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Basal Cell Carcinoma (BCC) is the most common type of cancer among the white population. Individuals with fair skin have an average lifetime risk of around 30% for developing BCC, and there is a noticeable upward trend in its incidence rate. The principal treatment objectives for BCC involve achieving the total excision of the tumor while maximizing the preservation of function and cosmesis. Surgery is considered the treatment of choice for BCC for two main reasons: it allows for the highest cure rates and facilitates histological control of resection margins. However, in the subgroup of patients with low-risk recurrence or medical contraindications for surgery, new non-surgical treatment alternatives can provide an excellent oncological and cosmetic outcome. An evident and justified instance of these local therapies occurred during the COVID-19 pandemic, a period when surgical interventions carried out in hospital settings were not a viable option.
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Interactions between algae and bacteria are ubiquitous and play fundamental roles in nutrient cycling and biomass production. Recent studies have shown that the plant auxin indole acetic acid (IAA) can mediate chemical crosstalk between algae and bacteria, resembling its role in plant-bacterial associations. Here, we report a mechanism for algal extracellular IAA production from L-tryptophan mediated by the enzyme L-amino acid oxidase (LAO1) in the model Chlamydomonas reinhardtii. High levels of IAA inhibit algal cell multiplication and chlorophyll degradation, and these inhibitory effects can be relieved by the presence of the plant-growth-promoting bacterium (PGPB) Methylobacterium aquaticum, whose growth is mutualistically enhanced by the presence of the alga. These findings reveal a complex interplay of microbial auxin production and degradation by algal-bacterial consortia and draws attention to potential ecophysiological roles of terrestrial microalgae and PGPB in association with land plants.
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A naturally occurring multispecies bacterial community composed of Bacillus cereus and two novel bacteria (Microbacterium forte sp. nov. and Stenotrophomonas goyi sp. nov.) has been identified from a contaminated culture of the microalga Chlamydomonas reinhardtii. When incubated in mannitol- and yeast extract-containing medium, this bacterial community can promote and sustain algal hydrogen production up to 313 mL H2·L-1 for 17 days and 163.5 mL H2·L-1 for 25 days in high-cell (76.7 µg·mL-1 of initial chlorophyll) and low-cell density (10 µg·mL-1 of initial chlorophyll) algal cultures, respectively. In low-cell density algal cultures, hydrogen production was compatible with algal growth (reaching up to 60 µg·mL-1 of chlorophyll). Among the bacterial community, M. forte sp. nov. was the sole responsible for the improvement in hydrogen production. However, algal growth was not observed in the Chlamydomonas-M. forte sp. nov. consortium during hydrogen-producing conditions (hypoxia), suggesting that the presence of B. cereus and S. goyi sp. nov. could be crucial to support the algal growth during hypoxia. Still, under nonhydrogen producing conditions (aerobiosis) the Chlamydomonas-M. forte sp. nov. consortium allowed algal growth (up to 40 µg·mL-1 of chlorophyll) and long-term algal viability (>45 days). The genome sequence and growth tests of M. forte sp. nov. have revealed that this bacterium is auxotroph for biotin and thiamine and unable to use sulfate as sulfur source; it requires S-reduced forms such as cysteine and methionine. Cocultures of Chlamydomonas reinhardtii and M. forte sp. nov. established a mutualistic association: the alga complemented the nutrient deficiencies of the bacterium, while the bacterium released ammonium (0.19 mM·day-1) and acetic acid (0.15 mM·day-1) for the alga. This work offers a promising avenue for photohydrogen production concomitant with algal biomass generation using nutrients not suitable for mixotrophic algal growth.
