Deletion of the PHO13 gene in Saccharomyces cerevisiae improves ethanol production from lignocellulosic hydrolysate in the presence of acetic and formic acids, and furfural

Fujitomi, Keisuke; Sanda, Tomoya; Hasunuma, Tomohisa; Kondo, Akihiko

doi:10.1016/j.biortech.2012.01.161

Cited by 76 publications

(40 citation statements)

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“…On the other hand, three independent studies have found that pho13⌬ improves xylose metabolism in strains engineered with a heterologous xylose-consuming pathway (11)(12)(13). Although the phenotypic improvement was significant, the mechanism was difficult to explain because xylose metabolism itself is not clearly understood in the engineered strains.…”

Section: Discussionmentioning

confidence: 99%

“…Although little is known about the biochemical and physiological functions of S. cerevisiae Pho13, deletion of the PHO13 gene (pho13⌬) results in significant and industrially relevant phenotypic changes in engineered strains capable of fermenting xylose (11)(12)(13)(14)(15)(16). Introduction of a heterologous metabolic pathway consisting of either xylose reductase/xylitol dehydrogenase/xylulokinase [X(R/X)D(H/X)K] or xylose isomerase/xylulokinase [X(I/ X)K] allows S. cerevisiae to metabolize xylose (17).…”

mentioning

confidence: 99%

“…In both cases, the loss of Pho13 function (pho13⌬) was shown to be responsible for the improvement in xylose metabolism (12,14). Moreover, pho13⌬ in S. cerevisiae strains overexpressing X(R/X)D(H/X)K improved tolerance to common fermentation inhibitors (weak acids and sugar degradation products) (13,15). In addition to its effects in strains overexpressing X(R/X)D(H/X)K, pho13⌬ in a strain overexpressing X(I/X)K improved the growth rate on xylose 8-fold (16).…”

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confidence: 99%

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Deletion of PHO13 , Encoding Haloacid Dehalogenase Type IIA Phosphatase, Results in Upregulation of the Pentose Phosphate Pathway in Saccharomyces cerevisiae

Kim

Lesmana

et al. 2015

Appl Environ Microbiol

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eThe haloacid dehalogenase (HAD) superfamily is one of the largest enzyme families, consisting mainly of phosphatases. Although intracellular phosphate plays important roles in many cellular activities, the biological functions of HAD enzymes are largely unknown. Pho13 is 1 of 16 putative HAD enzymes in Saccharomyces cerevisiae. Pho13 has not been studied extensively, but previous studies have identified PHO13 to be a deletion target for the generation of industrially attractive phenotypes, namely, efficient xylose fermentation and high tolerance to fermentation inhibitors. In order to understand the molecular mechanisms underlying the improved xylose-fermenting phenotype produced by deletion of PHO13 (pho13⌬), we investigated the response of S. cerevisiae to pho13⌬ at the transcriptomic level when cells were grown on glucose or xylose. Transcriptome sequencing analysis revealed that pho13⌬ resulted in upregulation of the pentose phosphate (PP) pathway and NADPH-producing enzymes when cells were grown on glucose or xylose. We also found that the transcriptional changes induced by pho13⌬ required the transcription factor Stb5, which is activated specifically under NADPH-limiting conditions. Thus, pho13⌬ resulted in the upregulation of the PP pathway and NADPH-producing enzymes as a part of an oxidative stress response mediated by activation of Stb5. Because the PP pathway is the primary pathway for xylose, its upregulation by pho13⌬ might explain the improved xylose metabolism. These findings will be useful for understanding the biological function of S. cerevisiae Pho13 and the HAD superfamily enzymes and for developing S. cerevisiae strains with industrially attractive phenotypes.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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Deletion of PHO13 , Encoding Haloacid Dehalogenase Type IIA Phosphatase, Results in Upregulation of the Pentose Phosphate Pathway in Saccharomyces cerevisiae

