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1.
Methods Mol Biol ; 2314: 1-58, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34235647

RESUMO

Building upon the foundational research of Robert Koch, who demonstrated the ability to grow Mycobacterium tuberculosis for the first time in 1882 using media made of coagulated bovine serum, microbiologists have continued to develop new and more efficient ways to grow mycobacteria. Presently, all known mycobacterial species can be grown in the laboratory using either axenic culture techniques or in vivo passage in laboratory animals. This chapter provides conventional protocols to grow mycobacteria for diagnostic purposes directly from clinical specimens, as well as in research laboratories for scientific purposes. Detailed protocols used for production of M. tuberculosis in large scale (under normoxic and hypoxic conditions) in bioreactors and for production of obligate intracellular pathogens such as Mycobacterium leprae and "Mycobacterium lepromatosis" using athymic nude mice and armadillos are provided.


Assuntos
Técnicas Bacteriológicas , Infecções por Mycobacterium/microbiologia , Mycobacterium/crescimento & desenvolvimento , Animais , Tatus , Técnicas Bacteriológicas/instrumentação , Reatores Biológicos , Modelos Animais de Doenças , Humanos , Camundongos Nus , Viabilidade Microbiana , Mycobacterium/isolamento & purificação , Mycobacterium leprae/crescimento & desenvolvimento , Mycobacterium leprae/isolamento & purificação , Fatores de Tempo
2.
Int J Mol Sci ; 22(14)2021 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-34299217

RESUMO

The mycobacterial cell wall is composed of large amounts of lipids with varying moieties. Some mycobacteria species hijack host cells and promote lipid droplet accumulation to build the cellular environment essential for their intracellular survival. Thus, lipids are thought to be important for mycobacteria survival as well as for the invasion, parasitization, and proliferation within host cells. However, their physiological roles have not been fully elucidated. Recent studies have revealed that mycobacteria modulate the peroxisome proliferator-activated receptor (PPAR) signaling and utilize host-derived triacylglycerol (TAG) and cholesterol as both nutrient sources and evasion from the host immune system. In this review, we discuss recent findings that describe the activation of PPARs by mycobacterial infections and their role in determining the fate of bacilli by inducing lipid metabolism, anti-inflammatory function, and autophagy.


Assuntos
Infecções por Mycobacterium/microbiologia , Mycobacterium/metabolismo , Receptores Ativados por Proliferador de Peroxissomo/metabolismo , Animais , Autofagia/fisiologia , Colesterol/metabolismo , Humanos , Metabolismo dos Lipídeos , Mycobacterium/crescimento & desenvolvimento , Mycobacterium/imunologia , Infecções por Mycobacterium/imunologia , Infecções por Mycobacterium/metabolismo , Receptores Ativados por Proliferador de Peroxissomo/genética , Transdução de Sinais
3.
Microbiol Spectr ; 7(2)2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-31025625

RESUMO

How do mycobacteria divide? Cell division has been studied extensively in the model rod-shaped bacteria Escherichia coli and Bacillus subtilis, but much less is understood about cell division in mycobacteria, a genus that includes the major human pathogens M. tuberculosis and M. leprae. In general, bacterial cell division requires the concerted effort of many proteins in both space and time to elongate the cell, replicate and segregate the chromosome, and construct and destruct the septum - processes which result in the creation of two new daughter cells. Here, we describe these distinct stages of cell division in B. subtilis and follow with the current knowledge in mycobacteria. As will become apparent, there are many differences between mycobacteria and B. subtilis in terms of both the broad outline of cell division and the molecular details. So, while the fundamental challenge of spatially and temporally organizing cell division is shared between these rod-shaped bacteria, they have solved these challenges in often vastly different ways.


