Structural, Physiological and Regulatory Analysis of Maltose Transporter Genes in Saccharomyces eubayanus CBS 12357T

Frontiers in Microbiology
Anja BrickweddeJean-Marc Daran

Abstract

Saccharomyces pastorianus lager brewing yeasts are domesticated hybrids of Saccharomyces cerevisiae and cold-tolerant Saccharomyces eubayanus. To understand the contribution of both parental genomes to maltose metabolism in brewing wort, this study focuses on maltose transport in the S. eubayanus type strain CBS 12357T/FM1318. To obtain complete sequences of the MAL loci of this strain, a near-complete genome assembly was generated using the Oxford Nanopore Technology MinION sequencing platform. Except for CHRXII, all sixteen chromosomes were assembled as single contigs. Four loci harboring putative maltose transporter genes (SeMALT1-4), located in subtelomeric regions of CHRII, CHRV, CHRXIII, and CHRXVI, were completely resolved. The near-identical loci on CHRV and CHRXVI strongly resembled canonical S. cerevisiae MAL loci, while those on CHRII and CHRXIII showed different structures suggestive of gene loss. Overexpression of SeMALT1-4 in a maltose-transport-deficient S. cerevisiae strain restored growth on maltose, but not on maltotriose, indicating maltose-specific transport functionality of all four transporters. Simultaneous CRISPR-Cas9-assisted deletion of only SeMALT2 and SeMALT4, which shared 99.7% sequence identity, el...Continue Reading

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Citations

Dec 6, 2018·Nucleic Acids Research·Arthur R Gorter de VriesJean-Marc G Daran
Sep 26, 2019·FEMS Yeast Research·Arthur R Gorter de VriesJean-Marc G Daran
Jan 18, 2020·Annual Review of Food Science and Technology·B GibsonV Vidgren
Feb 26, 2020·Microbial Biotechnology·Wladimir MardonesFrancisco A Cubillos
Apr 20, 2019·Frontiers in Genetics·Arthur R Gorter de VriesJean-Marc G Daran
Jun 26, 2020·Frontiers in Genetics·Arthur R Gorter de VriesJean-Marc G Daran
Nov 27, 2020·Frontiers in Genetics·Jennifer Molinet, Francisco A Cubillos
Mar 13, 2021·International Journal of Food Microbiology·Melisa González FloresChristian Ariel Lopes

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Datasets Mentioned

BETA
PRJNA450912
GSE117246

Methods Mentioned

BETA
SGA
PCR
Illumina sequencing
RNA seq

Software Mentioned

BLASTX
DESeq
EdgeR package
MAKER2
Augustus
SNAP
htseq
MinKNOW
BLASTP
Albacore

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