Assuntos
Chlamydomonas reinhardtii , Chlamydomonas , Microbacterium , Clorofila , Ácido Acético , Bactérias , Hipóxia , HidrogênioRESUMO
BACKGROUND: The efficiency of blood products (BP) requisition in elective non-cardiac surgeries is inherently complex. Moreover, it is aggravated in the pediatric population. This study aimed to identify the factors associated with using less than the requested BP during the transoperative period in pediatric patients undergoing elective non-cardiac surgery. METHODS: We conducted a cross-sectional comparative study including 320 patients undergoing elective non-cardiac surgery for whom BPs were requested. Low requirements were considered when less than 50% of the requested amount or no BPs were used, and high requirements when more than the requested amount was used. The Mann-Whitney's U test was applied for comparative analysis, and multiple logistic regression was used to adjust for factors associated with lower requirements. RESULTS: The median age of the patients was 3 years. From 320 patients, 68.1% (n = 218) received less than the requested amount of BP, while only 1.25% (n = 4) received more than the requested amount of BP. Factors associated with transfusion of less than the requested BPs were prolonged clotting time (odds ratio (OR) = 2.66) and anemia (OR = 0.43). CONCLUSIONS: Factors associated with lower than requested BP transfusion were prolonged clotting time and anemia.
INTRODUCCIÓN: La eficiencia de la solicitud de productos sanguíneos (PS) en las cirugías electivas no cardiacas es, de por sí, compleja. No obstante, se agrava para la población pediátrica. El objetivo de este estudio fue identificar los factores asociados con la utilización de una cantidad de PS menor a la solicitada durante el transoperatorio en pacientes pediátricos sometidos a cirugía electiva no cardiaca. MÉTODOS: Se realizó un estudio transversal comparativo donde se incluyeron 320 pacientes sometidos a cirugía electiva no cardiaca para quienes se solicitaron PS. Los requerimientos de hemoderivados se consideraron como menores cuando no se utilizaron o se utilizó menos del 50% de lo solicitado y como mayores cuando se utilizó una cantidad mayor a la solicitada. Se aplicó la prueba U de Mann-Whitney para el análisis comparativo y regresión logística múltiple para ajustar los factores asociados a la presencia de menores requerimientos. RESULTADOS: La mediana para la edad de los pacientes fue de 3 años. Se transfundió una cantidad de PS menor a la solicitada en el 68.1% (n = 218) de los pacientes, mientras que se transfundió una cantidad mayor a la solicitada solo en el 1.25% de los pacientes (n = 4). Los factores asociados con la transfusión de una cantidad de PS menor a la solicitada fueron tiempos de coagulación alargados (TCA) (razón de momios (RM) = 2.66) y anemia (RM = 0.43). CONCLUSIONES: Los factores asociados a una transfusión de PS inferior a la solicitada fueron el tiempo de coagulación prolongado y la anemia.
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Transfusão de Sangue , Cuidados Intraoperatórios , Criança , Pré-Escolar , Humanos , Estudos Transversais , Procedimentos Cirúrgicos EletivosRESUMO
The stability and harmony of ecological niches rely on intricate interactions between their members. During evolution, organisms have developed the ability to thrive in different environments, taking advantage of each other. Among these organisms, microalgae are a highly diverse and widely distributed group of major primary producers whose interactions with other organisms play essential roles in their habitats. Understanding the basis of these interactions is crucial to control and exploit these communities for ecological and biotechnological applications. The green microalga Chlamydomonas reinhardtii, a well-established model, is emerging as a model organism for studying a wide variety of microbial interactions with ecological and economic significance. In this review, we unite and discuss current knowledge that points to C. reinhardtii as a model organism for studying microbial interactions.
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Abstract Background: The efficiency of blood products (BP) requisition in elective non-cardiac surgeries is inherently complex. Moreover, it is aggravated in the pediatric population. This study aimed to identify the factors associated with using less than the requested BP during the transoperative period in pediatric patients undergoing elective non-cardiac surgery. Methods: We conducted a cross-sectional comparative study including 320 patients undergoing elective non-cardiac surgery for whom BPs were requested. Low requirements were considered when less than 50% of the requested amount or no BPs were used, and high requirements when more than the requested amount was used. The Mann-Whitney's U test was applied for comparative analysis, and multiple logistic regression was used to adjust for factors associated with lower requirements. Results: The median age of the patients was 3 years. From 320 patients, 68.1% (n = 218) received less than the requested amount of BP, while only 1.25% (n = 4) received more than the requested amount of BP. Factors associated with transfusion of less than the requested BPs were prolonged clotting time (odds ratio (OR) = 2.66) and anemia (OR = 0.43). Conclusions: Factors associated with lower than requested BP transfusion were prolonged clotting time and anemia.
