RESUMEN
An actinobacteria strain was isolated from an olive waste mill and tested for protease production on skimmed milk media. The strain identification was achieved through both 16 S rDNA sequencing and phenotypic characterization. The enzyme was purified using the ammonium sulfate/t-butanol three-phase partitioning (TPP) method, followed by characterization to investigate the effect of pH, temperature, and various chemical agents. Subsequently, the enzyme was assessed for its milk coagulation activity. The strain belonging to the Streptomyces genera, exhibits significant phylogenetic and phenotypic differences from the aligned species, suggesting its novelty as a new strain. The enzyme was best separated in the TPP aqueous phase with a 5.35 fold and 56.25% yield. Optimal activity was observed at pH 9.0 and 60 °C, with more than half of the activity retained within the pH range of 7-10 over one hour. The protease demonstrated complete stability between 30 and 60 °C. While metallic ions enhanced enzyme activity, EDTA acted as an inhibitor. The enzyme displayed resistance to H2O2, SDS, Tween 80, and Triton X-100. Notably, it was activated in organic solvents (ethyl acetate, petroleum ether, and xylene), maintaining > 75% of its original activity in butanol, ethanol, and methanol. Additionally, the enzyme yielded high milk coagulant activity of 11,478 SU/mL. The new Streptomyces sp. protease revealed high activity and stability under a wide range of biochemical conditions. Its use in the dairy industry appears particularly promising. Further industrial process investigations will be valuable in determining potential uses for this enzyme.
Asunto(s)
Estabilidad de Enzimas , Leche , Péptido Hidrolasas , Filogenia , Streptomyces , Temperatura , Streptomyces/aislamiento & purificación , Streptomyces/enzimología , Streptomyces/genética , Streptomyces/clasificación , Leche/microbiología , Animales , Concentración de Iones de Hidrógeno , Péptido Hidrolasas/metabolismo , Péptido Hidrolasas/aislamiento & purificación , Péptido Hidrolasas/química , Péptido Hidrolasas/genética , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/química , Proteínas Bacterianas/aislamiento & purificación , ARN Ribosómico 16S/genéticaRESUMEN
BACKGROUND: The native potatoes (Solanum tuberosum subsp. tuberosum L.) grown in Chile (Chiloé) represent a new, unexplored source of endophytes to find potential biological control agents for the prevention of bacterial diseases, like blackleg and soft rot, in potato crops. RESULT: The objective of this study was the selection of endophytic actinobacteria from native potatoes for antagonistic activity against Pectobacterium carotovorum subsp. carotovorum and Pectobacterium atrosepticum, and their potential to suppress tissue maceration symptoms in potato tubers. This potential was determined through the quorum quenching activity using a Chromobacterium violaceaum ATCC 12472 Wild type (WT) bioassay and its colonization behavior of the potato plant root system (S. tuberosum) by means of the Double labeling of oligonucleotide probes for fluorescence in situ hybridization (DOPE-FISH) targeting technique. The results showed that although Streptomyces sp. TP199 and Streptomyces sp. A2R31 were able to inhibit the growth of the pathogens, only the Streptomyces sp. TP199 isolate inhibited Pectobacterium sp. growth and diminished tissue maceration in tubers (p ≤ 0.05). Streptomyces sp. TP199 had metal-dependent acyl homoserine lactones (AHL) quorum quenching activity in vitro and was able to colonize the root endosphere 10 days after inoculation. CONCLUSIONS: We concluded that native potatoes from southern Chile possess endophyte actinobacteria that are potential agents for the disease management of soft rot and blackleg.
