Targets of oxidative stress in yeast sod mutants

Pereira, Marcos D.; Herdeiro, R.S.; Fernandes, Patricia N.; Eleuthério, Elis C. A.; Panek, Anita D.

doi:10.1016/s0304-4165(03)00003-5

Cited by 60 publications

(48 citation statements)

References 30 publications

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“…After 18 h of growth, the pH of each flask was adjusted to 5.0 with KOH-H 2 SO 4 , and 30 ml of each cell suspension was then transferred to 30 ml of potassium phosphate buffer (pH 5.0). Next, 50 g of the ROS-sensitive fluorescent dye 5-(and -6)-chloromethyl-2Ј,7Ј-dichlorodihydrofluorescein diacetate acetyl ester (CM-H 2 DCFDA; Invitrogen) dissolved in 100 l of absolute ethanol was added to each culture (29,39). Cells were incubated in a shaking incubator at 30°C for 2 h to allow diffusional entry of the dye and reaction with intracellular ROS.…”

Section: Methodsmentioning

confidence: 99%

Catalase Overexpression Reduces Lactic Acid-Induced Oxidative Stress in Saccharomyces cerevisiae

Abbott

Suir

Duong

et al. 2009

Appl Environ Microbiol

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Industrial production of lactic acid with the current pyruvate decarboxylase-negative Saccharomyces cerevisiae strains requires aeration to allow for respiratory generation of ATP to facilitate growth and, even under nongrowing conditions, cellular maintenance. In the current study, we observed an inhibition of aerobic growth in the presence of lactic acid. Unexpectedly, the cyb2⌬ reference strain, used to avoid aerobic consumption of lactic acid, had a specific growth rate of 0.25 h ؊1 in anaerobic batch cultures containing lactic acid but only 0.16 h ؊1 in identical aerobic cultures. Measurements of aerobic cultures of S. cerevisiae showed that the addition of lactic acid to the growth medium resulted in elevated levels of reactive oxygen species (ROS). To reduce the accumulation of lactic acid-induced ROS, cytosolic catalase (CTT1) was overexpressed by replacing the native promoter with the strong constitutive TPI1 promoter. Increased activity of catalase was confirmed and later correlated with decreased levels of ROS and increased specific growth rates in the presence of high lactic acid concentrations. The increased fitness of this genetically modified strain demonstrates the successful attenuation of additional stress that is derived from aerobic metabolism and may provide the basis for enhanced (micro)aerobic production of organic acids in S. cerevisiae.Lactic acid is an organic acid with a wide range of applications. In the food industry, lactic acid has traditionally been used as an antimicrobial as well as a flavor enhancer. Besides having applications in textile, cosmetic, and pharmaceutical industries (5), lactic acid has been applied for the manufacture of lactic acid polymers (11,40). These polymers have properties that are similar to those of petroleum-derived plastics. Skyrocketing oil prices caused by dwindling fossil fuel reserves coupled with pressures to tackle environmental issues are creating increased demand for bioderived, and often biodegradable, polymers, such as poly-lactic acid.Current industrial lactic acid fermentations are based on different species of lactic acid bacteria. These bacteria have complex nutrient requirements due to their limited ability to synthesize B vitamins and amino acids (8) and are intolerant to acidic conditions with a pH between 5.5 and 6.5 required for growth (40). Acidification of the growth medium during lactic acid fermentation is typically counteracted by the addition of neutralizing agents (e.g., CaCO 3 ), resulting in the formation of large quantities of insoluble salts, such as gypsum, during downstream processing.Saccharomyces cerevisiae has received attention as a possible alternative biocatalyst. This organism is relatively tolerant to low pH and has simple nutrient requirements. The production of lactic acid with metabolically engineered S. cerevisiae was achieved by introducing a NAD ϩ -dependent lactate dehydrogenase, leading to the simultaneous formation of both ethanol and lactate (1a, 12, 31, 32, 36). Further improvements were made by constr...

