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1.
Antonie Van Leeuwenhoek ; 112(9): 1297-1305, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30993571

RESUMO

Four bacterial strains, with the capability of inhibiting Pseudogymnoascus destructans, the causative agent of white-nose syndrome, were isolated from male Townsend's big-eared bats (Corynorhinus townsendii, Family: Vespertilionidae) in New Mexico. Isolates AC161, AC162, AC208, and AC230T were characterised as a novel clade using morphological, phenotypic and phylogenetic analysis. A draft genome of the type strain was completed to determine its taxonomy and secondary metabolite biosynthetic potential. Multi-locus sequence analysis nests AC230T with neighbours Streptomyces scopuliridis (NRRL B-24574T), Streptomyces lushanensis (NRRL B-24994T), Streptomyces odonnellii (NRRL B-24891T) and Streptomyces niveus (NRRL 2466T). Further phylogenetic analysis showed the MLSA distances between AC230T and its near neighbours are much greater than the generally accepted threshold (> 0.007) for bacterial species delineation. DNA-DNA relatedness between AC230T and its near neighbours ranged between 25.7 ± 2.1 and 29.9 ± 2.4%. The DNA G+C content of the genomic DNA of the type strain is 71.7 mol%. Isolate AC230T presents a white to ivory hue on most ISP media and its micromorphology exhibits ovoid spores with smooth surfaces in flexuous chains. Based on our study of AC230T, the strain warrants the assignment to a novel species, for which the name Streptomyces corynorhini sp. nov. is proposed. The type strain is AC230T (= JCM 33171T, = ATCC TSD155T).


Assuntos
Quirópteros/microbiologia , Streptomyces/classificação , Streptomyces/isolamento & purificação , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , Redes e Vias Metabólicas/genética , Microscopia Eletrônica de Varredura , Tipagem de Sequências Multilocus , New Mexico , Hibridização de Ácido Nucleico , Filogenia , Esporos Bacterianos/ultraestrutura , Streptomyces/genética , Streptomyces/fisiologia , Sequenciamento Completo do Genoma
2.
Antonie Van Leeuwenhoek ; 111(6): 897-904, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29170835

RESUMO

We report the development of a publicly accessible, curated nucleotide sequence database of hypocrealean entomopathogenic fungi. The goal is to provide a platform for users to easily access sequence data from taxonomic reference strains. The database can be used to accurately identify unknown entomopathogenic fungi based on sequence data for a variety of phylogenetically informative loci. The database provides full multi-locus sequence alignment capabilities. The initial release contains data compiled for 525 strains covering the phylogenetic diversity of three important entomopathogenic families: Clavicipitaceae, Cordycipitaceae, and Ophiocordycipitaceae. Furthermore, Entomopathogen ID can be expanded to other fungal clades of insect pathogens, as sequence data becomes available. The database will allow isolate characterisation and evolutionary analyses. We contend that this freely available, web-accessible database will facilitate the broader community to accurately identify fungal entomopathogens, which will allow users to communicate research results more effectively.


Assuntos
Hypocreales/genética , Insetos/microbiologia , Animais , DNA Fúngico/genética , Hypocreales/classificação , Filogenia
3.
Proc Natl Acad Sci U S A ; 112(39): 12175-80, 2015 Sep 29.
Artigo em Inglês | MEDLINE | ID: mdl-26324907

RESUMO

Although natural products have been a particularly rich source of human medicines, activity-based screening results in a very high rate of rediscovery of known molecules. Based on the large number of natural product biosynthetic genes in microbial genomes, many have proposed "genome mining" as an alternative approach for discovery efforts; however, this idea has yet to be performed experimentally on a large scale. Here, we demonstrate the feasibility of large-scale, high-throughput genome mining by screening a collection of over 10,000 actinomycetes for the genetic potential to make phosphonic acids, a class of natural products with diverse and useful bioactivities. Genome sequencing identified a diverse collection of phosphonate biosynthetic gene clusters within 278 strains. These clusters were classified into 64 distinct groups, of which 55 are likely to direct the synthesis of unknown compounds. Characterization of strains within five of these groups resulted in the discovery of a new archetypical pathway for phosphonate biosynthesis, the first (to our knowledge) dedicated pathway for H-phosphinates, and 11 previously undescribed phosphonic acid natural products. Among these compounds are argolaphos, a broad-spectrum antibacterial phosphonopeptide composed of aminomethylphosphonate in peptide linkage to a rare amino acid N(5)-hydroxyarginine; valinophos, an N-acetyl l-Val ester of 2,3-dihydroxypropylphosphonate; and phosphonocystoximate, an unusual thiohydroximate-containing molecule representing a new chemotype of sulfur-containing phosphonate natural products. Analysis of the genome sequences from the remaining strains suggests that the majority of the phosphonate biosynthetic repertoire of Actinobacteria has been captured at the gene level. This dereplicated strain collection now provides a reservoir of numerous, as yet undiscovered, phosphonate natural products.