Asunto(s)
Actinobacteria/fisiología , Antibiosis/fisiología , Endófitos/fisiología , Solanum tuberosum/microbiología , Actinobacteria/clasificación , Actinobacteria/genética , Actinobacteria/aislamiento & purificación , Agentes de Control Biológico/aislamiento & purificación , Chile , Endófitos/clasificación , Endófitos/genética , Endófitos/aislamiento & purificación , Pectobacterium/fisiología , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/prevención & control , Tubérculos de la Planta/microbiología , Percepción de Quorum , Streptomyces/clasificación , Streptomyces/genética , Streptomyces/aislamiento & purificación , Streptomyces/fisiologíaRESUMEN
Streptomyces thermoautotrophicus UBT1T has been suggested to merit generic status due to its phylogenetic placement and distinctive phenotypes among Actinomycetia. To evaluate whether 'S. thermoautotrophicus' represents a higher taxonomic rank, 'S. thermoautotrophicus' strains UBT1T and H1 were compared to Actinomycetia using 16S rRNA gene sequences and comparative genome analyses. The UBT1T and H1 genomes each contain at least two different 16S rRNA sequences, which are closely related to those of Acidothermus cellulolyticus (order Acidothermales). In multigene-based phylogenomic trees, UBT1T and H1 typically formed a sister group to the Streptosporangiales-Acidothermales clade. The Average Amino Acid Identity, Percentage of Conserved Proteins, and whole-genome Average Nucleotide Identity (Alignment Fraction) values were ≤58.5%, ≤48%, ≤75.5% (0.3) between 'S. thermoautotrophicus' and Streptosporangiales members, all below the respective thresholds for delineating genera. The values for genomics comparisons between strains UBT1T and H1 with Acidothermales, as well as members of the genus Streptomyces, were even lower. A review of the 'S. thermoautotrophicus' proteomic profiles and KEGG orthology demonstrated that UBT1T and H1 present pronounced differences, both tested and predicted, in phenotypic and chemotaxonomic characteristics compared to its sister clades and Streptomyces. The distinct phylogenetic position and the combination of genotypic and phenotypic characteristics justify the proposal of Carbonactinospora gen. nov., with the type species Carbonactinospora thermoautotrophica comb. nov. (type strain UBT1T, = DSM 100163T = KCTC 49540T) belonging to Carbonactinosporaceae fam. nov. within Actinomycetia.
Asunto(s)
Filogenia , Streptomyces , Actinobacteria , Técnicas de Tipificación Bacteriana , ADN Bacteriano/genética , Proteómica , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Streptomyces/clasificaciónRESUMEN
Resorting to a One Strain Many Compounds (OSMAC) approach, the marine Streptomyces sp. BRB081 strain was grown in six different media settings over 1, 2, 3 or 7 days. Extractions of mycelium and broth were conducted separately for each media and cultivation period by sonication using methanol/acetone 1:1 and agitation with ethyl acetate, respectively. All methanol/acetone and ethyl acetate crude extracts were analysed by HPLC-MS/MS and data treatment was performed through GNPS platform using MZmine 2 software. In parallel, the genome was sequenced, assembled and mined to search for biosynthetic gene clusters (BGC) of secondary metabolites using the AntiSMASH 5.0 software. Spectral library search tool allowed the annotation of desferrioxamines, fatty acid amides, diketopiperazines, xanthurenic acid and, remarkably, the cyclic octapeptides surugamides. Molecular network analysis allowed the observation of the surugamides cluster, where surugamide A and the protonated molecule corresponding to the B-E isomers, as well as two potentially new analogues, were detected. Data treatment through MZmine 2 software allowed to distinguish that the largest amount of surugamides was obtained by cultivating BRB081 in SCB medium during 7 days and extraction of culture broth. Using the same data treatment, a chemical barcode was created for easy visualization and comparison of the metabolites produced overtime in all media. By genome mining of BRB081 four regions of biosynthetic gene clusters of secondary metabolites were detected supporting the metabolic data. Cytotoxic evaluation of all crude extracts using MTT assay revealed the highest bioactivity was also observed for extracts obtained in the optimal conditions as those for surugamides production, suggesting these to be the main active compounds herein. This method allowed the identification of compounds in the crude extracts and guided the selection of best conditions for production of bioactive compounds.
