Similarities and differences in the biochemical and enzymological properties of the four isomaltases from <i>Saccharomyces cerevisiae</i>

Deng, Xu; Petitjean, Marjorie; Teste, Marie-Ange; Kooli, Wafa M.; Tranier, S.; François, Jean; Parrou, Jean-Luc

doi:10.1016/j.fob.2014.02.004

Cited by 29 publications

(31 citation statements)

References 63 publications

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“…While in this study overexpression of IMA1 was shown to result in faster growth on sucrose, IMA5 was shown to be predominantly an isomaltase and overexpression resulted in only very slow growth and low intracellular sucrose hydrolysis activity (Table 4) (Deng et al. 2014). It therefore seems likely that the increased expression of IMA5 is collateral to the upregulation of MAL23-C .…”

Section: Discussionmentioning

confidence: 99%

Elimination of sucrose transport and hydrolysis in Saccharomyces cerevisiae: a platform strain for engineering sucrose metabolism

Marques

Mans

Marella

et al. 2017

FEMS Yeast Research

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Many relevant options to improve efficacy and kinetics of sucrose metabolism in Saccharomyces cerevisiae and, thereby, the economics of sucrose-based processes remain to be investigated. An essential first step is to identify all native sucrose-hydrolysing enzymes and sucrose transporters in this yeast, including those that can be activated by suppressor mutations in sucrose-negative strains. A strain in which all known sucrose-transporter genes (MAL11, MAL21, MAL31, MPH2, MPH3) were deleted did not grow on sucrose after 2 months of incubation. In contrast, a strain with deletions in genes encoding sucrose-hydrolysing enzymes (SUC2, MAL12, MAL22, MAL32) still grew on sucrose. Its specific growth rate increased from 0.08 to 0.25 h−1 after sequential batch cultivation. This increase was accompanied by a 3-fold increase of in vitro sucrose-hydrolysis and isomaltase activities, as well as by a 3- to 5-fold upregulation of the isomaltase-encoding genes IMA1 and IMA5. One-step Cas9-mediated deletion of all isomaltase-encoding genes (IMA1-5) completely abolished sucrose hydrolysis. Even after 2 months of incubation, the resulting strain did not grow on sucrose. This sucrose-negative strain can be used as a platform to test metabolic engineering strategies and for fundamental studies into sucrose hydrolysis or transport.

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Section: Discussionmentioning

confidence: 99%

Elimination of sucrose transport and hydrolysis in Saccharomyces cerevisiae: a platform strain for engineering sucrose metabolism

Marques

Mans

Marella

et al. 2017

FEMS Yeast Research

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“…Sc has two types of α ‐glucosidase: α‐ 1,4‐glucosidase (EC 3.2.1.20, maltase) and oligo‐ α‐ 1,6‐glucosidase (EC 3.2.1.10, isomaltase) (Yamamoto et al , ), both belong to family 13 of the glycoside hydrolases (GH13), according to the CAZy classification (Lombard et al ., ). Maltases use maltose, maltulose, turanose and maltotriose as substrates; isomaltases use isomaltose, α ‐methylglucoside ( α ‐MG) and palatinose; whereas both use sucrose and p ‐nitrophenyl‐ α ‐ d ‐glucopyranoside ( α ‐ p NPG) (Yamamoto et al ., ; Teste et al , ; Voordeckers et al ., ; Deng et al ., ). Maltase genes and the respective proteins (Krakenaĭte and Glemzha, ; review by Needleman, ) were described earlier than those of isomaltase.…”

Section: Introductionmentioning

confidence: 97%

Maltase protein of Ogataea (Hansenula) polymorpha is a counterpart to the resurrected ancestor protein ancMALS of yeast maltases and isomaltases