Assuntos
Actinobacteria/química , Actinobacteria/genética , Produtos Biológicos/química , Descoberta de Drogas/tendências , Genoma Bacteriano/genética , Genômica/métodos , Ácidos Fosforosos/análise , Sequência de Bases , Descoberta de Drogas/métodos , Biblioteca Gênica , Genômica/tendências , Dados de Sequência Molecular , Análise de Sequência de DNA
4.
Appl Environ Microbiol ; 83(5)2017 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-27986729

RESUMO

At least two-thirds of commercial antibiotics today are derived from Actinobacteria, more specifically from the genus Streptomyces Antibiotic resistance and new emerging diseases pose great challenges in the field of microbiology. Cave systems, in which actinobacteria are ubiquitous and abundant, represent new opportunities for the discovery of novel bacterial species and the study of their interactions with emergent pathogens. White-nose syndrome is an invasive bat disease caused by the fungus Pseudogymnoascus destructans, which has killed more than six million bats in the last 7 years. In this study, we isolated naturally occurring actinobacteria from white-nose syndrome (WNS)-free bats from five cave systems and surface locations in the vicinity in New Mexico and Arizona, USA. We sequenced the 16S rRNA region and tested 632 isolates from 12 different bat species using a bilayer plate method to evaluate antifungal activity. Thirty-six actinobacteria inhibited or stopped the growth of P. destructans, with 32 (88.9%) actinobacteria belonging to the genus Streptomyces Isolates in the genera Rhodococcus, Streptosporangium, Luteipulveratus, and Nocardiopsis also showed inhibition. Twenty-five of the isolates with antifungal activity against P. destructans represent 15 novel Streptomyces spp. based on multilocus sequence analysis. Our results suggest that bats in western North America caves possess novel bacterial microbiota with the potential to inhibit P. destructansIMPORTANCE This study reports the largest collection of actinobacteria from bats with activity against Pseudogymnoascus destructans, the fungal causative agent of white-nose syndrome. Using multigene analysis, we discovered 15 potential novel species. This research demonstrates that bats and caves may serve as a rich reservoir for novel Streptomyces species with antimicrobial bioactive compounds.


Assuntos
Antifúngicos/farmacologia , Ascomicetos/efeitos dos fármacos , Quirópteros/microbiologia , Streptomyces/metabolismo , Actinobacteria/classificação , Actinobacteria/genética , Actinobacteria/isolamento & purificação , Actinobacteria/metabolismo , Doenças dos Animais/microbiologia , Animais , Arizona , Ascomicetos/crescimento & desenvolvimento , Ascomicetos/patogenicidade , DNA Bacteriano , Genes Bacterianos , Testes de Sensibilidade Microbiana , Microbiota , Tipagem de Sequências Multilocus , Micoses/microbiologia , Micoses/prevenção & controle , Micoses/veterinária , América do Norte , Nariz/microbiologia , Filogenia , RNA Ribossômico 16S , Streptomyces/classificação , Streptomyces/genética , Streptomyces/isolamento & purificação
5.
PLoS Biol ; 12(8): e1001920, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25093819

RESUMO

Microbes hold the key to life. They hold the secrets to our past (as the descendants of the earliest forms of life) and the prospects for our future (as we mine their genes for solutions to some of the planet's most pressing problems, from global warming to antibiotic resistance). However, the piecemeal approach that has defined efforts to study microbial genetic diversity for over 20 years and in over 30,000 genome projects risks squandering that promise. These efforts have covered less than 20% of the diversity of the cultured archaeal and bacterial species, which represent just 15% of the overall known prokaryotic diversity. Here we call for the funding of a systematic effort to produce a comprehensive genomic catalog of all cultured Bacteria and Archaea by sequencing, where available, the type strain of each species with a validly published name (currently∼11,000). This effort will provide an unprecedented level of coverage of our planet's genetic diversity, allow for the large-scale discovery of novel genes and functions, and lead to an improved understanding of microbial evolution and function in the environment.