Asunto(s)
Antineoplásicos/aislamiento & purificación , Metabolómica/métodos , Metabolismo Secundario , Streptomyces/crecimiento & desarrollo , Proteínas Bacterianas/genética , Técnicas Bacteriológicas , Vías Biosintéticas , Biología Marina , Familia de Multigenes , Filogenia , Streptomyces/química , Streptomyces/clasificación , Secuenciación Completa del GenomaRESUMEN
Streptomyces venezuelae is well known to produce various secondary metabolites, including chloramphenicol, jadomycin, and pikromycin. Although many strains have been classified as S. venezuelae species, only a limited number of strains have been explored extensively for their genomic contents. Moreover, genomic differences and diversity in secondary metabolite production between the strains have never been compared. Here, we report complete genome sequences of three S. venezuelae strains (ATCC 10712, ATCC 10595, and ATCC 21113) harboring chloramphenicol and jadomycin biosynthetic gene clusters (BGC). With these high-quality genome sequences, we revealed that the three strains share more than 85% of total genes and most of the secondary metabolite biosynthetic gene clusters (smBGC). Despite such conservation, the strains produced different amounts of chloramphenicol and jadomycin, indicating differential regulation of secondary metabolite production at the strain level. Interestingly, antagonistic production of chloramphenicol and jadomycin was observed in these strains. Through comparison of the chloramphenicol and jadomycin BGCs among the three strains, we found sequence variations in many genes, the non-coding RNA coding regions, and binding sites of regulators, which affect the production of the secondary metabolites. We anticipate that these genome sequences of closely related strains would serve as useful resources for understanding the complex secondary metabolism and for designing an optimal production process using Streptomyces strains.
Asunto(s)
Cloranfenicol/biosíntesis , Genómica , Isoquinolinas/metabolismo , Streptomyces/clasificación , Streptomyces/metabolismo , Cloranfenicol/química , Cloranfenicol/metabolismo , Isoquinolinas/química , Estructura Molecular , Streptomyces/química , Streptomyces/genéticaRESUMEN
Streptomyces spp. are Gram-positive bacteria well-known for their ability to produce antibiotics and other metabolites, but few studies on bacteriocins produced by these bacteria have been reported. We tested eight Streptomyces strains against different pathogenic bacteria, and selected S. griseus, S. nigrescens, S. bottroprensis, and S. violaceoruber for further study based on their inhibitory effects against bacteria, including human pathogens. S. bottropensis reached its highest activity at 312 h and was higher than the activities of S. violaceoruber and S. nigrescens. The best condition for bacteriocin precipitation was using diammonium sulfate at 50% saturation. Bacteriocins were susceptible to proteinase treatments and stable at high temperature (up to 100 °C). The highest inhibitory activities were observed between pH 5 and 6. Cross-activity assays indicated that each Streptomyces strain produced different bacteriocins. When preparations of S. griseus and S. nigrescens were subjected to SDS-PAGE, bands of inhibition were observed in the gel overlay assay at a position corresponding to ~ 2 and 3 kDa, respectively, suggesting that both strains are potential sources for novel bacteriocins.