Assuntos
Divisão Celular/fisiologia , Mycobacterium/crescimento & desenvolvimento , Mycobacterium/metabolismo , Bacillus subtilis/genética , Bacillus subtilis/crescimento & desenvolvimento , Bacillus subtilis/metabolismo , Proteínas de Bactérias/metabolismo , Proteínas de Ciclo Celular/metabolismo , Divisão Celular/genética , Parede Celular , Replicação do DNA , Escherichia coli/genética , Escherichia coli/crescimento & desenvolvimento , Escherichia coli/metabolismo , Mycobacterium/genética , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/crescimento & desenvolvimento , Mycobacterium tuberculosis/metabolismo
4.
Int J Mol Sci ; 20(5)2019 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-30818787

RESUMO

In dealing with Mycobacterium tuberculosis, the causative agent of the deadliest human disease-tuberculosis (TB)-utilization of cholesterol as a carbon source indicates the possibility of using cholesterol catabolic genes/proteins as novel drug targets. However, studies on cholesterol catabolism in mycobacterial species are scarce, and the number of mycobacterial species utilizing cholesterol as a carbon source is unknown. The availability of a large number of mycobacterial species' genomic data affords an opportunity to explore and predict mycobacterial species' ability to utilize cholesterol employing in silico methods. In this study, comprehensive comparative analysis of cholesterol catabolic genes/proteins in 93 mycobacterial species was achieved by deducing a comprehensive cholesterol catabolic pathway, developing a software tool for extracting homologous protein data and using protein structure and functional data. Based on the presence of cholesterol catabolic homologous proteins proven or predicted to be either essential or specifically required for the growth of M. tuberculosis H37Rv on cholesterol, we predict that among 93 mycobacterial species, 51 species will be able to utilize cholesterol as a carbon source. This study's predictions need further experimental validation and the results should be taken as a source of information on cholesterol catabolism and genes/proteins involved in this process among mycobacterial species.


Assuntos
Proteínas de Bactérias/genética , Colesterol/metabolismo , Genes Bacterianos , Mycobacterium/genética , Animais , Proteínas de Bactérias/metabolismo , Colesterol/química , Genes Essenciais , Macrófagos/metabolismo , Macrófagos/microbiologia , Redes e Vias Metabólicas , Camundongos , Viabilidade Microbiana/genética , Mycobacterium/crescimento & desenvolvimento , Infecções por Mycobacterium/genética , Infecções por Mycobacterium/microbiologia , Especificidade da Espécie
5.
Chem Phys Lipids ; 194: 125-38, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26256431

RESUMO

2-Alkynoic acids have bactericidal activity against Mycobacterium smegmatis but their activity fall sharply as the length of the carbon chain increased. In this study, derivatives of 2-alkynoic acids were synthesized and tested against fast- and slow-growing mycobacteria. Their activity was first evaluated in M. smegmatis against their parental 2-alkynoic acids, as well as isoniazid, a first-line antituberculosis drug. The introduction of additional unsaturation or heteroatoms into the carbon chain enhanced the antimycobacterial activity of longer chain alkynoic acids (more than 19 carbons long). In contrast, although the modification of the carboxylic group did not improve the antimycobacterial activity, it significantly reduced the toxicity of the compounds against eukaryotic cells. Importantly, 4-(alkylthio)but-2-ynoic acids, had better bactericidal activity than the parental 2-alkynoic acids and on a par with isoniazid against the slow-grower Mycobacterium bovis BCG. These compounds had also low toxicity against eukaryotic cells, suggesting that they could be potential therapeutic agents against other types of topical mycobacterial infections causing skin diseases including Mycobacterium abscessus, Mycobacterium ulcerans, and Mycobacterium leprae. Moreover, they provide a possible scaffold for future drug development.


