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1.
Int J Mol Sci ; 24(2)2023 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-36674628

RESUMO

Streptomyces lunaelactis strains have been isolated from moonmilk deposits, which are calcium carbonate speleothems used for centuries in traditional medicine for their antimicrobial properties. Genome mining revealed that these strains are a remarkable example of a Streptomyces species with huge heterogeneity regarding their content in biosynthetic gene clusters (BGCs) for specialized metabolite production. BGC 28a is one of the cryptic BGCs that is only carried by a subgroup of S. lunaelactis strains for which in silico analysis predicted the production of nonribosomal peptide antibiotics containing the non-proteogenic amino acid piperazic acid (Piz). Comparative metabolomics of culture extracts of S. lunaelactis strains either holding or not holding BGC 28a combined with MS/MS-guided peptidogenomics and 1H/13C NMR allowed us to identify the cyclic hexapeptide with the amino acid sequence (D-Phe)-(L-HO-Ile)-(D-Piz)-(L-Piz)-(D-Piz)-(L-Piz), called lunaemycin A, as the main compound synthesized by BGC 28a. Molecular networking further identified 18 additional lunaemycins, with 14 of them having their structure elucidated by HRMS/MS. Antimicrobial assays demonstrated a significant bactericidal activity of lunaemycins against Gram-positive bacteria, including multi-drug resistant clinical isolates. Our work demonstrates how an accurate in silico analysis of a cryptic BGC can highly facilitate the identification, the structural elucidation, and the bioactivity of its associated specialized metabolites.


Assuntos
Anti-Infecciosos , Streptomyces , Antibacterianos/farmacologia , Antibacterianos/metabolismo , Espectrometria de Massas em Tandem , Anti-Infecciosos/metabolismo , Streptomyces/genética , Streptomyces/metabolismo , Família Multigênica
2.
Molecules ; 28(2)2023 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-36677936

RESUMO

Pseudorabies virus (PRV) is a pathogen that causes Aujeszky's disease (AD) in animals, leading to huge economic losses to swine farms. In order to discover anti-PRV compounds, we studied the extracts of the strain Streptomyces jiujiangensis NBERC-24992, which showed significant anti-PRV activity. Eight benzoheterocyclic secondary metabolites, including three new compounds (1-3, virantmycins D-G) and five known compounds (4-8, virantmycin, A-503451 D, A-503451 D acetylate, A-503451 A, and A-503451 B), were isolated from the broth of NBERC-24992. The structures of the new compounds were identified by using extensive spectroscopic data, including mass spectrometry (MS), nuclear magnetic resonance (NMR), and electronic circular dichroism (ECD). Compound 1 was found to be a novel heterocyclic compound with a tricyclic skeleton from natural product. All compounds were tested for antiviral activity, and 4 (virantmycin) showed an excellent effect against PRV and was better than ribavirin and acyclovir. Our study revealed that chlorine atom and tetrahydroquinoline skeleton were important active moiety for antiviral activity. Virantmycin could be a suitable leading compound for an antiviral drug against PRV.


Assuntos
Herpesvirus Suídeo 1 , Pseudorraiva , Streptomyces , Suínos , Animais , Antivirais/uso terapêutico , Pseudorraiva/tratamento farmacológico , Streptomyces/metabolismo
3.
Sci Rep ; 13(1): 4, 2023 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-36593229

