Workshop Enologia em Vindima Impacto sensorial de la fermentacíon maloláctica en el vino Portugal June 2011 - Dr. Sibylle Krieger-Weber
Wine An experience Colour Aroma & flavour
Eveline Bartowsky, AWRI, 2009 Microbial metabolism
Microbial metabolism - flavour-active compounds From Swiegers, Bartowsky, Henschke & Pretorius, 2005 Eveline Bartowsky, AWRI, 2009
MLF & sensory interactions Malolactic fermentation ph, TA Softer mouthfeel L-malic acid L-lactic acid CO 2 + Microbial stability Potential microbial spoilage Sensory Fruity characters Buttery characters Vegetative characters MLF & certain strains of O. oeni can enhance the berry fruit aroma of red wine No MLF R1105 Strain A Spont Herbaceous Caramel Leathery Reduced Floral Fruit Flavour Intensity Plum Acidity Eveline Bartowsky, WAC Beaune 2011 Cabernet Sauvignon Berri 2002 Raspberry 2 4 6 Brown Hue Purple Hue Colour Intensity Bitterness Coarse texture after expectoration Coarse texture in mouth
Buttery aroma - Diacetyl O. oeni during MLF Derived from citric acid metabolism Aroma buttery, nutty, butterscotch CH 3 C=O C=O CH 3 1-4 mg/l = enhance flavour complexity > 5-7 mg/l = undesirable buttery aroma Eveline Bartowsky, AWRI, 2009
Diacetyl - strain VP41 VP41 BETA BETA Clare Valley Adelaide Hills Eveline Bartowsky, AWRI, Neustadt 2010
Diacetyl Cabernet Sauvignon - vineyard Adelaide Clare Valley Adelaide Hills ph 3.2 3.7 ph 3.2 3.7 Limestone Coast Clare Valley Langhorne Creek Padthaway Limestone Coast Diacetyl (mg/l) Eveline Bartowsky, AWRI, Neustadt 2010 Clare Valley Adelaide Hills Langhorne Creek Padthaway
Diacetyl - management during winemaking Diacetyl conc n Diacetyl conc n O. oeni strain variable temperature 18 C - higher 25 C - lower wine type white - lower red - higher SO 2 binds to diacetyl - sensorially inactive inoculation rate 10 4 - higher 10 6 -lower aeration air - higher anaerobic - lower fermentation time longer MLF - higher contact with yeast lees ph long contact- lower lower ph may favour Eveline Bartowsky, AWRI, Trier 2008
glucose citrate lyase citric acid oxaloacetic acid acetic acid aspartate aminotransferase aspartic acid lactic acid NAD NADH lactate dehydrogenase oxaloacetate decarboxylase CO 2 pyruvate dehydrogenase complex pyruvic acid acetylphosphate acetic acid α-acetolactic acid α-acetolactate decarboxylase CO 2 TTP pyruvate decarboxylase CO 2 acetaldehyde-ttp α-acetolactate synthase non-enzymatic decarboxylation diacetyl reductase CO 2 acetate kinase ATP DIACETYL NAD(P)H NAD(P) acetoin acetoin reductase NAD(P) NAD(P)H 2,3-butanediol Eveline Bartowsky, AWRI, 2004
Optional time points for MLF inoculation Pre AF Co-inoculation mid-af At pressing Post AF Grape sugar (%) 100 50 AF No MLF Grape vinification Eveline Bartowsky, AWRI, 2008
Fresh dark fruit aroma Raspberry 4 Smoky Savoury 3 2 1 0 Fresh dark fruit Cooked dark fruit Mixed spice Floral Cabernet Sauvignon 2006 Bordertown Fresh green Confectionary Total ester concentration correlates with sensory Eveline Bartowsky, AWRI, Neustadt 2010
Bacterial Metabolism of Acetaldehyde and other SO 2 binding compounds Ramón Mira de Orduña Dept. of Food Science & Technology, Cornell University
Current issues Legal SO 2 limits Organic wines Public perception Levels of carbonyls
Acetaldehyde and cancer
SO 2 Binding Compounds Microbial Acetaldehyde Glucuronic acid Alpha Ketoglutarate 5 oxofructose Pyruvate Gluconolactone Acetoin Glyoxal Glyceraldehyde Grape Galacturonic acid Glucose Fructose