Resumen Introducción: La eficiencia de la solicitud de productos sanguíneos (PS) en las cirugías electivas no cardiacas es, de por sí, compleja. No obstante, se agrava para la población pediátrica. El objetivo de este estudio fue identificar los factores asociados con la utilización de una cantidad de PS menor a la solicitada durante el transoperatorio en pacientes pediátricos sometidos a cirugía electiva no cardiaca. Métodos: Se realizó un estudio transversal comparativo donde se incluyeron 320 pacientes sometidos a cirugía electiva no cardiaca para quienes se solicitaron PS. Los requerimientos de hemoderivados se consideraron como menores cuando no se utilizaron o se utilizó menos del 50% de lo solicitado y como mayores cuando se utilizó una cantidad mayor a la solicitada. Se aplicó la prueba U de Mann-Whitney para el análisis comparativo y regresión logística múltiple para ajustar los factores asociados a la presencia de menores requerimientos. Resultados: La mediana para la edad de los pacientes fue de 3 años. Se transfundió una cantidad de PS menor a la solicitada en el 68.1% (n = 218) de los pacientes, mientras que se transfundió una cantidad mayor a la solicitada solo en el 1.25% de los pacientes (n = 4). Los factores asociados con la transfusión de una cantidad de PS menor a la solicitada fueron tiempos de coagulación alargados (TCA) (razón de momios (RM) = 2.66) y anemia (RM = 0.43). Conclusiones: Los factores asociados a una transfusión de PS inferior a la solicitada fueron el tiempo de coagulación prolongado y la anemia.
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Nitrous oxide (N2O) is a powerful greenhouse gas and an ozone-depleting compound whose synthesis and release have traditionally been ascribed to bacteria and fungi. Although plants and microalgae have been proposed as N2O producers in recent decades, the proteins involved in this process have been only recently unveiled. In the green microalga Chlamydomonas reinhardtii, flavodiiron proteins (FLVs) and cytochrome P450 (CYP55) are two nitric oxide (NO) reductases responsible for N2O synthesis in the chloroplast and mitochondria, respectively. However, the molecular mechanisms feeding these NO reductases are unknown. In this work, we use cavity ring-down spectroscopy to monitor N2O and CO2 in cultures of nitrite reductase mutants, which cannot grow on nitrate or nitrite and exhibit enhanced N2O emissions. We show that these mutants constitute a very useful tool to study the rates and kinetics of N2O release under different conditions and the metabolism of this greenhouse gas. Our results indicate that N2O production, which was higher in the light than in the dark, requires nitrate reductase as the major provider of NO as substrate. Finally, we show that the presence of nitrate reductase impacts CO2 emissions in both light and dark conditions, and we discuss the role of NO in the balance between CO2 fixation and release.
Assuntos
Chlamydomonas reinhardtii , Gases de Efeito Estufa , Microalgas , Dióxido de Carbono/metabolismo , Chlamydomonas reinhardtii/genética , Chlamydomonas reinhardtii/metabolismo , Microalgas/metabolismo , Nitrato Redutase/genética , Nitrato Redutase/metabolismo , Óxido Nítrico/metabolismo , Nitritos/metabolismo , Óxido Nitroso/metabolismoRESUMO
In the context of algal wastewater bioremediation, this study has identified a novel consortium formed by the bacterium Methylobacterium oryzae and the microalga Chlamydomonas reinhardtii that greatly increase biomass generation (1.22 g L-1·d-1), inorganic nitrogen removal (>99%), and hydrogen production (33 mL·L-1) when incubated in media containing ethanol and methanol. The key metabolic aspect of this relationship relied on the bacterial oxidation of ethanol to acetate, which supported heterotrophic algal growth. However, in the bacterial monocultures the acetate accumulation inhibited bacterial growth. Moreover, in the absence of methanol, ethanol was an unsuitable carbon source and its incomplete oxidation to acetaldehyde had a toxic effect on both the alga and the bacterium. In cocultures, both alcohols were used as carbon sources by the bacteria, the inhibitory effects were overcome and both microorganisms mutually benefited. Potential biotechnological applications in wastewater treatment, biomass generation and hydrogen production are discussed.