Asunto(s)
Antibacterianos/metabolismo , Bacteriocinas/metabolismo , Streptomyces/metabolismo , Sulfato de Amonio/química , Antibacterianos/aislamiento & purificación , Antibacterianos/farmacología , Bacteriocinas/genética , Bacteriocinas/aislamiento & purificación , Bacteriocinas/farmacología , Humanos , Concentración de Iones de Hidrógeno , Pruebas de Sensibilidad Microbiana , Estabilidad Proteica , Especificidad de la Especie , Streptomyces/clasificación , Streptomyces/genética , Streptomyces/crecimiento & desarrollo , TemperaturaRESUMEN
Bacterial strains isolated from attine ants showed activity against the insect specialized fungal pathogen Escovopsis and also against the human protozoan parasite Leishmania donovani. The bioassay guided fractionation of extracts from cultures of Streptomyces sp. ICBG292, isolated from the exoskeleton of Cyphomyrmex workers, led to the isolation of Mer-A2026B (1), piericidin-A1 (2) and nigericin (3). Nigericin (3) presented high activity against intracellular amastigotes of L. donovani (IC50 0.129 ± 0.008 µM). Streptomyces puniceus ICBG378, isolated from workers of Acromyrmex rugosus rugosus, produced dinactin (4) with potent anti-L. donovani activity against intracellular amastigotes (IC50 0.018 ± 0.003 µM). Compounds 3 and 4 showed good selectivity indexes, 88.91 and 656.11 respectively, and were more active than positive control, miltefosine. Compounds 1-4 were also active against some Escovopsis strains. Compounds 1 and 2 were also produced by Streptomyces sp. ICBG233, isolated from workers of Atta sexdens, and detected in ants' extracts by mass spectrometry, suggesting they are produced in the natural environment as defensive compounds involved in the symbiotic interaction.
Asunto(s)
Antiinfecciosos/metabolismo , Hormigas/microbiología , Hypocreales/efectos de los fármacos , Leishmania donovani/efectos de los fármacos , Streptomyces/aislamiento & purificación , Streptomyces/metabolismo , Animales , Antiinfecciosos/química , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Streptomyces/clasificaciónRESUMEN
A novel actinobacterium, designated strain 1AS2cT, was isolated from the wheat rhizosphere collected from the Brazilian Cerrado biome. A polyphasic approach, including phenotypic characterization and phylogenetic multilocus sequence analysis (MLSA), was used to determine the taxonomic position of this strain. Analysis of the 16S rRNA gene sequence indicated that the novel strain is closely related to Streptomyces cavourensis NBRC 13026T, Streptomyces albolongus NBRC 13465T and Streptomyces araujoniae ASBV-1T, sharing a similarity value of 98.2, 98.1 and 97.9â%, respectively. Additionally, MLSA of five housekeeping genes (atpD, gyrB, recA, rpoB and trpB) showed evolutionary distances beyond the 0.007 threshold, as well as low DNA-DNA relatedness between strain 1AS2cT and its closest phylogenetic neighbours (S. cavourensis NBRC 13026T, S. albolongus NBRC 13465T and S. araujoniae ASBV-1T: 56, 62.5 and 63.0â%, respectively), indicating a new phylogenetic lineage. The phylogenetic, chemotaxonomic and phenotypic characteristics support the assignment of strain 1AS2cT to the genus Streptomyces, representing a novel species. It is concluded that strain 1AS2cT (=CMAA 1679T=NRRL B-65479T=DSM 105299T) can be classified as the type strain of a novel species of the genus Streptomyces, for which the name Streptomyces rhizosphaericola sp. nov. is proposed.
