Disruption of multiple genes whose deletion causes lactic-acid resistance improves lactic-acid resistance and productivity in Saccharomyces cerevisiae

Suzuki, Toshihiro; Sakamoto, Takatoshi; Sugiyama, Minetaka; Ishida, Nobuhiro; Kambe, Hiromi; Obata, Seiji; Kaneko, Yoshinobu; Takahashi, Hitomi; Harashima, Satoshi

doi:10.1016/j.jbiosc.2012.11.014

Cited by 33 publications

(23 citation statements)

References 42 publications

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“…Accordingly, conferring higher acid tolerance to S . cerevisiae would improve both ethanol and lactic acid production under acidic conditions [25]. …”

Section: Introductionmentioning

confidence: 99%

Identification and Characterization of a Novel Issatchenkia orientalis GPI-Anchored Protein, IoGas1, Required for Resistance to Low pH and Salt Stress

et al. 2016

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The use of yeasts tolerant to acid (low pH) and salt stress is of industrial importance for several bioproduction processes. To identify new candidate genes having potential roles in low-pH tolerance, we screened an expression genomic DNA library of a multiple-stress-tolerant yeast, Issatchenkia orientalis (Pichia kudriavzevii), for clones that allowed Saccharomyces cerevisiae cells to grow under highly acidic conditions (pH 2.0). A genomic DNA clone containing two putative open reading frames was obtained, of which the putative protein-coding gene comprising 1629 bp was retransformed into the host. This transformant grew significantly at pH 2.0, and at pH 2.5 in the presence of 7.5% Na2SO4. The predicted amino acid sequence of this new gene, named I. orientalis GAS1 (IoGAS1), was 60% identical to the S. cerevisiae Gas1 protein, a glycosylphosphatidylinositol-anchored protein essential for maintaining cell wall integrity, and 58–59% identical to Candida albicans Phr1 and Phr2, pH-responsive proteins implicated in cell wall assembly and virulence. Northern hybridization analyses indicated that, as for the C. albicans homologs, IoGAS1 expression was pH-dependent, with expression increasing with decreasing pH (from 4.0 to 2.0) of the medium. These results suggest that IoGAS1 represents a novel pH-regulated system required for the adaptation of I. orientalis to environments of diverse pH. Heterologous expression of IoGAS1 complemented the growth and morphological defects of a S. cerevisiae gas1Δ mutant, demonstrating that IoGAS1 and the corresponding S. cerevisiae gene play similar roles in cell wall biosynthesis. Site-directed mutagenesis experiments revealed that two conserved glutamate residues (E161 and E262) in the IoGas1 protein play a crucial role in yeast morphogenesis and tolerance to low pH and salt stress. Furthermore, overexpression of IoGAS1 in S. cerevisiae remarkably improved the ethanol fermentation ability at pH 2.5, and at pH 2.0 in the presence of salt (5% Na2SO4), compared to that of a reference strain. Our results strongly suggest that constitutive expression of the IoGAS1 gene in S. cerevisiae could be advantageous for several fermentation processes under these stress conditions.

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“…Accordingly, conferring higher acid tolerance to S . cerevisiae would improve both ethanol and lactic acid production under acidic conditions [25]. …”

Section: Introductionmentioning

confidence: 99%

Identification and Characterization of a Novel Issatchenkia orientalis GPI-Anchored Protein, IoGas1, Required for Resistance to Low pH and Salt Stress

et al. 2016

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“…Still, below pH values of 2.8, lactic acid production with the engineered yeast strain dropped (7). Therefore, conferring higher lactic acid resistance on S. cerevisiae would improve lactic acid productivity under nonneutralized conditions (10).…”

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confidence: 99%

Nuclear Localization of Haa1, Which Is Linked to Its Phosphorylation Status, Mediates Lactic Acid Tolerance in Saccharomyces cerevisiae