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Chlamydomonas , Acetatos , Biomassa , Carbono , Chlamydomonas/metabolismo , Desnitrificação , Etanol , Hidrogênio , Metanol , Methylobacterium , Nitrogênio/metabolismoRESUMO
Nitrogen (N) is an essential constituent of all living organisms and the main limiting macronutrient. Even when dinitrogen gas is the most abundant form of N, it can only be used by fixing bacteria but is inaccessible to most organisms, algae among them. Algae preferentially use ammonium (NH4+) and nitrate (NO3-) for growth, and the reactions for their conversion into amino acids (N assimilation) constitute an important part of the nitrogen cycle by primary producers. Recently, it was claimed that algae are also involved in denitrification, because of the production of nitric oxide (NO), a signal molecule, which is also a substrate of NO reductases to produce nitrous oxide (N2O), a potent greenhouse gas. This review is focused on the microalga Chlamydomonas reinhardtii as an algal model and its participation in different reactions of the N cycle. Emphasis will be paid to new actors, such as putative genes involved in NO and N2O production and their occurrence in other algae genomes. Furthermore, algae/bacteria mutualism will be considered in terms of expanding the N cycle to ammonification and N fixation, which are based on the exchange of carbon and nitrogen between the two organisms.
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Biological hydrogen production by microalgae is a potential sustainable, renewable and clean source of energy. However, many barriers limiting photohydrogen production in these microorganisms remain unsolved. In order to explore this potential and make biohydrogen industrially affordable, the unicellular microalga Chlamydomonas reinhardtii is used as a model system to solve barriers and identify new approaches that can improve hydrogen production. Recently, Chlamydomonas-bacteria consortia have opened a new window to improve biohydrogen production. In this study, we review the different consortia that have been successfully employed and analyze the factors that could be behind the improved H2 production.
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Bactérias/metabolismo , Chlamydomonas reinhardtii/metabolismo , Hidrogênio/metabolismo , Absorção de Radiação , Oxigênio/metabolismo , Luz SolarRESUMO
The mitogen activated protein kinases (MAPKs) form part of a signaling cascade through phosphorylation reactions conserved in all eukaryotic organisms. The MAPK cascades are mainly composed by three proteins, MAPKKKs, MAPKKs and MAPKs. Some signals induce MAPKKK-mediated phosphorylation and activation of MAPKK that phosphorylate and activate MAPK. Afterward, MAPKs can act either in the cytoplasm or be imported into the nucleus to activate other proteins or transcription factors. In the green microalga Chlamydomonas reinhardtii the pathway for nitrogen (N) assimilation is well characterized, yet its regulation still has many unknown features. Nitric oxide (NO) is a fundamental signal molecule for N regulation, where nitrate reductase (NR) plays a central role in its synthesis. The MAPK cascades could be regulating N assimilation, since it has been described that the phosphorylation of NR by MAPK6 promotes NO production in Arabidopsis thaliana. We have identified the proteins involved in the MAPK cascades in Chlamydomonas reinhardtii, finding 17 MAPKs, 2 MAPKKs and 108 MAPKKKs (11 MEKK-, 94 RAF- and 3 ZIK-type) that have been structurally and phylogenetically characterized. The genetic expressions of MAPKs and the MAPKK were slightly regulated by N. However, the genetic expressions of MAPKKKs RAF14 and RAF79 showed a very strong repression by ammonium, which suggests that they may have a key role in the regulation of N assimilation, encouraging to further analyze in detail the role of MAPK cascades in the regulation of N metabolism.
