Information of commercial enzyme preparations (Bio-Laffort, France) used in

Transcription

1 Supporting Information Supplementary Table 1. Information of commercial enzyme preparations (Bio-Laffort, France) used in this study ( Commercial enzyme preparation Properties Application LafaseXL Extraction Lafase HE Grand Cru Lafase Fruit Liquid enzymatic preparation for red wine maceration and clarification Pectolytic enzyme preparation, purified in CE for the production red wines that are rich in colouring matter and structured tannins, destined for ageing. Purified pectolytic enzyme preparation for the production of fruity, colourful and round red wines. Extraction and clarification Maceration Maceration 1

2 Supplementary Table 2. General wine parameters of the final wine from the fermentation treatments. The values for each parameter are the mean values from two technical repeats performed by the FOSS Winescan. (note: succinic acid, tartaric acid and acetic acid in mg/l, all other comound in g/l except the ethanol is expressed as volume to volume, v/v) Sample Id ph VolatileAcidTotalAcid MalicAcid LacticAcid Glucose Fructose Ethanol Glycerol Succinic Acid Tartaric acid Acetic Acid U U U U Cru Cru Cru Cru Sample Id ph VolatileAcid TotalAcid MalicAcid LacticAcid Glucose Fructose Ethanol Glycerol Succinic Acid Tartaric acid Acetic Acid U U U U XL XL XL XL Sample Id ph Volatile Acid TotalAcidMalicAcidLacticAcid Glucose Fructose Ethanol Glycerol Succinic Acid Tartaric acid Acetic Acid U U U U Fruit Fruit Fruit Fruit

3 Supplementary Figure 1. Total phenolics, color, anthocyanins and tannin values obtained from the final wine (at least 1 year stored) acquired from the fermentations of different treatments. The values for each parameter (absorbance units) are the mean values from three technical repeats. (note: references for the methods are provided after figures) Phenolics Total color (A 420,520,620 )

4 25 Anthocyanins (A 520 ) Tannins composition (mg/l) in the wine Boulton, R.B., Singleton, V.L., Bisson, L.F. & Kunkee, R.E., Principles and practices of Winemaking. Chapman and Hall, New York, NY. Sarneckis, C.J., Dambergs, R.G., Jones, P., Mercuric, M., Herderich, M.F. & Smith, R.A. Quantification of condensed tannins by precipitation with methyl cellulose: development and validation of an optimised tool for grape and wine analysis. Aus. J. Grape Wine Res. 2006, 12: Somers, T.C., & Evans, M.E. Wine Quality: Correlation with colour density and anthocyanin equilibria in a group of young red wines. J. Sci. Food Agric. 1974, 25:

5 Supplementary Figure 2. PCA score plots of wine parameters from different fermentations (untreated and enzyme-treated). A. U1-4, Cru1-4; B. U5-8, Fruit1-4; C. U9-12, XL1-4. The value for each variable (wine parameter) is mean-value from two technical repeats performed using the FOSS Winescan. PCA-X: General Oenological Parameters A PC2=22.6% U Cru Fructose Volatile acid ph Succinic acid Malic acid PC1=60.7% B PC2=29.4% U Fruit Malic acid Tartaric acid ph Volatile acid PC1=48.8% C PC2=17.2% Succinic acid U XL ph Volatile acid PC1=61.7% 5

6 Supplementary File 1. Briefly, the sample was hydrolyzed using 2 M TFA (2 h, 110 o C) to monosaccharides, which were then converted to their methoxy derivatives using methanol/methanol HCl (16 h, 80 o C), followed by the silylation with HMDS/TMCS/pyridine (3:1:9, Sylon HTP kit, Sigma-Aldrich, MO, USA). The separation and analysis of these sugar derivatives were performed using a gas chromatograph (Agilent 6890 N, Agilent Technologies, CA, USA) coupled to an Agilent 5975 MS mass spectrometry fitted with a polar (95% dimethylpolysiloxane) ZB-Semivolatiles Guardian GC column (30 m, 0.25 mm ID, 0.25 µm film thickness). The nine main cell wall monosaccharides were analysed: arabinose (Ara), fucose (Fuc), rhamnose (Rha), xylose (Xyl), mannose (Man), galacturonic acid (GalA), galactose (Gal), glucose (Glc) and glucuronic acid (GlcA). 6

7 Supplementary File 2. To assess the variation of whole berry cell wall samples on the polymer level, AIR (10 mg) sourced from grape pomace (with and without enzyme treatment) was sequentially extracted using CDTA (diamino-cyclo-hexane-tetra-acetic acid) and NaOH to obtain the pectin-rich and hemicellulose-rich fractions respectively. 18 These fractions were then spotted onto nitrocellulose membranes and then probed with a number of monoclonal antibodies (mabs) and carbohydrate binding modules (CBMs). The raw data of the signal values were recorded and used for the multivariate data analysis; whereas the mean readings were scaled and then presented as a microarray heatmap, the highest signal was set as 100, and others were normalized according to the highest signal, a cutoff value of 5 was applied. 7

8 Supplementary File 3. The data generated from monosaccharide compositional analysis was evaluated by univariate statistical analyses (ANOVA, with P = 0.05) under the guidance of the Centre for Statistical Consultation at Stellenbosch University (Prof. Martin Kidd) using Statistica 10 (StatSoft Southern Africa -Analytics, Sandton, South Africa). Multivariate analysis was performed using the SIMCA 14 software package (MKS, Umea, Sweden) to perform the PCA (principal component analysis) advised by Professor Johan Trygg (Umea, Sweden). Data were prior to PCA modeling column centered and scaled to unit variance (UV). Cross-validation was used to assess model significance. 8