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1.
Int J Mol Sci ; 24(21)2023 Oct 27.
Artigo em Inglês | MEDLINE | ID: mdl-37958645

RESUMO

The co-culturing of microorganisms is a well-known strategy to study microbial interactions in the laboratory. This approach facilitates the identification of new signals and molecules produced by one species that affects other species' behavior. In this work, we have studied the effects of the interaction of nine Streptomyces species (S. albidoflavus, S. ambofaciens, S. argillaceus, S. griseus, S. lividans, S. olivaceus, S. parvulus, S. peucetius, and S. rochei) with the predator bacteria Myxococcus xanthus, five of which (S. albidoflavus, S. griseus, S. lividans, S. olivaceus, and S. argillaceus) induce mound formation of M. xanthus on complex media (Casitone Yeast extract (CYE) and Casitone tris (CTT); media on which M. xanthus does not form these aggregates under normal culture conditions. An in-depth study on S. griseus-M. xanthus interactions (the Streptomyces strain producing the strongest effect) has allowed the identification of two siderophores produced by S. griseus, demethylenenocardamine and nocardamine, responsible for this grouping effect over M. xanthus. Experiments using pure commercial nocardamine and different concentrations of FeSO4 show that iron depletion is responsible for the behavior of M. xanthus. Additionally, it was found that molecules, smaller than 3 kDa, produced by S. peucetius can induce the production of DK-xanthenes by M. xanthus.


Assuntos
Myxococcus xanthus , Myxococcus , Streptomyces , Interações Microbianas , Ferro
2.
Int J Mol Sci ; 23(23)2022 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-36499414

RESUMO

Bacteria of the Streptomyces genus constitute an authentic biotech gold mine thanks to their ability to produce a myriad of compounds and enzymes of great interest at various clinical, agricultural, and industrial levels. Understanding the physiology of these organisms and revealing their regulatory mechanisms is essential for their manipulation and application. Two-component systems (TCSs) constitute the predominant signal transduction mechanism in prokaryotes, and can detect a multitude of external and internal stimuli and trigger the appropriate cellular responses for adapting to diverse environmental conditions. These global regulatory systems usually coordinate various biological processes for the maintenance of homeostasis and proper cell function. Here, we review the multiple TCSs described and characterized in Streptomyces coelicolor, one of the most studied and important model species within this bacterial group. TCSs are involved in all cellular processes; hence, unravelling the complex regulatory network they form is essential for their potential biotechnological application.


Assuntos
Streptomyces coelicolor , Streptomyces , Streptomyces coelicolor/genética , Streptomyces coelicolor/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Streptomyces/metabolismo , Transdução de Sinais , Regulação Bacteriana da Expressão Gênica
3.
Emerg Infect Dis ; 27(6): 1756-1758, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34014154

RESUMO

We describe an Oropouche orthobunyavirus infection in a women 28 years of age in Colombia. We confirmed the diagnosis by viral isolation, quantitative reverse transcription PCR, and phylogenetic analysis of the small, medium, and large genomic segments. The virus is related to a strain isolated in Ecuador in 2016.


Assuntos
Infecções por Bunyaviridae , Orthobunyavirus , Colômbia , Equador , Feminino , Humanos , Orthobunyavirus/genética , Filogenia , RNA Viral
4.
Microb Cell Fact ; 16(1): 164, 2017 Sep 26.
Artigo em Inglês | MEDLINE | ID: mdl-28950904