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Proteínas de Algas/metabolismo , Chlamydomonas reinhardtii/metabolismo , Sistema de Sinalização das MAP Quinases , Nitrogênio/metabolismo , Proteínas de Algas/genética , Compostos de Amônio/metabolismo , Chlamydomonas reinhardtii/genética , Regulação da Expressão Gênica de Plantas , MAP Quinase Quinase Quinases/genética , MAP Quinase Quinase Quinases/metabolismo , Quinases de Proteína Quinase Ativadas por Mitógeno/genética , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Transdução de Sinais/genéticaRESUMO
The conservation of orthologs of most subunits of the origin recognition complex (ORC) has served to propose that the whole complex is common to all eukaryotes. However, various uncertainties have arisen concerning ORC subunit composition in a variety of lineages. Also, it is unclear whether the ancestral diversification of ORC in eukaryotes was accompanied by the neofunctionalization of some subunits, for example, role of ORC1 in centriole homeostasis. We have addressed these questions by reconstructing the distribution and evolutionary history of ORC1-5/CDC6 in a taxon-rich eukaryotic data set. First, we identified ORC subunits previously undetected in divergent lineages, which allowed us to propose a series of parsimonious scenarios for the origin of this multiprotein complex. Contrary to previous expectations, we found a global tendency in eukaryotes to increase or decrease the number of subunits as a consequence of genome duplications or streamlining, respectively. Interestingly, parasites show significantly lower number of subunits than free-living eukaryotes, especially those with the lowest genome size and gene content metrics. We also investigated the evolutionary origin of the ORC1 role in centriole homeostasis mediated by the PACT region in human cells. In particular, we tested the consequences of reducing ORC1 levels in the centriole-containing green alga Chlamydomonas reinhardtii. We found that the proportion of centrioles to flagella and nuclei was not dramatically affected. This, together with the PACT region not being significantly more conserved in centriole-bearing eukaryotes, supports the notion that this neofunctionalization of ORC1 would be a recent acquisition rather than an ancestral eukaryotic feature.
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Complexo de Reconhecimento de Origem/metabolismo , Animais , Replicação do DNA/genética , Replicação do DNA/fisiologia , Eucariotos , Células Eucarióticas/metabolismo , Evolução Molecular , Duplicação Gênica/genética , Duplicação Gênica/fisiologia , Genoma/genética , Humanos , Imuno-Histoquímica , Complexo de Reconhecimento de Origem/genética , Filogenia , Ligação Proteica/genética , Ligação Proteica/fisiologiaRESUMO
Nitric oxide is a gaseous secondary messenger that is critical for proper cell signaling and plant survival when exposed to stress. Nitric oxide (NO) synthesis in plants, under standard phototrophic oxygenic conditions, has long been a very controversial issue. A few algal strains contain NO synthase (NOS), which appears to be absent in all other algae and land plants. The experimental data have led to the hypothesis that molybdoenzyme nitrate reductase (NR) is the main enzyme responsible for NO production in most plants. Recently, NR was found to be a necessary partner in a dual system that also includes another molybdoenzyme, which was renamed NO-forming nitrite reductase (NOFNiR). This enzyme produces NO independently of the molybdenum center of NR and depends on the NR electron transport chain from NAD(P)H to heme. Under the circumstances in which NR is not present or active, the existence of another NO-forming system that is similar to the NOS system would account for NO production and NO effects. PII protein, which senses and integrates the signals of the Câ»N balance in the cell, likely has an important role in organizing cell responses. Here, we critically analyze these topics.