Assuntos
Genoma Arqueal/genética , Genoma Bacteriano/genética , Genômica , Análise de Sequência de DNA , Archaea/classificação , Archaea/genética , Bactérias/classificação , Bactérias/genética , Bases de Dados Genéticas , Filogenia
6.
Int J Syst Evol Microbiol ; 67(12): 5211-5215, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29087276

RESUMO

A novel streptomycete, strain 594T, isolated from Brazilian soil collected under cerrado (savanna) vegetation cover is described. Strain 594T produced thermophilic chitinolytic proteases in assays containing feather meal and corn steep liquor as sole sources of carbon and nitrogen. The strain produced white to grey aerial mycelium and spiral chains of spiny-surfaced spores on the aerial mycelium and did not produce diffusible pigments. The ll-isomer of diaminopimelic acid was present in the cell wall and menaquinones were predominantly MK-9(H6) (52 %) and MK-9(H8) (30 %) with 6 % MK-9(H4) and slightly less than 1 % MK-9(H2). Polar lipids present were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and an unknown phospholipid. The major fatty acids were anteiso-C15 : 0, anteiso-C16 : 0, anteiso-C14 : 0 and anteiso-C17 : 0. The G+C content of the genomic DNA was 70.4 mol%. Phylogenetic analysis of the nearly complete 16S rRNA gene sequence indicated that it differed from described Streptomyces species. Multilocus sequence analysis (MLSA) using five housekeeping genes (atpD, gyrB, rpoB, recA and trpB) comparing Streptomyces type strains showed that the MLSA distance of strain 594T to the most closely related species was greater than the 0.007 threshold. The in silico DNA-DNA relatedness between the genome sequence of strain 594T and that of the phylogenetically nearest species was well below the species level recommendation. There was thus multiple evidence justifying the description of this strain as representing a novel species, for which the name Streptomyces odonnellii sp. nov. is proposed. The type strain is 594T (=IMPPG 594T=DSM 41949T=NRRL B-24891T).


Assuntos
Pradaria , Filogenia , Microbiologia do Solo , Streptomyces/classificação , Técnicas de Tipagem Bacteriana , Composição de Bases , Brasil , DNA Bacteriano/genética , Ácido Diaminopimélico/química , Ácidos Graxos/química , Genes Bacterianos , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Streptomyces/genética , Vitamina K 2/análogos & derivados , Vitamina K 2/química
7.
Antonie Van Leeuwenhoek ; 110(4): 563-583, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28039547

RESUMO

The family Streptomycetaceae, notably species in the genus Streptomyces, have long been the subject of investigation due to their well-known ability to produce secondary metabolites. The emergence of drug resistant pathogens and the relative ease of producing genome sequences has renewed the importance of Streptomyces as producers of new natural products and resulted in revived efforts in isolating and describing strains from novel environments. A previous large study of the phylogeny in the Streptomycetaceae based on 16S rRNA gene sequences provided a useful framework for the relationships among species, but did not always have sufficient resolution to provide definitive identification. Multi-locus sequence analysis of 5 house-keeping genes has been shown to provide improved taxonomic resolution of Streptomyces species in a number of previous reports so a comprehensive study was undertaken to evaluate evolutionary relationships among species within the family Streptomycetaceae where type strains are available in the ARS Culture Collection or genome sequences are available in GenBank. The results of the analysis supported the distinctiveness of Kitasatospora and Streptacidiphilus as validly named genera since they cluster outside of the phylogenetic radiation of the genus Streptomyces. There is also support for the transfer of a number of Streptomyces species to the genus Kitasatospora as well for reducing at least 31 species clusters to a single taxon. The multi-locus sequence database resulting from the study is a useful tool for identification of new isolates and the phylogenetic analysis presented also provides a road map for planning future genome sequencing efforts in the Streptomycetaceae.