RESUMO

BACKGROUND: The industrial use of enzymes produced by microorganisms is continuously growing due to the need for sustainable solutions. Nevertheless, many of the plasmids used for recombinant production of proteins in bacteria are based on the use of antibiotic resistance genes as selection markers. The safety concerns and legal requirements surrounding the increased use of antibiotic resistance genes have made the development of new antibiotic-free approaches essential. RESULTS: In this work, a system completely free of antibiotic resistance genes and useful for the production of high yields of proteins in Streptomyces is described. This system is based on the separation of the two components of the yefM/yoeBsl (antitoxin/toxin) operon; the toxin (yoeBsl) gene, responsible for host death, is integrated into the genome and the antitoxin gene (yefMsl), which inactivates the toxin, is located in the expression plasmid. To develop this system, the toxin gene was integrated into the genome of a strain lacking the complete operon, and the antibiotic resistance gene integrated along with the toxin was eliminated by Cre recombinase to generate a final host strain free of any antibiotic resistance marker. In the same way, the antibiotic resistance gene from the final expression plasmid was removed by Dre recombinase. The usefulness of this system was analysed by checking the production of two hydrolases from different Streptomyces. Production of both proteins, with potential industrial use, was high and stable over time after strain storage and after serial subcultures. These results support the robustness and stability of the positive selection system developed. CONCLUSIONS: The total absence of antibiotic resistance genes makes this system a powerful tool for using Streptomyces as a host to produce proteins at the industrial level. This work is the first Streptomyces antibiotic marker-free system to be described. Graphical abstract Antibiotic marker-free platform for protein expression in Streptomyces. The antitoxin gene present in the expression plasmid counteracts the effect of the toxin gene in the genome. In absence of the expression plasmid, the toxin causes cell death ensuring that only plasmid-containing cells persist.


Assuntos
Proteínas Recombinantes/genética , Streptomyces/genética , Antibacterianos/farmacologia , Antitoxinas/genética , Antitoxinas/metabolismo , Engenharia Genética , Plasmídeos/genética , Plasmídeos/metabolismo , Proteínas Recombinantes/metabolismo , Streptomyces/efeitos dos fármacos , Streptomyces/metabolismo , Toxinas Biológicas/genética , Toxinas Biológicas/metabolismo
5.
Microb Cell Fact ; 15: 28, 2016 Feb 04.
Artigo em Inglês | MEDLINE | ID: mdl-26846788

RESUMO

BACKGROUND: Actinomycetes are saprophytic soil bacteria, and a rich source of industrial enzymes. While some of these enzymes can be produced using well-characterized production platforms such as Escherichia coli or Bacillus subtilis, Streptomyces lividans may be the preferred host for proper folding and efficient secretion of active enzymes. A combination of promoters, signal peptides and hosts were tested in order to obtain the best protein expression in this actinomycete. The xylanase, Xys1, from S. halstedii, the α-amylase, Amy, from S. griseus and the small laccase, SLAC, from S. coelicolor were used as reporters. RESULTS: The promoters xysAp from S. halstedii JM8 and pstSp from S. lividans were the most efficient among those tested. An improvement of 17 % was obtained in xylanase activity when the signal peptide of the α-amylase protein (Amy) of S. griseus IMRU3570 was used to direct its secretion. Enhanced expression of SsgA, a protein that plays a role in processes that require cell-wall remodelling, resulted in a improvement of 40 and 70 % of xylanase and amylase production, respectively. Deletion of genes SLI7232 and SLI4452 encoding putative repressors of xysAp provided improvement of production up to 70 % in the SLI7232 deletion strain. However, full derepression of this promoter activity was not obtained under the conditions assayed. CONCLUSIONS: Streptomyces lividans is a frequently used platform for industrial enzyme production and a rational strain-development approach delivered significant improvement of protein production by this host.


Assuntos
Amilases/biossíntese , Engenharia Genética/métodos , Streptomyces lividans/enzimologia , Xilosidases/biossíntese , Proteínas de Bactérias/metabolismo , Deleção de Genes , Plasmídeos/metabolismo , Regiões Promotoras Genéticas , Sinais Direcionadores de Proteínas , Transporte Proteico , Proteínas Repressoras/metabolismo
6.
Appl Environ Microbiol ; 80(8): 2417-28, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24509929