How much SO 2 is bound? Frequency 1.0 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0 Estimation of bound fraction using dissociation constants F I M R W 82.8% ACHO 82.8% 1 Mean(Glucose % of Bound SO2) 2 Mean(Galacturonic Acid % of Bound SO2) 3 Mean(Alpha Ketoglutarate % of Bound SO2) 4 Mean(Pyruvate % of Bound SO2) 5 Mean(Acetoin % of Bound SO2) 6 Mean(Acetaldehyde % Bound SO2) Acetaldehyde Pyruvate Alpha-Ketoglutarate Galacturonic Acid 10.1% Pyr 10.1% 4.91% α-kg 4.91% 2.11% 2.11% G.acid Wine Type (White, Red, Mead, Ice, Fruit)
Acetaldehyde produced by different yeast species 120 90 60 30 1.2 0.8 0.4 120 80 40 0 A 0 3 6 9 12 15 120 90 60 30 ACHD (mg l -1 ) ACHD (mg l -1 ) B 0 3 6 9 12 15 0 1.2 0.8 0.4 OD600nm OD600nm Time (days) 120 80 40 Hexoses (g l -1 ) Hexoses (g l -1 ) Time (days) 5 4 3 2 1 A Yield coefficient (mg g -1 sugar) 120 80 40 80 60 S. cerevisiae C. stellata C. vini H. anomola H. uvarum M. pulcher Z. bailli S. pombe 40 20 Final (mg l -1 ) 0 B Peak (mg l -1 ) C Yeast species
3500 3000 2500 Acetaldehyde levels during MLF in Riesling Malate (mg/l) Acetaldehyde (mg/l) 2000 1500 1000 500 0 100 80 60 40 20 10 20 30 40 50 1032 1054 1075 1076 1077 1098 1101 1105 1106 1108 1118 1124 Control 0 0 10 20 30 40 50 Time (Days)
Metabolism of SO 2 Binders During MLF SO 2 Binding Compounds (g l -1 ) 4 3 2 0.12 0.10 0.08 0.06 0.04 0.02 0 20 40 60 160 Time (Days) Galacturonate α-kg Pyruvate Acetoin Acetaldehyde Malate
Acetaldehyde Kinetics During AF and MLF Alcoholic fermentation Malolactic fermentation Acetaldehyde [mg l -1 ] 100 80 60 40 20 0 5 10 15 20 25 30 35 40 Time [d] 0.3 0.2 0.1 OD 650 nm
Reduction of SO 2 Binding Compounds after MLF Acetoin Alpha-Ketoglutarate Pyruvate Acetaldehyde 1000 800 600 400 100 80 60 40 20 0 1000 800 600 400 Pre-MLF Post-MLF 100 80 60 40 20 Glucose Galacturonic Acid Acetoin Alpha-Ketoglutarate Pyruvate Acetaldehyde SO 2 Binding Compound SO 2 Binding Compound 0 Glucose Galacturonic Acid (mgl -1 ) (mgl -1 )
Acetaldyde Pinot Noir Sequential Coinoculation 40 Acetaldehyde [mg/l] 35 30 25 20 15 10 ph 3.4 ph 3.7 ph 4.0 ph 3.4 ph 3.7 ph 4.0 5 0 10 20 30 40 50 60 70 0 10 20 30 40 50 60 70 80 time [d] time [d] Ramón Mira de Orduña, Alba Maggio 2011
Acetaldehyde and bound SO 2 Final Values ph 3.2 ph 3.4 ph 3.7 ph 4.0 Acetaldehyde mg l 1 Sequential 29.6 ±0 30.4±0.5 Coinoculation 19.0 ±1 12.5±0.1 * 16.0± 4 12.6 ±0 15.4±0.1 7.3 ±0.4 * Bound SO 2 mg l 1 Sequential 71.5 ±15 84.5 ±11 64.5 ±4 64 ±2 Coinoculation 59.5 ±7 57 ±7 59 ±4 45 ±6 * statistically significant difference at a confidence interval of 0.01 Ramón Mira de Orduña, Alba Maggio 2011
Inoculation Regime Grape sugar (%) 100 50 Pre AF Co-inoculation mid-af At pressing Post AF AF Co-inoculation Can shorten length of AF+MLF Can enhance fruity characters Time point of MLF inoculation Different wine composition Eveline Bartowsky, AWRI, Neustadt 2010
Red wine aroma Fruit Esters Esters associated with berry fruit attributes ethyl butanoate, hexanoate, octanoate & propanoate ethyl-2-methyl butanoate & propanoate ethyl 3-methyl butanoate 3-methyl butyl acetate Escudero et al, 2007; Pineau et al 2009 Eveline Bartowsky, WAC Beaune 2011
