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1.
Antonie Van Leeuwenhoek ; 112(9): 1297-1305, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-30993571

RESUMEN

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).


Asunto(s)
Quirópteros/microbiología , Streptomyces/clasificación , Streptomyces/aislamiento & purificación , Animales , Técnicas de Tipificación Bacteriana , Composición de Base , Redes y Vías Metabólicas/genética , Microscopía Electrónica de Rastreo , Tipificación de Secuencias Multilocus , New Mexico , Hibridación de Ácido Nucleico , Filogenia , Esporas Bacterianas/ultraestructura , Streptomyces/genética , Streptomyces/fisiología , Secuenciación Completa del Genoma
2.
Antonie Van Leeuwenhoek ; 111(6): 897-904, 2018 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-29170835

RESUMEN

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.


Asunto(s)
Hypocreales/genética , Insectos/microbiología , Animales , ADN de Hongos/genética , Hypocreales/clasificación , Filogenia
3.
Proc Natl Acad Sci U S A ; 112(39): 12175-80, 2015 Sep 29.
Artículo en Inglés | MEDLINE | ID: mdl-26324907

RESUMEN

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.


Asunto(s)
Actinobacteria/química , Actinobacteria/genética , Productos Biológicos/química , Descubrimiento de Drogas/tendencias , Genoma Bacteriano/genética , Genómica/métodos , Ácidos Fosforosos/análisis , Secuencia de Bases , Descubrimiento de Drogas/métodos , Biblioteca de Genes , Genómica/tendencias , Datos de Secuencia Molecular , Análisis de Secuencia de ADN
4.
Appl Environ Microbiol ; 83(5)2017 03 01.
Artículo en Inglés | MEDLINE | ID: mdl-27986729

RESUMEN

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.


Asunto(s)
Antifúngicos/farmacología , Ascomicetos/efectos de los fármacos , Quirópteros/microbiología , Streptomyces/metabolismo , Actinobacteria/clasificación , Actinobacteria/genética , Actinobacteria/aislamiento & purificación , Actinobacteria/metabolismo , Enfermedades de los Animales/microbiología , Animales , Arizona , Ascomicetos/crecimiento & desarrollo , Ascomicetos/patogenicidad , ADN Bacteriano , Genes Bacterianos , Pruebas de Sensibilidad Microbiana , Microbiota , Tipificación de Secuencias Multilocus , Micosis/microbiología , Micosis/prevención & control , Micosis/veterinaria , América del Norte , Nariz/microbiología , Filogenia , ARN Ribosómico 16S , Streptomyces/clasificación , Streptomyces/genética , Streptomyces/aislamiento & purificación
5.
Int J Syst Evol Microbiol ; 67(12): 5211-5215, 2017 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-29087276

RESUMEN

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).


Asunto(s)
Pradera , Filogenia , Microbiología del Suelo , Streptomyces/clasificación , Técnicas de Tipificación Bacteriana , Composición de Base , Brasil , ADN Bacteriano/genética , Ácido Diaminopimélico/química , Ácidos Grasos/química , Genes Bacterianos , Fosfolípidos/química , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Streptomyces/genética , Vitamina K 2/análogos & derivados , Vitamina K 2/química
6.
Antonie Van Leeuwenhoek ; 110(4): 563-583, 2017 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-28039547

RESUMEN

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.


Asunto(s)
ADN Bacteriano/genética , Genes Esenciales/genética , Tipificación de Secuencias Multilocus/métodos , Streptomyces/clasificación , Streptomyces/genética , Secuencia de Bases , ADN Ribosómico/genética , Filogenia , Análisis de Secuencia de ADN
7.
Antonie Van Leeuwenhoek ; 110(5): 705-717, 2017 May.
Artículo en Inglés | MEDLINE | ID: mdl-28185026

RESUMEN

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.