Asunto(s)
Filogenia , Rizosfera , Microbiología del Suelo , Streptomyces/clasificación , Triticum/microbiología , Técnicas de Tipificación Bacteriana , Composición de Base , Brasil , ADN Bacteriano/genética , Ácidos Grasos/química , Genes Bacterianos , Tipificación de Secuencias Multilocus , Fosfolípidos/química , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Streptomyces/aislamiento & purificaciónRESUMEN
A polyphasic approach was used for evaluating the taxonomic status of strain HST21T isolated from Salar de Huasco in the Atacama Desert. The results of 16S rRNA gene and multilocus sequence phylogenetic analyses assigned strain HST21T to the genus Streptomyceswith Streptomyces albidochromogenes DSM 41800Tand Streptomyces flavidovirens DSM 40150T as its nearest neighbours. Digital DNA-DNA hydridization (dDDH) and average nucleotide identity (ANI) values between the genome sequences of strain HST21T and S. albidochromogenes DSM 41800T (35.6 and 88.2â%) and S. flavidovirens DSM 40105T (47.2 and 88.8â%) were below the thresholds of 70â and 95-96â% for prokaryotic conspecific assignation. Phenotypic, chemotaxonomic and genetic results distinguished strain HST21T from its closest neighbours. Strain HST21T is characterized by the presence of ll-diaminopimelic acid in its peptidoglycan layer; glucose and ribose as whole cell sugars; diphosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylethanolamine, phosphatidylinositol, glycophospholipids, unknown lipids and phospholipids as polar lipids; and anteiso-C15â:â0 (21.6â%) and anteiso-C17â:â0 (20.5â%) as major fatty acids (>15â%). Based on these results, strain HST21T merits recognition as a novel species, for which the name Streptomyces altiplanensis sp. nov. is proposed. The type strain is HST21T=DSM 107267T=CECT 9647T. While analysing the phylogenies of strain HST21T, Streptomyces chryseus DSM 40420T and Streptomyces helvaticus DSM 40431T were found to have 100â% 16S rRNA gene sequence similarity with digital DNA-DNA hydridization (dDDH) and average nucleotide identity (ANI) values of 95.3 and 99.4â%, respectively. Therefore, S. helvaticus is considered as a later heterotypic synonym of S. chryseus and, consequently, we emend the description of S. chryseus.
Asunto(s)
Clima Desértico , Filogenia , Microbiología del Suelo , Streptomyces/clasificación , Álcalis , Altitud , Técnicas de Tipificación Bacteriana , Composición de Base , Chile , ADN Bacteriano/genética , Ácido Diaminopimélico/química , Ácidos Grasos/química , Concentración de Iones de Hidrógeno , Hibridación de Ácido Nucleico , Peptidoglicano/química , Fosfolípidos/química , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Streptomyces/aislamiento & purificaciónRESUMEN
Streptomyces strain HST28T isolated from the Salar de Huasco, an athalassohaline and poly-extreme high altitude saline wetland located in northern Chile, was the subject of a polyphasic taxonomic study. Strain HST28T showed morphological and chemotaxonomic features in line with its classification in the genus Streptomyces. Optimal growth of strain HST28T was obtained at 28 °C, pH 8-9 and up to 10â% (w/v) NaCl. Single (16S rRNA) and multi-locus gene sequence analyses showed that strain HST28T had a distinct phylogenetic position from its closest relatives, the type strains of Steptomyces aureus and Streptomyces kanamyceticus. Digital DNA-DNA hybridization (23.3 and 31.0â%) and average nucleotide identity (79.3 and 85.6â%) values between strain HST28T and its corresponding relatives mentioned above were below the threshold of 70 and 96â%, respectively, defined for assigning a prokaryotic strains to the same species. Strain HST28T was characterised by the presence of ll-diaminopimelic acid in its peptidoglycan layer; galactose, glucose, ribose and traces of arabinose and mannose as whole-cell sugars; phosphatidylmethylethanolamine, phosphatidylinositol, aminolipid, glycophospholipid and an unidentified lipid as polar lipids; and the predominating menaquinones MK-9(H6), MK-9(H8) and MK-9(H4) (>20â%) as well as anteiso-C15â:â0 and anteiso-C17â:â0 as major fatty acids (>15â%). Based on the phenotypic and genetic results, strain HST28T (DSM 107268T=CECT 9648T) merits recognition as a new species named Streptomyces huasconensis sp. nov.