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Section: Methodsmentioning

confidence: 99%

Catalase Overexpression Reduces Lactic Acid-Induced Oxidative Stress in Saccharomyces cerevisiae

Abbott

Suir

Duong

et al. 2009

Appl Environ Microbiol

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“…This experiment was carried out by the method of Steels et al (1994), Pereira et al (2003) and Silva et al (2009). The S. cerevisiae wild type strain BY4741 (MATa;his3;leu2;met15;ura3) used in the in vivo antioxidant assays was acquired from Euroscarf, Frankfurt, Germany.…”

Section: In Vivo Antioxidant Analysismentioning

confidence: 99%

“…cereviseae cells growing exponentially on glucose are very sensitive to an oxidative stress, because under these conditions they only ferment and do not oxidize glucose (Pereira et al, 2003). However, the use of antioxidants can help protect against oxidative stress, increasing cellular tolerance (Dai et al, 2006).…”

Section: In Vivo and In Vitro Antioxidant Assay Of S Cayennensismentioning

confidence: 99%

Evaluation of antimicrobial, antioxidant and phototoxic activities of extracts and isolated compounds from Stachytarpheta cayennensis (Rich.) Vahl, Verbenaceae

Souza¹,

Silva²,

Machado³

et al. 2010

Rev. bras. farmacogn.

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RESUMO: "Avalição da atividade antimicrobiana, antioxidante e fototóxica dos extratos e compostos isolados de Stachytarpheta cayennensis (Rich.) Vahl, Verbenaceae". Stachytarpheta cayennensis (Rich.) Vahl, Verbenaceae, é uma planta utilizada na medicina popular brasileira contra úlceras e lesões de pele. Internamente é usada contra inflamações, febre, doenças renais e aterosclerose. Essa planta foi estudada com o objetivo de identificar os compostos bioativos majoritários que possam justificar seu uso terapêutico contra lesões de pele e arteriosclerose. A atividade antioxidante do extrato bruto etanólico, partições e os compostos majoritários isolados das raízes de S. cayennesis foi avaliada através de testes in vivo e in vitro. In vitro a atividade antioxidante foi avaliada pelo teste fotocolorimétrico do radical 2,2-difenil-1-picrilidrazil (DPPH•). In vivo, Saccharomyces cerevisiae, um modelo de célula eucariótica, foi utilizado tanto para avaliar a fototoxicidade quanto a capacidade antioxidante contra as espécies reativas de oxigênio (EROS) menadiona e peróxido de hidrogênio. O extrato, partições e os dois compostos majoritários isolados, verbascosídeo e ácido betulínico foram capazes de aumentar a sobrevivência e diminuir a peroxidação lipídica de S. cerevisiae contra EROS. Verbascosídeo apresentou atividade antimicrobiana moderada contra Streptococcus pyogenes, S.epidermidis e Staphylococcus aureus. O extrato etanólico e as partições testadas não apresentaram fototoxicidade, indicando que S. cayennensis é uma planta segura para o tratamento de lesões de pele e como possível ingrediente em cosméticos.Unitermos: Stachytarpheta cayennensis, antioxidante, antimicrobial, fototoxicidade, arilpropanoide glicosilado, ácido betulínico.ABSTRACT: Stachytarpheta cayennensis (Rich.) Vahl, Verbenaceae, plant extract, is a Brazilian medicinal plant externally used in folk medicine for purulent ulcers, skin lesions and internally for inflammations, fever, renal disorders and atherosclerosis. S. cayennesis was studied to identify potential bioactive compounds that may justify their therapeutic use against skin lesions and atherosclerosis. The antioxidant, antimicrobial and phototoxicity capacities of the crude ethanolic extract, fractions and isolated compounds from roots of S. cayennesis were evaluated through in vivo and in vitro tests. Strains of Saccharomyces cerevisiae, an eukaryotic cell model, were used to assess both the phototoxicity and the capacity to protect against the lethal oxidative stress caused by menadione and hydrogen peroxide. The extract, fractions and the two major isolated compounds, verbascoside and betulinic acid, of S. cayennensis were able to increase the tolerance and decrease the lipid peroxidation of S. cerevisiae to reactive oxygen species (ROS). The antioxidant activity was also evaluated by scavenging of 2,2-diphenyl-1-picrylhydrazyl (DPPH•). Verbascoside exhibited a moderate antimicrobial activity against Streptococcus pyogenes, S.epidermidis and Staphylococcus aureus. Neither the ethanol...