Assuntos
DNA Bacteriano/genética , Genes Essenciais/genética , Tipagem de Sequências Multilocus/métodos , Streptomyces/classificação , Streptomyces/genética , Sequência de Bases , DNA Ribossômico/genética , Filogenia , Análise de Sequência de DNA
8.
Antonie Van Leeuwenhoek ; 110(5): 705-717, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28185026

RESUMO

A polyphasic study was undertaken to determine the taxonomic status of a Streptomyces strain which had been isolated from a high altitude Atacama Desert soil and shown to have bioactive properties. The strain, isolate H9T, was found to have chemotaxonomic, cultural and morphological properties that place it in the genus Streptomyces. 16S rRNA gene sequence analyses showed that the isolate forms a distinct branch at the periphery of a well-delineated subclade in the Streptomyces 16S rRNA gene tree together with the type strains of Streptomyces crystallinus, Streptomyces melanogenes and Streptomyces noboritoensis. Multi-locus sequence analysis (MLSA) based on five house-keeping gene alleles showed that isolate H9T is closely related to the latter two type strains and to Streptomyces polyantibioticus NRRL B-24448T. The isolate was distinguished readily from the type strains of S. melanogenes, S. noboritoensis and S. polyantibioticus using a combination of phenotypic properties. Consequently, the isolate is considered to represent a new species of Streptomyces for which the name Streptomyces aridus sp. nov. is proposed; the type strain is H9T (=NCIMB 14965T=NRRL B65268T). In addition, the MLSA and phenotypic data show that the S. melanogenes and S. noboritoensis type strains belong to a single species, it is proposed that S. melanogenes be recognised as a heterotypic synonym of S. noboritoensis for which an emended description is given.


Assuntos
Microbiologia do Solo , Streptomyces/classificação , Streptomyces/isolamento & purificação , Altitude , Técnicas de Tipagem Bacteriana , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Clima Desértico , Ácidos Graxos/análise , Microscopia Eletrônica de Varredura , Tipagem de Sequências Multilocus , Filogenia , RNA Ribossômico 16S/genética , Streptomyces/genética , Streptomyces/fisiologia
9.
Antonie Van Leeuwenhoek ; 110(9): 1133-1148, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28589342

RESUMO

A polyphasic study was undertaken to establish the taxonomic status of Streptomyces strains isolated from hyper-arid Atacama Desert soils. Analysis of the 16S rRNA gene sequences of the isolates showed that they formed a well-defined lineage that was loosely associated with the type strains of several Streptomyces species. Multi-locus sequence analysis based on five housekeeping gene alleles showed that the strains form a homogeneous taxon that is closely related to the type strains of Streptomyces ghanaensis and Streptomyces viridosporus. Representative isolates were shown to have chemotaxonomic and morphological properties consistent with their classification in the genus Streptomyces. The isolates have many phenotypic features in common, some of which distinguish them from S. ghanaensis NRRL B-12104T, their near phylogenetic neighbour. On the basis of these genotypic and phenotypic data it is proposed that the isolates be recognised as a new species within the genus Streptomyces, named Streptomyces asenjonii sp. nov. The type strain of the species is KNN35.1bT (NCIMB 15082T = NRRL B-65050T). Some of the isolates, including the type strain, showed antibacterial activity in standard plug assays. In addition, MLSA, average nucleotide identity and phenotypic data show that the type strains of S. ghanaensis and S. viridosporus belong to the same species. Consequently, it is proposed that the former be recognised as a heterotypic synonym of the latter and an emended description is given for S. viridosporus.


Assuntos
Filogenia , Microbiologia do Solo , Streptomyces/classificação , Streptomyces/genética , Aminoácidos/metabolismo , Antibacterianos/farmacologia , Chile , Clima Desértico , Ácido Diaminopimélico/química , Ácidos Graxos/química , Genoma Bacteriano/genética , Tipagem de Sequências Multilocus , Fenótipo , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Especificidade da Espécie , Streptomyces/química , Streptomyces/efeitos dos fármacos , Açúcares/metabolismo
10.
Nat Chem Biol ; 10(11): 963-8, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25262415

RESUMO

Actinobacteria encode a wealth of natural product biosynthetic gene clusters, whose systematic study is complicated by numerous repetitive motifs. By combining several metrics, we developed a method for the global classification of these gene clusters into families (GCFs) and analyzed the biosynthetic capacity of Actinobacteria in 830 genome sequences, including 344 obtained for this project. The GCF network, comprising 11,422 gene clusters grouped into 4,122 GCFs, was validated in hundreds of strains by correlating confident mass spectrometric detection of known small molecules with the presence or absence of their established biosynthetic gene clusters. The method also linked previously unassigned GCFs to known natural products, an approach that will enable de novo, bioassay-free discovery of new natural products using large data sets. Extrapolation from the 830-genome data set reveals that Actinobacteria encode hundreds of thousands of future drug leads, and the strong correlation between phylogeny and GCFs frames a roadmap to efficiently access them.