Assuntos
Antibacterianos/síntese química , Antibacterianos/farmacologia , Ácidos Graxos Insaturados/farmacologia , Mycobacterium/efeitos dos fármacos , Antibacterianos/química , Ácidos Graxos Insaturados/síntese química , Ácidos Graxos Insaturados/química , Testes de Sensibilidade Microbiana , Estrutura Molecular , Mycobacterium/crescimento & desenvolvimento
6.
FEMS Microbiol Lett ; 313(1): 68-74, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21039782

RESUMO

ATP synthase is a validated drug target for the treatment of tuberculosis, and ATP synthase inhibitors are promising candidate drugs for the treatment of infections caused by other slow-growing mycobacteria, such as Mycobacterium leprae and Mycobacterium ulcerans. ATP synthase is an essential enzyme in the energy metabolism of Mycobacterium tuberculosis; however, no biochemical data are available to characterize the role of ATP synthase in slow-growing mycobacterial strains. Here, we show that inverted membrane vesicles from the slow-growing model strain Mycobacterium bovis BCG are active in ATP synthesis, but ATP synthase displays no detectable ATP hydrolysis activity and does not set up a proton-motive force (PMF) using ATP as a substrate. Treatment with methanol as well as PMF activation unmasked the ATP hydrolysis activity, indicating that the intrinsic subunit ɛ and inhibitory ADP are responsible for the suppression of hydrolytic activity. These results suggest that the enzyme is needed for the synthesis of ATP, not for the maintenance of the PMF. For the development of new antimycobacterial drugs acting on ATP synthase, screening for ATP synthesis inhibitors, but not for ATP hydrolysis blockers, can be regarded as a promising strategy.


Assuntos
Trifosfato de Adenosina/metabolismo , ATPases Mitocondriais Próton-Translocadoras/metabolismo , Mycobacterium/enzimologia , Mycobacterium/crescimento & desenvolvimento
7.
PLoS One ; 4(7): e6263, 2009 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-19606228

RESUMO

BACKGROUND: Mycobacterium indicus pranii (MIP), popularly known as Mw, is a cultivable, non-pathogenic organism, which, based on its growth and metabolic properties, is classified in Runyon Group IV along with M. fortuitum, M. smegmatis and M. vaccae. The novelty of this bacterium was accredited to its immunological ability to undergo antigen driven blast transformation of leukocytes and delayed hypersensitivity skin test in leprosy patients, a disease endemic in the Indian sub-continent. Consequently, MIP has been extensively evaluated for its biochemical and immunological properties leading to its usage as an immunomodulator in leprosy and tuberculosis patients. However, owing to advances in sequencing and culture techniques, the citing of new strains with almost 100% similarity in the sequences of marker genes like 16S rRNA, has compromised the identity of MIP as a novel species. Hence, to define its precise taxonomic position, we have carried out polyphasic taxonomic studies on MIP that integrate its phenotypic, chemotaxonomic and molecular phylogenetic attributes. METHODOLOGY/PRINCIPAL FINDINGS: The comparative analysis of 16S rRNA sequence of MIP by using BLAST algorithm at NCBI (nr database) revealed a similarity of > or =99% with M. intracellulare, M. arosiense, M. chimaera, M. seoulense, M. avium subsp. hominissuis, M. avium subsp. paratuberculosis and M. bohemicum. Further analysis with other widely used markers like rpoB and hsp65 could resolve the phylogenetic relationship between MIP and other closely related mycobacteria apart from M. intracellulare and M. chimaera, which shares > or =99% similarity with corresponding MIP orthologues. Molecular phylogenetic analysis, based on the concatenation of candidate orthologues of 16S rRNA, hsp65 and rpoB, also substantiated its distinctiveness from all the related organisms used in the analysis excluding M. intracellulare and M. chimaera with which it exhibited a close proximity. This necessitated further analysis of MIP with more sensitive and segregating parameters to ascertain its precise taxonomic position as a new species. The analysis of MIP and its comparison with other mycobacterial reference strains based on cellular and biochemical features, growth characteristics and chemotaxonomic studies like FAME profiling confirmed that MIP is uniquely endowed with diverse metabolic attributes that effectively distinguishes it from all the closely related mycobacteria including M. intracellulare and M. chimaera. CONCLUSION: The results presented in this study coupled with the non-pathogenic nature and different biochemical and immunomodulatory properties of MIP affirm it as a distinct species belonging to M. avium complex (MAC). It is further proposed to use an earlier suggested name Mycobacterium indicus pranii for this newly established mycobacterial species. This study also exemplifies the growing need for a uniform, consensus based broader polyphasic frame work for the purpose of taxonomy and speciation, particularly in the genus Mycobacterium.