Asunto(s)
Microbiología del Suelo , Streptomyces/clasificación , Streptomyces/aislamiento & purificación , Altitud , Técnicas de Tipificación Bacteriana , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Clima Desértico , Ácidos Grasos/análisis , Microscopía Electrónica de Rastreo , Tipificación de Secuencias Multilocus , Filogenia , ARN Ribosómico 16S/genética , Streptomyces/genética , Streptomyces/fisiología
8.
Antonie Van Leeuwenhoek ; 110(9): 1133-1148, 2017 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-28589342

RESUMEN

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.


Asunto(s)
Filogenia , Microbiología del Suelo , Streptomyces/clasificación , Streptomyces/genética , Aminoácidos/metabolismo , Antibacterianos/farmacología , Chile , Clima Desértico , Ácido Diaminopimélico/química , Ácidos Grasos/química , Genoma Bacteriano/genética , Tipificación de Secuencias Multilocus , Fenotipo , Fosfolípidos/química , ARN Ribosómico 16S/genética , Especificidad de la Especie , Streptomyces/química , Streptomyces/efectos de los fármacos , Azúcares/metabolismo
9.
Nat Chem Biol ; 10(11): 963-8, 2014 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-25262415

RESUMEN

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.


Asunto(s)
Actinobacteria/genética , Actinobacteria/metabolismo , Productos Biológicos/metabolismo , Descubrimiento de Drogas/métodos , Genómica , Metabolómica , Productos Biológicos/química , Productos Biológicos/aislamiento & purificación , Estructura Molecular , Familia de Multigenes/genética , Metabolismo Secundario
10.
Int J Syst Evol Microbiol ; 66(6): 2444-2450, 2016 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-26971011

RESUMEN

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.


Asunto(s)
Filogenia , Streptomyces/clasificación , Técnicas de Tipificación Bacteriana , ADN Bacteriano/genética , Genes Bacterianos , Hibridación de Ácido Nucleico , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN
11.
Antonie Van Leeuwenhoek ; 109(3): 349-56, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-26739474

RESUMEN

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.


Asunto(s)
Código de Barras del ADN Taxonómico , Tipificación de Secuencias Multilocus , Streptomyces/clasificación , Streptomyces/genética , Técnicas Bacteriológicas , Filogenia
12.
Proc Natl Acad Sci U S A ; 110(51): 20759-64, 2013 Dec 17.
Artículo en Inglés | MEDLINE | ID: mdl-24297932

RESUMEN

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.


Asunto(s)
Actinobacteria , Proteínas Bacterianas , Metagenoma , Organofosfonatos/metabolismo , Fosfotransferasas (Fosfomutasas) , Microbiología del Suelo , Actinobacteria/enzimología , Actinobacteria/genética , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Secuencia de Bases , Datos de Secuencia Molecular , Fosfotransferasas (Fosfomutasas)/genética , Fosfotransferasas (Fosfomutasas)/metabolismo
13.
Antonie Van Leeuwenhoek ; 105(5): 849-61, 2014 May.
Artículo en Inglés | MEDLINE | ID: mdl-24604690

RESUMEN

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.


Asunto(s)
Antibacterianos/metabolismo , Macrólidos/metabolismo , Rifamicinas/metabolismo , Streptomyces/clasificación , Streptomyces/metabolismo , Técnicas de Tipificación Bacteriana , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Clima Desértico , Microscopía Electrónica de Rastreo , Datos de Secuencia Molecular , Tipificación de Secuencias Multilocus , Filogenia , ARN Ribosómico 16S/genética , Microbiología del Suelo , Streptomyces/genética , Streptomyces/aislamiento & purificación
14.
J Proteome Res ; 11(1): 85-94, 2012 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-21978092

RESUMEN

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.


