Moderate concentrations of ethanol inhibit endocytosis of the yeast maltose transporter

Lucero, Pilar; Peñalver, Élida; Moreno, Eulalia; Lagunas, Rosario

doi:10.1128/aem.63.10.3831-3836.1997

Cited by 44 publications

(28 citation statements)

References 40 publications

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“…Maltose transport capacity is regulated by glucoserepression and maltose-induction of genes encoding maltose transporters and also by glucose-triggered catabolite inactivation of existing maltose transporter proteins 19,27 . The present results emphasise also a third factor, namely the lipid composition, in particular the sterol content of the plasma membrane.…”

Section: Discussionmentioning

confidence: 99%

“…Standard maltose transport assays were made essentially as described 19 . Yeast were harvested by centrifugation (10 min at 5000 × g), washed with ice-cold distilled water and suspended to 200 mg fresh yeast · mL -1 in icecold 0.1 M tartrate /Tris buffer pH 4.2.…”

Section: Maltose Transportmentioning

confidence: 99%

“…In the present work we manipulated the sterol and fatty acid composition of yeast by growing it, with maltose as sole carbon source, under strictly anaerobic conditions in the presence or absence of lipid supplements. The rates of maltose and maltotriose transport and subsequent fermentation are known to be strongly regulated at transcription and post-translation levels by glucose repression of ␣-glucoside transporters and maltases and by glucose-triggered catabolite inactivation of the transporters 19,27 . Our objective was to investigate whether the rate of maltose transport is further limited by other physiological factors of practical importance during brewery fermentations.…”

Section: Introductionmentioning

confidence: 99%

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Direct Evidence That Maltose Transport Activity Is Affected by the Lipid Composition of Brewer's Yeast

Guimarães

Virtanen

Londesborough

2006

Journal of the Institute of Brewing

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A brewer's yeast strain was grown with maltose as sole carbon source under strictly anaerobic conditions with and without ergosterol and /or unsaturated fatty acid (Tween 80) supplements. Under all these conditions the MALx1 genes for maltose transporters were strongly expressed during growth. The fatty acid unsaturation indices of growing and stationary phase yeast were increased from about 20% to 56-69% by supplementation with Tween 80. Ergosterol contents were increased up to at least 4-fold by supplementation with ergosterol and Tween 80. Maltose transport activity measured at 20°C was not increased by supplementation with Tween 80 alone, but was increased 2-fold and 3-fold, respectively, in growing and stationary phase yeast by supplementation with ergosterol together with Tween 80. The stimulation of maltose transport by ergosterol was greater when the transport was measured at temperatures (10°C and 0°C) lower than 20°C. The results show that proper function of maltose transporters requires adequate amounts of ergosterol in the yeast. This effect may partly explain the low maltose (and maltotriose) uptake rates both in the second half of brewery fermentations, when the sterol content of yeast has fallen, and when fresh wort is pitched with sterol-deficient cropped yeast.

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Section: Discussionmentioning

confidence: 99%

Section: Maltose Transportmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Direct Evidence That Maltose Transport Activity Is Affected by the Lipid Composition of Brewer's Yeast

Guimarães

Virtanen

Londesborough

2006

Journal of the Institute of Brewing

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“…The zero-trans maltotriose uptake rate was assayed using [U-14 C]-maltotriose as described by 120 Lucero et al (1997). Yeast strains were grown in YP-Maltose at 20 °C to an OD600 between 4 and 121…”

Section: Maltotriose Uptake Assay 119mentioning

confidence: 99%

A deletion in theSTA1promoter determines maltotriose and starch utilization inSTA1+Saccharomyces cerevisiaestrains

