Abstract:Endocarpon pusillum is a lichen-forming fungus with an outstanding stress resistance property closely related to its antioxidant system. In this study, thioredoxin (Trx), one of the main components of antioxidant defense systems in E. pusillum (EpTrx), was characterized and analyzed both in transgenic yeasts and in vitro. Our analyses identified that the heterologous expression of EpTrx in the yeast Pichia pastoris significantly enhanced its resistance to osmotic and oxidative stresses. Assays in vitro showed … Show more
“…An enhanced tolerance of strain TPX1 to high concentration of salt was just as expected, which showed similar results with some related Prxs [29, 30]. This finding is of great significance to reducing the costs for scale-up of ethanol production.…”
Section: Resultssupporting
confidence: 86%
“…However, when 3 mM of H 2 O 2 was added to the plates, TPX1 achieved a much better growth status than the strain 423, which indicated that the gene KmTPX1 might increase the tolerance of S. cerevisiae to H 2 O 2 . Similarly, a kind of 2-Cys Prxs from Oryza sativa [29] and the thioredoxin from Endocarpon pusillum [30] both have been proved to enhance the tolerance of S. cerevisiae to peroxides, including H 2 O 2 and 2-methyl-1,4-naphthoquinone (MD).Fig. 2Effects of KmTPX1 overexpression on the growth behavior under oxidative stress.…”
Section: Resultsmentioning
confidence: 99%
“…Finally, no significant effect of KmTPX1 in cell growth at high temperature was observed in spite of the relations between high temperature and intracellular ROS [31]. In contrast, another Prx family protein, thioredoxin from Endocarpon pusillum , showed a positive role in increasing yeast tolerance to high temperature [30]. …”
BackgroundBioethanol from lignocellulosic materials is of great significance to the production of renewable fuels due to its wide sources. However, multiple inhibitors generated from pretreatments represent great challenges for its industrial-scale fermentation. Despite the complex toxicity mechanisms, lignocellulose-derived inhibitors have been reported to be related to the levels of intracellular reactive oxygen species (ROS), which makes oxidoreductase a potential target for the enhancement of the tolerance of yeasts to these inhibitors.ResultsA typical 2-Cys peroxiredoxin from Kluyveromyces marxianus Y179 (KmTPX1) was identified, and its overexpression was achieved in Saccharomyces cerevisiae 280. Strain TPX1 with overexpressed KmTPX1 gene showed an enhanced tolerance to oxidative stresses. Serial dilution assay indicated that KmTPX1 gene contributed to a better cellular growth behavior, when the cells were exposed to multiple lignocellulose-derived inhibitors, such as formic acid, acetic acid, furfural, ethanol, and salt. In particular, KmTPX1 expression also possessed enhanced tolerance to a mixture of formic acid, acetic acid, and furfural (FAF) with a shorter lag period. The maximum glucose consumption rate and ethanol generation rate in KmTPX1-expressing strain were significantly improved, compared with the control. The mechanism of improved tolerance to FAF depends on the lower level of intracellular ROS for cell survival under stress.ConclusionA new functional gene KmTPX1 from K. marxianus is firstly associated with the enhanced tolerance to multiple lignocellulose-derived inhibitors in S. cerevisiae. We provided a possible detoxification mechanism of the KmTPX1 for further theoretical research; meanwhile, we provided a powerful potential for application of the KmTPX1 overexpressing strain in ethanol production from lignocellulosic materials.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-017-0766-4) contains supplementary material, which is available to authorized users.
“…An enhanced tolerance of strain TPX1 to high concentration of salt was just as expected, which showed similar results with some related Prxs [29, 30]. This finding is of great significance to reducing the costs for scale-up of ethanol production.…”
Section: Resultssupporting
confidence: 86%
“…However, when 3 mM of H 2 O 2 was added to the plates, TPX1 achieved a much better growth status than the strain 423, which indicated that the gene KmTPX1 might increase the tolerance of S. cerevisiae to H 2 O 2 . Similarly, a kind of 2-Cys Prxs from Oryza sativa [29] and the thioredoxin from Endocarpon pusillum [30] both have been proved to enhance the tolerance of S. cerevisiae to peroxides, including H 2 O 2 and 2-methyl-1,4-naphthoquinone (MD).Fig. 2Effects of KmTPX1 overexpression on the growth behavior under oxidative stress.…”
Section: Resultsmentioning
confidence: 99%
“…Finally, no significant effect of KmTPX1 in cell growth at high temperature was observed in spite of the relations between high temperature and intracellular ROS [31]. In contrast, another Prx family protein, thioredoxin from Endocarpon pusillum , showed a positive role in increasing yeast tolerance to high temperature [30]. …”
BackgroundBioethanol from lignocellulosic materials is of great significance to the production of renewable fuels due to its wide sources. However, multiple inhibitors generated from pretreatments represent great challenges for its industrial-scale fermentation. Despite the complex toxicity mechanisms, lignocellulose-derived inhibitors have been reported to be related to the levels of intracellular reactive oxygen species (ROS), which makes oxidoreductase a potential target for the enhancement of the tolerance of yeasts to these inhibitors.ResultsA typical 2-Cys peroxiredoxin from Kluyveromyces marxianus Y179 (KmTPX1) was identified, and its overexpression was achieved in Saccharomyces cerevisiae 280. Strain TPX1 with overexpressed KmTPX1 gene showed an enhanced tolerance to oxidative stresses. Serial dilution assay indicated that KmTPX1 gene contributed to a better cellular growth behavior, when the cells were exposed to multiple lignocellulose-derived inhibitors, such as formic acid, acetic acid, furfural, ethanol, and salt. In particular, KmTPX1 expression also possessed enhanced tolerance to a mixture of formic acid, acetic acid, and furfural (FAF) with a shorter lag period. The maximum glucose consumption rate and ethanol generation rate in KmTPX1-expressing strain were significantly improved, compared with the control. The mechanism of improved tolerance to FAF depends on the lower level of intracellular ROS for cell survival under stress.ConclusionA new functional gene KmTPX1 from K. marxianus is firstly associated with the enhanced tolerance to multiple lignocellulose-derived inhibitors in S. cerevisiae. We provided a possible detoxification mechanism of the KmTPX1 for further theoretical research; meanwhile, we provided a powerful potential for application of the KmTPX1 overexpressing strain in ethanol production from lignocellulosic materials.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-017-0766-4) contains supplementary material, which is available to authorized users.
