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Overcoming inefficient utilisation of fructose as a cause of problem fermentations (in collaboration with The Australian Wine Research Institute – AWR 05/06)

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Final Report
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Overview

This report is a background knowledge source on overcoming the inefficient use of fructose by wine yeast, which leaves fructose dominant in late, sluggish or stuck ferments. After screening 20 commercial strains, AWRI 796 was mutagenised and adaptively evolved over 350 generations of continuous fermentation. Of 378 isolates screened, two used fructose relatively faster than the parent in a defined grape juice medium, shortening fermentation from 153 to 117 hours. In Chardonnay juice and Merlot must, the evolved strains finished sooner than the parent but not faster than the commercial fructophilic strain Fermichamp. A significant amount of research has been conducted on this subject since publication; refer to more recent published research and other Wine Australia resources.

The glucophilic character of the yeast Saccharomyces cerevisiae has been reported in the literature (Berthels et al., 2004; Guillaume et al., 2007). By this attribute glucose and fructose (the two main sugars present in grape juice) are depleted at different rates during the alcoholic fermentation, with fructose predominating in the latter and more difficult stages. At this time when nutrients are low and alcohol concentration is high, the greater ratio of fructose compared to glucose can make attempts to reinvigorate sluggish fermentations or restart stuck fermentations very challenging. Lower ratios of glucose compare to fructose (e.g. ~1:10) have themselves been reported as a cause of arrested fermentation (Schutz and Gafner, 1993).

Problem fermentations delay grape processing, tie up valuable and limited fermentation space, demand greater resources to manage and rescue, and frequently result in a suboptimal product due to oxidation and spoilage. Ironically, the frequency of such fermentations means that they are widely accepted as an unavoidable downside of winemaking. Improved strains, however, are likely to offer a solution.

In seeking to produce strains which are better able to complete fermentation and specifically exhibit a greater ability to utilise fructose, a range of improvement strategies can be used. These can be broadly classed as either recombinant or nonrecombinant. Both have disadvantages: on one hand in terms of the degree of knowledge of the genetic basis of the sought property and the acceptability of recombinant products to consumers, and on the other, the limited extent of improvementthat is possible or likely.

Abstract

In this report are detailed the outcomes of a PhD project, aimed to improve the ability of wine yeast to consume fructose. This objective was sought using adaptive evolution as the improvement strategy, thereby also providing an evaluation of the use of adaptive evolution to improve wine yeast for commercial application.

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