Project description:This study aimed to investigate the variations in the protein composition of Streptomyces sp. PU10 when cultivated with either Impranil (polyestere-polyurethane) or glucose as the carbon source. We analyzed both the intracellular and extracellular protein fractions to gain insights into the intricate processes involving PU degradation, intermediate metabolic pathways in PU degradation, and the connection between primary and secondary metabolism within Streptomyces sp. PU10.
Project description:The terminal compartments of Streptomyces are less prone to transcription than the rest of the chromosome. Indeed, the expression of the highly variable regions enriched in those compartments is generally conditional and often requires an empirical approach to characterize the inducing conditions. For instance, in the context of identifying adequate antibiotic production conditions, an OSMAC (“One Strain Many Compounds”) approach is frequently implemented, based on strain cultivation in different environmental conditions (composition of the medium, growth time, temperature, co-cultures, etc.). Likewise, to find the expression conditions of a complete prophage of Streptomyces ambofaciens ATCC 23877 (named 'Samy' phage/prophage), we conducted a similar approach by analyzing the transcriptomes in five solid media (HT, SAF, ONA, MMM, MMM+NAG). The terminal compartments of Streptomyces are less prone to transcription than the rest of the chromosome. Indeed, the expression of the highly variable regions enriched in those compartments is generally conditional and often requires an empirical approach to characterize the inducing conditions. For instance, in the context of identifying adequate antibiotic production conditions, an OSMAC (“One Strain Many Compounds”) approach is frequently implemented, based on strain cultivation in different environmental conditions (composition of the medium, growth time, temperature, co-cultures, etc.). Likewise, to find the expression conditions of a complete prophage of Streptomyces ambofaciens ATCC 23877 (named 'Samy' phage/prophage), we conducted a similar approach by analyzing the transcriptomes in five solid media (HT, SAF, ONA, MMM, MMM+NAG).
Project description:Streptomyces sp. M7 has demonstrated ability to remove lindane from culture media and soils. In this study, we used MS-based label-free quantitative proteomic to understand lindane degradation and its metabolic context in Streptomyces sp. M7. We identified the proteins involved in the up-stream degradation pathway. Our results demonstrated that mineralization of lindane is feasible since proteins from an unusual down-stream degradation pathway were also identified. Degradative steps were supported by an active catabolism that supplied energy and reducing equivalents in the form of NADPH. This is the first study in which degradation steps of an organochlorine compound and metabolic context are elucidate in a biotechnological genus as Streptomyces. These results serve as basement to study other degradative actinobacteria and to improve the degradation processes of Streptomyces sp. M7.
Project description:Activating the cryptic secondary metabolic gene clusters is a vital research field in Streptomyces. The marine Streptomyces sp. FJNU027 strain which could produce tirandamycins was cultured in the oligotrophic medium. Compared with normal medium, a differential product in oligotrophic culture was found by HPLC assay. After mass fermentation, 2 mg of the differential product was obtained from 30 L fermentation broth by solvent extraction, column chromatography over sephadex LH-20 and reverse phase C18, and other methods. It was identified as 4,4',5,5'-tetramethyl-[1,1'- diphenyl]-2,2'-diol by NMR and MS data. The production of this compound was enhanced with the increment of cultural time. Transcriptome sequencing analysis showed that the highest upregulated genes under oligotrophic condition were glycosidase, TraR/DksA C4-type zinc finger protein and ribonuclease encoding genes, while the expression of a MarR family transcriptional regulator was most significantly decreased under oligotrophic condition. The results indicate that oligotrophic culture is an effective method for altering the secondary metabolism of Streptomyces.
Project description:In order to define the impact of phosphate (Pi) availability on cellular metabolism the project aimed to perform a comparative analysis of the proteomes of two Streptomyces strains with different abilities to produce antibiotics, S. coelicolor and S. lividans as well as of the pptA mutant of S. lividans, grown low (1mM) and high (5mM) phosphate (Pi) availability conditions. Interestingly, in contrast to most Streptomyces species, S. coelicolor produces more antibiotics in Pi proficiency than in Pi limitation, S. lividans does not produce antibiotics in any Pi conditions and the pptA mutant produces antibiotics only in Pi limitation. This in-depth proteomic comparison of three Streptomyces strains (S. coelicolor, S. lividans wt and pptA mutant), in different growth conditions (time and Pi concentration in the medium) was performed on four biological replicates. Protein abundance changes were determined using two label-free mass spectrometry based-quantification methods: spectral count (SC) and MS1 ion intensities named XIC (for eXtracted Ion Current). Our proteomic data reveal for the first time, the impact of Pi availability on the abundance of approximately 4000 proteins of these Streptomyces strains with different abilities to produce antibiotics. The most striking feature differentiating these strains was the much higher abundance of enzymes of the respiratory chain in both phosphate conditions in S. coelicolor compared to the S. lividans strains.
Project description:This project aims to discover novel bioactive compounds from Streptomyces isolated from the rhizosphere from wild medicinal plants from Hamedan province, Iran. Proteomics is used to assist in discovery and characterization of the compounds. Streptomyces isolates are grown on ISP-4 medium for three days, proteins were extracted and analysed by shotgun proteomics.
Project description:The objective of this study was to evaluate the effect of extracellular culture medium of Streptomyces AGN23, a strain inducing disease resistance in plants, on the Arabidopsis transcriptome.
Project description:This study aimed to characterize the non-targeted metabolite profile of Streptomyces sp. LZZY-S40 to explore its potential for producing bioactive secondary metabolites. The strain LZZY-S40 was cultivated on ISP 3 agar plates at 28°C for 7 days. Spores were harvested and inoculated into 250 mL Erlenmeyer flasks containing 50 mL of seed medium (pH 7.2–7.4) for 48 hours at 28°C with shaking at 250 rpm. The seed culture (8% v/v) was then transferred into fermentation medium and incubated under the same conditions for 7 days. After fermentation, the culture broth was centrifuged at 12,000 rpm for 10 minutes to separate the supernatant from the mycelial biomass. The supernatant was extracted three times with ethyl acetate, and the mycelial pellet was extracted with methanol. Organic extracts were concentrated under reduced pressure at temperatures below 40°C. The combined crude extracts were analyzed by untargeted liquid chromatography–mass spectrometry (LC–MS). Data were acquired in both positive and negative ionization modes across an m/z range of 50-1200, with chromatographic separation achieved using a gradient elution program at a flow rate of 400 μL/min and a column temperature of 40°C.
Project description:Gene expression in Streptomyces turgidiscabies pathogenicity island was studied after transferring cells into thaxtomin A inducing medium OBB using Agilent 60mer oligonucleotide array with probes designed for S.scabies and S.turgidiscabies. Gene expression study was used to confirm results of gene prediction.