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FEBS Open Bio Dec 2023Amino acid-based surfactants are valuable compounds for cosmetic formulations. The chemical synthesis of acyl amino acids is conventionally performed by the...
Amino acid-based surfactants are valuable compounds for cosmetic formulations. The chemical synthesis of acyl amino acids is conventionally performed by the Schotten-Baumann reaction using fatty acyl chlorides, but aminoacylases have also been investigated for use in biocatalytic synthesis with free fatty acids. Aminoacylases and their properties are diverse; they belong to different peptidase families and show differences in substrate specificity and biocatalytic potential. Bacterial aminoacylases capable of synthesis have been isolated from Burkholderia, Mycolicibacterium, and Streptomyces. Although several proteases and peptidases from S. griseus have been described, no aminoacylases from this species have been identified yet. In this study, we investigated two novel enzymes produced by S. griseus DSM 40236 . We identified and cloned the respective genes and recombinantly expressed an α-aminoacylase (EC3.5.1.14), designated SgAA, and an ε-lysine acylase (EC3.5.1.17), designated SgELA, in S. lividans TK23. The purified aminoacylase SgAA was biochemically characterized, focusing on its hydrolytic activity to determine temperature- and pH optima and stabilities. The aminoacylase could hydrolyze various acetyl amino acids at the N -position with a broad specificity regarding the sidechain. Substrates with longer acyl chains, like lauroyl amino acids, were hydrolyzed to a lesser extent. Purified aminoacylase SgELA specific for the hydrolysis of N -acetyl-l-lysine was unstable and lost its enzymatic activity upon storage for a longer period but could initially be characterized. The pH optimum of SgELA was pH 8.0. While synthesis of acyl amino acids was not observed with SgELA, SgAA catalyzed the synthesis of lauroyl-methionine.
Topics: Humans; Streptomyces lividans; Streptomyces griseus; Streptomyces; Amino Acids; Lysine
PubMed: 37879963
DOI: 10.1002/2211-5463.13723 -
Molecules (Basel, Switzerland) Jun 2021This study aimed to produce, purify, structurally elucidate, and explore the biological activities of metabolites produced by isolate KJ623766, a recovered soil...
BACKGROUND
This study aimed to produce, purify, structurally elucidate, and explore the biological activities of metabolites produced by isolate KJ623766, a recovered soil bacterium previously screened in our lab that showed promising cytotoxic activities against various cancer cell lines.
METHODS
Production of cytotoxic metabolites from isolate KJ623766 was carried out in a 14L laboratory fermenter under specified optimum conditions. Using a 3-(4,5-dimethylthazol-2-yl)-2,5-diphenyl tetrazolium-bromide assay, the cytotoxic activity of the ethyl acetate extract against 2 and cancer cell lines was determined. Bioassay-guided fractionation of the ethyl acetate extract using different chromatographic techniques was used for cytotoxic metabolite purification. Chemical structures of the purified metabolites were identified using mass, 1D, and 2D NMR spectroscopic analysis.
RESULTS
Bioassay-guided fractionation of the ethyl acetate extract led to the purification of two cytotoxic metabolites, R1 and R2, of reproducible amounts of 5 and 1.5 mg/L, respectively. The structures of R1 and R2 metabolites were identified as β- and γ-rhodomycinone with CD of 6.3, 9.45, 64.8 and 9.11, 9.35, 67.3 µg/mL against 2, and cell lines, respectively. Values were comparable to those of the positive control doxorubicin.
CONCLUSIONS
This is the first report about the production of β- and γ-rhodomycinone, two important scaffolds for synthesis of anticancer drugs, from .
Topics: Animals; Anthracyclines; Antibiotics, Antineoplastic; Biological Products; Caco-2 Cells; Chlorocebus aethiops; HeLa Cells; Humans; Streptomyces griseus; Vero Cells
PubMed: 34209170
DOI: 10.3390/molecules26134009 -
The Journal of Biological Chemistry May 2022The sugars streptose and dihydrohydroxystreptose (DHHS) are unique to the bacteria Streptomyces griseus and Coxiella burnetii, respectively. Streptose forms the central...