RESUMO

A potentially novel actinobacterium isolated from forest soil, Streptomyces sp. KSF103 was evaluated for its insecticidal effect against several mosquito species namely Aedes aegypti, Aedes albopictus, Anopheles cracens and Culex quinquefasciatus. Mosquito larvae and adults were exposed to various concentrations of the ethyl acetate (EA) extract for 24 h. Considerable mortality was evident after the EA extract treatment for all four important vector mosquitoes. Larvicidal activity of the EA extract resulted in LC50 at 0.045 mg/mL and LC90 at 0.080 mg/mL for Ae. aegypti; LC50 at 0.060 mg/mL and LC90 at 0.247 mg/mL for Ae. albopictus; LC50 at 2.141 mg/mL and LC90 at 6.345 mg/mL for An. cracens; and LC50 at 0.272 mg/mL and LC90 at 0.980 mg/mL for Cx. quinquefasciatus. In adulticidal tests, the EA extract was the most toxic to Ae. albopictus adults (LD50 = 2.445 mg/mL; LD90 = 20.004 mg/mL), followed by An. cracens (LD50 = 5.121 mg/mL; LD90 = 147.854 mg/mL) and then Ae. aegypti (LD50 = 28.873 mg/mL; LD90 = 274.823 mg/mL). Additionally, the EA extract exhibited ovicidal activity against Ae. aegypti (LC50 = 0.715 mg/mL; LC90 = 6.956 mg/mL), Ae. albopictus (LC50 = 0.715 mg/mL; LC90 = 6.956 mg/mL), and An. cracens (LC50 = 0.715 mg/mL; LC90 = 6.956 mg/mL), evaluated up to 168 h post-treatment. It displayed no toxicity on the freshwater microalga Chlorella sp. Beijerinck UMACC 313, marine microalga Chlorella sp. Beijerinck UMACC 258 and the ant Odontoponera denticulata. In conclusion, the EA extract showed promising larvicidal, adulticidal and ovicidal activity against Ae. aegypti, Ae. albopictus, An. cracens, and Cx. quinquefasciatus (larvae only). The results suggest that the EA extract of Streptomyces sp. KSF103 has the potential to be used as an environmental-friendly approach in mosquito control. The current study would serve as an initial step toward complementing microbe-based bioinsecticides for synthetic insecticides against medically important mosquitoes.


Assuntos
Aedes , Chlorella , Culex , Inseticidas , Streptomyces , Animais , Inseticidas/farmacologia , Extratos Vegetais/farmacologia , Mosquitos Vetores , Larva , Folhas de Planta
4.
Molecules ; 28(2)2023 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-36677698

RESUMO

Two new napyradiomycins derivatives, napyradiomycin A4 (1) and A80915 H (2), along with five known ones, were isolated from the ethyl acetate extract of fermentation culture of Streptomyces kebangsaanensis WS-68302. Their structures were elucidated by extensive spectroscopic analysis, including HR-MS, 1D and 2D NMR, CD spectrum, as well as comparison with literature data. Compound 1 exhibited significant antiviral activity against PRV (Pseudorabies virus) with an IC50 value of 2.056 µM and therapeutic ratio at 14.98, suggesting that it might have potential for development of an antiviral agent. Moreover, compound 1 displayed the strongest inhibition against PRV protein among the tested napyradiomycins in the indirect immunofuorescence assay. Compounds 3 and 4 showed higher activities against swine pathogenic Streptococcus suis than the positive control penicillin G sodium salt, with MIC values of 3.125 and 6.25 µg/mL, respectively. Compounds 1 and 3-6 exhibited moderate antibacterial activity against the swine pathogenic Erysipelothrix rhusiopathiae, with MIC values ranging from 25 to 50 µg/mL.


Assuntos
Antibacterianos , Streptomyces , Animais , Suínos , Antibacterianos/química , Streptomyces/química , Espectroscopia de Ressonância Magnética , Testes de Sensibilidade Microbiana
5.
Microb Cell Fact ; 22(1): 5, 2023 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-36609255