RESUMO

The atypical two-component system (TCS) AbrC1/C2/C3 (encoded by SCO4598, SCO4597, and SCO4596), comprising two histidine kinases (HKs) and a response regulator (RR), is crucial for antibiotic production in Streptomyces coelicolor and for morphological differentiation under certain nutritional conditions. In this study, we demonstrate that deletion of the RR-encoding gene, abrC3 (SCO4596), results in a dramatic decrease in actinorhodin (ACT) and undecylprodiginine (RED) production and delays morphological development. In contrast, the overexpression of abrC3 in the parent strain leads to a 33% increase in ACT production in liquid medium. Transcriptomic analysis and chromatin immunoprecipitation with microarray technology (ChIP-chip) analysis of the ΔabrC3 mutant and the parent strain revealed that AbrC3 directly controls ACT production by binding to the actII-ORF4 promoter region; this was independently verified by in vitro DNA-binding assays. This binding is dependent on the sequence 5'-GAASGSGRMS-3'. In contrast, the regulation of RED production is not due to direct binding of AbrC3 to either the redZ or redD promoter region. This study also revealed other members of the AbrC3 regulon: AbrC3 is a positive autoregulator which also binds to the promoter regions of SCO0736, bdtA (SCO3328), absR1 (SCO6992), and SCO6809. The direct targets share the 10-base consensus binding sequence and may be responsible for some of the phenotypes of the ΔabrC3 mutant. The identification of the AbrC3 regulon as part of the complex regulatory network governing antibiotic production widens our knowledge regarding TCS involvement in control of antibiotic synthesis and may contribute to the rational design of new hyperproducer host strains through genetic manipulation of such systems.


Assuntos
Antibacterianos/biossíntese , Regulação Bacteriana da Expressão Gênica , Regulon , Streptomyces coelicolor/genética , Fatores de Transcrição/metabolismo , Antraquinonas/metabolismo , Sítios de Ligação , Imunoprecipitação da Cromatina , DNA Bacteriano/metabolismo , Deleção de Genes , Perfilação da Expressão Gênica , Prodigiosina/análogos & derivados , Prodigiosina/biossíntese , Ligação Proteica , Streptomyces coelicolor/citologia
7.
Trop Anim Health Prod ; 46(4): 705-9, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24510223

RESUMO

Toxoplasma gondii infection in pigs has epidemiological concern for its contributing role in human infections. We determined seroprevalence of T. gondii infection in 402 domestic pigs raised in backyards in Veracruz State, Mexico using the modified agglutination test (MAT; cut off 1:25); 182 (45.3%) of the 402 pigs were seropositive with MAT titers of 1:25 in 28, 1:50 in 22, 1:100 in 18, 1:200 in 30, 1:400 in 35, 1:800 in 23, 1:1,600 in 11, and 1:3,200 or higher in 15. Seropositive pigs were found in 137 (53.3%) of 257 homes in all 7 municipalities surveyed. Multivariate analysis showed that T. gondii seropositivity in pigs was associated with tropical-humid climate (OR = 4.32; 95% CI 1.47-12.62; P = 0.007) of the raising municipalities, feeding with leftovers (OR = 2.83; 95% CI 1.01-7.91; P = 0.04), storing pig food in the owner's home (OR = 2.39; 95% CI 1.09-5.22; P = 0.02), and free ranging (OR = 3.48; 95% CI 1.49-8.15; P = 0.003). Results indicate that backyard pigs in Veracruz have the highest seroprevalence of T. gondii infection obtained by MAT in pigs studied in Mexico so far. The correlates of T. gondii infection found in the present study may be useful for an optimal planning of preventive measures against T. gondii infection in pigs. Results also remark the risk of T. gondii infection in humans by ingestion of raw or undercook pork in Mexico.


Assuntos
Doenças dos Suínos/epidemiologia , Toxoplasmose Animal/epidemiologia , Animais , Anticorpos Antiprotozoários/sangue , Feminino , Masculino , México/epidemiologia , Fatores de Risco , Estudos Soroepidemiológicos , Suínos , Doenças dos Suínos/parasitologia , Toxoplasma/imunologia
8.
Acta Parasitol ; 69(2): 1148-1156, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38592371

RESUMO

PURPOSE: Chagas disease (CD) a Neglected Tropical Diseases is an important public health issue in countries where is still endemic, included in the Sustainable Development Goals (SDG). Traditionally restricted to rural areas with diverse routes of transmissions from vectorial to oral with acute manifestations but being more common diagnosed in chronic stages. The aim of this investigation was to characterize the Knowledge, Attitudes and Practices (KAP) related to Chagas disease (CD) in two rural settlements of the Colombian Caribbean with previous records of the disease and/or the parasite. METHODS: A cross-sectional descriptive study was made in two rural settlements in Colombia and surveillance instrument was developed to measure Knowledge, Attitudes and Practices (KAP) related to Chagas disease (CD). RESULTS: In a population with > 60% women and access to social security around 66.5%; 81,6% were homeowners with access to water and electricity > 90% but only 9% of sewerage. The level of knowledge about CD was around 62% but lack of specificity about comprehension of transmission routes (74,6%), and symptoms (85,3%) were found; concluding that 86% of the surveyed sample had very poor level of knowledge about the disease despite preventive campaigns carried out in the two communities studied. CONCLUSIONS: Despite of a low frequency of CD in this Caribbean areas, the presence of vector, risk factors plus poor level of knowledge about the disease justify that public health intervention strategies should be implemented and monitored over time to maintain uninterrupted surveillance of Chagas Disease.


