Effect of ‘loss of function’ mutation in SER1 in wine yeast: fermentation outcomes in co-inoculation with non-Saccharomyces
Effect of ‘loss of function’ mutation in SER1 in wine yeast: fermentation outcomes in co-inoculation with non-Saccharomyces
In wine fermentation, improved wine complexity and sensorial properties can arise from the use of non-Saccharomyces yeast. Generally less alcohol tolerant, such strains often do not finish fermentation, therefore requiring a second inoculation with the more robust Saccharomyces cerevisiae, usually added on Day 3. This sequential approach affords non-Saccharomyces time to make an impact before being overtaken by S. cerevisiae. However, two inoculations are inconvenient; therefore the identification of a slow growing S. cerevisiae strain that can be used in a single co-inoculation with the non-Saccharomyces yeast is highly attractive. In this study we investigated the use of the naturally occurring ‘loss of function’ SER1 variant, identified in a Sake yeast, for the purposes of carrying out co-inoculated wine fermentations. The SER1-232(G > C; G78R) change was introduced into the commonly used wine strain, EC1118, via CRISPR/Cas9 editing. In a chemically defined grape juice medium, the SER1(G78R) mutant grew and fermented more slowly and increased acetic acid, succinic acid and glycerol concentrations. Simultaneous inoculation with the slower-growing mutant with a Metschnikowia pulcherrima or Lachancea thermotolerans strain in sterile Sauvignon blanc juice resulted in differences in sensorial compounds, most likely derived from the presence of non-Saccharomyces yeasts. The EC1118 SER1 (G78R) mutant completed fermentation with M. pulcherrima, MP2, and in fact improved the viability of MP2 compared to when it was used as a monoculture. The SER1 (G78R) mutant also promoted both the growth of the SO2 -sensitive L. thermotolerans strain, Viniflora® Concerto™, in a juice high in SO2 and its subsequent dominance during fermentation. In co-fermentations with wild-type EC1118, the Concerto™ population was substantially reduced with no significant changes in wine properties. This research adds to our understanding of the use of a novel slow-growing S. cerevisiae yeast in wine fermentations co-inoculated with non-Saccharomyces strains.
co-inoculation, CRISPR/Cas9, Lachancea thermotolerans, Metschnikowia pulcherrima, Saccharomyces cerevisiae, SER1, SO
47-61
Lang, Tom A.
483fd372-b7ce-45f4-9198-461ecf47907e
Walker, Michelle E.
5e8a98ce-9e08-409a-99e4-a0b96a490940
Boss, Paul K.
a4958f4e-9fa3-4b3b-9be5-f74494167d39
Jiranek, Vladimir
8e5a8dfd-f5b2-43e3-928b-11dff324abc7
Lang, Tom A.
483fd372-b7ce-45f4-9198-461ecf47907e
Walker, Michelle E.
5e8a98ce-9e08-409a-99e4-a0b96a490940
Boss, Paul K.
a4958f4e-9fa3-4b3b-9be5-f74494167d39
Jiranek, Vladimir
8e5a8dfd-f5b2-43e3-928b-11dff324abc7
Lang, Tom A., Walker, Michelle E., Boss, Paul K. and Jiranek, Vladimir
(2022)
Effect of ‘loss of function’ mutation in SER1 in wine yeast: fermentation outcomes in co-inoculation with non-Saccharomyces.
Oeno One, 56 (2), .
(doi:10.20870/oeno-one.2022.56.2.4908).