RESUMO

BACKGROUND: New antibiotics are urgently needed in clinical treatment of superdrug-resistant bacteria. Nonribosomal peptides (NRPs) are a major source of antibiotics because they exhibit structural diversity, and unique antibacterial mechanisms and resistance. Analysis of gene clusters of S. agglomeratus 5-1-3 showed that Clusters 3, 6, 12, 21, and 28 were used to synthesize NRPs. Here, we examined secondary metabolites of S. agglomeratus 5-1-3 isolated from soils in the Qinghai-Tibet Plateau, China, for NRPs with antibacterial activity. RESULTS: We isolated a total of 36 Streptomyces strains with distinct colony morphological characteristics from 7 soil samples. We screened 8 Streptomyces strains resistant to methicillin-resistant Staphylococcus aureus (MRSA). We then selected S. agglomeratus 5-1-3 for further study based on results of an antibacterial activity test. Here, we isolated three compounds from S. agglomeratus 5-1-3 and characterized their properties. The crude extract was extracted with ethyl acetate and purified with column chromatography and semipreparative high-performance liquid chromatography (HPLC). We characterized the three compounds using NMR analyses as echinomycin (1), 5,7,4'-trihydroxy-3.3',5'-trimethoxy flavone (2), and 2,6,2', 6'-tetramethoxy-4,4-bis(2,3-epoxy-1-hydroxypropyl)-biphenyl (3). We tested the antibacterial activity of pure compounds from strain 5-1-3 with the Oxford cup method. NRP echinomycin (1) showed excellent anti-MRSA activity with a minimum inhibitory concentration (MIC) of 2.0 µg/mL. Meanwhile, MIC of compound 2 and 3 was 128.0 µg/mL for both. In addition, 203 mg of echinomycin was isolated from 10 L of the crude extract broth of strain 5-1-3. CONCLUSION: In this study, S. agglomeratus 5-1-3 with strong resistance to MRSA was isolated from the soils in the Qinghai-Tibet Plateau. Strain 5-1-3 had a high yield of echinomycin (1) an NRP with a MIC of 2 µg/mL against MRSA. We propose that echinomycin derived from S. agglomeratus 5-1-3 may be a potent antibacterial agent for pharmaceutical use.


Assuntos
Equinomicina , Staphylococcus aureus Resistente à Meticilina , Streptomyces , Tibet , Antibacterianos/química , Streptomyces/química , Testes de Sensibilidade Microbiana , Misturas Complexas , Solo
6.
World J Microbiol Biotechnol ; 39(3): 70, 2023 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-36617604

RESUMO

The effect of Streptomyces spp. on organic matter degradation was investigated in the present study. Streptomyces spp. isolated from compost systems were eliminated based on the results of cellulose, starch, xylan degradation tests, morphological inspection, and 16S rRNA analysis. The eliminated strains were re-given to compost systems to determine their effect on organic matter degradation and maturation. Sample analyses indicated that 15 days of composting had been adequate to maintain maturation. The amounts of strains added to the system were high enough to create a detectable change such as inhibition of other microbiota members. Results also indicated a variant change in organic matter degradation due to the added strain. The difference in organic matter degradation between strains depended partially on the segregation of secondary metabolites. On the other hand, strains also inhibited each other in the case of their binary and triple utilization in compost. Another explanation for variant activity was provided based on the enzymatic activity of the strains validated by metagenomic counts evaluation. Metagenome count numbers revealed the tendency of compost microbiota toward degradation products of cellulose. Findings obtained from composting experiments and metagenome analyses indicated the presence of a different degradation route based on xylan activity. Results also implied a decrease in competition between the dominant strain and microbiota members in the case of sequential xylan and cellulose degradation. Meticulous evaluation of results obtained from metagenome analysis also provided some insights on certain conditions regarding the progress of composting along with storage conditions of manure before use.


Assuntos
Compostagem , Streptomyces , Metagenoma , Solo , Streptomyces/genética , RNA Ribossômico 16S/genética , Xilanos , Celulose , Esterco
7.
Curr Microbiol ; 80(2): 71, 2023 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-36622468

RESUMO

Modern society has a great challenge to decrease waste and minimize the adverse effects of wastes on the economy, environment, and individual health. Thus, this study focuses on the use of eight agro-wastes (banana peel, barley straw, canola straw, pomegranate peel, orange peel, pumpkin pulp+seeds, maple leaf, and brewer's spent grains) by a novel bacterium (Streptomyces thermocarboxydus) for enzymes production. Further, the study explored the subsequent degradation of those wastes by the bacterium. This bacterium was isolated from forest soil and identified as Streptomyces thermocarboxydus by 16S rRNA sequence analysis. The biodegrading capability of S. thermocarboxydus was determined by observing the clear zone around the colony cultured on the agar plate containing the different biomasses as sole carbon sources and calculating the substrate degradation ratios. Furthermore, scanning electron microscopy images of eight agro-wastes before and after bacterial treatment and weight loss of agro-wastes revealed the bacterium degraded the biomasses. The different trends of enzyme activities were observed for various wastes, and the maximum activity depended on the type of agro-wastes. Overall, S. thermocarboxydus was found to be a potential candidate for pectinase and xylanase production. The enzyme production varies with the concentration of the biomasses.