Assuntos
Actinobacteria/genética , Actinobacteria/metabolismo , Produtos Biológicos/metabolismo , Descoberta de Drogas/métodos , Genômica , Metabolômica , Produtos Biológicos/química , Produtos Biológicos/isolamento & purificação , Estrutura Molecular , Família Multigênica/genética , Metabolismo Secundário
11.
Int J Syst Evol Microbiol ; 66(6): 2444-2450, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26971011

RESUMO

Previous phylogenetic analysis of species of the genus Streptomyces based on 16S rRNA gene sequences resulted in a statistically well-supported clade (100 % bootstrap value) containing eight species that exhibited very similar gross morphology in producing open looped (Retinaculum-Apertum) to spiral (Spira) chains of spiny- to hairysurfaced, dark green spores on their aerial mycelium. The type strains of the species in this clade, specifically Streptomyces bambergiensis, Streptomyces cyanoalbus, Streptomyces emeiensis, Streptomyces hirsutus, Streptomyces prasinopilosus and Streptomyces prasinus, were subjected to multi-locus sequence analysis (MLSA) utilizing partial sequences of the housekeeping genes atpD, gyrB, recA, rpoB and trpB to clarify their taxonomic status. The type strains of several recently described species with similar gross morphology, including Streptomyces chlorus, Streptomyces herbaceus, Streptomyces incanus, Streptomyces pratens and Streptomyces viridis, were also studied along with six unidentified green-spored Streptomyces strains from the ARS Culture Collection. The MLSAs suggest that three of the species under study (S. bambergiensis, S. cyanoalbus and S. emeiensis) represent synonyms of other previously described species (S. prasinus, S. hirsutus and S. prasinopilosus, respectively). These relationships were confirmed through determination of in silico DNA-DNA hybridization estimates based on draft genome sequences. The five recently described species appear to be phylogenetically distinct but the unidentified strains from the ARS Culture Collection could be identified as representatives of S. hirsutus, S. prasinopilosus or S. prasinus.


Assuntos
Filogenia , Streptomyces/classificação , Técnicas de Tipagem Bacteriana , DNA Bacteriano/genética , Genes Bacterianos , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
12.
Antonie Van Leeuwenhoek ; 109(3): 349-56, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26739474

RESUMO

Multi-locus sequence analysis has been demonstrated to be a useful tool for identification of Streptomyces species and was previously applied to phylogenetically differentiate the type strains of species pathogenic on potatoes (Solanum tuberosum L.). The ARS Culture Collection (NRRL) contains 43 strains identified as Streptomyces scabiei deposited at various times since the 1950s and these were subjected to multi-locus sequence analysis utilising partial sequences of the house-keeping genes atpD, gyrB, recA, rpoB and trpB. Phylogenetic analyses confirmed the identity of 17 of these strains as Streptomyces scabiei, 9 of the strains as the potato-pathogenic species Streptomyces europaeiscabiei and 6 strains as potentially new phytopathogenic species. Of the 16 other strains, 12 were identified as members of previously described non-pathogenic Streptomyces species while the remaining 4 strains may represent heretofore unrecognised non-pathogenic species. This study demonstrated the value of this technique for the relatively rapid, simple and sensitive molecular identification of Streptomyces strains held in culture collections.