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All eukaryotic molybdenum (Mo) enzymes contain in their active site a Mo Cofactor (Moco), which is formed by a tricyclic pyranopterin with a dithiolene chelating the Mo atom. Here, the eukaryotic Moco biosynthetic pathway and the eukaryotic Moco enzymes are overviewed, including nitrate reductase (NR), sulfite oxidase, xanthine oxidoreductase, aldehyde oxidase, and the last one discovered, the moonlighting enzyme mitochondrial Amidoxime Reducing Component (mARC). The mARC enzymes catalyze the reduction of hydroxylated compounds, mostly N-hydroxylated (NHC), but as well of nitrite to nitric oxide, a second messenger. mARC shows a broad spectrum of NHC as substrates, some are prodrugs containing an amidoxime structure, some are mutagens, such as 6-hydroxylaminepurine and some others, which most probably will be discovered soon. Interestingly, all known mARC need the reducing power supplied by different partners. For the NHC reduction, mARC uses cytochrome b5 and cytochrome b5 reductase, however for the nitrite reduction, plant mARC uses NR. Despite the functional importance of mARC enzymatic reactions, the structural mechanism of its Moco-mediated catalysis is starting to be revealed. We propose and compare the mARC catalytic mechanism of nitrite versus NHC reduction. By using the recently resolved structure of a prokaryotic MOSC enzyme, from the mARC protein family, we have modeled an in silico three-dimensional structure of a eukaryotic homologue.
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Coenzimas/metabolismo , Enzimas/metabolismo , Metaloproteínas/metabolismo , Pteridinas/metabolismo , Animais , Miosinas Cardíacas/metabolismo , Coenzimas/biossíntese , Enzimas/química , Enzimas/genética , Células Eucarióticas/metabolismo , Mamíferos , Redes e Vias Metabólicas , Metaloproteínas/biossíntese , Molibdênio/metabolismo , Cofatores de Molibdênio , Cadeias Leves de Miosina/metabolismo , Nitrato Redutase/metabolismo , Nitritos/metabolismo , Oxirredutases/genética , Oxirredutases/metabolismoRESUMO
Nitrogen assimilation and metabolism are essential processes for all living organisms, yet there is still much to be learnt on how they are regulated. The use of Chlamydomonas reinhardtii as a model system has been instrumental not only in identifying conserved regulation mechanisms that control the nitrogen assimilation pathway, but also in understanding how the intracellular nitrogen status regulates metabolic processes of industrial interest such as the synthesis of biolipids. While the genetic regulators that control the nitrogen pathway are successfully being unravelled, other layers of regulation have received less attention. Amino acids, for example, regulate nitrogen assimilation in certain organisms, but their role in Chlamydomonas has not thoroughly been explored. Previous results had suggested that arginine might repress key genes of the nitrogen assimilation pathway by acting within the ammonium negative signalling cascade, upstream of the nitric oxide (NO) inducible guanylate cyclase CYG56. We tested this hypothesis with a combination of genetic and chemical approaches. Antagonising the effects of arginine with an arginine biosynthesis mutant or with two chemical analogues released gene expression from ammonium mediated repression. The cyg56 and related non1 mutants, which are partially insensitive to ammonium repression, were also partially insensitive to repression by arginine. Finally, we show that the addition of arginine to the medium leads to an increase in intracellular NO. Our data reveal that arginine acts as a negative signal for the assimilation of nitrogen within the ammonium-CYG56 negative signalling cascade, and provide a connection between amino acid metabolism and nitrogen assimilation in microalgae.