Assuntos
Frutas , Streptomyces , Biomassa , RNA Ribossômico 16S/genética , Streptomyces/genética
8.
J Am Chem Soc ; 145(3): 1886-1896, 2023 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-36634356

RESUMO

The logical and effective discovery of macrolactams, structurally unique natural molecules with diverse biological activities, has been limited by a lack of targeted search methods. Herein, a targeted discovery method for natural macrolactams was devised by coupling genomic signature-based PCR screening of a bacterial DNA library with spectroscopic signature-based early identification of macrolactams. DNA library screening facilitated the efficient selection of 43 potential macrolactam-producing strains (3.6% of 1,188 strains screened). The PCR amplicons of the amine-deprotecting enzyme-coding genes were analyzed to predict the macrolactam type (α-methyl, α-alkyl, or ß-methyl) produced by the hit strains. 1H-15N HSQC-TOCSY NMR analysis of 15N-labeled culture extracts enabled macrolactam detection and structural type assignment without any purification steps. This method identified a high-titer Micromonospora strain producing salinilactam (1), a previously reported α-methyl macrolactam, and two Streptomyces strains producing new α-alkyl and ß-methyl macrolactams. Subsequent purification and spectroscopic analysis led to the structural revision of 1 and the discovery of muanlactam (2), an α-alkyl macrolactam with diene amide and tetraene chromophores, and concolactam (3), a ß-methyl macrolactam with a [16,6,6]-tricyclic skeleton. Detailed genomic analysis of the strains producing 1-3 identified putative biosynthetic gene clusters and pathways. Compound 2 displayed significant cytotoxicity against various cancer cell lines (IC50 = 1.58 µM against HCT116), whereas 3 showed inhibitory activity against Staphylococcus aureus sortase A. This genomic and spectroscopic signature-based method provides an efficient search strategy for new natural macrolactams and will be generally applicable for the discovery of nitrogen-bearing natural products.


Assuntos
Streptomyces , Estrutura Molecular , Lactamas Macrocíclicas/farmacologia , Lactamas Macrocíclicas/química , Streptomyces/metabolismo , Genômica , Reação em Cadeia da Polimerase , Família Multigênica
9.
J Nat Prod ; 86(1): 94-102, 2023 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-36599087

RESUMO

Heterologous expression of a cdps-p450 locus from Streptomyces sp. NRRL S-1521 led to the identification of guanitrypmycin D1, a new guaninylated diketopiperazine. The cytochrome P450 GutD1521 catalyzed the regiospecific transfer of guanine to C-2 of the indole ring of cyclo-(l-Trp-l-Tyr) via a C-C linkage and represents a new chemical transformation within this enzyme class. Furthermore, GutD1521 efficiently accepts several other tryptophan-containing cyclodipeptides or derivatives for regiospecific coupling with guanine, thus generating different guanitrypmycin analogs.


Assuntos
Streptomyces , Streptomyces/metabolismo , Sistema Enzimático do Citocromo P-450/metabolismo , Guanina/metabolismo
10.
J Nat Prod ; 86(1): 176-181, 2023 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-36634313

RESUMO

Six new azoxy-aromatic compounds (o-alkylazoxymycins A-F, 1-6) and two new nitrogen-bearing phenylvaleric/phenylheptanoic acid derivatives (o-alkylphemycins A and B, 7 and 8) were isolated from Streptomyces sp. Py50. Their structures were elucidated based on HRESIMS, NMR, UV spectroscopic analyses, and X-ray crystallographic data. O-Alkylazoxymycins A-F (1-6) are the first natural examples of azoxy compounds with the azoxy bond attached to the ortho-position of the phenylheptanoic acid or phenylvaleric acid moiety. Compounds 1, 5, and 6 were active against Epidermophyton floccosum with MIC50 values ranging from 10.1 to 51.2 µM. A plausible biosynthetic pathway of 2 and 3 was proposed.