Krogerus

Magalhães

Kuivanen

et al. 2019

Preprint

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12Diastatic strains of Saccharomyces cerevisiae are common contaminants in beer fermentations and 13 are capable of producing an extracellular STA1-encoded glucoamylase. Recent studies have 14 revealed variable diastatic ability in strains tested positive for STA1, and here we elucidate genetic 15 determinants behind this variation. We show that poorly diastatic strains have a 1162 bp deletion in 16 the promoter of STA1. With CRISPR/Cas9-aided reverse engineering, we show that this deletion 17 greatly decreases the ability to grow in beer and consume dextrin, and the expression of STA1. New 18 PCR primers were designed for differentiation of highly and poorly diastatic strains based on the 19 presence of the deletion in the STA1 promoter. In addition, using publically available whole genome 20 sequence data, we show that the STA1 gene is prevalent in among the 'Beer 2'/'Mosaic Beer' 21 brewing strains. These strains utilize maltotriose efficiently, but the mechanisms for this have been 22 unknown. By deleting STA1 from a number of highly diastatic strains, we show here that 23 extracellular hydrolysis of maltotriose through STA1 appears to be the dominant mechanism 24 enabling maltotriose use during wort fermentation in STA1+ strains. The formation and retention of 25 STA1 seems to be an alternative evolutionary strategy for efficient utilization of sugars present in 26 brewer's wort. The results of this study allow for the improved reliability of molecular detection 27 methods for diastatic contaminants in beer, and can be exploited for strain development where 28 maltotriose use is desired. 29 the SGA1-derived peptide, while the FLO11-derived peptide allows for extracellular secretion of the 49 protein (Adam et al., 2004). The upstream sequences of STA1 and FLO11 are also nearly identical, 50 meaning that these genes are largely co-regulated (Gagiano et al., 1999). 51Current detection methods for diastatic S. cerevisiae rely mainly on either the microbiological 52 detection through culturing on specialized selective growth media, or the molecular detection of the 53 4 STA1 gene through conventional or quantitative PCR (Brandl, 2006;Meier-Dörnberg et al., 2018; 54 van der Aa Kühle, 1998;Yamauchi et al., 1998). The main weakness of the microbiological 55 methods is that they are time-consuming. While the molecular methods are rapid, a recent study 56 (Meier-Dörnberg et al., 2018) has revealed that there is considerable variability in diastatic ability 57 and beer-spoilage potential in strains carrying the STA1 gene. In fact, some strains that carry the 58 STA1 gene do not show spoilage potential, and these benign strains would be erroneously flagged as 59 diastatic with the current molecular methods. 60In this study, we examined the diastatic ability of a range of STA1+ S. cerevisiae strains. We also 61 sequenced the open reading frame and upstream sequence of STA1 in these same strains to search 62 for polymorphisms that might explain the variable diastatic ability. Sequencing revealed that the 63 poorly dias...

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“…about 2 mg dry yeast mL À1 ) by centrifugation (10 min, 4000 g, 0 C), washed with ice-cold water and then with ice-cold 0.1 M tartrate-Tris (pH 4.2) and finally suspended in the same buffer to 200 mg of fresh yeast mL À1 . Zero-trans 14 C-maltose or -maltotriose transport rates were determined as described by Lucero et al (24) with modifications described previously (15). To estimate K m and V max , series of assays at five different concentrations of maltose (5, 25, 50, 75 and 100 mM) or maltotriose (2.5, 5, 10, 25 and 50 mM) were performed at 20 C. Median estimates of K m and V max were calculated by the direct linear plot method of Eisenthal and Cornish-Bowden (25).…”

Section: Transport Assaysmentioning

confidence: 99%

Characterization of the Saccharomyces bayanus-type AGT1 transporter of lager yeast

Vidgren

Londesborough

2012

J. Inst. Brew.

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Transport of maltose and maltotriose into the yeast cell is thought to be rate‐limiting in the utilization of these sugars. The maltose and maltotriose transporters Malx1, Agt1, Mtt1 and Mphx are present in different combinations in brewer's yeast strains, conferring different maltose and maltotriose transport characteristics on the strains. A new putative maltose/maltotriose transporter ORF was identified during whole genome sequencing of the lager strain WS34/70 (Y. Nakao et al., DNA Res., 2009, 16, 115–129). Sequence comparisons suggested that this putative α‐glucoside transporter might be a Saccharomyces bayanus counterpart of the Agt1 (Saccharomyces cerevisiae type) transporter. In the present work, the transporter coded by a SbAGT1 gene from a lager strain, A15 (and with the same sequence as the corresponding gene in WS34/70) was characterized. It is shown that this SbAGT1 encodes a functional α‐glucoside transporter with a wide‐substrate range, including maltose and maltotriose. Trehalose, α‐methylglucoside and sucrose were inhibitors, suggesting they are also substrates. The SbAgt1 transporter had similar affinities for maltose and maltotriose (17 ± 7 and 22 ± 2 m m, respectively) and a higher Vmax for maltose than maltotriose (21 ± 7 and 12 ± 2 µmol min−1 g dry yeast−1, respectively). Copyright © 2012 The Institute of Brewing & Distilling

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Moderate concentrations of ethanol inhibit endocytosis of the yeast maltose transporter

Cited by 44 publications

References 40 publications

Direct Evidence That Maltose Transport Activity Is Affected by the Lipid Composition of Brewer's Yeast

Direct Evidence That Maltose Transport Activity Is Affected by the Lipid Composition of Brewer's Yeast

A deletion in theSTA1promoter determines maltotriose and starch utilization inSTA1+Saccharomyces cerevisiaestrains

Characterization of the Saccharomyces bayanus-type AGT1 transporter of lager yeast

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