Asunto(s)
Altitud , Filogenia , Salinidad , Streptomyces/clasificación , Humedales , Técnicas de Tipificación Bacteriana , Composición de Base , Chile , ADN Bacteriano/genética , Ácido Diaminopimélico/química , Ácidos Grasos/química , Hibridación de Ácido Nucleico , Peptidoglicano/química , Fosfolípidos/química , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Streptomyces/aislamiento & purificación , Vitamina K 2/análogos & derivados , Vitamina K 2/químicaRESUMEN
Many insects have been associated with actinobacteria in protective symbiosis where antimicrobial metabolites inhibit host pathogens. However, the microbiota of neotropical insects such as the stingless-bee Tetragonisca angustula is poorly explored. T. angustula is a meliponid bee widely distributed in Latin America, its honey is traditionally exploited because of its ethno-pharmacological properties and its antimicrobial activity has been demonstrated. Also, the well-structured nest of this species allows exploration of the microbiota of its different components. Even though Streptomyces spp. have been cultured from stingless-bees, little is known about their role in this insect-microbe relationship. In this study, we examined the association between culturable actinobacteria and T. angustula, and evaluated the isolates' potential as antimicrobial producers. We isolated 51 actinobacteria from adult bees and different substrates of the hive of T. angustula (pollen and honey storage, garbage pellets and cerumen). We then performed a 16S rRNA phylogenetic analysis that clusters the bacteria to previously described lineages of host-associated Streptomyces. In addition, all the isolates were classified according to their antibacterial activity against human pathogens, measured by a growth inhibition test based on diffusion in agar. More than 50â% of our isolates exhibit antimicrobial activity, mainly to Gram-positive bacteria and fungi and only two against Gram-negative bacteria. Additionally, we obtained electron micrographs of adult bees with what appears to be patches of hyphae with Streptomyces-like cell morphology on their body surface. Our results suggest that T. angustula possibly uptakes and transfers actinobacteria from the environment, acting as vectors for these potentially beneficial organisms. This research provides new insights regarding the microbiota associated with T. angustula and justify future studies exploring the full diversity of the microbial community associated with the hive and the possible exchange of microbes with the crops they pollinate.
Asunto(s)
Antiinfecciosos/metabolismo , Abejas/microbiología , Filogenia , Streptomyces/clasificación , Streptomyces/metabolismo , Actinobacteria/clasificación , Actinobacteria/genética , Actinobacteria/metabolismo , Animales , Antiinfecciosos/farmacología , Bacterias/efectos de los fármacos , Abejas/ultraestructura , Hongos/efectos de los fármacos , Interacciones Microbiota-Huesped , Pruebas de Sensibilidad Microbiana , Microbiota/genética , ARN Ribosómico 16S/genética , Streptomyces/genéticaRESUMEN
ABSTRACT Fifty seven soil-borne actinomycete strains were assessed for the antibiotic production. Two of the most active isolates, designed as Streptomyces ST-13 and DK-15 exhibited a broad range of antimicrobial activity and therefore they were selected for HPLC fractionation against the most suppressed bacteria Staphylococcus aureus (ST-13) and Chromobacterium violaceum (DK-15). LC/MS analysis of extracts showed the presence of polyketides factumycin (DK15) and tetrangomycin (ST13). The taxonomic position of the antibiotic-producing actinomycetes was determined using a polyphasic approach. Phenotypic characterization and 16S rRNA gene sequence analysis of the isolates matched those described for members of the genus Streptomyces. DK-15 strain exhibited the highest 16S rRNA gene sequence similarity to Streptomyces globosus DSM-40815 (T) and Streptomyces toxytricini DSM-40178 (T) and ST-13 strain to Streptomyces ederensis DSM-40741 (T) and Streptomyces phaeochromogenes DSM-40073 (T). For the proper identification, MALDI-TOF/MS profile of whole-cell proteins led to the identification of S. globosus DK-15 (accession number: KX527570) and S. ederensis ST13 (accession number: KX527568). To our knowledge, there is no report about the production of these antibiotics by S.globosus and S. ederensis, thus isolates DK15 and ST13 identified as S. globosus DK-15 and S.ederensis ST-13 can be considered as new sources of these unique antibacterial metabolites.
