A deletion in the STA1 promoter determines maltotriose and starch utilization in STA1+ Saccharomyces cerevisiae strains

Kristoffer Krogerus (Corresponding Author), Frederico Magalhães, Joosu Kuivanen, Brian Gibson

Research output: Contribution to journalArticleScientificpeer-review

Abstract

Diastatic strains of Saccharomyces cerevisiae are common contaminants in beer fermentations and are capable of producing an extracellular STA1-encoded glucoamylase. Recent studies have revealed variable diastatic ability in strains tested positive for STA1, and here, we elucidate genetic determinants behind this variation. We show that poorly diastatic strains have a 1162-bp deletion in the promoter of STA1. With CRISPR/Cas9-aided reverse engineering, we show that this deletion greatly decreases the ability to grow in beer and consume dextrin, and the expression of STA1. New PCR primers were designed for differentiation of highly and poorly diastatic strains based on the presence of the deletion in the STA1 promoter. In addition, using publically available whole genome sequence data, we show that the STA1 gene is prevalent among the ‘Beer 2’/‘Mosaic Beer’ brewing strains. These strains utilize maltotriose efficiently, but the mechanisms for this have been unknown. By deleting STA1 from a number of highly diastatic strains, we show here that extracellular hydrolysis of maltotriose through STA1 appears to be the dominant mechanism enabling maltotriose use during wort fermentation in STA1+ strains. The formation and retention of STA1 seems to be an alternative evolutionary strategy for efficient utilization of sugars present in brewer’s wort. The results of this study allow for the improved reliability of molecular detection methods for diastatic contaminants in beer and can be exploited for strain development where maltotriose use is desired.

Original languageEnglish
Pages (from-to)7597-7615
Number of pages19
JournalApplied Microbiology and Biotechnology
Volume103
Issue number18
Early online date26 Jul 2019
DOIs
Publication statusPublished - 1 Sep 2019
MoE publication typeA1 Journal article-refereed

Fingerprint

Starch
Saccharomyces cerevisiae
Fermentation
Clustered Regularly Interspaced Short Palindromic Repeats
Glucan 1,4-alpha-Glucosidase
Hydrolysis
maltotriose
Genome
Polymerase Chain Reaction
Genes

Keywords

  • Beer
  • Dextrin
  • Diastatic
  • Genome
  • Starch
  • Yeast

Cite this

@article{b01c2ff336a74c4a8538d9caaa4a449a,
title = "A deletion in the STA1 promoter determines maltotriose and starch utilization in STA1+ Saccharomyces cerevisiae strains",
abstract = "Diastatic strains of Saccharomyces cerevisiae are common contaminants in beer fermentations and are capable of producing an extracellular STA1-encoded glucoamylase. Recent studies have revealed variable diastatic ability in strains tested positive for STA1, and here, we elucidate genetic determinants behind this variation. We show that poorly diastatic strains have a 1162-bp deletion in the promoter of STA1. With CRISPR/Cas9-aided reverse engineering, we show that this deletion greatly decreases the ability to grow in beer and consume dextrin, and the expression of STA1. New PCR primers were designed for differentiation of highly and poorly diastatic strains based on the presence of the deletion in the STA1 promoter. In addition, using publically available whole genome sequence data, we show that the STA1 gene is prevalent among the ‘Beer 2’/‘Mosaic Beer’ brewing strains. These strains utilize maltotriose efficiently, but the mechanisms for this have been unknown. By deleting STA1 from a number of highly diastatic strains, we show here that extracellular hydrolysis of maltotriose through STA1 appears to be the dominant mechanism enabling maltotriose use during wort fermentation in STA1+ strains. The formation and retention of STA1 seems to be an alternative evolutionary strategy for efficient utilization of sugars present in brewer’s wort. The results of this study allow for the improved reliability of molecular detection methods for diastatic contaminants in beer and can be exploited for strain development where maltotriose use is desired.",
keywords = "Beer, Dextrin, Diastatic, Genome, Starch, Yeast",
author = "Kristoffer Krogerus and Frederico Magalh{\~a}es and Joosu Kuivanen and Brian Gibson",
year = "2019",
month = "9",
day = "1",
doi = "10.1007/s00253-019-10021-y",
language = "English",
volume = "103",
pages = "7597--7615",
journal = "Applied Microbiology and Biotechnology",
issn = "0175-7598",
publisher = "Springer",
number = "18",

}

A deletion in the STA1 promoter determines maltotriose and starch utilization in STA1+ Saccharomyces cerevisiae strains. / Krogerus, Kristoffer (Corresponding Author); Magalhães, Frederico; Kuivanen, Joosu; Gibson, Brian.

