Extracellular maltotriose hydrolysis by<i>Saccharomyces cerevisiae</i>cells lacking the<i>AGT1</i>permease

Alves, Sérgio L.; Thevelein, Johan M.; Stambuk, Boris U.

doi:10.1111/lam.13048

Cited by 8 publications

(6 citation statements)

References 38 publications

(86 reference statements)

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“…It has previously been assumed that transport of maltotriose into the yeast cells is required for it to be consumed during wort fermentations (Day et al 2002; Rautio and Londesborough 2003; Alves et al 2008), but our results suggest that extracellular hydrolysis of maltotriose through STA1 allows for efficient consumption of maltotriose from the wort throughout fermentation. A recent study also reported extracellular hydrolysis of maltotriose by an α-glucosidase encoded by IMA5 (Alves et al 2018); however, in that case, maltotriose hydrolysis was only observed after an extensive lag phase. In brewing yeast where maltotriose use is enabled through the AGT1 -encoded transporter , maltotriose is typically consumed from the wort only towards the later parts of fermentation when maltose concentrations are low (Rautio and Londesborough 2003).…”

Section: Discussionmentioning

confidence: 97%

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

Krogerus

Magalhães

Kuivanen

et al. 2019

Appl Microbiol Biotechnol

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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. Electronic supplementary material The online version of this article (10.1007/s00253-019-10021-y) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 97%

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

Krogerus

Magalhães

Kuivanen

et al. 2019

Appl Microbiol Biotechnol

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“…The underlying reason for this molecule to be involved in the control of a complex important regulon is likely because it is a marker of the ecological niche of E. coli . To be sure, maltotriose is abundant in mixtures that result from incomplete fermentation of plant seed mixtures [this accounts for Saccharomyces cerevisiae being unable to ferment maltotriose efficiently (Alves et al ., )]. Maltotriose is thus expected to be present as a major catabolic by‐product of carbohydrate fibres in the gut of many mammals, where E. coli thrives (van Zanten et al ., ).…”

Section: An Open List Of Basic Cellular Maxwell's Demonsmentioning

confidence: 99%

Omnipresent Maxwell's demons orchestrate information management in living cells

Boël

Danot

Lorenzo

et al. 2019

Microbial Biotechnology

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Summary The development of synthetic biology calls for accurate understanding of the critical functions that allow construction and operation of a living cell. Besides coding for ubiquitous structures, minimal genomes encode a wealth of functions that dissipate energy in an unanticipated way. Analysis of these functions shows that they are meant to manage information under conditions when discrimination of substrates in a noisy background is preferred over a simple recognition process. We show here that many of these functions, including transporters and the ribosome construction machinery, behave as would behave a material implementation of the information‐managing agent theorized by Maxwell almost 150 years ago and commonly known as Maxwell's demon (MxD). A core gene set encoding these functions belongs to the minimal genome required to allow the construction of an autonomous cell. These MxDs allow the cell to perform computations in an energy‐efficient way that is vastly better than our contemporary computers.

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“…Strain CAT-1 has the AGT1 gene in chromosome VII [ 76 ], but this strain does not efficiently ferment maltotriose (another substrate of the AGT1 permease) [ 44 ] due to the presence of a divergent and non-functional promoter region, also found in other industrial yeast strains unable to ferment maltotriose [ 77 ]. Instead, strain CAT-1 uses maltotriose only after an extensive lag phase, and it metabolizes this sugar aerobically due to its extracellular hydrolysis mediated by the isomaltase encoded by the IMA5 gene [ 44 , 78 ]. Indeed, as can be seen in Table 3 , strain CAT-1 had no p NPαG transport activity, a measurement of this permease’s activity, indicating that it was also necessary to overexpress the AGT1 gene.…”

Section: Resultsmentioning

confidence: 99%

Improved Sugarcane-Based Fermentation Processes by an Industrial Fuel-Ethanol Yeast Strain

Muller,

de Godoy,

Dário

et al. 2023

JoF

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In Brazil, sucrose-rich broths (cane juice and/or molasses) are used to produce billions of liters of both fuel ethanol and cachaça per year using selected Saccharomyces cerevisiae industrial strains. Considering the important role of feedstock (sugar) prices in the overall process economics, to improve sucrose fermentation the genetic characteristics of a group of eight fuel-ethanol and five cachaça industrial yeasts that tend to dominate the fermentors during the production season were determined by array comparative genomic hybridization. The widespread presence of genes encoding invertase at multiple telomeres has been shown to be a common feature of both baker’s and distillers’ yeast strains, and is postulated to be an adaptation to sucrose-rich broths. Our results show that only two strains (one fuel-ethanol and one cachaça yeast) have amplification of genes encoding invertase, with high specific activity. The other industrial yeast strains had a single locus (SUC2) in their genome, with different patterns of invertase activity. These results indicate that invertase activity probably does not limit sucrose fermentation during fuel-ethanol and cachaça production by these industrial strains. Using this knowledge, we changed the mode of sucrose metabolism of an industrial strain by avoiding extracellular invertase activity, overexpressing the intracellular invertase, and increasing its transport through the AGT1 permease. This approach allowed the direct consumption of the disaccharide by the cells, without releasing glucose or fructose into the medium, and a 11% higher ethanol production from sucrose by the modified industrial yeast, when compared to its parental strain.

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Extracellular maltotriose hydrolysis bySaccharomyces cerevisiaecells lacking theAGT1permease

Cited by 8 publications

References 38 publications

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

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

Omnipresent Maxwell's demons orchestrate information management in living cells

Improved Sugarcane-Based Fermentation Processes by an Industrial Fuel-Ethanol Yeast Strain

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