Search

Halogenated phenazine meroterpenoids are a structurally unusual family of marine actinobacterial natural products that exhibit antibiotic, antibiofilm, and cytotoxic bioactivities. Despite a lack of established phenazine halogenation biochemistry, genomic analysis of Streptomyces sp. CNZ-289, a prolific lavanducyanin and C2-halogenated derivative producer, suggested the involvement of vanadium-dependent haloperoxidases. We subsequently discovered lavanducyanin halogenase (LvcH), characterized it in vitro as a regioselective vanadium-dependent chloroperoxidase, and applied it in late-stage chemoenzymatic synthesis.

Respirantins are 18-membered antimycin-type depsipeptides produced by Streptomyces sp. and Kitasatospora sp. These compounds have shown extraordinary anticancer activities against a panel of cancer cell lines with nanomolar levels of IC50 values. However, further investigation has been impeded by the low titers of the natural producers and the challenging chemical synthesis due to their structural complexity. The biosynthetic gene cluster (BGC) of respirantin was previously proposed based on a bioinformatic comparison of the four members of antimycin-type depsipeptides. In this study, we report the first successful reconstitution of respirantin in Streptomyces albus using a synthetic BGC. This heterologous system serves as an accessible platform for the production and diversification of respirantins. Through polyketide synthase pathway engineering, biocatalysis, and chemical derivatization, we generated nine respirantin compounds, including six new derivatives. Cytotoxicity screening against human MCF-7 and Hela cancer cell lines revealed a unique biphasic dose-response profile of respirantin. Furthermore, a structure-activity relationship study has elucidated the essential functional groups that contribute to its remarkable cytotoxicity. This work paves the way for respirantin-based anticancer drug discovery and development.

Marine-derived Streptomyces have long been recognized as a source of novel, pharmaceutically relevant natural products. Among these bacteria, the MAR4 clade within the genus Streptomyces has been identified as metabolically rich, yielding over 93 different compounds to date. MAR4 strains are particularly noteworthy for the production of halogenated hybrid isoprenoid natural products, a relatively rare class of bacterial metabolites that possess a wide range of biological activities. MAR4 genomes are enriched in vanadium haloperoxidase and prenyltransferase genes, thus accounting for the production of these compounds. Functional characterization of the enzymes encoded in MAR4 genomes has advanced our understanding of halogenated, hybrid isoprenoid biosynthesis. Despite the exceptional biosynthetic capabilities of MAR4 bacteria, the large body of research they have stimulated has yet to be compiled. Here we review 35 years of natural product research on MAR4 strains and update the molecular diversity of this unique group of bacteria.

Background: SepIVA has been reported to be an essential septation factor in Mycolicibacterium smegmatis and Mycobacterium tuberculosis. It is a coiled-coil protein with similarity to DivIVA, a protein necessary for polar growth in members of the phylum Actinomycetota. Orthologues of SepIVA are broadly distributed among actinomycetes, including in Streptomyces spp. Results: To clarify the role of SepIVA and its potential involvement in cell division in streptomycetes, we generated sepIVA deletion mutants in Streptomyces venezuelae and found that sepIVA is dispensable for growth, cell division and sporulation. Further, mNeonGreen-SepIVA fusion protein did not localize at division septa, and we found no evidence of involvement of SepIVA in cell division. Instead, mNeonGreen-SepIVA was accumulated at the tips of growing vegetative hyphae in ways reminiscent of the apical localization of polarisome components like DivIVA. Bacterial two-hybrid system analyses revealed an interaction between SepIVA and DivIVA. The results indicate that SepIVA is associated with polar growth. However, no phenotypic effects of sepIVA deletion could be detected, and no evidence was observed of redundancy with the other DivIVA-like coiled-coil proteins Scy and FilP that are also associated with apical growth in streptomycetes. Conclusions: We conclude that S. venezuelae SepIVA, in contrast to the situation in mycobacteria, is dispensable for growth and viability. The results suggest that it is associated with polar growth rather than septum formation. [ABSTRACT FROM AUTHOR]

Background: Uricase is a bio-drug used to reduce urate accumulation in gout disease. Thus, there is a continuous demand for screening soil samples derived from a variety of different sources in order to isolate a strain that possesses a high potential for producing uricase. Methods: Streptomyces sp. strain NEAE-5 demonstrated a significant capacity for uricase production was identified based on the physiological, morphological and biochemical characteristics, as well as 16S rDNA sequencing analysis. Using a Plackett–Burman statistical design, the impact of eighteen process factors on uricase production by Streptomyces griseorubens strain NEAE-5 was investigated. Using central composite design, the most important variables that had a favourable positive impact on uricase production by Streptomyces griseorubens strain NEAE-5 were further optimized. Results: It is clear that the morphological and chemotaxonomic features of Streptomyces sp. strain NEAE-5 are typical for the Streptomyces genus. Phylogenetic analysis indicated that Streptomyces sp. strain NEAE-5 belongs to the genus Streptomyces and closely related to Streptomyces griseorubens which it has a 95–96% identity in 16S rDNA gene sequencing. Accordingly, the strain is proposed to be identified as Streptomyces griseorubens strain NEAE-5. The three factors that had the significant positive impacts on uricase production were uric acid, hypoxanthine, and yeast extract. As a result, the best conditions for achieving the highest experimental uricase production by Streptomyces griseorubens strain NEAE-5 after central composite design were (g/L): uric acid 6.96, glycerol 5, hypoxanthine 5.51, MgSO4.7H2O 0.1, KNO3 2, CaCl2 0.5, K2HPO4 0.5, NaCl 0.5, yeast extract 1.08. In addition, the period of incubation is seven days, pH 7.5 and 37 °C with an inoculum size of 2 mL (105 cfu/mL) /100 mL medium. Conclusions: After optimization, the obtained uricase activity was 120.35 U/mL, indicating that the Streptomyces griseorubens strain NEAE-5 is a potent uricase producer and that the statistical approach used for optimization was appropriate. [ABSTRACT FROM AUTHOR]