Assuntos
Doença de Chagas , Conhecimentos, Atitudes e Prática em Saúde , Doenças Negligenciadas , População Rural , Doença de Chagas/prevenção & controle , Doença de Chagas/epidemiologia , Humanos , Colômbia/epidemiologia , Estudos Transversais , Feminino , Masculino , Adulto , Pessoa de Meia-Idade , Doenças Negligenciadas/prevenção & controle , Doenças Negligenciadas/epidemiologia , Adulto Jovem , Adolescente , Idoso , Inquéritos e Questionários , Região do Caribe/epidemiologia
9.
Microb Cell Fact ; 12: 39, 2013 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-23617558

RESUMO

BACKGROUND: Bacteria included in the genus Streptomyces exhibit several attractive characteristics that make them adequate hosts for the heterologous expression of proteins. One of them is that some of its species have a high secretion capacity and hence the protein of interest could be released to the culture supernatant, facilitating downstream processing. To date, all the expression vectors described for these bacteria contain antibiotic resistance genes as selection markers. However, the use of antibiotics to produce proteins at industrial level is currently becoming more restricted owing to the possibility of contamination of the final product. In this report, we describe the use of the S. lividans yefM/yoeBsl toxin-antitoxin system to develop a stable plasmid expression system. RESULTS: In order to use the yefM/yoeBsl system to stabilize expression plasmids in Streptomyces, a S. lividans mutant strain that contained only the toxin gene (yoeBsl) in its genome and the antitoxin gene (yefMsl) located in a temperature-sensitive plasmid was constructed and used as host. This strain was transformed with an expression plasmid harbouring both the antitoxin gene and the gene encoding the protein of interest. Thus, after elimination of the temperature-sensitive plasmid, only cells with the expression plasmid were able to survive. On using this system, two proteins - an α-amylase from S. griseus and a xylanase from S. halstedii - were overproduced without the addition of antibiotic to the culture medium. The production of both proteins was high, even after long incubations (8 days), and after serial subcultures, confirming the stability of the plasmids without antibiotic selection. CONCLUSIONS: This is the first report that describes the use of a toxin-antitoxin system to maintain high -copy plasmids in Streptomyces. This finding could be a valuable tool for using Streptomyces as a host to produce proteins at the industrial and pharmaceutical levels without the use of antibiotics in the production step.


Assuntos
Antitoxinas/genética , Proteínas de Bactérias/genética , Proteínas de Escherichia coli/genética , Plasmídeos/metabolismo , Streptomyces lividans/metabolismo , Toxinas Biológicas/genética , Clonagem Molecular , Endo-1,4-beta-Xilanases/genética , Endo-1,4-beta-Xilanases/metabolismo , Plasmídeos/genética , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética , Streptomyces/enzimologia , alfa-Amilases/genética , alfa-Amilases/metabolismo
10.
Microb Cell Fact ; 12: 127, 2013 Dec 19.
Artigo em Inglês | MEDLINE | ID: mdl-24354561

RESUMO

Streptomyces, the main antibiotic-producing bacteria, responds to changing environmental conditions through a complex sensing mechanism and two-component systems (TCSs) play a crucial role in this extraordinary "sensing" device.Moreover, TCSs are involved in the biosynthetic control of a wide range of secondary metabolites, among them commercial antibiotics. Increased knowledge about TCSs can be a powerful asset in the manipulation of bacteria through genetic engineering with a view to obtaining higher efficiencies in secondary metabolite production. In this review we summarise the available information about Streptomyces TCSs, focusing specifically on their connections to antibiotic production.