The sugars streptose and dihydrohydroxystreptose (DHHS) are unique to the bacteria Streptomyces griseus and Coxiella burnetii, respectively. Streptose forms the central moiety of the antibiotic streptomycin, while DHHS is found in the O-antigen of the zoonotic pathogen C. burnetii. Biosynthesis of these sugars has been proposed to follow a similar path to that of TDP-rhamnose, catalyzed by the enzymes RmlA, RmlB, RmlC, and RmlD, but the exact mechanism is unclear. Streptose and DHHS biosynthesis unusually requires a ring contraction step that could be performed by orthologs of RmlC or RmlD. Genome sequencing of S. griseus and C. burnetii has identified StrM and CBU1838 proteins as RmlC orthologs in these respective species. Here, we demonstrate that both enzymes can perform the RmlC 3'',5'' double epimerization activity necessary to support TDP-rhamnose biosynthesis in vivo. This is consistent with the ring contraction step being performed on a double epimerized substrate. We further demonstrate that proton exchange is faster at the 3''-position than the 5''-position, in contrast to a previously studied ortholog. We additionally solved the crystal structures of CBU1838 and StrM in complex with TDP and show that they form an active site highly similar to those of the previously characterized enzymes RmlC, EvaD, and ChmJ. These results support the hypothesis that streptose and DHHS are biosynthesized using the TDP pathway and that an RmlD paralog most likely performs ring contraction following double epimerization. This work will support the elucidation of the full pathways for biosynthesis of these unique sugars.
Topics: Antigens, Bacterial; Carbohydrate Epimerases; Coxiella burnetii; Nucleoside Diphosphate Sugars; Streptomyces griseus; Thymine Nucleotides
PubMed: 35398092
DOI: 10.1016/j.jbc.2022.101903 -
World Journal of Microbiology &... Nov 2022Following the discovery of streptomycin from Streptomyces griseus in the 1940s by Selman Waksman and colleagues, aminoglycosides were first used to treat tuberculosis... (Review)
Review
Following the discovery of streptomycin from Streptomyces griseus in the 1940s by Selman Waksman and colleagues, aminoglycosides were first used to treat tuberculosis and then numerous derivatives have since been used to combat a wide variety of bacterial infections. These bactericidal antibiotics were used as first-line treatments for several decades but were largely replaced by ß-lactams and fluoroquinolones in the 1980s, although widespread emergence of antibiotic-resistance has led to renewed interest in aminoglycosides. The primary site of action for aminoglycosides is the 30 S ribosomal subunit where they disrupt protein translation, which contributes to widespread cellular damage through a number of secondary effects including rapid uptake of aminoglycosides via elevated proton-motive force (PMF), membrane damage and breakdown, oxidative stress, and hyperpolarisation of the membrane. Several factors associated with aminoglycoside entry have been shown to impact upon bacterial killing, and more recent work has revealed a complex relationship between metabolic states and the efficacy of different aminoglycosides. Hence, it is imperative to consider the environmental conditions and bacterial physiology and how this can impact upon aminoglycoside entry and potency. This mini-review seeks to discuss recent advances in this area and how this might affect the future use of aminoglycosides.
Topics: Aminoglycosides; Anti-Bacterial Agents; Drug Resistance, Microbial; Bacteria; Streptomyces griseus
PubMed: 36350431
DOI: 10.1007/s11274-022-03445-8 -
Plants (Basel, Switzerland) Oct 2022With this research, we aimed to determine the impact of grafting and rootstock seed treated with ( () or shikimic acid (SA) at a 60 ppm concentration on tomato ( L.)...
With this research, we aimed to determine the impact of grafting and rootstock seed treated with ( () or shikimic acid (SA) at a 60 ppm concentration on tomato ( L.) production grown under low-temperature conditions. Two open-field trials were performed during both winter seasons of 2020 and 2021 at the Experimental Farm, Faculty of Agriculture, Cairo University, Giza, Egypt. A tomato cultivar (Peto 86) was used as a scion and two tomato phenotypes were employed as rootstocks ( L and GS hybrid), as well as self-grafted as a control. Effects of sub-optimal temperature on vegetative growth, yield, and fruit quality were tested. The results indicate that, under cold stress, rootstock seed priming, especially with , enhanced plant growth, total yield, and fruit quality properties. GS hybrid rootstock was more effective than that of rootstock in terms of mitigating the negative effect of cold stress. GS hybrid, inoculated with increased the total yield per plant by 10.5% and 5.7% in the first and second seasons, respectively. Higher levels of GA3 and mineral content were noticed in leaves that were grafted and treated with compared to the control treatment. Additionally, the great enhancing effects of all anatomical features of tomato plants were recorded with GS hybrid rootstock, inoculated by . These results prove that grafting on GS hybrid rootstock treated with is a potential choice to alleviate the cold stress of commercial tomato varieties.
PubMed: 36365275
DOI: 10.3390/plants11212822 -
Scientific Reports Jul 2015Marine actinobacteria are known to be a rich source for novel metabolites with diverse biological activities. In this study, a potential extracellular L-asparaginase was...