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Compostos de Amônio/metabolismo , Arginina/metabolismo , Chlamydomonas reinhardtii/crescimento & desenvolvimento , Redes Reguladoras de Genes , Nitrogênio/metabolismo , Chlamydomonas reinhardtii/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Guanilato Ciclase/genética , Guanilato Ciclase/metabolismo , Mutação , Óxido Nítrico/metabolismo , Proteínas de Plantas/genética , Transdução de SinaisRESUMO
Nitrogen is a key nutrient for land plants and phytoplankton in terrestrial and aquatic ecosystems. The model alga Chlamydomonas reinhardtii can grow efficiently on several inorganic nitrogen sources (e.g. ammonium, nitrate, nitrite) as well as many amino acids. In this study, we show that Chlamydomonas is unable to use proline, hydroxyproline and peptides that contain these amino acids. However, we discovered that algal growth on these substrates is supported in association with Methylobacterium spp., and that a mutualistic carbon-nitrogen metabolic exchange between Chlamydomonas and Methylobacterium spp. is established. Specifically, the mineralization of these amino acids and peptides by Methylobacterium spp. produces ammonium that can be assimilated by Chlamydomonas, and CO2 photosynthetically fixed by Chlamydomonas yields glycerol that can be assimilated by Methylobacterium. As Chlamydomonas is an algal ancestor to land plants and Methylobacterium is a plant growth-promoting bacterium, this new model of mutualism may facilitate insights into the ecology and evolution of plant-bacterial interactions and design principles of synthetic ecology.
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Aminoácidos/metabolismo , Chlamydomonas/metabolismo , Methylobacterium/metabolismo , Peptídeos/metabolismo , Carbono/metabolismo , Chlamydomonas/crescimento & desenvolvimento , Methylobacterium/crescimento & desenvolvimento , Nitratos/metabolismo , Nitritos/metabolismo , Fotossíntese , SimbioseRESUMO
BACKGROUND: A recent Commentary article entitled "On the pathways feeding the H2 production process in nutrient-replete, hypoxic conditions" by Dr. Scoma and Dr. Tóth, Biotechnology for Biofuels (2017), opened a very interesting debate about the H2 production photosynthetic-linked pathways occurring in Chlamydomonas cultures grown in acetate-containing media and incubated under hypoxia/anoxia conditions. This Commentary article mainly focused on the results of our previous article "Low oxygen levels contribute to improve photohydrogen production in mixotrophic non-stressed Chlamydomonas cultures," by Jurado-Oller et al., Biotechnology for Biofuels (7, 2015; 8:149). MAIN BODY: Here, we review some previous knowledge about the H2 production pathways linked to photosynthesis in Chlamydomonas, especially focusing on the role of the PSII-dependent and -independent pathways in acetate-containing nutrient-replete cultures. The potential contributions of these pathways to H2 production under anoxia/hypoxia are discussed. CONCLUSION: Despite the fact that the PSII inhibitor DCMU is broadly used to discern between the two different photosynthetic pathways operating under H2 production conditions, its use may lead to distinctive conclusions depending on the growth conditions. The different potential sources of reductive power needed for the PSII-independent H2 production in mixotrophic nutrient-replete cultures are a matter of debate and conclusive evidences are still missing.
RESUMO
Molybdenum (Mo) is present in the active center of eukaryotic enzymes as a tricyclic pyranopterin chelate compound forming the Mo Cofactor (Moco). Four Moco containing enzymes are known in eukaryotes, nitrate reductase (NR), sulfite oxidase (SO), xanthine oxidoreductase (XOR), and aldehyde oxidase (AO). A fifth Moco enzyme has been recently identified. Because of the ability of this enzyme to convert by reduction several amidoximes prodrugs into their active amino forms, it was named mARC (mitochondrial Amidoxime Reducing Component). This enzyme is also able to catalyze the reduction of a broad range of N-hydroxylated compounds (NHC) as the base analogue 6-hydroxylaminopurine (HAP), as well as nitrite to nitric oxide (NO). All the mARC proteins need reducing power that is supplied by other proteins. The human and plants mARC proteins require a Cytochrome b5 (Cytb5) and a Cytochrome b5 reductase (Cytb5-R) to form an electron transfer chain from NADH to the NHC. Recently, plant mARC proteins were shown to be implicated in the reduction of nitrite to NO, and it was proposed that the electrons required for the reaction were supplied by NR instead of Cytochrome b5 components. This newly characterized mARC activity was termed NO Forming Nitrite Reductase (NOFNiR). Moonlighting proteins form a special class of multifunctional enzymes that can perform more than one function; if the extra function is not physiologically relevant, they are called promiscuous enzymes. In this review, we summarize the current knowledge on the mARC protein, and we propose that mARC is a new moonlighting enzyme. © 2017 BioFactors, 43(4):486-494, 2017.