Assuntos
Antibacterianos/metabolismo , Streptomyces/metabolismo , Metabolismo Secundário , Streptomyces/genética
11.
Front Microbiol ; 14: 1217350, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37492264

RESUMO

Bacteria of the genus Streptomyces have a plethora of transcriptional regulators, among which the xenobiotic response element (XRE) plays an important role. In this organism, XRE regulators are often followed downstream by small proteins of unknown function containing a DUF397 domain. It has been proposed that XRE/DUF397 pairs constitute type II toxin-antitoxin (TA) systems. However, previous work carried out by our group has shown that one of these systems is a strong activator of antibiotic production in S. coelicolor and other Streptomyces species. In this work, we have studied the overexpression of fourteen XRE/DUF397 pairs present in the S. coelicolor genome and found that none behave as a type II TA system. Instead, they act as pleiotropic regulators affecting, in a dependent manner, antibiotic production and morphological differentiation on different culture media. After deleting, individually, six XRE/DUF397 pairs (those systems producing more notable phenotypic changes when overexpressed: SCO2246/45, SCO2253/52, SCO4176/77, SCO4678/79, SCO6236/35, and SCO7615/16), the pair SCO7615/16 was identified as producing the most dramatic differences as compared to the wild-type strain. The SCO7615/16 mutant had a different phenotype on each of the media tested (R2YE, LB, NMMP, YEPD, and MSA). In particular, on R2YE and YEPD media, a bald phenotype was observed even after 7 days, with little or no actinorhodin (ACT) production. Lower ACT production was also observed on LB medium, but the bacteria were able to produce aerial mycelium. On NMMP medium, the mutant produced a larger amount of ACT as compared with the wild-type strain.

12.
Mutat Res ; 742(1-2): 66-71, 2012 Feb 18.
Artigo em Inglês | MEDLINE | ID: mdl-22198330

RESUMO

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous pollutants presenting a public health risk, particularly to children, a vulnerable population. PAHs have genotoxic and carcinogenic properties, which depend on their metabolism. Many enzymes involved in PAH metabolism, including CYP1A1, CYP1B1, GSTM and GSTT are polymorphic, which may modulate the activation/deactivation of these compounds. We evaluated PAH exposure and DNA damage in children living in the vicinity of the main petrochemical complex located in the Gulf of Mexico, and explored the modulation by genetic polymorphisms of PAH excretion and related DNA damage. The participants (n=82) were children aged 6-10y attending schools near the industrial area. Urinary 1-hydroxypyrene (1-OHP; a biomarker of PAH exposure) was determined by reverse-phase-HPLC; DNA damage by the comet assay (Olive Tail Moment (OTM) parameter); CYP1A1*2C and CYP1B1*3 polymorphisms by real time-PCR; and GSTM1*0 and GSTT1*0 by multiplex PCR. The median value of 1-OHP was 0.37µmol/mol creatinine; 59% of children had higher 1-OHP concentrations than those reported in environmentally exposed adults (0.24µmol/mol creatinine). A stratified analysis showed increased DNA damage in children with 1-OHP concentrations greater than the median value. We observed higher 1-OHP concentrations in children with CYP1A1*2C or GSTM1*0 polymorphisms, and a positive influence of CYP1A1*2C on OTM values in children with the highest PAH exposure. The data indicate that children living in the surroundings of petrochemical industrial areas are exposed to high PAH levels, contributing to DNA damage and suggesting an increased health risk; furthermore, data suggest that polymorphisms affecting activation enzymes may modulate PAH metabolism and toxicity.


Assuntos
Dano ao DNA/efeitos dos fármacos , Exposição Ambiental , Hidrocarbonetos Policíclicos Aromáticos/toxicidade , Criança , Citocromo P-450 CYP1A1/genética , Feminino , Humanos , Masculino , México , Hidrocarbonetos Policíclicos Aromáticos/metabolismo , Polimorfismo Genético , Pirenos/farmacocinética
13.
Food Nutr Bull ; 33(4): 261-6, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23424892