Marine actinobacteria are known to be a rich source for novel metabolites with diverse biological activities. In this study, a potential extracellular L-asparaginase was characterised from the Streptomyces griseus NIOT-VKMA29. Box-Behnken based optimization was used to determine the culture medium components to enhance the L-asparaginase production. pH, starch, yeast extract and L-asparagine has a direct correlation for enzyme production with a maximum yield of 56.78 IU mL(-1). A verification experiment was performed to validate the experiment and more than 99% validity was established. L-Asparaginase biosynthesis gene (ansA) from Streptomyces griseus NIOT-VKMA29 was heterologously expressed in Escherichia coli M15 and the enzyme production was increased threefold (123 IU mL(-1)) over the native strain. The ansA gene sequences reported in this study encloses several base substitutions with that of reported sequences in GenBank, resulting in altered amino acid sequences of the translated protein.
Topics: Asparaginase; Bacterial Proteins; Escherichia coli; Recombinant Proteins; Streptomyces griseus
PubMed: 26206135
DOI: 10.1038/srep12404 -
Microbiology Resource Announcements Jan 2024We present the complete genome sequences of two viruses with siphovirus morphology, isolated from soils collected in Southwestern Indiana using the host . Spelly is a...
We present the complete genome sequences of two viruses with siphovirus morphology, isolated from soils collected in Southwestern Indiana using the host . Spelly is a BE2 cluster phage with a 131,347-bp genome. Phredrick is a BK1 cluster phage with a 128,873-bp genome.
PubMed: 38112473
DOI: 10.1128/mra.01049-23 -
Journal of Bacteriology Aug 2009Small noncoding RNAs (sRNAs) have been shown to control diverse cellular processes in prokaryotes. To identify and characterize novel bacterial sRNAs, a gram-positive,...
Small noncoding RNAs (sRNAs) have been shown to control diverse cellular processes in prokaryotes. To identify and characterize novel bacterial sRNAs, a gram-positive, soil-inhabiting, filamentous bacterium, Streptomyces griseus, was examined, on the assumption that Streptomyces should express sRNAs as important regulators of morphological and physiological differentiation. By bioinformatics investigation, 54 sRNA candidates, which were encoded on intergenic regions of the S. griseus chromosome and were highly conserved in those of both Streptomyces coelicolor A3(2) and Streptomyces avermitilis, were selected. Of these 54 sRNA candidates, 17 transcripts were detected by Northern blot analysis of the total RNAs isolated from cells grown on solid medium. Then, the direction of transcription of each sRNA candidate gene was determined by S1 nuclease mapping, followed by exclusion of four sRNA candidates that were considered riboswitches of their downstream open reading frames (ORFs). Finally, a further sRNA candidate was excluded because it was cotranscribed with the upstream ORF determined by reverse transcription-PCR. Thus, 12 sRNAs ranging in size from 40 to 300 nucleotides were identified in S. griseus. Seven of them were apparently transcribed in a growth phase-dependent manner. Furthermore, of the 12 sRNAs, the expression profiles of 7 were significantly influenced by a mutation of adpA, which encodes the central transcriptional regulator of the A-factor regulatory cascade involved in both morphological differentiation and secondary metabolism in S. griseus. However, disruption of all 12 sRNA genes showed no detectable phenotypic changes; all the disruptants grew and formed aerial mycelium and spores with the same time course as the wild-type strain on various media and produced streptomycin similarly to the wild-type strain.
Topics: Bacterial Proteins; Blotting, Northern; Electrophoretic Mobility Shift Assay; Gene Expression Regulation, Bacterial; RNA, Untranslated; Reverse Transcriptase Polymerase Chain Reaction; Streptomyces griseus
PubMed: 19465662
DOI: 10.1128/JB.00087-09 -
The Plant Pathology Journal Apr 2020S4-7, a well-characterized keystone taxon among strawberry microbial communities, shows exceptional disease-preventing ability. The whole-genome sequence, functional...
S4-7, a well-characterized keystone taxon among strawberry microbial communities, shows exceptional disease-preventing ability. The whole-genome sequence, functional genes, and bioactive secondary metabolites of the strain have been described in previous studies. However, proteomics studies of not only the S4-7 strain, but also the genus as a whole, remain limited to date. Therefore, in the present study, we created a proteomics reference map for S4-7. Additionally, analysis of differentially expressed proteins was performed against a 2 mutant, which was deficient in spore chain development and did not express an antifungal activity-regulatory transcription factor. We believe that our data provide a foundation for further in-depth studies of functional keystone taxa of the phytobiome and elucidation of the mechanisms underlying plant-microbe interactions, es-pecially those involving the genus.
PubMed: 32296298
DOI: 10.5423/PPJ.NT.02.2020.0037