Sugiyama

Akase

Nakanishi

et al. 2014

Appl Environ Microbiol

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Improvement of the lactic acid resistance of the yeast Saccharomyces cerevisiae is important for the application of the yeast in industrial production of lactic acid from renewable resources. However, we still do not know the precise mechanisms of the lactic acid adaptation response in yeast and, consequently, lack effective approaches for improving its lactic acid tolerance. To enhance our understanding of the adaptation response, we screened for S. cerevisiae genes that confer enhanced lactic acid resistance when present in multiple copies and identified the transcriptional factor Haa1 as conferring resistance to toxic levels of lactic acid when overexpressed. The enhanced tolerance probably results from increased expression of its target genes. When cells that expressed Haa1 only from the endogenous promoter were exposed to lactic acid stress, the main subcellular localization of Haa1 changed from the cytoplasm to the nucleus within 5 min. This nuclear accumulation induced upregulation of the Haa1 target genes YGP1, GPG1, and SPI1, while the degree of Haa1 phosphorylation observed under lactic acid-free conditions decreased. Disruption of the exportin gene MSN5 led to accumulation of Haa1 in the nucleus even when no lactic acid was present. Since Msn5 was reported to interact with Haa1 and preferentially exports phosphorylated cargo proteins, our results suggest that regulation of the subcellular localization of Haa1, together with alteration of its phosphorylation status, mediates the adaptation to lactic acid stress in yeast.

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“…Rational engineering has been used also for improving LA tolerance of a weak LA producer, that is, Saccharomyces cerevisiae . A gene deletion library indicated that several genes affect LA tolerance in this microorganism . Disruption of these genes increased LA resistance and LA productivity.…”

Section: Metabolic Engineering Strategies For Direct Production Of Lamentioning

confidence: 99%

“…Disruption of these genes increased LA resistance and LA productivity. Furthermore, multiple gene disruption had cumulative effects . Adaptive evolution approach was recently used to improve LA tolerance of Leuconostoc mesenteroides up to 70 g/L .…”

Section: Metabolic Engineering Strategies For Direct Production Of Lamentioning

confidence: 99%

Metabolic engineering strategies for consolidated production of lactic acid from lignocellulosic biomass

Mazzoli

2020

Biotech and App Biochem

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Lactic acid (LA) is one of the most desired molecules by the chemical industry. Current expansion of LA market is mainly driven by its application as building block for the synthesis of polylactide (PLA), that is, a family of biodegradable and biocompatible plastic polymers. PLA can potentially replace oil‐derived polymers as general purpose plastic, but current LA prices fails to make PLA cost‐competitive with traditional plastics. Nowadays, LA is mainly produced by fermentation of expensive starchy biomass. Hopefully, cheaper lignocellulosic feedstock could be used in future second‐generation biorefinery processes. However, most efficient natural LA producers cannot ferment lignocellulose without prior biomass saccharification. Metabolic engineering may develop improved microorganisms that feature both efficient biomass hydrolysis and LA production, thus supporting consolidated bioprocessing (CBP), that is, one‐pot fermentation, of lignocellulose to LA. CBP could dramatically reduce LA production cost, thus contributing to the expansion of more environmental sustainable plastics and commodity chemicals. This review presents an overview of “recombinant cellulolytic strategies”, mainly consisting in introducing cellulase systems in native producers of LA, and “native cellulolytic strategies” aimed at improving LA production in natural cellulolytic microorganisms. Issues and perspectives of these approaches will be discussed.

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Disruption of multiple genes whose deletion causes lactic-acid resistance improves lactic-acid resistance and productivity in Saccharomyces cerevisiae

Cited by 33 publications

References 42 publications

Identification and Characterization of a Novel Issatchenkia orientalis GPI-Anchored Protein, IoGas1, Required for Resistance to Low pH and Salt Stress

Identification and Characterization of a Novel Issatchenkia orientalis GPI-Anchored Protein, IoGas1, Required for Resistance to Low pH and Salt Stress

Nuclear Localization of Haa1, Which Is Linked to Its Phosphorylation Status, Mediates Lactic Acid Tolerance in Saccharomyces cerevisiae

Metabolic engineering strategies for consolidated production of lactic acid from lignocellulosic biomass

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