Sulfate transport mutants affect hydrogen sulfide and sulfite production during alcoholic fermentation
Sulfate transport mutants affect hydrogen sulfide and sulfite production during alcoholic fermentation
Hydrogen sulfide is a common wine fault, with a rotten-egg odour, which is directly related to yeast metabolism in response to nitrogen and sulfur availability. In grape juice, sulfate is the most abundant inorganic sulfur compound, which is taken up by yeast through two high-affinity sulfate transporters, Sul1p and Sul2p, and a low affinity transporter, Soa1p. Sulfate contributes to H2S production under nitrogen limitation, by being reduced via the Sulfur Assimilation Pathway (SAP). Therefore, yeast strains with limited H2S are highly desirable. We report on the use of toxic analogues of sulfate following ethyl methane sulfate treatment, to isolate six wine yeast mutants that produce no or reduced H2S and SO2 during fermentation in synthetic and natural juice. Four amino acid substitutions (A99V, G380R, N588K and E856K) in Sul1p were found in all strains except D25-1 which had heterozygous alleles. Two changes were also identified in Sul2p (L268S and A470T). The Sul1p (G380R) and Sul2p (A470T) mutations were chosen for further investigation as these residues are conserved amongst SLC26 membrane proteins (including sulfate permeases). The mutations were introduced into EC1118 using Crispr cas9 technology and shown to reduce accumulation of H2S and do not result in increased SO2 production during fermentation of model medium (chemically defined grape juice) or Riesling juice. The Sul1p (G380R) and Sul2p (A470T) mutations are newly reported as causal mutations. Our findings contribute to knowledge of the genetic basis of H2S production as well as the potential use of these strains for winemaking and in yeast breeding programmes.
hydrogen sulfide, Saccharomyces cerevisiae, Sul1p, Sul2p, sulfate assimilation pathway, sulfate transporters
367-381
Walker, Michelle E.
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Zhang, Jin
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Sumby, Krista M.
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Lee, Andrea
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Houlès, Anne
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Li, Sijing
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Jiranek, Vladimir
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1 June 2021
Walker, Michelle E.
5e8a98ce-9e08-409a-99e4-a0b96a490940
Zhang, Jin
6f92519d-9a26-46e5-bfe8-70734b9c5a8f
Sumby, Krista M.
f54b8f1c-c29e-488b-ab5e-dbb9b2576ba9
Lee, Andrea
dc098c0b-04c0-4492-ad52-b74f6612b5a2
Houlès, Anne
7f170ca0-d4f2-4a8f-a98e-2213629a88ab
Li, Sijing
54889e34-524e-4455-9d2b-f7852fee34d0
Jiranek, Vladimir
8e5a8dfd-f5b2-43e3-928b-11dff324abc7
Walker, Michelle E., Zhang, Jin, Sumby, Krista M., Lee, Andrea, Houlès, Anne, Li, Sijing and Jiranek, Vladimir
(2021)
Sulfate transport mutants affect hydrogen sulfide and sulfite production during alcoholic fermentation.
Yeast, 38 (6), .
(doi:10.1002/yea.3553).
Abstract
Hydrogen sulfide is a common wine fault, with a rotten-egg odour, which is directly related to yeast metabolism in response to nitrogen and sulfur availability. In grape juice, sulfate is the most abundant inorganic sulfur compound, which is taken up by yeast through two high-affinity sulfate transporters, Sul1p and Sul2p, and a low affinity transporter, Soa1p. Sulfate contributes to H2S production under nitrogen limitation, by being reduced via the Sulfur Assimilation Pathway (SAP). Therefore, yeast strains with limited H2S are highly desirable. We report on the use of toxic analogues of sulfate following ethyl methane sulfate treatment, to isolate six wine yeast mutants that produce no or reduced H2S and SO2 during fermentation in synthetic and natural juice. Four amino acid substitutions (A99V, G380R, N588K and E856K) in Sul1p were found in all strains except D25-1 which had heterozygous alleles. Two changes were also identified in Sul2p (L268S and A470T). The Sul1p (G380R) and Sul2p (A470T) mutations were chosen for further investigation as these residues are conserved amongst SLC26 membrane proteins (including sulfate permeases). The mutations were introduced into EC1118 using Crispr cas9 technology and shown to reduce accumulation of H2S and do not result in increased SO2 production during fermentation of model medium (chemically defined grape juice) or Riesling juice. The Sul1p (G380R) and Sul2p (A470T) mutations are newly reported as causal mutations. Our findings contribute to knowledge of the genetic basis of H2S production as well as the potential use of these strains for winemaking and in yeast breeding programmes.
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More information
Accepted/In Press date: 9 February 2021
e-pub ahead of print date: 1 March 2021
Published date: 1 June 2021
Additional Information:
Funding Information:
We thank Treasury Wine Estates, Barossa Valley, for donating the white juices used in this study. Packages of Distinction and PDM and EC1118 ADWY were gifted by Mauri Yeast Australia Pty. Ltd. and Lallemand Australia Pty. Ltd., respectively. We thank Nick van Holst for measurement of total SO2 by aspiration and Dr Jennie Gardner for help in editing this manuscript. Special thanks to Ms Stephanie Nguyen (University of Adelaide) for kindly helping with protein modelling using UCSF Chimera 1.13.1. UCSF Chimera was developed by the Resource for Biocomputing, Visualization, and Informatics at the University of California, San Francisco, with support from NIH P41-GM103311. This project was supported by funding from Wine Australia (UA1101, UA1302, 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. MW was a recipient of a Wine 2030 small grant (University of Adelaide, School of Economics). The University of Adelaide is a member of the Wine Innovation Cluster (http://www/thewaite.org/waite-partners/wine-innovation-cluster/).
Keywords:
hydrogen sulfide, Saccharomyces cerevisiae, Sul1p, Sul2p, sulfate assimilation pathway, sulfate transporters
Identifiers
Local EPrints ID: 482770
URI: http://eprints.soton.ac.uk/id/eprint/482770
ISSN: 0749-503X
PURE UUID: 6c154cb1-c66c-413c-b7c7-254f7793f8e8
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Date deposited: 12 Oct 2023 16:42
Last modified: 18 Mar 2024 04:12
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Contributors
Author:
Michelle E. Walker
Author:
Jin Zhang
Author:
Krista M. Sumby
Author:
Andrea Lee
Author:
Anne Houlès
Author:
Sijing Li
Author:
Vladimir Jiranek
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