“…The role of detoxification enzymes in cell protection has been well-documented in other plants (Sappl et al, 2009; Das and Roychoudhury, 2014), as well as the roles of proteins such as nucleoredoxin, multidomain thioredoxin, and ferritin (Kang and Udvardi, 2014; Li and Wei, 2016). In this study ferritins were found to be upregulated in the drought-stressed plants, potentially in order to sequester free iron that would otherwise catalyze the Fenton reaction and produce highly reactive hydroxyl radicals.…”
Prunus persica L. Batsch, or peach, is one of the most important crops and it is widely established in irrigated arid and semi-arid regions. However, due to variations in the climate and the increased aridity, drought has become a major constraint, causing crop losses worldwide. The use of drought-tolerant rootstocks in modern fruit production appears to be a useful method of alleviating water deficit problems. However, the transcriptomic variation and the major molecular mechanisms that underlie the adaptation of drought-tolerant rootstocks to water shortage remain unclear. Hence, in this study, high-throughput sequencing (RNA-seq) was performed to assess the transcriptomic changes and the key genes involved in the response to drought in root tissues (GF677 rootstock) and leaf tissues (graft, var. Catherina) subjected to 16 days of drought stress. In total, 12 RNA libraries were constructed and sequenced. This generated a total of 315 M raw reads from both tissues, which allowed the assembly of 22,079 and 17,854 genes associated with the root and leaf tissues, respectively. Subsets of 500 differentially expressed genes (DEGs) in roots and 236 in leaves were identified and functionally annotated with 56 gene ontology (GO) terms and 99 metabolic pathways, which were mostly associated with aminobenzoate degradation and phenylpropanoid biosynthesis. The GO analysis highlighted the biological functions that were exclusive to the root tissue, such as “locomotion,” “hormone metabolic process,” and “detection of stimulus,” indicating the stress-buffering role of the GF677 rootstock. Furthermore, the complex regulatory network involved in the drought response was revealed, involving proteins that are associated with signaling transduction, transcription and hormone regulation, redox homeostasis, and frontline barriers. We identified two poorly characterized genes in P. persica: growth-regulating factor 5 (GRF5), which may be involved in cellular expansion, and AtHB12, which may be involved in root elongation. The reliability of the RNA-seq experiment was validated by analyzing the expression patterns of 34 DEGs potentially involved in drought tolerance using quantitative reverse transcription polymerase chain reaction. The transcriptomic resources generated in this study provide a broad characterization of the acclimation of P. persica to drought, shedding light on the major molecular responses to the most important environmental stressor.
“…As dominant species, Endocarpon pusillum Hedw. has been intensively studied from many aspects, such as physiology under desiccation and starvation stress 10 , photosynthetic rate 11 , genome 12 , transcriptome 13 and stress resistance functional protein 14 . And all the above results supported that Endocarpon pusillum has good drought resistant ability, which could be one potential species used in the prospective ‘desert biocarpet engieering’ (DBCE) 15 , which has been proposed by Wei 16 , referring to developing during a short period into a dominant protection system instead of the naked desert with the help of the artificial inoculation of the microorganisms.…”
Endocarpon species are key components of biological soil crusts. Phenotypic and systematic molecular analyses were carried out to identify samples of Endocarpon collected from the southeast edge of the Tengger Desert in China. These morphological and molecular analyses revealed two previously undescribed species that form highly supported independent monophyletic clades within Endocarpon. The new taxa were named Endocarpon deserticola sp. nov. and E. unifoliatum sp. nov. Furthermore, our results indicated that the newly developed protein coding markers adenylate kinase (ADK) and ubiquitin-conjugating enzyme h (UCEH) are useful for assessing species boundaries in phylogenic analyses.
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