Rapid Methods for Detecting<i>Saccharomyces Diastaticus</i>, a Beer Spoilage Yeast, Using the Polymerase Chain Reaction

Yamauchi, Hirofumi; Yamamoto, Hiroshi; Shibano, Yuji; Amaya, Noriko; Saeki, Takeshi

doi:10.1094/asbcj-56-0058

Cited by 21 publications

(26 citation statements)

References 7 publications

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“…The polymerase chain reaction (PCR) for amplifying unique fragments of DNA or RNA has been suggested for detection of many different pathogens and spoilage bacteria in the food and drinks industries. Two recently published examples relating to yeasts are the detection of diastatic wild yeasts (formerly S. diastaticus) (Yamauchi et a/., 1998) and the rapid recognition of different strains of culture yeast (Pecar, Tonissen and Rogers, 1999). Also, like IF, which requires specific sera, PCR detection of a particular contaminant is possible only when the specific primer, reacting with the DNA fragments of the contaminant but not culture yeast, is available.…”

Section: Detection Of Wild Yeastsmentioning

confidence: 99%

Wild yeasts in brewing and distilling

Campbell¹

2003

Brewing Microbiology

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Section: Detection Of Wild Yeastsmentioning

confidence: 99%

Wild yeasts in brewing and distilling

Campbell¹

2003

Brewing Microbiology

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“…Another DMG, STA1 , detects the high attenuating S. cerevisiae var. diastaticus and distinguishes from top‐fermenting S. cerevisiae strains . STA1 encodes an extracellular glucoamylase , which hydrolyses α ‐D bonds as well as α ‐D linkages .…”

Section: Introductionmentioning

confidence: 99%

Novel diagnostic marker genes differentiateSaccharomyceswith respect to their potential application

et al. 2018

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Marker genes are widely used to differentiate and classify yeast at a species level. Diagnostic marker genes (DMGs) derived from a whole genome comparison allow the differentiation of strains of Saccharomyces cerevisiae according to their potential application. This approach, which has not been previously described, uses the software tool BIAst Diagnostic Gene Finder (BADGE). We compared the genome sequences of 25 brewing yeast strains to predict DMGs for the discrimination of Saccharomyces yeasts according to their brewing style. Two promising DMGs were chosen for PCR screening in order to test the presence of these genes in 83 yeast strains. A first PCR screening of 25 top‐fermenting S. cerevisiae strains, using the DMG‐specific primer pairs for wheat beer marker I (putative AMD2) and wheat beer marker II (GEX1), distinguished yeast strains of the wheat beer style from all other beer styles. The PCR screening of an extended set of 83 yeast strains displayed a more variable distribution of the selected DMGs to different potential applications. Furthermore, 60 out of 83 yeast strains were positive for at least one DMG. The diagnostic marker genes proved to be useful for the differentiation of wheat beer strains (100% positive identification for both genes) to German Alt‐Kölsch and ale yeasts (87.5% negative identification for both genes). © 2018 The Institute of Brewing & Distilling

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“…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.…”

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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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Rapid Methods for DetectingSaccharomyces Diastaticus, a Beer Spoilage Yeast, Using the Polymerase Chain Reaction

Cited by 21 publications

References 7 publications

Wild yeasts in brewing and distilling

Wild yeasts in brewing and distilling

Novel diagnostic marker genes differentiateSaccharomyceswith respect to their potential application

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

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