Asunto(s)
Actinobacteria/enzimología , Proteínas Bacterianas/biosíntesis , Biosíntesis de Péptidos Independientes de Ácidos Nucleicos , Péptido Sintasas/metabolismo , Sintasas Poliquetidas/metabolismo , Actinobacteria/genética , Actinobacteria/metabolismo , Secuencia de Aminoácidos , Familia de Multigenes , Péptido Sintasas/genética , Sintasas Poliquetidas/genética , Policétidos/metabolismo , Proteómica , Análisis de Secuencia de Proteína , Espectrometría de Masas en Tándem
15.
Int J Syst Evol Microbiol ; 62(Pt 12): 2978-2985, 2012 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-22286909

RESUMEN

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.


Asunto(s)
Filogenia , Streptomyces/clasificación , Girasa de ADN/genética , ADN Bacteriano/genética , Genes Bacterianos , Datos de Secuencia Molecular , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Streptomyces/genética
16.
J Bacteriol ; 193(24): 7021-2, 2011 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-22123769

RESUMEN

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.


Asunto(s)
Antibacterianos/metabolismo , Genoma Bacteriano , Streptomyces/genética , Tunicamicina/biosíntesis , Secuencia de Bases , Cefalosporinas/metabolismo , Datos de Secuencia Molecular , Streptomyces/metabolismo
17.
Int J Syst Evol Microbiol ; 61(Pt 10): 2525-2531, 2011 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-21112986

RESUMEN

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.


Asunto(s)
Técnicas de Tipificación Bacteriana/métodos , Tipificación de Secuencias Multilocus/métodos , Enfermedades de las Plantas/microbiología , Streptomyces/clasificación , Streptomyces/genética , Proteínas Bacterianas/genética , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Datos de Secuencia Molecular , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Streptomyces/aislamiento & purificación
18.
Int J Syst Evol Microbiol ; 61(Pt 2): 310-314, 2011 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-20305067

RESUMEN

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)).


Asunto(s)
Actinomycetales/clasificación , Filogenia , Actinomycetales/genética , Técnicas de Tipificación Bacteriana , ADN Bacteriano/genética , Genotipo , Micromonosporaceae/clasificación , Micromonosporaceae/genética , Datos de Secuencia Molecular , Fenotipo , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN
19.
Int J Syst Evol Microbiol ; 61(Pt 9): 2304-2310, 2011 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-20971832

RESUMEN

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)).


Asunto(s)
Bacillales/clasificación , Bacillales/aislamiento & purificación , Microbiología Ambiental , Bacillales/genética , Técnicas de Tipificación Bacteriana , Composición de Base , Pared Celular/química , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Ácido Diaminopimélico/análisis , Alemania , Concentración de Iones de Hidrógeno , Datos de Secuencia Molecular , Peptidoglicano/química , Fosfolípidos/análisis , Filogenia , Pigmentos Biológicos/metabolismo , Quinonas/análisis , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Esporas Bacterianas/citología , Temperatura
20.
ACS Chem Biol ; 13(4): 1029-1037, 2018 04 20.
Artículo en Inglés | MEDLINE | ID: mdl-29510029

RESUMEN

Natural products (NPs) are a rich source of medicines, but traditional discovery methods are often unsuccessful due to high rates of rediscovery. Genetic approaches for NP discovery are promising, but progress has been slow due to the difficulty of identifying unique biosynthetic gene clusters (BGCs) and poor gene expression. We previously developed the metabologenomics method, which combines genomic and metabolomic data to discover new NPs and their BGCs. Here, we utilize metabologenomics in combination with molecular networking to discover a novel class of NPs, the tyrobetaines: nonribosomal peptides with an unusual trimethylammonium tyrosine residue. The BGC for this unusual class of compounds was identified using metabologenomics and computational structure prediction data. Heterologous expression confirmed the BGC and suggests an unusual mechanism for trimethylammonium formation. Overall, the discovery of the tyrobetaines shows the great potential of metabologenomics combined with molecular networking and computational structure prediction for identifying interesting biosynthetic reactions and novel NPs.


Asunto(s)
Productos Biológicos/metabolismo , Descubrimiento de Drogas , Genómica , Metabolómica , Familia de Multigenes , Betaína/análogos & derivados , Vías Biosintéticas
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