Assuntos
Streptomyces , Streptomyces/química , Espectroscopia de Ressonância Magnética , Compostos Azo/química , Cristalografia por Raios X , Vias Biossintéticas , Estrutura Molecular
11.
World J Microbiol Biotechnol ; 39(3): 78, 2023 Jan 16.
Artigo em Inglês | MEDLINE | ID: mdl-36645528

RESUMO

The two-component system (TCS) found in various organisms is a regulatory system, which is involved in the response by the organism to stimuli, thereby regulating the internal behavior of the cell. It is commonly found in prokaryotes and is an important signaling system in bacteria. TCSs are involved in the regulation of physiological and morphological differentiation of the industrially important microbes from the genus Streptomyces, which produce a vast array of bioactive secondary metabolites (SMs). Genetic engineering of TCSs can substantially increase the yield of target SMs, which is valuable for industrial-scale production. Research on TCS has mainly been completed in the model strain Streptomyces coelicolor. In this review, we summarize the recent advances in the functional identification and elucidation of the regulatory mechanisms of various TCSs in S. coelicolor, with a focus on their roles in the biosynthesis of important SMs.


Assuntos
Streptomyces coelicolor , Streptomyces , Streptomyces coelicolor/genética , Streptomyces coelicolor/metabolismo , Streptomyces/metabolismo , Regulação Bacteriana da Expressão Gênica , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo
12.
ACS Synth Biol ; 12(1): 61-70, 2023 01 20.
Artigo em Inglês | MEDLINE | ID: mdl-36516042

RESUMO

The CRISPR/Cas9 system provides an efficient tool for engineering genomes. However, its application to Streptomyces genome engineering has been hampered by excessive toxicity associated with overexpression of Cas9 protein. As the level of Cas9 toxicity varies significantly between Streptomyces species, species-specific optimization of Cas9 expression is a strategy to mitigate its toxicity while maintaining sufficient double-strand break (DSB) activity for genome engineering. Using a pool of randomized constitutive promoters and a blue pigment indigoidine biosynthetic gene (IndC) as a reporter, we developed the CaExTun (Cas9 Expression Tuning) platform, which enables rapid screening of a large pool of promoter-Cas9 constructs to quickly recover the one with high DSB activity and no apparent toxicity. We demonstrate the utility of CaExTun using four model Streptomyces species. We also show that CaExTun can be applied to the CRISPRi system by allowing the construction of a library of promoter-dCas9 constructs that confer a wide range of gene repression levels. As demonstrated here, CaExTun is a versatile tool for the rapid optimization of the CRISPR/Cas9 system in a species-specific manner and thus will facilitate CRISPR/Cas9-based genome engineering efforts in Streptomyces.


Assuntos
Streptomyces , Streptomyces/genética , Streptomyces/metabolismo , Sistemas CRISPR-Cas/genética , Regiões Promotoras Genéticas/genética , Edição de Genes
13.
Appl Microbiol Biotechnol ; 107(1): 369-378, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36478282

RESUMO

Streptomyces avermitilis is a gram-positive bacterium that undergoes complex physiological and morphological differentiation during its life cycle, which has implications in secondary metabolite production. Avermectin, produced by S. avermitilis, is widely used as an anthelmintic and insecticidal agent. In this study, we have applied Raman microspectroscopic imaging to elucidate the correlation between production of avermectin and the morphological differentiation in S. avermitilis. We demonstrate distinctive variations in the localization of secondary metabolites at various stages of morphological differentiation. Under solid culture, avermectin was detected in the mycelia formed at the later stages of morphological differentiation (e.g., spore-bearing mycelium and spiral spore chains), but not in the early-stage substrate mycelium. On the contrary, under liquid culture condition, avermectin was found concentrated in the mycelial pellet formed at the early MII stage of differentiation. Furthermore, the chemical profiles of the mycelia were substantially different depending on the culture condition. Raman spectra corresponding to proteins, lipids, and cytochrome were observed in the mycelia irrespective of the stage of morphological differentiation, however, carotenoid was observed under solid culture condition particularly in spore-bearing mycelium and spiral spore chains. KEY POINTS: • Avermectin production is regulated during mycelial differentiation • Liquid and solid culture conditions affects mycelial differentiation • Raman microspectroscopic analysis reveals localization profiles of avermectin.