RESUMO

BACKGROUND: Corn tortilla is the staple food of Mexico and its fortification with zinc, iron, and other micronutrients is intended to reduce micronutrient deficiencies. However, no studies have been performed to determine the relative amount of zinc absorbed from the fortified product and whether zinc absorption is affected by the simultaneous addition of iron. OBJECTIVE: To compare zinc absorption from corn tortilla fortified with zinc oxide versus zinc sulfate and to determine the effect of simultaneous addition of two doses of iron on zinc bioavailability. METHODS: A randomized, double-blind, crossover design was carried out in two phases. In the first phase, 10 adult women received corn tortillas with either 20 mg/kg of zinc oxide added, 20 mg/kg of zinc sulfate added, or no zinc added. In the second phase, 10 adult women received corn tortilla with 20 mg/kg of zinc oxide added and either with no iron added or with iron added at one of two different levels. Zinc absorption was measured by the stable isotope method. RESULTS: The mean (+/- SEM) fractional zinc absorption from unfortified tortilla, tortilla fortified with zinc oxide, and tortilla fortified with zinc sulfate did not differ among treatments: 0.35 +/- 0.07, 0.36 +/- 0.05, and 0.37 +/- 0.07, respectively. The three treatment groups with 0, 30, and 60 mg/kg of added iron had similar fractional zinc absorption (0.32 +/- 0.04, 0.33 +/- 0.02, and 0.32 +/- 0.05, respectively) and similar amounts of zinc absorbed (4.8 +/- 0.7, 4.5 +/- 0.3, and 4.8 +/- 0.7 mg/day, respectively). CONCLUSIONS: Since zinc oxide is more stable and less expensive and was absorbed equally as well as zinc sulfate, we suggest its use for corn tortilla fortification. Simultaneous addition of zinc and iron to corn tortilla does not modify zinc bioavailability at iron doses of 30 and 60 mg/kg of corn flour.


Assuntos
Ferro da Dieta/administração & dosagem , Óxido de Zinco/administração & dosagem , Óxido de Zinco/farmacocinética , Absorção , Adulto , Disponibilidade Biológica , Índice de Massa Corporal , Estudos Cross-Over , Método Duplo-Cego , Feminino , Farinha , Alimentos Fortificados , Humanos , Ferro da Dieta/sangue , Modelos Lineares , Micronutrientes/administração & dosagem , Adulto Jovem , Zea mays , Zinco/administração & dosagem , Zinco/sangue , Óxido de Zinco/sangue , Sulfato de Zinco/administração & dosagem
14.
Antibiotics (Basel) ; 11(5)2022 May 17.
Artigo em Inglês | MEDLINE | ID: mdl-35625316

RESUMO

Streptomyces bacteria produce a wide number of antibiotics and antitumor compounds that have attracted the attention of pharmaceutical and biotech companies. In this study, we provide evidence showing that the xylem sap from grapevines has a positive effect on the production of different antibiotics by several Streptomyces species, including S. ambofaciens ATCC 23877 and S. argillaceus ATCC 12596 among others. The production of several already known compounds was induced: actinomycin D, chromomycin A3, fungichromin B, mithramycin A, etc., and four compounds with molecular formulas not included in the Dictionary of Natural Products (DNP v28.2) were also produced. The molecules present in the xylem sap that acts as elicitors were smaller than 3 kDa and soluble in water and insoluble in ether, ethyl acetate, or methanol. A combination of potassium citrate and di-D-fructose dianhydrides (related to levanbiose or inulobiose) seemed to be the main effectors identified from the active fraction. However, the level of induction obtained in the presence of these compounds mix was weaker and delayed with respect to the one got when using the whole xylem sap or the 3 kDa sap fraction, suggesting that another, not identified, elicitor must be also implied in this induction.

15.
J Ind Microbiol Biotechnol ; 38(9): 1419-26, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21181427

RESUMO

Xylanases are very often modular enzymes composed of one or more catalytic domains and carbohydrate-binding modules (CBMs) connected by a flexible linker region. Usually, when these proteins are processed they lose their carbohydrate-binding capacity. Here, the role of the linker regions and cellulose- or xylan-binding domains in the processing of Xys1L from Streptomyces halstedii JM8 and Xyl30L from Streptomyces avermitilis UAH30 was studied. Xys1 variants with different linker lengths were tested, these being unable to avoid protein processing. Moreover, several fusion proteins between the Xys1 and Xyl30 domains were obtained and their proteolytic stability was studied. We demonstrate that CBM processing takes place even in the complete absence of the linker sequence. We also show that the specific carbohydrate module determines this cleavage in the proteins studied.