In: Applied Microbiology and Biotechnology, Vol. 103, No. 18, 01.09.2019, p. 7597-7615.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - A deletion in the STA1 promoter determines maltotriose and starch utilization in STA1+ Saccharomyces cerevisiae strains

AU - Krogerus, Kristoffer

AU - Magalhães, Frederico

AU - Kuivanen, Joosu

AU - Gibson, Brian

PY - 2019/9/1

Y1 - 2019/9/1

N2 - Diastatic strains of Saccharomyces cerevisiae are common contaminants in beer fermentations and are capable of producing an extracellular STA1-encoded glucoamylase. Recent studies have revealed variable diastatic ability in strains tested positive for STA1, and here, we elucidate genetic determinants behind this variation. We show that poorly diastatic strains have a 1162-bp deletion in the promoter of STA1. With CRISPR/Cas9-aided reverse engineering, we show that this deletion greatly decreases the ability to grow in beer and consume dextrin, and the expression of STA1. New PCR primers were designed for differentiation of highly and poorly diastatic strains based on the presence of the deletion in the STA1 promoter. In addition, using publically available whole genome sequence data, we show that the STA1 gene is prevalent among the ‘Beer 2’/‘Mosaic Beer’ brewing strains. These strains utilize maltotriose efficiently, but the mechanisms for this have been unknown. By deleting STA1 from a number of highly diastatic strains, we show here that extracellular hydrolysis of maltotriose through STA1 appears to be the dominant mechanism enabling maltotriose use during wort fermentation in STA1+ strains. The formation and retention of STA1 seems to be an alternative evolutionary strategy for efficient utilization of sugars present in brewer’s wort. The results of this study allow for the improved reliability of molecular detection methods for diastatic contaminants in beer and can be exploited for strain development where maltotriose use is desired.

AB - Diastatic strains of Saccharomyces cerevisiae are common contaminants in beer fermentations and are capable of producing an extracellular STA1-encoded glucoamylase. Recent studies have revealed variable diastatic ability in strains tested positive for STA1, and here, we elucidate genetic determinants behind this variation. We show that poorly diastatic strains have a 1162-bp deletion in the promoter of STA1. With CRISPR/Cas9-aided reverse engineering, we show that this deletion greatly decreases the ability to grow in beer and consume dextrin, and the expression of STA1. New PCR primers were designed for differentiation of highly and poorly diastatic strains based on the presence of the deletion in the STA1 promoter. In addition, using publically available whole genome sequence data, we show that the STA1 gene is prevalent among the ‘Beer 2’/‘Mosaic Beer’ brewing strains. These strains utilize maltotriose efficiently, but the mechanisms for this have been unknown. By deleting STA1 from a number of highly diastatic strains, we show here that extracellular hydrolysis of maltotriose through STA1 appears to be the dominant mechanism enabling maltotriose use during wort fermentation in STA1+ strains. The formation and retention of STA1 seems to be an alternative evolutionary strategy for efficient utilization of sugars present in brewer’s wort. The results of this study allow for the improved reliability of molecular detection methods for diastatic contaminants in beer and can be exploited for strain development where maltotriose use is desired.

KW - Beer

KW - Dextrin

KW - Diastatic

KW - Genome

KW - Starch

KW - Yeast

UR - http://www.scopus.com/inward/record.url?scp=85069663027&partnerID=8YFLogxK

U2 - 10.1007/s00253-019-10021-y

DO - 10.1007/s00253-019-10021-y

M3 - Article

VL - 103

SP - 7597

EP - 7615

JO - Applied Microbiology and Biotechnology

JF - Applied Microbiology and Biotechnology

SN - 0175-7598

IS - 18

ER -