Assuntos
Regulação Bacteriana da Expressão Gênica , Streptomyces , Streptomyces/metabolismo , Ivermectina , Micélio/metabolismo
14.
J Nat Prod ; 86(1): 34-44, 2023 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-36535025

RESUMO

Sixteen new biisoflavones, bisoflavolins A-N (1-16), were discovered from cultures of the Takla Makan desert-derived strain Streptomyces sp. HDN154127. The chemical structures, including axial chirality, were elucidated by NMR, MS, and ECD analyses. Antibacterial activity of dimerized compounds was tested against seven different bacteria. The dimerized compounds showed better activity (MIC from 0.8 to 50.0 µM) than the corresponding monomers (daidzein and genistein, MIC > 50.0 µM). The rare dimeric and chlorinated structures in 1-16 were proved to be biotransformation products obtained from soy isoflavones and sodium chloride, which constituted the culture medium. This is the first report of an actinomycete that promotes both dimerization and chlorination utilizing natural isoflavones as skeletons sources.


Assuntos
Isoflavonas , Streptomyces , Streptomyces/química , Halogenação , Dimerização , Isoflavonas/farmacologia , Isoflavonas/química , Genisteína
15.
J Antibiot (Tokyo) ; 76(2): 93-100, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36564595

RESUMO

A polyphasic approach was used to determine the taxonomic position of a marine actinomycete, designated isolate CWH03T, which we previously reported to produce new linear azole-containing peptides spongiicolazolicins A and B. Strain CWH03T is mesophilic, neutrophilic, and halotolerant streptomycete that forms spiral spore chains on aerial mycelium. Comparative 16S rRNA gene sequencing showed that CWH03T was most closely related to Streptomyces tirandamycinicus HNM0039T (99.7%), Streptomyces spongiicola HNM0071T (99.4%), 'Streptomyces marianii' ICN19T (99.1%) and Streptomyces wuyuanensis CGMCC4.7042T (99.0%). The phylogenetic tree prepared using the 16S rRNA gene, as well as the phylogenomic tree using the genome BLAST distance phylogeny method and 81 core housekeeping genes, respectively, showed that the closest relative of strain CWH03T was S. spongiicola HNM0071T. The average nucleotide identity and digital DNA-DNA hybridization values between strains CWH03T and S. spongiicola HNM0071T were 91.46% and 44.2%, respectively, which were below the thresholds of 96% and 70% for prokaryotic conspecific assignation. The G+C content of the genomic DNA of strain CWH03T was 72.3%. Whole-cell hydrolysates of strain CWH03T contained LL-diaminopimelic acid. The predominant menaquinone was MK-9(H8) (88.3%), and the major fatty acids were iso-C16:0 (28.4%), anteiso-C15:0 (15.0%) and iso-C15:0 (12.9%). The major phospholipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and an unidentified phospholipid. Based on data obtained from phenotypic, phylogenetic, genomic, and chemotaxonomic analyses, strain CWH03T represents a novel species of the genus Streptomyces, for which the proposed name is Streptomyces pacificus sp. nov. The type strain is CWH03T ( = NBRC 114659T = TBRC 15780T).


Assuntos
Actinobacteria , Streptomyces , Filogenia , RNA Ribossômico 16S/genética , Actinobacteria/genética , Análise de Sequência de DNA , Fosfolipídeos/química , Ácidos Graxos/química , DNA Bacteriano/genética , Vitamina K 2/química
16.
Mol Plant Pathol ; 24(2): 167-178, 2023 02.
Artigo em Inglês | MEDLINE | ID: mdl-36478143