Assuntos
Mycobacterium/classificação , Algoritmos , Dados de Sequência Molecular , Mycobacterium/genética , Mycobacterium/crescimento & desenvolvimento , Filogenia , RNA Ribossômico 16S/genética , Especificidade da Espécie
8.
Microbiol Mol Biol Rev ; 72(1): 126-56, table of contents, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18322037

RESUMO

The genus Mycobacterium is best known for its two major pathogenic species, M. tuberculosis and M. leprae, the causative agents of two of the world's oldest diseases, tuberculosis and leprosy, respectively. M. tuberculosis kills approximately two million people each year and is thought to latently infect one-third of the world's population. One of the most remarkable features of the nonsporulating M. tuberculosis is its ability to remain dormant within an individual for decades before reactivating into active tuberculosis. Thus, control of cell division is a critical part of the disease. The mycobacterial cell wall has unique characteristics and is impermeable to a number of compounds, a feature in part responsible for inherent resistance to numerous drugs. The complexity of the cell wall represents a challenge to the organism, requiring specialized mechanisms to allow cell division to occur. Besides these mycobacterial specializations, all bacteria face some common challenges when they divide. First, they must maintain their normal architecture during and after cell division. In the case of mycobacteria, that means synthesizing the many layers of complex cell wall and maintaining their rod shape. Second, they need to coordinate synthesis and breakdown of cell wall components to maintain integrity throughout division. Finally, they need to regulate cell division in response to environmental stimuli. Here we discuss these challenges and the mechanisms that mycobacteria employ to meet them. Because these organisms are difficult to study, in many cases we extrapolate from information known for gram-negative bacteria or more closely related GC-rich gram-positive organisms.


Assuntos
Divisão Celular , Mycobacterium/citologia , Mycobacterium/crescimento & desenvolvimento , Parede Celular/metabolismo , Parede Celular/ultraestrutura
9.
Artigo em Alemão | MEDLINE | ID: mdl-16283122

RESUMO

A characteristic feature of mycobacteria is their slow growth rate, which in addition strongly varies in different species of the genus. All highly pathogenic species such as M. tuberculosis and M. leprae causing tuberculosis and leprosy, respectively, belong to the slow growing mycobacteria, while the apathogenic and opportunistic species are members of the fast growing mycobacteria. This suggests that the question be posed whether there is causality between mycobacterial growth rate and virulence. We discuss possible reasons for the slow and variable growth rates of mycobacteria and the current state of knowledge concerning the significance of slow growth for mycobacterial pathogenicity.


Assuntos
Infecções por Mycobacterium/microbiologia , Mycobacterium/crescimento & desenvolvimento , Mycobacterium/patogenicidade , Animais , Proliferação de Células , Humanos , Tuberculose/microbiologia
11.
Arch Microbiol ; 176(5): 381-5, 2001 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-11702081

RESUMO

Mycobacterium sp. Pyr-1 produces an enzyme with nitroreductase activity that reduces 1-nitropyrene and 4-nitrobenzoic acid to the corresponding aromatic amines. This enzyme was constitutive and required NADH; and its activity was enhanced by FAD. It was inhibited by antimycin A, dicumarol, and o-iodosobenzoic acid; and it was inactivated by ammonium sulfate precipitation. After purification to homogeneity, the protein produced a single band on native and SDS-polyacrylamide gels and had a single amino-terminal sequence. The N-terminal amino acid sequence was identical to the corresponding sequences of the lipoamide dehydrogenases of M. leprae, M. tuberculosis and Corynebacterium glutamicum. The amino-terminal sequence was also similar to lipoamide dehydrogenases from M. smegmatis and several other bacteria. The amino acid sequence of an internal peptide (12 of 13 amino acids) was nearly identical to the corresponding sequences of lipoamide dehydrogenases from M. leprae and M. tuberculosis and was similar to those of C. glutamicum, Streptomyces coelicolor and S. seoulensis. The data show that a unique lipoamide dehydrogenase in Mycobacterium sp. Pyr-1, which differs from classic (Type I) bacterial nitroreductases, reduces aromatic nitro compounds to aromatic amines.