Assuntos
Coenzimas/metabolismo , Metaloproteínas/metabolismo , Pteridinas/metabolismo , Aldeído Oxidase/metabolismo , Animais , Citocromos b5/metabolismo , Humanos , Cofatores de Molibdênio , Nitrato Redutase/metabolismo , Sulfito Oxidase/metabolismo , Xantina Desidrogenase/metabolismoRESUMO
The mARC (mitochondrial Amidoxime Reducing Component) proteins are recently discovered molybdenum (Mo) Cofactor containing enzymes. They are involved in the reduction of several N-hydroxylated compounds (NHC) and nitrite. Some NHC are prodrugs containing an amidoxime structure or mutagens such as 6-hydroxylaminopurine (HAP). We have studied this protein in the green alga Chlamydomonas reinhardtii (crARC). Interestingly, all the ARC proteins need the reducing power supplied by other proteins. It is known that crARC requires a cytochrome b5 (crCytb5-1) and a cytochrome b5 reductase (crCytb5-R) that form an electron transport chain from NADH to the substrates. Here, we have investigated NHC reduction by crARC, the interaction with its partners and the function of important conserved amino acids. Interactions among crARC, crCytb5-1 and crCytb5-R have been studied by size-exclusion chromatography. A protein complex between crARC, crCytb5-1 and crCytb5-R was identified. Twelve conserved crARC amino acids have been substituted by alanine by in vitro mutagenesis. We have determined that the amino acids D182, F210 and R276 are essential for NHC reduction activity, R276 is important and F210 is critical for the Mo Cofactor chelation. Finally, the crARC C-termini were shown to be involved in protein aggregation or oligomerization.
Assuntos
Coenzimas/metabolismo , Citocromos b5/metabolismo , Metaloproteínas/metabolismo , Pteridinas/metabolismo , Substituição de Aminoácidos , Sítios de Ligação , Chlamydomonas reinhardtii/enzimologia , Chlamydomonas reinhardtii/metabolismo , Coenzimas/química , Coenzimas/genética , Citocromos b5/química , Citocromos b5/genética , Metaloproteínas/química , Metaloproteínas/genética , Cofatores de Molibdênio , Ligação Proteica , Multimerização Proteica , Pteridinas/químicaRESUMO
The green alga Chlamydomonas is a valuable model system capable of assimilating different forms of nitrogen (N). Nitrate (NO3-) has a relevant role in plant-like organisms, first as a nitrogen source for growth and second as a signalling molecule. Several modules are necessary for Chlamydomonas to handle nitrate, including transporters, nitrate reductase (NR), nitrite reductase (NiR), GS/GOGAT enzymes for ammonium assimilation, and regulatory protein(s). Transporters provide a first step for influx/efflux, homeostasis, and sensing of nitrate; and NIT2 is the key transcription factor (RWP-RK) for mediating the nitrate-dependent activation of a number of genes. Here, we review how NR participates in the cycle NO3- âNO2- âNO âNO3-. NR uses the partner protein amidoxime-reducing component/nitric oxide-forming nitrite reductase (ARC/NOFNiR) for the conversion of nitrite (NO2-) into nitric oxide (NO). It also uses the truncated haemoglobin THB1 in the conversion of nitric oxide to nitrate. Nitric oxide is a negative signal for nitrate assimilation; it inhibits the activity and expression of high-affinity nitrate/nitrite transporters and NR. During this cycle, the positive signal of nitrate is transformed into the negative signal of nitric oxide, which can then be converted back into nitrate. Thus, NR is back in the spotlight as a strategic regulator of the nitric oxide cycle and the nitrate assimilation pathway.