Assuntos
Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Processamento de Proteína Pós-Traducional , Streptomyces/enzimologia , Xilosidases/química , Xilosidases/metabolismo , Sequência de Aminoácidos , Proteínas de Bactérias/genética , Domínio Catalítico , Celulose/metabolismo , Dados de Sequência Molecular , Estrutura Terciária de Proteína , Proteínas Recombinantes de Fusão/metabolismo , Xilanos/metabolismo , Xilosidases/genética
16.
Phys Rev E ; 104(6-1): 064309, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-35030861

RESUMO

This report is concerned with the relevance of the microscopic rules that implement individual neuronal activation, in determining the collective dynamics, under variations of the network topology. To fix ideas we study the dynamics of two cellular automaton models, commonly used, rather in-distinctively, as the building blocks of large-scale neuronal networks. One model, due to Greenberg and Hastings (GH), can be described by evolution equations mimicking an integrate-and-fire process, while the other model, due to Kinouchi and Copelli (KC), represents an abstract branching process, where a single active neuron activates a given number of postsynaptic neurons according to a prescribed "activity" branching ratio. Despite the apparent similarity between the local neuronal dynamics of the two models, it is shown that they exhibit very different collective dynamics as a function of the network topology. The GH model shows qualitatively different dynamical regimes as the network topology is varied, including transients to a ground (inactive) state, continuous and discontinuous dynamical phase transitions. In contrast, the KC model only exhibits a continuous phase transition, independently of the network topology. These results highlight the importance of paying attention to the microscopic rules chosen to model the interneuronal interactions in large-scale numerical simulations, in particular when the network topology is far from a mean-field description. One such case is the extensive work being done in the context of the Human Connectome, where a wide variety of types of models are being used to understand the brain collective dynamics.

17.
Front Microbiol ; 12: 674214, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34421839

RESUMO

Ocean currents, multiple fecal bacteria input sources, and jurisdictional boundaries can complicate pollution source tracking and associated mitigation and management efforts within the nearshore coastal environment. In this study, multiple microbial source tracking tools were employed to characterize the impact and reach of an ocean wastewater treatment facility discharge in Mexico northward along the coast and across the Southwest United States- Mexico Border. Water samples were evaluated for fecal indicator bacteria (FIB), Enterococcus by culture-based methods, and human-associated genetic marker (HF183) and Enterococcus by droplet digital polymerase chain reaction (ddPCR). In addition, 16S rRNA gene sequence analysis was performed and the SourceTracker algorithm was used to characterize the bacterial community of the wastewater treatment plume and its contribution to beach waters. Sampling dates were chosen based on ocean conditions associated with northern currents. Evidence of a gradient in human fecal pollution that extended north from the wastewater discharge across the United States/Mexico border from the point source was observed using human-associated genetic markers and microbial community analysis. The spatial extent of fecal contamination observed was largely dependent on swell and ocean conditions. These findings demonstrate the utility of a combination of molecular tools for understanding and tracking specific pollutant sources in dynamic coastal water environments.

18.
Front Microbiol ; 11: 587750, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33162964

RESUMO

Antibiotic resistance currently presents one of the biggest threats to humans. The development and implementation of strategies against the spread of superbugs is a priority for public health. In addition to raising social awareness, approaches such as the discovery of new antibiotic molecules and the elucidation of resistance mechanisms are common measures. Accordingly, the two-component system (TCS) of Streptomyces coelicolor AbrB1/B2, offer amenable ways to study both antibiotic production and resistance. Global transcriptomic comparisons between the wild-type strain S. coelicolor M145 and the mutant ΔabrB, using RNA-Seq, showed that the AbrB1/B2 TCS is implicated in the regulation of different biological processes associated with stress responses, primary and secondary metabolism, and development and differentiation. The ΔabrB mutant showed the up-regulation of antibiotic biosynthetic gene clusters and the down-regulation of the vancomycin resistance gene cluster, according to the phenotypic observations of increased antibiotic production of actinorhodin and undecylprodigiosin, and greater susceptibility to vancomycin. The role of AbrB1/B2 in vancomycin resistance has also been shown by an in silico analysis, which strongly indicates that AbrB1/B2 is a homolog of VraR/S from Staphylococcus aureus and LiaR/S from Enterococcus faecium/Enterococcus faecalis, both of which are implied in vancomycin resistance in these pathogenic organisms that present a serious threat to public health. The results obtained are interesting from a biotechnological perspective since, on one hand, this TCS is a negative regulator of antibiotic production and its high degree of conservation throughout Streptomyces spp. makes it a valuable tool for improving antibiotic production and the discovery of cryptic metabolites with antibiotic action. On the other hand, AbrB1/B2 contributes to vancomycin resistance and is a homolog of VraR/S and LiaR/S, important regulators in clinically relevant antibiotic-resistant bacteria. Therefore, the study of AbrB1/B2 could provide new insight into the mechanism of this type of resistance.