Assuntos
Di-Hidrolipoamida Desidrogenase/química , Mycobacterium/enzimologia , Nitrorredutases/isolamento & purificação , Nitrorredutases/metabolismo , Sequência de Aminoácidos , Di-Hidrolipoamida Desidrogenase/metabolismo , Dados de Sequência Molecular , Mycobacterium/crescimento & desenvolvimento , Nitrorredutases/química
12.
Epidemiol Infect ; 126(3): 379-87, 2001 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-11467795

RESUMO

More than 36000 individuals living in rural Malawi were skin tested with antigens derived from 12 different species of environmental mycobacteria. Most were simultaneously tested with RT23 tuberculin, and all were followed up for both tuberculosis and leprosy incidence. Skin test results indicated widespread sensitivity to the environmental antigens, in particular to Mycobacterium scrofulaceum, M. intracellulare and one strain of M. fortuitum. Individuals with evidence of exposure to 'fast growers' (i.e. with induration to antigens from fast growers which exceeded their sensitivity to tuberculin), but not those exposed to 'slow growers', were at reduced risk of contracting both tuberculosis and leprosy, compared to individuals whose indurations to the environmental antigen were less than that to tuberculin. This evidence for cross protection from natural exposure to certain environmental mycobacteria may explain geographic distributions of mycobacterial disease and has important implications for the mechanisms and measurement of protection by mycobacterial vaccines.


Assuntos
Antígenos de Bactérias/imunologia , Exposição Ambiental/estatística & dados numéricos , Hanseníase/etiologia , Mycobacterium/crescimento & desenvolvimento , Mycobacterium/imunologia , Saúde da População Rural/estatística & dados numéricos , Pele/microbiologia , Microbiologia do Solo , Teste Tuberculínico , Tuberculose/etiologia , Microbiologia da Água , Adolescente , Adulto , Distribuição por Idade , Criança , Feminino , Seguimentos , Humanos , Incidência , Hanseníase/epidemiologia , Hanseníase/prevenção & controle , Malaui/epidemiologia , Masculino , Mycobacterium/classificação , Mycobacterium/patogenicidade , Vigilância da População , Fatores de Risco , Distribuição por Sexo , Inquéritos e Questionários , Tuberculose/epidemiologia , Tuberculose/prevenção & controle
13.
Acta Cient Venez ; 52 Suppl 1: 42-4, 2001.
Artigo em Espanhol | MEDLINE | ID: mdl-11899705

RESUMO

Variation in growth rate has provided a basis for the broad classification of Mycobacterium: (i) the slow-growing class includes the human pathogens Mycobacterium tuberculosis and M. leprae, and (ii) the fast-growing class includes saprophytic nonpathogens, such as M. smegmatis. An intimate association between DNA chromosomal replication and growth rate have been suggested. However, the molecular basis of this relation is unknown. In this article, we summarise our recent work on the study of the origin replication region of some species of mycobacteria, determination of the factors regulating the initiation of DNA replication and the characterisation of the main factor of the replicative machinery, the DnaA protein.