Assuntos
Código de Barras de DNA Taxonômico , Tipagem de Sequências Multilocus , Streptomyces/classificação , Streptomyces/genética , Técnicas Bacteriológicas , Filogenia
13.
Proc Natl Acad Sci U S A ; 110(51): 20759-64, 2013 Dec 17.
Artigo em Inglês | MEDLINE | ID: mdl-24297932

RESUMO

Phosphonates, molecules containing direct carbon-phosphorus bonds, compose a structurally diverse class of natural products with interesting and useful biological properties. Although their synthesis in protozoa was discovered more than 50 y ago, the extent and diversity of phosphonate production in nature remains poorly characterized. The rearrangement of phosphoenolpyruvate (PEP) to phosphonopyruvate, catalyzed by the enzyme PEP mutase (PepM), is shared by the vast majority of known phosphonate biosynthetic pathways. Thus, the pepM gene can be used as a molecular marker to examine the occurrence and abundance of phosphonate-producing organisms. Based on the presence of this gene, phosphonate biosynthesis is common in microbes, with ~5% of sequenced bacterial genomes and 7% of genome equivalents in metagenomic datasets carrying pepM homologs. Similarly, we detected the pepM gene in ~5% of random actinomycete isolates. The pepM-containing gene neighborhoods from 25 of these isolates were cloned, sequenced, and compared with those found in sequenced genomes. PEP mutase sequence conservation is strongly correlated with conservation of other nearby genes, suggesting that the diversity of phosphonate biosynthetic pathways can be predicted by examining PEP mutase diversity. We used this approach to estimate the range of phosphonate biosynthetic pathways in nature, revealing dozens of discrete groups in pepM amplicons from local soils, whereas hundreds were observed in metagenomic datasets. Collectively, our analyses show that phosphonate biosynthesis is both diverse and relatively common in nature, suggesting that the role of phosphonate molecules in the biosphere may be more important than is often recognized.


Assuntos
Actinobacteria , Proteínas de Bactérias , Metagenoma , Organofosfonatos/metabolismo , Fosfotransferases (Fosfomutases) , Microbiologia do Solo , Actinobacteria/enzimologia , Actinobacteria/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Sequência de Bases , Dados de Sequência Molecular , Fosfotransferases (Fosfomutases)/genética , Fosfotransferases (Fosfomutases)/metabolismo
14.
Antonie Van Leeuwenhoek ; 105(5): 849-61, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24604690

RESUMO

A polyphasic study was carried out to establish the taxonomic status of an Atacama Desert isolate, Streptomyces strain C34(T), which synthesises novel antibiotics, the chaxalactins and chaxamycins. The organism was shown to have chemotaxonomic, cultural and morphological properties consistent with its classification in the genus Streptomyces. Analysis of 16S rRNA gene sequences showed that strain C34(T) formed a distinct phyletic line in the Streptomyces gene tree that was very loosely associated with the type strains of several Streptomyces species. Multilocus sequence analysis based on five house-keeping gene alleles underpinned the separation of strain C34(T) from all of its nearest phylogenetic neighbours, apart from Streptomyces chiangmaiensis TA-1(T) and Streptomyces hyderabadensis OU-40(T) which are not currently in the MLSA database. Strain C34(T) was distinguished readily from the S. chiangmaiensis and S. hyderabadensis strains by using a combination of cultural and phenotypic data. Consequently, strain C34(T) is considered to represent a new species of the genus Streptomyces for which the name Streptomyces leeuwenhoekii sp. nov. is proposed. The type strain is C34(T) (= DSM 42122(T) = NRRL B-24963(T)). Analysis of the whole-genome sequence of S. leeuwenhoekii, with 6,780 predicted open reading frames and a total genome size of around 7.86 Mb, revealed a high potential for natural product biosynthesis.


Assuntos
Antibacterianos/metabolismo , Macrolídeos/metabolismo , Rifamicinas/metabolismo , Streptomyces/classificação , Streptomyces/metabolismo , Técnicas de Tipagem Bacteriana , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Clima Desértico , Microscopia Eletrônica de Varredura , Dados de Sequência Molecular , Tipagem de Sequências Multilocus , Filogenia , RNA Ribossômico 16S/genética , Microbiologia do Solo , Streptomyces/genética , Streptomyces/isolamento & purificação
15.
J Proteome Res ; 11(1): 85-94, 2012 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-21978092