Kim

Lesmana

et al. 2015

Appl Environ Microbiol

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“…Some examples of successful rational engineering efforts include: engineering of the redox state by enhancing the intracellular glutathione by GSH1 overexpression; co-expression of transaldolase and aldehyde dehydrogenase in the presence of furfural; over expression of aldehyde dehydrogenase to regenerate NAD + and relieve redox imbalance; over expression of RNA binding protein lsm6 for improved tolerance to furfural acetic acid and sulfur compounds; and combining the NADH consuming acetic acid consumption pathway and NADH producing xylose utilization pathway (Fujitomi et al 2012;Gao and Xia 2012;Tanaka et al 2012;Kim and Hahn 2013). Evolutionary engineering was carried out both by mutagenesis or adaptation to lignocellulose derived inhibitors.…”

Section: Inhibitor Tolerance Of Genetically Engineered Microbial Strainsmentioning

confidence: 99%

Can Microbially Derived Advanced Biofuels Ever Compete with Conventional Bioethanol? A Critical Review

Veettil

Kumar

Koukoulas³

2016

BioResources

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“…The estimated dry cell weight (DCW) was approximately 0.15-fold the wet cell weight. All fermentations were performed at 30°C under oxygen limited conditions at an agitation speed of 500 rpm in 100-mL closed bottles equipped with a bubbling CO 2 outlet and a stir bar, as described previously [32]. Cell growth was monitored by absorbance at 600 nm.…”

Section: Xylose Fermentation Under Oxygen-limited Conditionsmentioning

confidence: 99%

Development of a GIN11/FRT-based multiple-gene integration technique affording inhibitor-tolerant, hemicellulolytic, xylose-utilizing abilities to industrial Saccharomyces cerevisiae strains for ethanol production from undetoxified lignocellulosic hemicelluloses

et al. 2014

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Background: Bioethanol produced by the yeast Saccharomyces cerevisiae is currently one of the most promising alternatives to conventional transport fuels. Lignocellulosic hemicelluloses obtained after hydrothermal pretreatment are important feedstock for bioethanol production. However, hemicellulosic materials cannot be directly fermented by yeast: xylan backbone of hemicelluloses must first be hydrolyzed by heterologous hemicellulases to release xylose, and the yeast must then ferment xylose in the presence of fermentation inhibitors generated during the pretreatment. Results: A GIN11/FRT-based multiple-gene integration system was developed for introducing multiple functions into the recombinant S. cerevisiae strains engineered with the xylose metabolic pathway. Antibiotic markers were efficiently recycled by a novel counter selection strategy using galactose-induced expression of both FLP recombinase gene and GIN11 flanked by FLP recombinase recognition target (FRT) sequences. Nine genes were functionally expressed in an industrial diploid strain of S. cerevisiae: endoxylanase gene from Trichoderma reesei, xylosidase gene from Aspergillus oryzae, β-glucosidase gene from Aspergillus aculeatus, xylose reductase and xylitol dehydrogenase genes from Scheffersomyces stipitis, and XKS1, TAL1, FDH1 and ADH1 variant from S. cerevisiae. The genes were introduced using the homozygous integration system and afforded hemicellulolytic, xylose-assimilating and inhibitor-tolerant abilities to the strain. The engineered yeast strain demonstrated 2.7-fold higher ethanol titer from hemicellulosic material than a xylose-assimilating yeast strain. Furthermore, hemicellulolytic enzymes displayed on the yeast cell surface hydrolyzed hemicelluloses that were not hydrolyzed by a commercial enzyme, leading to increased sugar utilization for improved ethanol production.

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Deletion of the PHO13 gene in Saccharomyces cerevisiae improves ethanol production from lignocellulosic hydrolysate in the presence of acetic and formic acids, and furfural

Cited by 76 publications

References 24 publications

Deletion of PHO13 , Encoding Haloacid Dehalogenase Type IIA Phosphatase, Results in Upregulation of the Pentose Phosphate Pathway in Saccharomyces cerevisiae

Deletion of PHO13 , Encoding Haloacid Dehalogenase Type IIA Phosphatase, Results in Upregulation of the Pentose Phosphate Pathway in Saccharomyces cerevisiae

Can Microbially Derived Advanced Biofuels Ever Compete with Conventional Bioethanol? A Critical Review

Development of a GIN11/FRT-based multiple-gene integration technique affording inhibitor-tolerant, hemicellulolytic, xylose-utilizing abilities to industrial Saccharomyces cerevisiae strains for ethanol production from undetoxified lignocellulosic hemicelluloses

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