Asunto(s)
Streptomyces/aislamiento & purificación , Streptomyces/metabolismo , Antibacterianos/biosíntesis , Filogenia , Piridonas/metabolismo , Microbiología del Suelo , Streptomyces/clasificación , Streptomyces/genética , Benzo(a)Antracenos/metabolismo , ADN Bacteriano/genética , Técnicas de Tipificación BacterianaRESUMEN
The present study aimed to identify novel microbial producers of bioemulsificant compounds from Antarctic soils. Fifty-nine microbial strains were isolated from five different locations at South Shetland Islands, Antarctica, and screened for biosurfactant production by ß-hemolytic activity. Strain So 3.2 was determined as bioemulsifier-producer and identified by phenotypic and molecular characterization as Streptomyces luridus. Emulsification activity, oil displacement method and drop-collapsing test were performed to evaluate the biosurfactant activity with different oils and hydrocarbons using two different culture media (Luria Bertani and Bushnell Haas in the presence of different carbon sources: glucose, glycerol, olive oil and n-Hexadecane). Cell free supernatant of Bushnell Haas culture supplemented with n-Hexadecane showed the best results for all tests. Emulsification of hydrocarbons exceeded 60%, reaching up to 90% on oil with high API grade, while displacement tests ranged from 8 cm to 4 cm in diameter according the culture media and tested oils. Our results revealed that Streptomyces luridus So3.2 is able to produce bioemulsifiers capable of emulsifying hydrocarbons and oils, which could be used in different biotechnological applications, particularly for bioremediation of environments contaminated by oil leaks.
Asunto(s)
Emulsionantes/metabolismo , Hidrocarburos/química , Streptomyces/aislamiento & purificación , Regiones Antárticas , Biodegradación Ambiental , Sistema Libre de Células , Medios de Cultivo/química , Microbiología del Suelo , Streptomyces/clasificación , Streptomyces/crecimiento & desarrollo , Streptomyces/metabolismoRESUMEN
Fifty seven soil-borne actinomycete strains were assessed for the antibiotic production. Two of the most active isolates, designed as Streptomyces ST-13 and DK-15 exhibited a broad range of antimicrobial activity and therefore they were selected for HPLC fractionation against the most suppressed bacteria Staphylococcus aureus (ST-13) and Chromobacterium violaceum (DK-15). LC/MS analysis of extracts showed the presence of polyketides factumycin (DK15) and tetrangomycin (ST13). The taxonomic position of the antibiotic-producing actinomycetes was determined using a polyphasic approach. Phenotypic characterization and 16S rRNA gene sequence analysis of the isolates matched those described for members of the genus Streptomyces. DK-15 strain exhibited the highest 16S rRNA gene sequence similarity to Streptomyces globosus DSM-40815 (T) and Streptomyces toxytricini DSM-40178 (T) and ST-13 strain to Streptomyces ederensis DSM-40741 (T) and Streptomyces phaeochromogenes DSM-40073 (T). For the proper identification, MALDI-TOF/MS profile of whole-cell proteins led to the identification of S. globosus DK-15 (accession number: KX527570) and S. ederensis ST13 (accession number: KX527568). To our knowledge, there is no report about the production of these antibiotics by S.globosus and S. ederensis, thus isolates DK15 and ST13 identified as S. globosus DK-15 and S.ederensis ST-13 can be considered as new sources of these unique antibacterial metabolites.
Asunto(s)
Antibacterianos/biosíntesis , Streptomyces/aislamiento & purificación , Streptomyces/metabolismo , Técnicas de Tipificación Bacteriana , Benzo(a)Antracenos/metabolismo , ADN Bacteriano/genética , Filogenia , Piridonas/metabolismo , Microbiología del Suelo , Streptomyces/clasificación , Streptomyces/genéticaRESUMEN
ABSTRACT As the largest genus in Actinobacteria family, Streptomyces species have the ability to synthesize numerous compounds of diverse structures with bioactivities. Streptomyces mangrovisoli MUSC 149T was previously isolated as a novel streptomycete from mangrove forest in east coast of Peninsular Malaysia. The high quality draft genome of MUSC 149T comprises 9,165,825 bp with G + C content of 72.5%. Through bioinformatics analysis, 21 gene clusters identified in the genome were associated with the production of bioactive secondary metabolites. The presence of these biosynthetic gene clusters in MUSC 149T suggests the potential exploitation of the strain for production of medically important compounds.