Viigand

Visnapuu

Mardo

et al. 2016

Yeast

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Saccharomyces cerevisiae maltases use maltose, maltulose, turanose and maltotriose as substrates, isomaltases use isomaltose, α‐methylglucoside and palatinose and both use sucrose. These enzymes are hypothesized to have evolved from a promiscuous α‐glucosidase ancMALS through duplication and mutation of the genes. We studied substrate specificity of the maltase protein MAL1 from an earlier diverged yeast, Ogataea polymorpha (Op), in the light of this hypothesis. MAL1 has extended substrate specificity and its properties are strikingly similar to those of resurrected ancMALS. Moreover, amino acids considered to determine selective substrate binding are highly conserved between Op MAL1 and ancMALS. Op MAL1 represents an α‐glucosidase in which both maltase and isomaltase activities are well optimized in a single enzyme. Substitution of Thr200 (corresponds to Val216 in S. cerevisiae isomaltase IMA1) with Val in MAL1 drastically reduced the hydrolysis of maltose‐like substrates (α‐1,4‐glucosides), confirming the requirement of Thr at the respective position for this function. Differential scanning fluorimetry (DSF) of the catalytically inactive mutant Asp199Ala of MAL1 in the presence of its substrates and selected monosaccharides suggested that the substrate‐binding pocket of MAL1 has three subsites (–1, +1 and +2) and that binding is strongest at the –1 subsite. The DSF assay results were in good accordance with affinity (K m) and inhibition (K i) data of the enzyme for tested substrates, indicating the power of the method to predict substrate binding. Deletion of either the maltase (MAL1) or α‐glucoside permease (MAL2) gene in Op abolished the growth of yeast on MAL1 substrates, confirming the requirement of both proteins for usage of these sugars. © 2016 The Authors. Yeast published by John Wiley & Sons, Ltd.

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“…), which was later on attributed to its very high sensitivity to temperature in vitro (Deng et al . ). These last authors have shown that the IMA5‐ encoded enzyme is not able to hydrolyse maltose, has high activity with isomaltose and palatinose, and presents low, yet detectable activity with maltotriose.…”

Section: Resultsmentioning

confidence: 97%

“…It is worth noting that while the IMA1 gene is in close proximity to the MAL1 loci, all other IMA genes cluster with hexose transporters, including the IMA5 gene found between the HXT8 and HXT9 genes on chromosome X of S. cerevisiae. Initial attempts to characterize the specificity of this enzyme in cell extracts failed (Brown et al 2010;Teste et al 2010;Voordeckers et al 2012), which was later on attributed to its very high sensitivity to temperature in vitro (Deng et al 2014). These last authors have shown that the IMA5-encoded enzyme is not able to hydrolyse maltose, has high activity with isomaltose and palatinose, and presents low, yet detectable activity with maltotriose.…”

Section: Resultsmentioning

confidence: 99%

Extracellular maltotriose hydrolysis bySaccharomyces cerevisiaecells lacking theAGT1permease

Alves

Thevelein

Stambuk

2018

Lett Appl Microbiol

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Maltotriose is the second most abundant sugar present in brewing. However, many yeast strains have difficulties to consume maltotriose, mainly because of its low uptake rate by the yeast cells when compared to glucose and maltose uptake. The AGT1 permease is required for efficient maltotriose fermentation, but some strains deleted in this gene are still able to grow on maltotriose after an extensive lag phase. This manuscript shows that such delayed growth on maltotriose is a consequence of extracellular hydrolysis of the sugar. Our results also indicate that the IMA5-encoded α-glucosidase is likely the enzyme responsible for this phenotype.

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Similarities and differences in the biochemical and enzymological properties of the four isomaltases from Saccharomyces cerevisiae

Cited by 29 publications

References 63 publications

Elimination of sucrose transport and hydrolysis in Saccharomyces cerevisiae: a platform strain for engineering sucrose metabolism

Elimination of sucrose transport and hydrolysis in Saccharomyces cerevisiae: a platform strain for engineering sucrose metabolism

Maltase protein of Ogataea (Hansenula) polymorpha is a counterpart to the resurrected ancestor protein ancMALS of yeast maltases and isomaltases

Extracellular maltotriose hydrolysis bySaccharomyces cerevisiaecells lacking theAGT1permease

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