Abstract
In wine fermentation, improved wine complexity and sensorial properties can arise from the use of non-Saccharomyces yeast. Generally less alcohol tolerant, such strains often do not finish fermentation, therefore requiring a second inoculation with the more robust Saccharomyces cerevisiae, usually added on Day 3. This sequential approach affords non-Saccharomyces time to make an impact before being overtaken by S. cerevisiae. However, two inoculations are inconvenient; therefore the identification of a slow growing S. cerevisiae strain that can be used in a single co-inoculation with the non-Saccharomyces yeast is highly attractive. In this study we investigated the use of the naturally occurring ‘loss of function’ SER1 variant, identified in a Sake yeast, for the purposes of carrying out co-inoculated wine fermentations. The SER1-232(G > C; G78R) change was introduced into the commonly used wine strain, EC1118, via CRISPR/Cas9 editing. In a chemically defined grape juice medium, the SER1(G78R) mutant grew and fermented more slowly and increased acetic acid, succinic acid and glycerol concentrations. Simultaneous inoculation with the slower-growing mutant with a Metschnikowia pulcherrima or Lachancea thermotolerans strain in sterile Sauvignon blanc juice resulted in differences in sensorial compounds, most likely derived from the presence of non-Saccharomyces yeasts. The EC1118 SER1 (G78R) mutant completed fermentation with M. pulcherrima, MP2, and in fact improved the viability of MP2 compared to when it was used as a monoculture. The SER1 (G78R) mutant also promoted both the growth of the SO2 -sensitive L. thermotolerans strain, Viniflora® Concerto™, in a juice high in SO2 and its subsequent dominance during fermentation. In co-fermentations with wild-type EC1118, the Concerto™ population was substantially reduced with no significant changes in wine properties. This research adds to our understanding of the use of a novel slow-growing S. cerevisiae yeast in wine fermentations co-inoculated with non-Saccharomyces strains.
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Accepted/In Press date: 29 March 2022
e-pub ahead of print date: 20 April 2022
Additional Information:
Funding Information:
This project was supported by funding from Wine Australia [UA1803_2.1] and The Australian Research Council Training Centre for Innovative Wine Production (www.ARCwinecentre.org.au; project number IC170100008), which is funded by the Australian Government with additional support from Wine Australia and industry partners. Wine Australia invests in and manages research, development and extension on behalf of Australia’s winegrowers and winemakers and the Australian Government. Tom Lang is a recipient of a University of Adelaide Faculty of Sciences Divisional Scholarship and Wine Australia Supplementary Scholarship (AGW Ph1603).
Funding Information:
The authors would like to thank Nick van Holst for his technical assistance and Tom Ellis and William Shaw from the Imperial College London for providing plasmids pWS173 and pWS082. We also thank Emily Nicholson and Sue Maffei at CSIRO for their assistance with the GC-MS analysis of the wine volatile compounds. This project was supported by funding from Wine Australia [UA1803_2.1] and The Australian Research Council Training Centre for Innovative Wine Production (www.ARCwinecentre.org.au; project number IC170100008), which is funded by the Australian Government with additional support from Wine Australia and industry partners. Wine Australia invests in and manages research, development and extension on behalf of Australia?s winegrowers and winemakers and the Australian Government. Tom Lang is a recipient of a University of Adelaide Faculty of Sciences Divisional Scholarship and Wine Australia Supplementary Scholarship (AGW Ph1603). The University of Adelaide and CSIRO are members of the Wine Innovation Cluster (http://www/thewaite.org/waite-partners/wine-innovation-cluster/).
Publisher Copyright:
© 2022, International Viticulture and Enology Society. All rights reserved.
Keywords:
co-inoculation, CRISPR/Cas9, Lachancea thermotolerans, Metschnikowia pulcherrima, Saccharomyces cerevisiae, SER1, SO
Identifiers
Local EPrints ID: 482790
URI: http://eprints.soton.ac.uk/id/eprint/482790
ISSN: 2494-1271
PURE UUID: 5a7399a2-ab54-47ba-aa31-2944b584bc0a
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Date deposited: 12 Oct 2023 16:45
Last modified: 18 Mar 2024 04:12
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Contributors
Author:
Tom A. Lang
Author:
Michelle E. Walker
Author:
Paul K. Boss
Author:
Vladimir Jiranek
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