RESUMO

Streptomyces scabies is the best-characterized plant-pathogenic streptomycete, which is a special species among the large genus Streptomyces. The pathogenicity of S. scabies relies on the production of the secondary metabolite thaxtomin A. Little is known about the molecular mechanisms underlying the regulation of thaxtomin biosynthesis in S. scabies beyond the pathway-specific activator TxtR and the cellulose utilization repressor CebR. The leucine-responsive regulatory protein (Lrp) family modulates secondary metabolism in nonpathogenic streptomycetes. However, the regulatory relationship between the Lrp and pathogenic streptomycetes remains unknown. In this study, we demonstrated that SCAB_Lrp (SCAB_77931) from S. scabies significantly affects thaxtomin biosynthesis, pathogenicity, and morphological development. SCAB_Lrp deletion resulted in a dramatic decline in thaxtomin A production and a low-virulence phenotype of S. scabies. An in-depth dissection of the regulatory mechanism of SCAB_Lrp revealed that it positively regulates the transcription of the thaxtomin biosynthetic gene cluster by directly binding to the promoter of the cluster-situated regulator gene txtR. SCAB_Lrp also controls the morphological development of S. scabies by directly activating the transcription of amfC, whiB, and ssgB. SCAB_Lrp directly controls the transcription of its own gene by binding a specific sequence (5'-GGACAGTCGCCGTGCTACG-3'). Moreover, phenylalanine and methionine have been characterized as SCAB_Lrp effectors by strengthening the binding affinity and complex status between SCAB_Lrp and DNA. Our findings characterize a multifunctional regulatory protein, SCAB_Lrp, that controls secondary metabolism, pathogenicity, and sporulation in S. scabies and provide new insights into the complex regulatory network that modulates thaxtomin phytotoxins in pathogenic Streptomyces.


Assuntos
Escabiose , Solanum tuberosum , Streptomyces , Virulência/genética , Proteína Reguladora de Resposta a Leucina/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Streptomyces/genética , Streptomyces/metabolismo , Doenças das Plantas , Solanum tuberosum/metabolismo
17.
Appl Microbiol Biotechnol ; 107(2-3): 881-896, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36585512

RESUMO

INTRODUCTION: Epsilon-poly-L-lysine (ε-PL) is produced by Streptomyces species in acidic and aerobic conditions, which inevitably induces rapid generation of reactive oxygen species (ROS). The devastating effects of ROS on biomolecules and cell vitality have been well-studied, while the positive effects of ROS are rarely reported. RESULTS: In this study, we found that a proper dose of intracellular ROS (about 3.3 µmol H2O2 /g DCW) could induce a physiological modification to promote the ε-PL production (from 1.2 to 1.5 g/L). It resulted in larger sizes of colony and mycelial pellets as well as vibrant, aggregated, and more robust mycelia, which were of high capability of ROS detoxication. Physiological studies showed that appropriate doses of ROS activated the metabolism of the pentose phosphate pathway at both transcriptional and enzymatic levels, which was beneficial for biomass accumulation. The biosynthesis of lysine was also promoted in terms of transcriptional regulatory overexpression, increased transcription and enzymatic activity of key genes, larger pools of metabolites in the TCA cycle, replenishment pathway, and diaminoheptanedioic acid pathway. In addition, energy provision was ensured by activated metabolism of the TCA cycle, a larger pool of NADH, and higher activity of the electron transport system. Increased transcription of HrdD and pls further accelerated the ε-PL biosynthesis. SIGNIFICANCE: These results indicated that ROS at proper intracellular dose could act as an inducing signal to activate the ε-PL biosynthesis, which laid a foundation for further process regulation to maintain optimal ROS dose in industrial ε-PL production and was of theoretical and practical significance. KEY POINTS: • A proper dose of intracellular ROS positively influences the ε-PL production. • Proper dose of ROS enhanced the mycelial activity and antioxidative capability. • ROS increased lysine synthesis metabolism, energy provision and pls expression.


Assuntos
Polilisina , Streptomyces , Espécies Reativas de Oxigênio/metabolismo , Peróxido de Hidrogênio/metabolismo , Streptomyces/genética , Ciclo do Ácido Cítrico
18.
Curr Opin Microbiol ; 71: 102257, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36565538

RESUMO

Streptomyces are ubiquitous terrestrial bacteria that are renowned for their robust metabolic capabilities and their behavioral flexibility. In competing for environmental niches, these bacteria can employ novel growth and dispersal behaviors. They also wield their diverse metabolic repertoire for everything from maximizing nutrient uptake, to preventing phage replication or inhibiting bacterial and fungal growth. Increasingly, they are found to live in association with plants and insects, often conferring protective benefits to their host courtesy of their ability to produce pathogen-inhibitory antimicrobial compounds. Here, we highlight recent advances in understanding the competitive and cooperative interactions between Streptomyces and phage, microbes, and higher organisms in their environment.