IMPACT OF ML BACTERIA STRAIN AROMA AND MOUTHFEEL IN CAB. SAUVIGNON WINE Australia collaboration AWRI (Bartowsky/Costello) 2006-2010 To demonstrate the role of MLF in affecting wine mouthfeel properties other than due to diacetyl or ph/acidity To demonstrate the role of MLF in affecting the varietal aroma (red beery fruit) Establish the chemical functional component(s) of the major effect Establish the nature of mechanism involved Define winemaking conditions that promote mouthfeel affects especially with regard to the concept of coinoculation of bacteria and yeast
ph & aroma in wine Wines Stabilised Bottled Analysis Clare Valley AF ~ 550 kg L2056 18-20 C MLF, 20 C R1105 R1106 ph 3.3 R1118 Non-MLF Control R1105 R1106 ph 3.7 R1118 Non-MLF Control 2006 vintage Post MLF: ph adjustment to ph 3.5 Eveline Bartowsky, WAC Beaune 2011
Malic acid metabolism 2.0 Limestone Coast (13.8% alc) Clare Valley (14.7% alc) 1.5 1.0 ph 3.3 L-malic acid (g/l) 0.5 0.0 2.0 1.5 1.0 0 10 20 30 40 50 60 70 80 0 10 20 30 40 50 60 70 80 90 100 ph 3.7 0.5 0.0 0 10 20 30 40 50 60 70 80 0 10 20 30 40 50 60 70 80 Time (days) after inoculation Eveline Bartowsky, AWRI, 2009
Wine ph affects O. oeni metabolism 3-Methyl butyl acetate 2-Methyl butyl acetate 2-Phenyl ethyl acetate Ethyl dodecanoate Hexyl acetate 2-Methyl butanoic acid 2-Methyl propanoate PC2 23.5% 2-Methyl propyl acetate Diacetyl Ethyl propanoate Savoury aroma Ethyl 3-methyl butanoate Ethyl 2-methyl propanoate Ethyl 2-methyl butanoate Hexanol Savoury flavour Butanol Dark fruit aroma Overall green flavour Octanoic acid Overall fruit flavour PC1 42.6% 2006 Cab. Sauvignon, Limestone Coast ph 3.3 No MLF MLF ph 3.7 No MLF MLF Ethyl octanoate Ethyl acetate Ethyl lactate Acetic acid Eveline Bartowsky, WAC Beaune 2011
Sensory Clare Valley ph 3.3 ph 3.7 Savoury Raspberry 4 3 Fresh dark fruit Savoury Raspberry 4 3 Fresh dark fruit Smoky 2 1 Cooked dark fruit Smoky 2 1 Cooked dark fruit Coffee/choc 0 Floral Coffee/choc 0 Floral Licorice Confectionary Licorice Confectionary Mixed spice Fresh green Mixed spice Fresh green 1105 1106 1118 no-mlf Eveline Bartowsky, AWRI, 2009
What happens after 3 years storage? Esters At bottling after 3 years Sensory Relative changed compared with No-MLF 2006 Clare Valley R1105 R1106 R1118 No MLF Esters 100% Sensory 0.00 Eveline Bartowsky, WAC Beaune 2011
2009 : Red wines Clare Valley Malic acid (g/l) Adelaide McLaren Vale Kangaroo Island Adelaide Hills Langhorne Creek Padthaway Limestone Coast Days after ML strain Inoculation Eveline Bartowsky, WAC Beaune 2011
2009 : Red wines Relative % change in sum of esters 24 28 45-24 24 52 Time to - complete MLF Clare Valley McLaren Vale Eveline Bartowsky, WAC Beaune 2011
Cabernet Sauvignon 2006-3.3 2008-3.5 Malic acid (g/l) 2006-3.7 2009-3.5 Clare Valley Adelaide Clare Valley Days after ML strain Inoculation Eveline Bartowsky, WAC Beaune 2011
Cabernet Sauvignon - Clare Valley 2009 2008 2006 No MLF R1105 R1106 R1118 No MLF R1105 R1106 No MLF R1105 R1106 R1118 Change in sensory rating (vs. Non-MLF) Total Red Fruit Esters (Relative %) O all fruit ar Red berries ar Dark fruit ar Dark berries ar Cooked fruit ar Dark berries fl O all fruit fl Fruit at Raspberry Dark fruit ar Cooked dark fruit ar O all fruit fl Eveline Bartowsky, WAC Beaune 2011
O. oeni & Lb. plantarum 2010 vintage Clare Valley O. oeni Lb. plantarum Eveline Bartowsky, WAC Beaune 2011
2010 Cabernet Sauvignon ALPHA Lalvin 31 PN4 O. oeni Lb. plantarum V22 Eveline Bartowsky, WAC Beaune 2011
Trial 2010. Sensorial impact different bacteria strains Vindima 2010. Alentejo (Portugal)