Assuntos
Cromossomos Bacterianos/fisiologia , Replicação do DNA/fisiologia , DNA Bacteriano/fisiologia , Mycobacterium/genética , Proteínas de Bactérias/genética , Cromossomos Bacterianos/genética , Replicação do DNA/genética , DNA Bacteriano/genética , Proteínas de Ligação a DNA/genética , Mycobacterium/crescimento & desenvolvimento
15.
Probl Tuberk ; (5): 49-52, 2000.
Artigo em Russo | MEDLINE | ID: mdl-11077855

RESUMO

The paper presents the results of studied of cultural biochemical and immunochemical properties of the cultured mycobacteria isolated from patients with leprous lesions of (M.01, M.011) and the soil (M. lufu) in compared to other well-known mycobacteria and between them. M.01, M.011, and M. lufu differ from other representatives of the genus Mycobacterium, but they are close to each other in their cultural biochemical and immunochemical properties, antigenic composition, and protein spectra. The findings are an additional characteristic of the biological properties of M.01, M.011, and M. lufu.


Assuntos
Hanseníase/microbiologia , Mycobacterium leprae/isolamento & purificação , Mycobacterium/isolamento & purificação , Microbiologia do Solo , Animais , Antígenos de Bactérias/análise , Meios de Cultura , Resistência Microbiana a Medicamentos , Cobaias , Humanos , Imunoquímica , Mycobacterium/efeitos dos fármacos , Mycobacterium/crescimento & desenvolvimento , Mycobacterium/imunologia , Mycobacterium/metabolismo , Mycobacterium leprae/efeitos dos fármacos , Mycobacterium leprae/crescimento & desenvolvimento , Mycobacterium leprae/imunologia , Mycobacterium leprae/metabolismo
17.
FEMS Microbiol Lett ; 182(2): 297-301, 2000 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-10620682

RESUMO

Pathogenic species of Mycobacterium are slowly growing intracellular bacteria. Slow growth is important for the parasitism of these organisms and chronicity of the disease, but its precise mechanism has not been elucidated. Recently, we found that a novel DNA-binding protein (MDPI) was expressed (7-10% in total protein) in mycobacteria, such as Mycobacterium bovis bacillus Calmette-Guérin, Mycobacterium tuberculosis, and Mycobacterium leprae. In this study, we observed that MDPI interfered with replication, transcription, and translation in the analysis in in vitro E. coli cell-free macromolecular biosynthesizing systems. Furthermore, MDPI inhibited the rapid growth of both Escherichia coli and Mycobacterium smegmatis, and NH(2)-terminal second amino acid, asparagine, was observed to be important in terms of this function. These data suggest an important role of MDPI for suppression of growth rates of mycobacteria.


Assuntos
Proteínas de Bactérias , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/fisiologia , Mycobacterium/crescimento & desenvolvimento , Mycobacterium/genética , DNA Bacteriano/biossíntese , Escherichia coli/genética , Escherichia coli/metabolismo , Regulação Bacteriana da Expressão Gênica , Mycobacterium/metabolismo , Biossíntese de Proteínas , Transcrição Genética , Transformação Bacteriana
18.
Int J Antimicrob Agents ; 13(2): 133-5, 1999 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-10595573

RESUMO

The resistance of mycobacteria to beta-lactam antibiotics is attributed to their ability to synthesize beta-lactamase. In our previous studies, beta-lactam/beta-lactamase-inhibitor combinations suppressed the growth of several mycobacteria in axenic cultures and ampicillin/sulbactam was bactericidal to Mycobacterium tuberculosis H37Rv in vitro, and to Mycobacterium leprae multiplying in mouse foot-pads. Since both these organisms multiply in phagocytic cells in the host, it is important to know whether the drug combination is active against mycobacteria multiplying in macrophages. We tested the action of ampicillin/sulbactam against four potentially pathogenic (to humans or to animals) mycobacteria, M. simiae, M. haemophilum, M. avium, M. microti, when phagocytosed by mouse macrophages. Bacteria were exposed to monolayers of peritoneal macrophages harvested from BALB/c mice. Unphagocytosed bacilli were removed and three concentrations of ampicillin/sulbactam were tested. Optimum activity was observed at 100 mg/l which killed 58-97% of the mycobacteria within macrophages, as determined by the CFU. beta-Lactam/beta-lactamase-inhibitors, especially ampicillin/sulbactam, might provide an effective alternative therapy against infections caused by mycobacteria resistant to other drugs.


