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1.
Biochem Biophys Res Commun ; 687: 149186, 2023 12 20.
Artigo em Inglês | MEDLINE | ID: mdl-37931420

RESUMO

FtsZ, a major cytoskeletal protein in all bacteria and archaea, forms a ring that directs cytokinesis. Bacterial FtsZ is considered the ancestral homolog of the eukaryotic microtubule (MT)-forming tubulins, sharing GTPase activity and the ability to assemble into protofilaments, rings, and sheets, but not MTs. Previous studies from our laboratory demonstrated that structures of isolated brain MTs spontaneously generate electrical oscillations and bursts of electrical activity similar to action potentials. No information about whether the prokaryotic tubulins may share similar properties is available. Here, we obtained by ammonium sulfate precipitation an enriched protein fraction of the endogenous FtsZ from wild-type Escherichia coli ATCC 25922 without any transfection or overexpression of the protein. As revealed by electron microscopy, FtsZ was detected by dot blot analysis and immunofluorescence that assembled into filaments and sheets in a polymerization buffer. We used the patch-clamp technique to explore the electrical properties of sheets of FtsZ and bacterial cells. Electrical recordings at various holding potentials ranging from ±200 mV showed a complex oscillatory behavior, with several peak frequencies between 12 and 110 Hz in the power spectra and a linear mean current response. To confirm the oscillatory electrical behavior of FtsZ we also conducted experiments with commercial recombinant FtsZ, with similar results. We also detected, by local field potentials, similar electrical oscillations in K+-depolarized pellets of E. coli cultures. FtsZ oscillations had a wider range of frequency peaks than MT sheets from eukaryotic origin. The findings indicate that the bacterial cytoskeleton generates electrical oscillators that may play a relevant role in cell division and unknown signaling mechanisms in bacterial populations.


Assuntos
Escherichia coli , Tubulina (Proteína) , Tubulina (Proteína)/metabolismo , Escherichia coli/metabolismo , Proteínas de Bactérias/metabolismo , Proteínas do Citoesqueleto/metabolismo , Bactérias/metabolismo
2.
BMC Microbiol ; 16(1): 240, 2016 Oct 13.
Artigo em Inglês | MEDLINE | ID: mdl-27737643

RESUMO

BACKGROUND: Despite the fact that most vitamins are present in a variety of foods, malnutrition, unbalanced diets or insufficient intake of foods are still the cause of vitamin deficiencies in humans in some countries. Vitamin B12 (Cobalamin) is a complex compound that is only naturally produced by bacteria and archea. It has been reported that certain strains belonging to lactic acid bacteria group are capable of synthesized water-soluble vitamins such as those included in the B-group, as vitamin B12. In this context, the goal of the present paper was to evaluate and characterize the production of vitamin B12 in Lactobacillus coryniformis CRL 1001, a heterofermentative strain isolated from silage. RESULTS: Cell extract of L. coryniformis CRL 1001, isolated from silage, is able to correct the coenzyme B12 requirement of Salmonella enterica serovar Typhimurium AR 2680 in minimal medium. The chemical characterization of the corrinoid-like molecule isolated from CRL 1001 cell extract using HPLC and mass spectrometry is reported. The majority of the corrinoid produced by this strain has adenine like Coα-ligand instead 5,6-dimethylbenzimidazole. Genomic studies revealed the presence of the complete machinery of the anaerobic biosynthesis pathway of coenzyme B12. The detected genes encode all proteins for the corrin ring biosynthesis and for the binding of upper (ß) and lower (α) ligands in one continuous stretch of the chromosome. CONCLUSIONS: The results here described show for the first time that L. coryniformis subsp. coryniformis CRL 1001 is able to produce pseudocobalamin containing adenine instead of 5,6-dimethlbenzimidazole in the Coα-ligand. Genomic analysis allowed the identification and characterization of the complete de novo biosynthetic pathway of the corrinoid produced by the CRL 1001 strain.


Assuntos
Lactobacillus/genética , Lactobacillus/metabolismo , Família Multigênica , Vitamina B 12/biossíntese , Vitamina B 12/genética , Adenina/metabolismo , Anaerobiose , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Sequência de Bases , Benzimidazóis/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Cobamidas/genética , Cobamidas/metabolismo , Genes Bacterianos , Lactobacillus/isolamento & purificação , Espectrometria de Massas/métodos , Fases de Leitura Aberta , Salmonella enterica/genética , Salmonella enterica/metabolismo , Salmonella typhimurium/genética , Análise de Sequência de Proteína , Silagem/microbiologia , Vitamina B 12/química , Vitamina B 12/isolamento & purificação
3.
Cell Genom ; 4(8): 100609, 2024 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-39019033

RESUMO

Little is known about the role of non-coding regions in the etiology of autism spectrum disorder (ASD). We examined three classes of non-coding regions: human accelerated regions (HARs), which show signatures of positive selection in humans; experimentally validated neural VISTA enhancers (VEs); and conserved regions predicted to act as neural enhancers (CNEs). Targeted and whole-genome analysis of >16,600 samples and >4,900 ASD probands revealed that likely recessive, rare, inherited variants in HARs, VEs, and CNEs substantially contribute to ASD risk in probands whose parents share ancestry, which enriches for recessive contributions, but modestly contribute, if at all, in simplex family structures. We identified multiple patient variants in HARs near IL1RAPL1 and in VEs near OTX1 and SIM1 and showed that they change enhancer activity. Our results implicate both human-evolved and evolutionarily conserved non-coding regions in ASD risk and suggest potential mechanisms of how regulatory changes can modulate social behavior.


