Engineering of an endogenous hexose transporter into a specific D-xylose transporter facilitates glucose-xylose co-consumption in Saccharomyces cerevisiae

Nijland, Jeroen G.; Shin, Hyun Yong; Jong, René M. de; Waal, Paul P. de; Klaassen, Paul; Driessen, Arnold J. M.

doi:10.1186/s13068-014-0168-9

Cited by 108 publications

(139 citation statements)

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“…For effective sugar assimilation, sugar-specific transporters have significant roles and are commonly regulated at both the transcriptional and enzymatic levels (18). Recent characterization of conserved structural motifs and amino acids responsible for xylose-and cellodextrin-specific transporters in S. cerevisiae provides useful insights into complex sugar utilization (19)(20)(21)(22).…”

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confidence: 99%

Activating and Elucidating Metabolism of Complex Sugars in Yarrowia lipolytica

Ryu

Hipp

Trinh

2016

Appl Environ Microbiol

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The oleaginous yeast Yarrowia lipolytica is an industrially important host for production of organic acids, oleochemicals, lipids, and proteins with broad biotechnological applications. Albeit known for decades, the unique native metabolism of Y. lipolytica for using complex fermentable sugars, which are abundant in lignocellulosic biomass, is poorly understood. In this study, we activated and elucidated the native sugar metabolism in Y. lipolytica for cell growth on xylose and cellobiose as well as their mixtures with glucose through comprehensive metabolic and transcriptomic analyses. We identified 7 putative glucose-specific transporters, 16 putative xylose-specific transporters, and 4 putative cellobiose-specific transporters that are transcriptionally upregulated for growth on respective single sugars. Y. lipolytica is capable of using xylose as a carbon source, but xylose dehydrogenase is the key bottleneck of xylose assimilation and is transcriptionally repressed by glucose. Y. lipolytica has a set of 5 extracellular and 6 intracellular ␤-glucosidases and is capable of assimilating cellobiose via extra-and intracellular mechanisms, the latter being dominant for growth on cellobiose as a sole carbon source. Strikingly, Y. lipolytica exhibited enhanced sugar utilization for growth in mixed sugars, with strong carbon catabolite activation for growth on the mixture of xylose and cellobiose and with mild carbon catabolite repression of glucose on xylose and cellobiose. The results of this study shed light on fundamental understanding of the complex native sugar metabolism of Y. lipolytica and will help guide inverse metabolic engineering of Y. lipolytica for enhanced conversion of biomass-derived fermentable sugars to chemicals and fuels.

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confidence: 99%

Activating and Elucidating Metabolism of Complex Sugars in Yarrowia lipolytica

Ryu

Hipp

Trinh

2016

Appl Environ Microbiol

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“…Previous studies demonstrated that the conserved asparagine at position 366 of Hxt11 is a key determinant for the sugar specificity (Nijland et al, 2014; Shin et al, 2015). To improve on the xylose transport specificity of the chimeric Hxt11/2 transporter, the HXT2 gene was mutagenized at the corresponding position N361.…”

Section: Resultsmentioning

confidence: 99%

The amino‐terminal tail of Hxt11 confers membrane stability to the Hxt2 sugar transporter and improves xylose fermentation in the presence of acetic acid

Shin

Nijland

Waal

et al. 2017

Biotech & Bioengineering

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Hxt2 is a glucose repressed, high affinity glucose transporter of the yeast Saccharomyces cerevisiae and is subjected to high glucose induced degradation. Hxt11 is a sugar transporter that is stably expressed at the membrane irrespective the sugar concentration. To transfer this property to Hxt2, the N‐terminal tail of Hxt2 was replaced by the corresponding region of Hxt11 yielding a chimeric Hxt11/2 transporter. This resulted in the stable expression of Hxt2 at the membrane and improved the growth on 8% d‐glucose and 4% d‐xylose. Mutation of N361 of Hxt11/2 into threonine reversed the specificity for d‐xylose over d‐glucose with high d‐xylose transport rates. This mutant supported efficient sugar fermentation of both d‐glucose and d‐xylose at industrially relevant sugar concentrations even in the presence of the inhibitor acetic acid which is normally present in lignocellulosic hydrolysates. Biotechnol. Bioeng. 2017;114: 1937–1945. © 2017 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.

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“…Engineered membrane transporters can be of use to improve uptake of nutrients in production strains, like sugars for microbial bioethanol production (Ren et al, 2009;Nijland et al, 2014), or to enhance solute efflux that yields tolerance for cytotoxic products or allows secretion of desired products from the cell, thus boosting carbon flux of a metabolic pathway toward the desired product (Foo and Leong, 2013). However, membrane protein engineering is not easy due to the limited structural information available and difficulties to establish activity assays that can be performed in a high-throughput manner (Lian et al, 2014).…”

Section: Pharmaceutical and Biotechnological Relevance Of Pnu Transpomentioning

confidence: 99%

Structure, function, evolution and application of bacterial Pnu-type vitamin transporters

Jaehme

Slotboom

2015

Biological Chemistry

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Many bacteria can take up vitamins from the environment via specific transport machineries. Uptake is essential for organisms that lack complete vitamin biosynthesis pathways, but even in the presence of biosynthesis routes uptake is likely preferred, because it is energetically less costly. Pnu transporters represent a class of membrane transporters for a diverse set of B-type vitamins. They were identified 30 years ago and catalyze transport by the mechanism of facilitated diffusion, without direct coupling to ATP hydrolysis or transport of coupling ions. Instead, directionality is achieved by metabolic trapping, in which the vitamin substrate is converted into a derivative that cannot be transported, for instance by phosphorylation. The recent crystal structure of the nicotinamide riboside transporter PnuC has provided the first insights in substrate recognition and selectivity. Here, we will summarize the current knowledge about the function, structure, and evolution of Pnu transporters. Additionally, we will highlight their role for potential biotechnological and pharmaceutical applications.

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Engineering of an endogenous hexose transporter into a specific D-xylose transporter facilitates glucose-xylose co-consumption in Saccharomyces cerevisiae

Cited by 108 publications

References 28 publications

Activating and Elucidating Metabolism of Complex Sugars in Yarrowia lipolytica

Activating and Elucidating Metabolism of Complex Sugars in Yarrowia lipolytica

The amino‐terminal tail of Hxt11 confers membrane stability to the Hxt2 sugar transporter and improves xylose fermentation in the presence of acetic acid

Structure, function, evolution and application of bacterial Pnu-type vitamin transporters

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