Assuntos
Ampicilina/farmacologia , Inibidores Enzimáticos/farmacologia , Mycobacterium/efeitos dos fármacos , Penicilinas/farmacologia , Sulbactam/farmacologia , Inibidores de beta-Lactamases , Animais , Contagem de Colônia Microbiana , Interações Medicamentosas , Humanos , Técnicas In Vitro , Macrófagos Peritoneais/efeitos dos fármacos , Macrófagos Peritoneais/microbiologia , Camundongos , Camundongos Endogâmicos BALB C , Mycobacterium/crescimento & desenvolvimento , Fagocitose
19.
Proc Natl Acad Sci U S A ; 95(15): 8916-21, 1998 Jul 21.
Artigo em Inglês | MEDLINE | ID: mdl-9671779

RESUMO

Viable cells of Micrococcus luteus secrete a factor, which promotes the resuscitation and growth of dormant, nongrowing cells of the same organism. The resuscitation-promoting factor (Rpf) is a protein, which has been purified to homogeneity. In picomolar concentrations, it increases the viable cell count of dormant M. luteus cultures at least 100-fold and can also stimulate the growth of viable cells. Rpf also stimulates the growth of several other high G+C Gram-positive organisms, including Mycobacterium avium, Mycobacterium bovis (BCG), Mycobacterium kansasii, Mycobacterium smegmatis, and Mycobacterium tuberculosis. Similar genes are widely distributed among high G+C Gram-positive bacteria; genome sequencing has uncovered examples in Mycobacterium leprae and Mb. tuberculosis and others have been detected by hybridization in Mb. smegmatis, Corynebacterium glutamicum, and Streptomyces spp. The mycobacterial gene products may provide different targets for the detection and control of these important pathogens. This report is thus a description of a proteinaceous autocrine or paracrine bacterial growth factor or cytokine.


Assuntos
Proteínas de Bactérias , Citocinas/isolamento & purificação , Micrococcus luteus/química , Sequência de Aminoácidos , Sequência de Bases , Citocinas/química , Citocinas/genética , Primers do DNA , Genes Bacterianos , Micrococcus luteus/genética , Dados de Sequência Molecular , Mycobacterium/crescimento & desenvolvimento , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Homologia de Sequência de Aminoácidos
20.
Vaccine ; 16(13): 1344-8, 1998 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-9682400

RESUMO

The growth of Mycobacterium w, a candidate strain for leprosy vaccine in submerged culture, was inhibited by the presence of over 40% oxygen saturation in the medium. Intracellular levels of superoxide dismutase and catalase were very low in the beginning. However, under controlled oxygenation, these levels increased with time. The augmentations of these antioxidant enzymes were associated with the elevated oxygen consumption by the culture. By maintaining the oxygen level below 20% during 6-day culture, it was possible to grow Mycobacterium w in five production batches up to a cell density of 3.7 +/- 0.70 x 10(9) bacilli ml-1. The shelf life of the vaccine produced in different batches was more than 2 years, both at 4 degrees C and at 26 degrees C. This provides a cost-effective, unit culture technology for the production of this candidate leprosy vaccine from a nonpathogenic organism, which will facilitate the widespread use of the vaccine.


Assuntos
Vacinas Bacterianas , Hanseníase/prevenção & controle , Mycobacterium/metabolismo , Consumo de Oxigênio , Vacinas Bacterianas/imunologia , Vacinas Bacterianas/uso terapêutico , Técnicas Bacteriológicas , Reatores Biológicos , Catalase/metabolismo , Estabilidade de Medicamentos , Armazenamento de Medicamentos , Mycobacterium/enzimologia , Mycobacterium/crescimento & desenvolvimento , Superóxido Dismutase/metabolismo
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