Assuntos
Transtorno do Espectro Autista , Humanos , Transtorno do Espectro Autista/genética , Transtorno do Espectro Autista/epidemiologia , Predisposição Genética para Doença , Elementos Facilitadores Genéticos/genética , Masculino , Evolução Molecular , Feminino
4.
medRxiv ; 2023 Sep 22.
Artigo em Inglês | MEDLINE | ID: mdl-37790480

RESUMO

Little is known about the role of noncoding regions in the etiology of autism spectrum disorder (ASD). We examined three classes of noncoding regions: Human Accelerated Regions (HARs), which show signatures of positive selection in humans; experimentally validated neural Vista Enhancers (VEs); and conserved regions predicted to act as neural enhancers (CNEs). Targeted and whole genome analysis of >16,600 samples and >4900 ASD probands revealed that likely recessive, rare, inherited variants in HARs, VEs, and CNEs substantially contribute to ASD risk in probands whose parents share ancestry, which enriches for recessive contributions, but modestly, if at all, in simplex family structures. We identified multiple patient variants in HARs near IL1RAPL1 and in a VE near SIM1 and showed that they change enhancer activity. Our results implicate both human-evolved and evolutionarily conserved noncoding regions in ASD risk and suggest potential mechanisms of how changes in regulatory regions can modulate social behavior.

5.
Food Res Int ; 107: 289-296, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29580488

RESUMO

The aim of this work was to evaluate the effect of meat curing agents on the bioprotective activity of the bacteriocinogenic strain, Enterococcus (E.) mundtii CRL35 against Listeria (L.) monocytogenes during meat fermentation. The ability of E. mundtii CRL35 to grow, acidify and produce bacteriocin in situ was assayed in a meat model system in the presence of curing additives (CA). E. mundtii CRL35 showed optimal growth and acidification rates in the presence of CA. More importantly, the highest bacteriocin titer was achieved in the presence of these food agents. In addition, the CA produced a statistical significant enhancement of the enterocin CRL35 activity. This positive effect was demonstrated in vitro in a meat based culture medium, by time-kill kinetics and finally by using a beaker sausage model with a challenge experiment with the pathogenic L. monocytogenes FBUNT strain. E. mundtii CRL35 was found to be a promising strain of use as a safety adjunct culture in meat industry and a novel functional supplement for sausage fermentation, ensuring hygiene and quality of the final product.


Assuntos
Bacteriocinas/metabolismo , Enterococcus/metabolismo , Microbiologia de Alimentos/métodos , Listeria monocytogenes/efeitos dos fármacos , Produtos da Carne/microbiologia , Carne/microbiologia , Bacteriocinas/farmacologia , Fermentação , Conservação de Alimentos/métodos , Listeria monocytogenes/crescimento & desenvolvimento
6.
DNA Res ; 24(1): 11-24, 2017 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-27773878

RESUMO

The study of enterococcal genomes has grown considerably in recent years. While special attention is paid to comparative genomic analysis among clinical relevant isolates, in this study we performed an exhaustive comparative analysis of enterococcal genomes of food origin and/or with potential to be used as probiotics. Beyond common genetic features, we especially aimed to identify those that are specific to enterococcal strains isolated from a certain food-related source as well as features present in a species-specific manner. Thus, the genome sequences of 25 Enterococcus strains, from 7 different species, were examined and compared. Their phylogenetic relationship was reconstructed based on orthologous proteins and whole genomes. Likewise, markers associated with a successful colonization (bacteriocin genes and genomic islands) and genome plasticity (phages and clustered regularly interspaced short palindromic repeats) were investigated for lifestyle specific genetic features. At the same time, a search for antibiotic resistance genes was carried out, since they are of big concern in the food industry. Finally, it was possible to locate 1617 FIGfam families as a core proteome universally present among the genera and to determine that most of the accessory genes code for hypothetical proteins, providing reasonable hints to support their functional characterization.


Assuntos
Enterococcus/genética , Microbiologia de Alimentos , Genoma Bacteriano , Probióticos , Bacteriocinas/genética , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas , Resistência Microbiana a Medicamentos/genética , Enterococcus/classificação , Transferência Genética Horizontal , Marcadores Genéticos , Filogenia
7.
Genome Announc ; 2(3)2014 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-24855298

RESUMO

Enterococcus mundtii CRL35 is a bacteriocinogenic strain isolated from an artisanal cheese of northwestern Argentina. Here we report its draft genome sequence, consisting of 82 contigs. In silico genomic analysis of biotechnological properties was performed to determine the potential of this microorganism to be used in a food model system.

8.
Genome Announc ; 2(6)2014 Dec 11.
Artigo em Inglês | MEDLINE | ID: mdl-25502678

RESUMO

Lactobacillus mucosae CRL573, isolated from child fecal samples, efficiently converts fructose and/or sucrose into the low-calorie sugar mannitol when cultured in modified MRS medium at pH 5.0. Also, the strain is capable of producing bacteriocin. The draft genome sequence of this strain with potential industrial applications is presented here.

9.
Genome Announc ; 1(4)2013 Jul 18.
Artigo em Inglês | MEDLINE | ID: mdl-23868133

RESUMO

We report the draft genome sequence of the bacteriocin producer Enterococcus faecium strain CRL 1879, isolated from a northwestern Argentinian artisanal cheese. The draft genome sequence is composed of 73 contigs for 2,886,747 bp, with 3,140 protein-coding genes. Six biosynthetic clusters for bacteriocin class II production were found. Typical virulence determinants, which have relevance in food safety, were not present.

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