RESUMO

Actinobacteria such as streptomycetes are renowned for their ability to produce bioactive natural products including nonribosomal peptides (NRPs) and polyketides (PKs). The advent of genome sequencing has revealed an even larger genetic repertoire for secondary metabolism with most of the small molecule products of these gene clusters still unknown. Here, we employed a "protein-first" method called PrISM (Proteomic Investigation of Secondary Metabolism) to screen 26 unsequenced actinomycetes using mass spectrometry-based proteomics for the targeted detection of expressed nonribosomal peptide synthetases or polyketide synthases. Improvements to the original PrISM screening approach (Nat. Biotechnol. 2009, 27, 951-956), for example, improved de novo peptide sequencing, have enabled the discovery of 10 NRPS/PKS gene clusters from 6 strains. Taking advantage of the concurrence of biosynthetic enzymes and the secondary metabolites they generate, two natural products were associated with their previously "orphan" gene clusters. This work has demonstrated the feasibility of a proteomics-based strategy for use in screening for NRP/PK production in actinomycetes (often >8 Mbp, high GC genomes) versus the bacilli (2-4 Mbp genomes) used previously.


Assuntos
Actinobacteria/enzimologia , Proteínas de Bactérias/biossíntese , Biossíntese de Peptídeos Independentes de Ácido Nucleico , Peptídeo Sintases/metabolismo , Policetídeo Sintases/metabolismo , Actinobacteria/genética , Actinobacteria/metabolismo , Sequência de Aminoácidos , Família Multigênica , Peptídeo Sintases/genética , Policetídeo Sintases/genética , Policetídeos/metabolismo , Proteômica , Análise de Sequência de Proteína , Espectrometria de Massas em Tandem
16.
Int J Syst Evol Microbiol ; 62(Pt 12): 2978-2985, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22286909

RESUMO

The 16S rRNA and gyrB genes of 22 Streptomyces strains belonging to the Streptomyces griseus cluster were sequenced, and their taxonomic positions were re-evaluated. For correct analysis, all of the publicly available sequences of the species were collected and compared with those obtained in this study. Species for which no consensus sequence could be identified were excluded from the phylogenetic analysis. The levels of 16S rRNA gene sequence similarity within the cluster ranged from 98.6 to 100% with a mean value of 99.6 ± 0.3%, and those of the gyrB gene ranged from 93.6 to 99.9% with a mean value of 96.3 ± 1.5%. The observed average nucleotide substitution rate of the gyrB gene was ten times higher than that of the 16S rRNA gene, showing a far higher degree of variation. Strains sharing 99.3% or more gyrB sequence similarity (corresponding to an evolutionary distance of 0.0073) always formed monophyletic groups in both trees. Through the combined analysis of the two genes, clear cases of synonymy could be identified and, according to the priority rule, the assertion of the status of Streptomyces setonii as a distinct species and the reclassification of Streptomyces fimicarius as a later synonym of S. setonii and Streptomyces albovinaceus as a later synonym of Streptomyces globisporus are proposed. Emended descriptions of S. setonii and S. globisporus are provided.


Assuntos
Filogenia , Streptomyces/classificação , DNA Girase/genética , DNA Bacteriano/genética , Genes Bacterianos , Dados de Sequência Molecular , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Streptomyces/genética
17.
J Bacteriol ; 193(24): 7021-2, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22123769

RESUMO

We announce the sequencing of Streptomyces chartreusis NRRL 12338 and NRRL 3882 and Streptomyces lysosuperificus ATCC 31396. These are producers of tunicamycins, chartreusins, cephalosporins, holomycins, and calcimycin. The announced genomes, together with the published Streptomyces clavuligerus genome, will facilitate data mining of these secondary metabolites.


Assuntos
Antibacterianos/metabolismo , Genoma Bacteriano , Streptomyces/genética , Tunicamicina/biossíntese , Sequência de Bases , Cefalosporinas/metabolismo , Dados de Sequência Molecular , Streptomyces/metabolismo
18.
Int J Syst Evol Microbiol ; 61(Pt 10): 2525-2531, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21112986