Assuntos
Anti-Infecciosos , Streptomyces , Animais , Anti-Infecciosos/metabolismo , Meio Ambiente , Plantas , Insetos
19.
World J Microbiol Biotechnol ; 39(1): 32, 2022 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-36462123

RESUMO

Given the environmental burden of textile industry, especially of dyeing processes and the volume of synthetic dyes and surfactants, the intensive development of the greener approaches is under way. Herein, an environmentaly-friendly dyeing of polyamide (PA) and PA/Elastane (PA/EA) knits using live bacterial approach in water environment, completely eliminating usage of textile auxiliaries is described. A total of 12 pigment-producing Streptomyces strains were isolated and purified from soil and rizoshere or bark of smoke tree Cotinus coggygria samples. The antibacterial, antifungal and cytotoxic effects of crude bacterial extracts were tested. Antimicrobial effect was obtained by the majority of extracts but only two streptomycetes extracts, 11-5 and BPS51, showed moderate cytotoxicity against HaCaT human cell line. This was the reason to select 11-5 and BPS51 strains for the dyeing of the textile materials. Excellent properties of dyeing wool, silk and PA are achieved initially using live cultures, and the bioprocess is optimized on commercial PA and PA/EA knits used for stockings production. Satisfactory coloration of both knits is achieved with dynamic conditions (culture shaking at 180 rpm over 5-14 days at 30 ºC) giving the best coloration results, except in the case of the PA sample dyed with a bacterial strain 11-5. The prolongation of dyeing time leads to higher color yields independently of fabric and bacteria strain. Although the color differences between the samples before and after washing are observed, washing fastness after three washing cycles can be considered as satisfactory.


Assuntos
Nylons , Streptomyces , Humanos , Animais , Nylons/farmacologia , Corantes , Poliuretanos , Extratos Vegetais
20.
Sci Rep ; 12(1): 21851, 2022 12 17.
Artigo em Inglês | MEDLINE | ID: mdl-36528632

RESUMO

Microbial-based strategy in nanotechnology offers economic, eco-friendly, and biosafety advantages over traditional chemical and physical protocols. The current study describes a novel biosynthesis protocol for chitosan nanoparticles (CNPs), employing a pioneer Streptomyces sp. strain NEAE-83, which exhibited a significant potential for CNPs biosynthesis. It was identified as Streptomyces microflavus strain NEAE-83 based on morphological, and physiological properties as well as the 16S rRNA sequence (GenBank accession number: MG384964). CNPs were characterized by SEM, TEM, EDXS, zeta potential, FTIR, XRD, TGA, and DSC. CNPs biosynthesis was maximized using a mathematical model, face-centered central composite design (CCFCD). The highest yield of CNPs (9.41 mg/mL) was obtained in run no. 27, using an initial pH of 5.5, 1% chitosan, 40 °C, and a 12 h incubation period. Innovatively, the artificial neural network (ANN), was used for validating and predicting CNPs biosynthesis based on the trials data of CCFCD. Despite the high precision degree of both models, ANN was supreme in the prediction of CNPs biosynthesis compared to CCFCD. ANN had a higher prediction efficacy and, lower error values (RMSE, MDA, and SSE). CNPs biosynthesized by Streptomyces microflavus strain NEAE-83 showed in-vitro antibacterial activity against Pectobacterium carotovorum, which causes the potato soft rot. These results suggested its potential application for controlling the destructive potato soft rot diseases. This is the first report on the biosynthesis of CNPs using a newly isolated; Streptomyces microflavus strain NEAE-83 as an eco-friendly approach and optimization of the biosynthesis process by artificial intelligence.


Assuntos
Quitosana , Nanopartículas , Solanum tuberosum , Streptomyces , Pectobacterium carotovorum/genética , RNA Ribossômico 16S/genética , Inteligência Artificial , Streptomyces/genética , Solanum tuberosum/genética
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