Trial 2010. Sensorial impact different bacteria strains. Variety: Aragonês (Tempranillo). AF Ganimede Technique. Quick maceration and fermentation. Yeast: YSEO ICV GRE (20g/HL). Nutrition: Fermaid E (20+20g/Hl). Racking to 7 tanks (200 litres capacity). Bacteria inoculation timing: Around 10g/l residual sugar/ 25ºC /ph:3,55. FML Temperature at 18ºC. Bacteria Strains: ALPHA / BETA / VP41 / ELIOS1 / V22 /PN4 / CONTROL
Sensory Analysis of wines by tasting to standard ISO 11035 The descriptive sensory analysis employed is in accordance with ISO standard 11035. The use of this method means that a panel of professional wine tasters can identify and select descriptors for creating a sensory profile of a wine. This tasting panel was formed by 5 international qualified tasters previously trained by this method and wines to taste. The tasting was done by Excell Ibérica Labs.
1. Visual phase. Representation of the variables (previously defined descriptors) and findings (wine samples) for the visual phase. The axes reflect a variance of 100%. Important differences between the samples, showing more intensity and purple tonality in VP41 and Elios1 samples.
2. Aromas or olfactive phase. Representation of the variables (previously defined descriptors) and findings (wine samples) for the olfactive phase. The axes reflect a variance of 55,82%. We can perceive 4 different groups, VP41 and V22 as more varietal samples with high aromatic intensity, floral notes, mints, licorice, candy notes and fruit in syrup. A second group formed by Beta, Elios1 and PN4 more spiciness, with dry fruit notes, lactic and some chemical. One group with the control sample with meat aromas (umami), green fruit, herbaceous and yeasty character.
3. Taste and mouthfeel or gustative phase. Representation of the variables (previously defined descriptors) and findings (wine samples) for the gustatory phase. The axes reflect a variance of 79,90%. In the gustatory phase, once again we perceive different groups. In this case, bacteria V22 is distinguished by the rest because its different mouthfeel descriptors, concentration, large, sweet and balanced. Close to it, there are VP41 and Alpha, with similar descriptors but less intensity. With Elios1 and Beta we perceive more acidity perception. PN4 and Control more vegetal character, bitterness and astringency.
4. Retronasal phase. Representation of the variables (previously defined descriptors) and findings (wine samples) for the retronasal phase. The axes reflect a variance of 74,30%. We see that in retronasal phase the samples are grouped by the same way than the gustatory phase, noticed this coincidence. V22 has the best descriptors in this phase, complex, fruity, persistence and licorous. Following by Alpha (with some lactic notes) and VP41. Beta and Elios1 with herbaceous aromatic notes. Control and PN4 with reduced notes and hot in retronasal.
Summary Changes in ester concentration is dependent upon several factors O. oeni strain Wine composition Viticultural region MLF conditions Vintage Adelaide Increase in total fruit berry compounds translates to an increase in berry related sensory descriptor terms Escudero et al, 2007; Pineau et al 2009 ML strains are exhibiting consistent characteristics Extended to Lb. plantarum MLF can be used to enhance the fruity berry characteristics of Cabernet Sauvignon ML strains behave similarly in other red varieties regarding fruity berry characters