RESUMO

The identification and classification of species within the genus Streptomyces is difficult because there are presently 576 species with validly published names and this number increases every year. The value of multilocus sequence analysis applied to the systematics of Streptomyces species has been well demonstrated in several recently published papers. In this study the sequence fragments of four housekeeping genes, atpD, recA, rpoB and trpB, were determined for the type strains of 10 known phytopathogenic species of the genus Streptomyces, including Streptomyces scabiei, Streptomyces acidiscabies, Streptomyces europaeiscabiei, Streptomyces luridiscabiei, Streptomyces niveiscabiei, Streptomyces puniciscabiei, Streptomyces reticuliscabiei, Streptomyces stelliscabiei, Streptomyces turgidiscabies and Streptomyces ipomoeae, as well as six uncharacterized phytopathogenic Streptomyces isolates. The type strains of 52 other species, including 19 species observed to be phylogenetically closely related to these, based on 16S rRNA gene sequence analysis, were also included in the study. Phylogenetic analysis of single gene alignments and a concatenated four-gene alignment demonstrated that the phytopathogenic species are taxonomically distinct from each other in spite of high 16S rRNA gene sequence similarities and provided a tool for the identification of unknown putative phytopathogenic Streptomyces strains at the species level.


Assuntos
Técnicas de Tipagem Bacteriana/métodos , Tipagem de Sequências Multilocus/métodos , Doenças das Plantas/microbiologia , Streptomyces/classificação , Streptomyces/genética , Proteínas de Bactérias/genética , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Dados de Sequência Molecular , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Streptomyces/isolamento & purificação
19.
Int J Syst Evol Microbiol ; 61(Pt 2): 310-314, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-20305067

RESUMO

In the course of a polyphasic study it was observed that 'Dactylosporangium variesporum' NRRL B-16296 is misclassified in the genus Dactylosporangium as it exhibits properties consistent with its assignment to the genus Saccharothrix. Phylogenetic analyses based on 16S rRNA gene sequences show that the strain falls within the evolutionary radiation of the genus Saccharothrix, a result which is supported by corresponding chemotaxonomic and morphological markers. The strain is phylogenetically most closely, albeit loosely, related to Saccharothrix espanaensis, but can be readily distinguished from this and other species of the genus Saccharothrix with validly described names by using a range of phenotypic properties. The combined genotypic and phenotypic data demonstrate conclusively that this strain should be classified as a new species in the genus Saccharothrix for which the name Saccharothrix variisporea sp. nov. is proposed. The type strain is NRRL B-16296(T) (=ATCC 31203(T) =DSM 43911(T) =JCM 3273(T) =NBRC 14104(T)).


Assuntos
Actinomycetales/classificação , Filogenia , Actinomycetales/genética , Técnicas de Tipagem Bacteriana , DNA Bacteriano/genética , Genótipo , Micromonosporaceae/classificação , Micromonosporaceae/genética , Dados de Sequência Molecular , Fenótipo , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
20.
Int J Syst Evol Microbiol ; 61(Pt 9): 2304-2310, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-20971832

RESUMO

A Gram-positive, spore-forming, aerobic, filamentous bacterium, strain JFMB-ATE(T), was isolated in 2008 during environmental screening of a plastic surface in grade C in a contract manufacturing organization in southern Germany. The isolate grew at temperatures of 25-50 °C and at pH 5.0-8.5, forming ivory-coloured colonies with sparse white aerial mycelia. Chemotaxonomic and molecular characteristics of the isolate matched those described for members of the family Thermoactinomycetaceae, except that the cell-wall peptidoglycan contained LL-diaminopimelic acid, while all previously described members of this family display this diagnostic diamino acid in meso-conformation. The DNA G+C content of the novel strain was 54.6 mol%, the main polar lipids were diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol, and the major menaquinone was MK-7. The major fatty acids had saturated C14-C16 branched chains. No diagnostic sugars were detected. Based on the chemotaxonomic results and 16S rRNA gene sequence analysis, the isolate is proposed to represent a novel genus and species, Kroppenstedtia eburnea gen. nov. sp. nov. The type strain is JFMB-ATE(T) ( = DSM 45196(T)  = NRRL B-24804(T)  = CCUG 59226(T)).


Assuntos
Bacillales/classificação , Bacillales/isolamento & purificação , Microbiologia Ambiental , Bacillales/genética , Técnicas de Tipagem Bacteriana , Composição de Bases , Parede Celular/química , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Ácido Diaminopimélico/análise , Alemanha , Concentração de Íons de Hidrogênio , Dados de Sequência Molecular , Peptidoglicano/química , Fosfolipídeos/análise , Filogenia , Pigmentos Biológicos/metabolismo , Quinonas/análise , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Esporos Bacterianos/citologia , Temperatura
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