impact on Wine Ageing Impact of adding glutathione (15 mg/L) at bottling on the aldehyde-thiol aromas of Sauvignon Blanc after 4 years of ageing Control with GSH (15 mg/L added) Oxidative evolution (waxy and honey) Sotolon (µg/L) 19 6 Phenylacetaldehyde (ng/L) 216 125 Fruity Aromas 4MMPOH (ng/L) 15 350 3 MH (ng/L) 28 444 Grape GSH can be rapidly lost by oxidative juice handling - GRP GSH in yeast can be supplemented with a timely nutritional addition 15 ppm+ GSH is needed in wine for optimal protection

ARC Projects - 2015 Impact of FRESHArom on Redox Status in White Wines Experimental Design – Sampling – Measurables Control Treatment – Winery Standard Experimental Treatment – Modified Protocol (Harmonized) Must chemistry, Finished wine chemistry, Glutathione, A420, A280, Sensory Evaluation Complete procedure/processes used in producing the wines, storage conditions Must: End Primary Fermentation: Cellaring Glutathione Glutathione Glutathione Fe Fe A280 Phenolics A420 Color Sensory/Aroma Evaluation

Post-Fermentation Glutathione (mg/L) ARC Projects - 2015 Impact of FRESHArom on Redox Status in White Wines Preliminary Data – Glutathione Impact on Wine Ageing Post-Fermentation Glutathione (mg/L) Winery Varietal Yeast Control Treatment Fold Increase Sonoma-Cutrer Chardonnay VL2 5.4 15.6 2.9 5.6 16.6 3.0 X-Pure 5.7 15.0 2.6 6.6 15.5 2.4 J Lohr - Monterey Riesling VL1 9.2 11.6 1.3 Fresno State Cab Sauv Rose’ Rose' 10.7 22.9 2.1 Schug Carneros Sauvignon Blanc QA 23 17.8 21.4 1.2 Benziger - Sonoma VL3 24.0 30.8

ARC 2015 - FRESHArom Winery Type ID Fe Benziger Sauv Blanc Control Post F 0.6 Exp Post F 0.5 CSU Fresno Cab Sauv 15CSROS- Post F 0.4 15CSROS+ Post F J Lohr Riesling 15WRFG Control Post F 2.0 15WRFG Exp Post F 6.8 Schug SBHVV15 CTRL Post F SBHVV15200 Post F SBHVV15300 Post F 0.7 Sonoma Cutrer Chardonnay 15CHKTV025V No KMBS Post F 15CHKTV025VF No KMBS Post F 15CHKTV)25V KMBS Post F 15CHKTV025VF KMBS Post F 15CHKTV025x No KMBS Post F 15CHKTV025XF No KMBS Post F 15CHKTV025X KMBS Post F 15CHKTV025XF KMBS Post F

ARC 2015 - FRESHArom Winery Type ID ABS 420nm ABS 280nm Benziger Sauv Blanc Control Post F 0.055 0.855 Exp Post F 0.054 0.861 CSU Fresno Cab Sauv 15CSROS- Post F 0.107 0.918 15CSROS+ Post F 0.223 0.900 J Lohr Riesling 15WRFG Control Post F 0.067 0.897 15WRFG Exp Post F 0.056 0.875 Schug SBHVV15 CTRL Post F 0.073 SBHVV15200 Post F 0.074 0.856 SBHVV15300 Post F 0.075 0.866 Sonoma Cutrer Chardonnay 15CHKTV025V No KMBS Post F 0.105 0.792 15CHKTV025VF No KMBS Post F 0.106 0.789 15CHKTV)25V KMBS Post F 0.062 0.754 15CHKTV025VF KMBS Post F 0.064 0.753 15CHKTV025x No KMBS Post F 0.115 0.913 15CHKTV025XF No KMBS Post F 0.113 0.925 15CHKTV025X KMBS Post F 0.066 0.891 15CHKTV025XF KMBS Post F 0.071 0.886

GROUP 1 – ZYMAFLORE X-Pure ARC 2015 - FRESHArom Sensory Evaluation of FRESHArom Trial - Chardonnay Wines – Sonoma - Cara Morrison, Zidanelia Arcidiacono, Lindsey Otis, Peter Salamone @Sonoma Cutrer Winery April 7, 2016 Forced Rank Evaluation for Aroma, Mouthfeel/Flavors, Color/Other GROUP 1 – ZYMAFLORE VL2 Trial points rank 025 VF 7 1 025 V 7 2 028VTF 10 3 028 VT 16 4 GROUP 1 – ZYMAFLORE X-Pure 025 XF 6 1 028 XTF 9 2 025 X 11 3 028 XT 14 4 V = Zymaflore VL2 X = Zymaflore X-Pure F = Fresharom T = Tanin Galalcool

Sample Winemakers Acute Tasters ARC 2015 - FRESHArom Sensory Evaluation of FRESHArom Trials - ARC Meeting - Petaluma - 23 Tasters - 20 Winemakers – 8 Acute Tasters (fail false positive, 2 differences) Pairwise Difference and Preference Testing Sample Winemakers Acute Tasters S-C CH-25 VL2F 73% 88% S-C CH-25 VL2 27% 12% FSU SEM VL2F 81% 86% FSU SEM VL2 19% 14% S-C CH-25 XF 47% 57% S-C CH-25 X 53% 43% JLohr WR VL1F 37% 57% JLohr WR VL1 63% 43%

FRESHArom effectiveness spans varietal and yeast strain ARC 2015 - FRESHArom FRESHArom Trials Summary - Conclusions Proper FRESHArom application can significantly increase the glutathione concentration in finished wines FRESHArom effectiveness spans varietal and yeast strain FRESHArom sensory difference in finished wines is noticeable and preferred especially by experienced wine tasters Next Question: How does FRESHArom application impact SO2 management in the cellar?