Isolation and characterization of xylitol-assimilating mutants of recombinant Saccharomyces cerevisiae

Tani, Tatsunori; Taguchi, Hisataka; Fujimori, Kazuhiro E.; Sahara, Takehiko; Ohgiya, Satoru; Kamagata, Yoichi; Akamatsu, Takashi

doi:10.1016/j.jbiosc.2016.03.008

Cited by 7 publications

(8 citation statements)

References 51 publications

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“… 2001 ; Tani et al. 2016 ). Expression of the high-affinity l -arabinose transporter Pc AraT from P. chrysogenum enables l -arabinose uptake in the presence of glucose (Bracher et al.…”

Section: Resultsmentioning

confidence: 99%

“… 2001 ), deregulated expression of the Gal2 galactose/ l -arabinose transporter (Tani et al. 2016 ; Wang et al. 2017 ), here accomplished by deletion of GAL80 (Torchia et al.…”

Section: Discussionmentioning

confidence: 99%

“… 2001 ; Tani et al. 2016 ) and expression of the fungal l -arabinose transporter Pc AraT (Bracher et al 2018 ) yielded a strain that, in aerobic shake-flask cultures, exhibited a specific growth rate on l -arabinose of 0.26 h −1 . Moreover, deletion of all four genes encoding enzymes with glucose-phosphorylating activity rendered this ‘arabinose-specialist’ strain S. cerevisiae IMX728 unable to grow on glucose.…”

Section: Discussionmentioning

confidence: 99%

“…Since transcription of GAL2 is both induced by d -galactose and repressed by d -glucose, design of l -arabinose-fermenting S. cerevisiae strains often includes expression of GAL2 behind a strong constitutive promoter (Tani et al. 2016 ; Wang et al. 2017 ).…”

Section: Introductionmentioning

confidence: 99%

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Laboratory evolution of a glucose-phosphorylation-deficient, arabinose-fermenting S. cerevisiae strain reveals mutations in GAL2 that enable glucose-insensitive l-arabinose uptake

Verhoeven

Bracher

Nijland

et al. 2018

FEMS Yeast Research

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Cas9-assisted genome editing was used to construct an engineered glucose-phosphorylation-negative S. cerevisiae strain, expressing the Lactobacillus plantaruml-arabinose pathway and the Penicillium chrysogenum transporter PcAraT. This strain, which showed a growth rate of 0.26 h−1 on l-arabinose in aerobic batch cultures, was subsequently evolved for anaerobic growth on l-arabinose in the presence of d-glucose and d-xylose. In four strains isolated from two independent evolution experiments the galactose-transporter gene GAL2 had been duplicated, with all alleles encoding Gal2N376T or Gal2N376I substitutions. In one strain, a single GAL2 allele additionally encoded a Gal2T89I substitution, which was subsequently also detected in the independently evolved strain IMS0010. In 14C-sugar-transport assays, Gal2N376S, Gal2N376T and Gal2N376I substitutions showed a much lower glucose sensitivity of l-arabinose transport and a much higher Km for d-glucose transport than wild-type Gal2. Introduction of the Gal2N376I substitution in a non-evolved strain enabled growth on l-arabinose in the presence of d-glucose. Gal2N376T, T89I and Gal2T89I variants showed a lower Km for l-arabinose and a higher Km for d-glucose than wild-type Gal2, while reverting Gal2N376T, T89I to Gal2N376 in an evolved strain negatively affected anaerobic growth on l-arabinose. This study indicates that optimal conversion of mixed-sugar feedstocks may require complex ‘transporter landscapes’, consisting of sugar transporters with complementary kinetic and regulatory properties.

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“… 2001 ; Tani et al. 2016 ). Expression of the high-affinity l -arabinose transporter Pc AraT from P. chrysogenum enables l -arabinose uptake in the presence of glucose (Bracher et al.…”

Section: Resultsmentioning

confidence: 99%

“… 2001 ), deregulated expression of the Gal2 galactose/ l -arabinose transporter (Tani et al. 2016 ; Wang et al. 2017 ), here accomplished by deletion of GAL80 (Torchia et al.…”

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Laboratory evolution of a glucose-phosphorylation-deficient, arabinose-fermenting S. cerevisiae strain reveals mutations in GAL2 that enable glucose-insensitive l-arabinose uptake

Verhoeven

Bracher

Nijland

et al. 2018

FEMS Yeast Research

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show abstract

“…Various genes from the galactose pathway have been used to improve D-xylose utilization. For instance, the galactose transporter Gal2p, the expression of which is controlled at the gene level by the GAL regulon, has been shown to transport D-xylose [111,253] and xylitol [254] Gal2p variants have been used in several studies to increase utilization of D-xylose by improving its transport inside the cell [255,256]. Phosphoglucomutase is encoded by PGM2 and catalyzes the interconversion between glucose-1-phosphate and glucose-6-phosphate, which is the last step in the Leloir pathway.…”

Section: Effect Of D-xylose On Other D-glucose-responsive Signaling Pathwaysmentioning

confidence: 99%

D-Xylose Sensing in Saccharomyces cerevisiae: Insights from D-Glucose Signaling and Native D-Xylose Utilizers

Brink

Borgström

Persson

et al. 2021

IJMS

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Extension of the substrate range is among one of the metabolic engineering goals for microorganisms used in biotechnological processes because it enables the use of a wide range of raw materials as substrates. One of the most prominent examples is the engineering of baker’s yeast Saccharomyces cerevisiae for the utilization of d-xylose, a five-carbon sugar found in high abundance in lignocellulosic biomass and a key substrate to achieve good process economy in chemical production from renewable and non-edible plant feedstocks. Despite many excellent engineering strategies that have allowed recombinant S. cerevisiae to ferment d-xylose to ethanol at high yields, the consumption rate of d-xylose is still significantly lower than that of its preferred sugar d-glucose. In mixed d-glucose/d-xylose cultivations, d-xylose is only utilized after d-glucose depletion, which leads to prolonged process times and added costs. Due to this limitation, the response on d-xylose in the native sugar signaling pathways has emerged as a promising next-level engineering target. Here we review the current status of the knowledge of the response of S. cerevisiae signaling pathways to d-xylose. To do this, we first summarize the response of the native sensing and signaling pathways in S. cerevisiae to d-glucose (the preferred sugar of the yeast). Using the d-glucose case as a point of reference, we then proceed to discuss the known signaling response to d-xylose in S. cerevisiae and current attempts of improving the response by signaling engineering using native targets and synthetic (non-native) regulatory circuits.

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Analysis of metabolisms and transports of xylitol using xylose- and xylitol-assimilating Saccharomyces cerevisiae

Tani

Taguchi

Akamatsu

2017

Journal of Bioscience and Bioengineering

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Isolation and characterization of xylitol-assimilating mutants of recombinant Saccharomyces cerevisiae

Cited by 7 publications

References 51 publications

Laboratory evolution of a glucose-phosphorylation-deficient, arabinose-fermenting S. cerevisiae strain reveals mutations in GAL2 that enable glucose-insensitive l-arabinose uptake

Laboratory evolution of a glucose-phosphorylation-deficient, arabinose-fermenting S. cerevisiae strain reveals mutations in GAL2 that enable glucose-insensitive l-arabinose uptake

D-Xylose Sensing in Saccharomyces cerevisiae: Insights from D-Glucose Signaling and Native D-Xylose Utilizers

Analysis of metabolisms and transports of xylitol using xylose- and xylitol-assimilating Saccharomyces cerevisiae

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