Grape and Wine Proteins: Their Fractionation by Hydrophobic Interaction Chromatography and Identification by Chromatographic and Proteomic Analysis

Abstract

A method to fractionate grape and wine proteins by hydrophobic interaction chromatography (HIC) was developed. This method allowed the isolation of a thaumatin-like protein in a single step with high yield and >90% purity and has potential to purify several other proteins. In addition, by separating HIC fractions by reverse phase HPLC and by collecting the obtained peaks, the grape juice proteins were further separated, by SDS-PAGE, into 24 bands. The bands were subjected to nanoLC-MS/MS analysis, and the results were matched against a database and characterized as various Vitis vinifera proteins. Moreover, either directly or by homology searching, identity or function was attributed to all of the gel bands identified, which mainly consisted of grape chitinases and thaumatin-like proteins but also included vacuolar invertase, PR-4 type proteins, and a lipid transfer protein from grapes.

The challenging SO2-mediated chemical build-up of protein aggregates in wines

“…The resulting HIC chromatogram (Fig. 2) is similar to the chromatograms published elsewhere for similar protein fractions (Marangon, Van Sluyter, Haynes, & Waters, 2009), with slight differences in protein quantity ratios, most likely derived from the use of different biological samples. Fractions H1-H7, referring to peaks 1-7 in Fig.…”

Identification and Partial Characterization of Extracellular Aspartic Protease Genes from Metschnikowia pulcherrima IWBT Y1123 and Candida apicola IWBT Y1384

“…Lane a shows the profile of the grape proteins after extraction from grape juice by acetone precipitation. The band at ϳ60 kDa may be grape invertase (27,30), and the intense band just beneath 25 kDa may be the common grape proteins thaumatin-like protein (22 kDa) and/or chitinase (25 kDa) (49,50). These proteins are known to be very stable due to their conformation, and thus degradation due to incubation con- ditions was not expected (51).…”

Influence of skin contact and different extractants on extraction of proteins and phenolic substances in Sauvignon Blanc grape skin

“…Samples (50 μL) were loaded at 1 mL/min onto a C8 column (4.6 mm × 250 mm, Vydac 208TP54, PhaseSep, Melbourne, Vic., Australia), fitted with a C8 guard column kit (4.6 mm × 5 mm, Vydac 208GK54) which was equilibrated in a mixture of 83% (v/v) solvent B [0.1% trifluoroacetic acid (TFA) in 8% acetonitrile] and 17% solvent A [80% acetonitrile, 0.1% (v/v) TFA] and held at 35°C. Proteins were eluted by a gradient of solvent A, from 17% solvent A to 49% solvent A in the first 7 min, from 49 to 57% in 7 to 15 min, from 57 to 65% in 15 to 16 min, from 65 to 81% in 16 to 30 min, and then held at 81% for 5 min before re‐equilibrating the column in the starting conditions for an additional 6 min (Marangon et al ). Elution was monitored by absorbance at 210, 220, 260, 280 and 320 nm.…”

“…Samples (50 μL (Marangon et al 2009). Elution was monitored by absorbance at 210, 220, 260, 280 and 320 nm.…”