Dissecting the polysaccharide-rich grape cell wall changes during winemaking using combined high-throughput and fractionation methods.

Limited information is available on grape wall-derived polymeric structure/composition and how this changes during fermentation. Commercial winemaking operations use enzymes that target the polysaccharide-rich polymers of the cell walls of grape tissues to clarify musts and extract pigments during the fermentations. In this study, we have assessed changes in polysaccharide composition/turnover throughout the winemaking process by applying recently developed cell wall profiling approaches for monosaccharide composition (GC-MS), infra-red (IR) spectroscopy and comprehensive microarray polymer profiling (CoMPP). CoMPP performed on the concentrated soluble wine polysaccharides showed a fraction rich in rhamnogalacturonan I (RGI), homogalacturonan (HG) and arabinogalactan proteins (AGPs). We also used chemical and enzymatic fractionation techniques in addition to CoMPP to understand the berry deconstruction process more in-depth. CoMPP and gravimetric analysis of the fractionated pomace used aqueous buffers and CDTA solutions to obtain a pectin-rich fraction (pulp tightly-bound to skins) containing HG, RGI and AGPs; and then alkali (sodium carbonate and potassium hydroxide), liberating a xyloglucan-rich fraction (mainly skins). Interestingly this fraction was found to include pectins consisting of tightly associated and highly methyl-esterified HG and RGI networks. This was supported by enzymatic fractionation targeting pectin and xyloglucan polymers. A unique aspect is datasets suggesting that enzyme-resistant pectin polymers 'coat' the inner xyloglucan-rich skin cells. This data has important implications for developing effective strategies for efficient release of favorable compounds (pigments, tannins, aromatics, etc.) from the berry tissues during winemaking. This study provides a framework to understand the complex interactions between the grape matrix and carbohydrate-active enzymes to produce wine of desired quality and consistency.