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“…tert-Butyl hydroperoxide (t-BOOH) is a lipid-soluble hydroperoxide that has been used to induce lipid peroxidation in yeast and animal cells (Masaki et al, 1989;Pereira et al, 2003). Dose-response curves indicated growth of both the wild-type and slr1736 mutant strains was negatively impacted at 150 mM t-BOOH/HL while 200 mM was lethal (data not shown).…”

Section: Sensitivity Of Tocopherol-deficient Mutants To Compounds Thamentioning

confidence: 99%

“…To assess this proposed function, the tocopheroldeficient mutants and wild type were subjected to treatments known to induce lipid peroxidation. t-BOOH is an alkyl peroxide routinely used to induce lipid peroxidation in other systems (Masaki et al, 1989;Pereira et al, 2003). Surprisingly, t-BOOH did not differentially impact the slr1736 mutant and wild type (Fig.…”

mentioning

confidence: 92%

Tocopherols Protect Synechocystis sp. Strain PCC 6803 from Lipid Peroxidation

et al. 2005

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Tocopherols (vitamin E) are lipid-soluble antioxidants synthesized only by photosynthetic eukaryotes and some cyanobacteria, and have been assumed to play important roles in protecting photosynthetic membranes from oxidative stress. To test this hypothesis, tocopherol-deficient mutants of Synechocystis sp. strain PCC 6803 (slr1736 and slr1737 mutants) were challenged with a series of reactive oxygen species-generating and lipid peroxidation-inducing chemicals in combination with high-light (HL) intensity stress. The tocopherol-deficient mutants and wild type were indistinguishable in their growth responses to HL in the presence and absence of superoxide and singlet oxygen-generating chemicals. However, the mutants showed enhanced sensitivity to linoleic or linolenic acid treatments in combination with HL, consistent with tocopherols playing a crucial role in protecting Synechocystis sp. strain PCC 6803 cells from lipid peroxidation. The tocopherol-deficient mutants were also more susceptible to HL treatment in the presence of sublethal levels of norflurazon, an inhibitor of carotenoid synthesis, suggesting carotenoids and tocopherols functionally interact or have complementary or overlapping roles in protecting Synechocystis sp. strain PCC 6803 from lipid peroxidation and HL stress.Oxygenic photosynthetic organisms continuously produce oxygen in the presence of light, and as such cellular damage from various reactive oxygen species (ROS), including singlet oxygen (2 ), hydrogen peroxide (H 2 O 2 ), and the hydroxyl radical (OH ), is a constant threat. Photosynthetic organisms have therefore evolved extensive detoxifying and protective mechanisms, which both limit the production of and potential damage by ROS. Examples include superoxide dismutase (SOD), which reduces O 2 2 to H 2 O 2 ; ascorbate peroxidase, which reduces H 2 O 2 to H 2 O; and nonphotochemical quenching that quenches singlet state chlorophylls ( 1 Chl*) and harmlessly dissipates excessive excitation energy as heat, thereby reducing 1 O 2 production (Asada, 1999;Muller et al., 2001). ROS, such as OH , can trigger a lipid peroxidation chain reaction by abstracting an allylic hydrogen from polyunsaturated fatty acid (PUFA)-containing lipids producing lipid radicals that are converted to lipid peroxyl radicals (LOO ) upon O 2 addition. LOO can subsequently attack another PUFA generating a second LOO and propagating a chain reaction of lipid peroxidation that perturbs membrane structure and function (Porter, 1986). Given the susceptibility of PUFAs to ROS damage, it seems counterintuitive that the PUFA-enriched thylakoid membranes would house the photosynthetic machinery, a potential ROS generator. In contrast to the well-studied mechanisms of water-soluble ROS detoxification in photosynthetic organisms (Asada, 1999), the mechanisms preventing or limiting oxidative damage in photosynthetic membranes are less well understood. Several peroxiredoxins have been implicated in reducing lipid hydroperoxides (LOOH) to the less toxic lipid hydroxides (LOH) in ...

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Targets of oxidative stress in yeast sod mutants

Cited by 60 publications

References 30 publications

Catalase Overexpression Reduces Lactic Acid-Induced Oxidative Stress in Saccharomyces cerevisiae

Catalase Overexpression Reduces Lactic Acid-Induced Oxidative Stress in Saccharomyces cerevisiae

Evaluation of antimicrobial, antioxidant and phototoxic activities of extracts and isolated compounds from Stachytarpheta cayennensis (Rich.) Vahl, Verbenaceae

Tocopherols Protect Synechocystis sp. Strain PCC 6803 from Lipid Peroxidation

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