Asunto(s)
Streptomyces/aislamiento & purificación , Genoma Bacteriano , Sedimentos Geológicos/microbiología , Filogenia , Streptomyces/clasificación , Streptomyces/genética , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Composición de Base , ADN Bacteriano/genética , Datos de Secuencia Molecular , Secuencia de Bases , MalasiaRESUMEN
Streptomyces leeuwenhoekii strains C34T, C38, C58 and C79 were isolated from a soil sample collected from the Chaxa Lagoon, located in the Salar de Atacama in northern Chile. These streptomycetes produce a variety of new specialised metabolites with antibiotic, anti-cancer and anti-inflammatory activities. Moreover, genome mining performed on two of these strains has revealed the presence of biosynthetic gene clusters with the potential to produce new specialised metabolites. This review focusses on this new clade of Streptomyces strains, summarises the literature and presents new information on strain C34T.
Asunto(s)
Streptomyces/clasificación , Streptomyces/fisiología , Antibacterianos/biosíntesis , Antibacterianos/química , Chile , Genoma Bacteriano/genética , Estructura Molecular , Familia de Multigenes/genética , Filogenia , Microbiología del Suelo , Streptomyces/genética , Streptomyces/metabolismoRESUMEN
As the largest genus in Actinobacteria family, Streptomyces species have the ability to synthesize numerous compounds of diverse structures with bioactivities. Streptomyces mangrovisoli MUSC 149T was previously isolated as a novel streptomycete from mangrove forest in east coast of Peninsular Malaysia. The high quality draft genome of MUSC 149T comprises 9,165,825bp with G+C content of 72.5%. Through bioinformatics analysis, 21 gene clusters identified in the genome were associated with the production of bioactive secondary metabolites. The presence of these biosynthetic gene clusters in MUSC 149T suggests the potential exploitation of the strain for production of medically important compounds.
Asunto(s)
Genoma Bacteriano , Sedimentos Geológicos/microbiología , Streptomyces/aislamiento & purificación , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Composición de Base , Secuencia de Bases , ADN Bacteriano/genética , Malasia , Datos de Secuencia Molecular , Filogenia , Streptomyces/clasificación , Streptomyces/genéticaRESUMEN
ABSTRACT Here, we show the draft genome sequence of Streptomyces sp. F1, a strain isolated from soil with great potential for secretion of hydrolytic enzymes used to deconstruct cellulosic biomass. The draft genome assembly of Streptomyces sp. strain F1 has 69 contigs with a total genome size of 8,142,296 bp and G + C 72.65%. Preliminary genome analysis identified 175 proteins as Carbohydrate-Active Enzymes, being 85 glycoside hydrolases organized in 33 distinct families. This draft genome information provides new insights on the key genes encoding hydrolytic enzymes involved in biomass deconstruction employed by soil bacteria.
Asunto(s)
Proteínas Bacterianas/genética , Genoma Bacteriano , Glicósido Hidrolasas/genética , Microbiología del Suelo , Streptomyces/enzimología , Streptomyces/aislamiento & purificación , Proteínas Bacterianas/metabolismo , Composición de Base , Brasil , Glicósido Hidrolasas/metabolismo , Familia de Multigenes , Filogenia , Streptomyces/clasificación , Streptomyces/genéticaRESUMEN
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) (52â%) and MK-9(H8) (30â%) with 6â% MK-9(H4) and slightly less than 1â% MK-9(H2). Polar lipids present were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and an unknown phospholipid. The major fatty acids were anteiso-C15â:â0, anteiso-C16â:â0, anteiso-C14â:â0 and anteiso-C17â:â0. 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/químicaRESUMEN
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.