19.
Microorganisms ; 8(12)2020 Dec 16.
Artigo em Inglês | MEDLINE | ID: mdl-33339339

RESUMO

Actinomycetes constitute a large group of Gram-positive bacteria present in different habitats. One of these habitats involves the association of these bacteria with insects. In this work, we have studied twenty-four actinomycetes strains isolated from the intestinal tract and feces from larvae of the xylophagous coleopteran Cerambyx welensii and have shown that seventeen strains present hydrolytic activity of some of the following substrates: cellulose, hemicellulose, starch and proteins. Fourteen of the isolates produce antimicrobial molecules against the Gram-positive bacteria Micrococcus luteus. Analysis of seven strains led us to identify the production of a wide number of compounds including streptanoate, alpiniamide A, alteramides A and B, coproporphyrin III, deferoxamine, demethylenenocardamine, dihydropicromycin, nocardamine, picromycin, surugamides A, B, C, D and E, tirandamycins A and B, and valinomycin. A significant number of other compounds, whose molecular formulae are not included in the Dictionary of Natural Products (DNP), were also present in the extracts analyzed, which opens up the possibility of identifying new active antibiotics. Molecular identification of ten of the isolated bacteria determined that six of them belong to the genus Streptomyces, two of them are included in the genus Amycolatopsis and two in the genus Nocardiopsis.

20.
J Environ Health Sci Eng ; 18(2): 1189-1205, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33312634

RESUMO

PURPOSE: The aim of the present work was to assess the electrogenic activity of bacteria from hydrothermal vent sediments achieved under sulfate reducing (SR) conditions in a microbial fuel cell design with acetate, propionate and butyrate as electron donors. METHODS: Two different mixtures of volatile fatty acids (VFA) were evaluated as the carbon source at two chemical oxygen demand (COD) proportions. The mixtures of VFA used were: acetate, propionate and butyrate COD: 3:0.5:0.5 (stage 1) and acetate - butyrate COD: 3.5:0.5 (stage 2). Periodical analysis of sulfate (SO4 -2), sulfide (HS-) and COD were conducted to assess sulfate reduction (SR) and COD removal along with measurements of voltage and current to assess the global performance of the consortium in the system. RESULTS: Percentage of SR was of 97.5 ± 0.7 and 74.3 ± 1.5% for stage 1 and 2, respectively. The % COD removal was of 91 ± 2.1 and 75.3 ± 9.6 for stage 1 and 2, respectively. Although SR and COD removal were higher at stage 1, in regards of energy, stage 2 presented higher current and power densities and Coulombic efficiency as follows: 741.7 ± 30.5 µA/m2, 376 ± 34.4 µW/m2 and 5 ± 2.7%, whereas for stage 1 these values were: 419 ± 71 µA/m2, 52.7 ± 18 µW/m2 and 0.02%, respectively. A metagenomic analysis - stage 2 - in the anodic chamber, demonstrated that SR was due to Dethiosulfovibrionaceae (HA73), Desulfobacter and Desulfococcus and the electrogenic microorganisms were Planococcus, SHD-231, Proteiniclasticum, vadinCA02, and families Porphyromonadacea and Pseudomonadaceae. CONCLUSIONS: It was demonstrated that microorganisms prevenient from hydrothermal vent sediments adapted to a microbial fuel cell system are able to generate electricity coupled to 74.3 ± 1.5 and 75.3 ± 9.6% of SR and COD removal respectively, with a mixture of acetate - butyrate.

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