Physicochemical properties of the bacterial surface are involved in several interfacial phenomena, such as microbial adhesion. Ecology Soil salinity is a crucial parameter for the distribution of Streptomyces. The objective of this study was to investigate the impact of NaCl on the hydrophobicity and electron donor/acceptor characteristics of the cell surface of fourteen Streptomyces strains isolated from soils of the Beni Amir region (Morocco) with different salinities. The physicochemical properties of the surface were evaluated using the MATS (microbial adhesion to solvents) method at two concentrations of NaCl (0.1 M and 1.2 M). The results obtained show a significant change from hydrophilic to hydrophobic character. In particular, the Streptomyces lilaceus A53 strain showed the lowest variation (4.21%). On the other hand, the Streptomyces albogriseolus A65 strain presented the greatest variation (86.15%). These changes were observed when the salt concentration increased significantly from 0.1 M to 1.2 M NaCl. The electron donor/acceptor character systematically decreases and even becomes null for the majority of strains. Furthermore, a strong correlation between cell surface hydrophobicity and salinity of the original soil was observed with MATS at 1.2 M NaCl. This study highlighted the crucial importance of the NaCl concentration in the modulation of the physicochemical properties of the surface of Streptomyces bacteria. [ABSTRACT FROM AUTHOR]

This study aimed to isolate Streptomyces sp. from Moroccan terrestrial ecosystems and identify bioactive compounds through GC–MS analysis. Antimicrobial activity was assessed against various pathogenic microorganisms including Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853, Escherichia coli ATCC 25922 and Candida albicans ATCC 60193, and multi-drug resistant strains comprising Listeria monocytogenes, Klebsiella pneumoniae 19K 929, Proteus sp. 19K1313, Klebsiella pneumoniae 20B1572, Proteus vulgaris 16C1737, and Klebsiella pneumoniae 20B1572. Based on the results of the gene sequencing of gene 16S rRNA and phylogenetic analysis, the E2 isolate belongs to the genus Streptomyces with the highest degree of resemblance (97.51%) to the Streptomyces africanus strain NBRC 101005 (NR_112600.1). The isolate exhibited broad-spectrum antibacterial activity, with maximum efficacy against Klebsiella pneumoniae 20B1572 indicated by an inhibition zone diameter of 22.5 ± 0.71mm and a minimum inhibitory concentration (MIC) of 0.0625 mg/mL. The in vitro antioxidant potential of E2 strain was determined through screening of its ethyl acetate extract against sets of antioxidant assays. The results were indicative of E2 strain displaying strong antioxidant activity against ABTS, DPPH free radicals, and FRAP. Furthermore, there was a high significant correlation (p < 0.0001) between the total phenolic and flavonoid content and antioxidant activities. The GC–MS analysis of the extract identified six volatile compounds, with Eugenol (96%) and Maltol (93%) being the most prominent. Additionally, the HPLC–UV/vis analysis revealed six phenolic compounds: gallic acid, chlorogenic acid, vanillic acid, trans-ferulic acid, ellagic acid, and cinnamic acid. Overall, the study highlights Streptomyces sp. strain E2 as a potential source of potent antimicrobial and antioxidant metabolites, offering promise in addressing antibiotic resistance and oxidative stress-related conditions. [ABSTRACT FROM AUTHOR]

Streptomyces produces diverse secondary metabolites of biopharmaceutical importance, yet the rate of biosynthesis of these metabolites is often hampered by complex transcriptional regulation. Therefore, a fundamental understanding of transcriptional regulation in Streptomyces is key to fully harness its genetic potential. Here, independent component analysis (ICA) of 454 high‐quality gene expression profiles of the model species Streptomyces coelicolor is performed, of which 249 profiles are newly generated for S. coelicolor cultivated on 20 different carbon sources and 64 engineered strains with overexpressed sigma factors. ICA of the transcriptome dataset reveals 117 independently modulated groups of genes (iModulons), which account for 81.6% of the variance in the dataset. The genes in each iModulon are involved in specific cellular responses, which are often transcriptionally controlled by specific regulators. Also, iModulons accurately predict 25 secondary metabolite biosynthetic gene clusters encoded in the genome. This systemic analysis leads to reveal the functions of previously uncharacterized genes, putative regulons for 40 transcriptional regulators, including 30 sigma factors, and regulation of secondary metabolism via phosphate‐ and iron‐dependent mechanisms in S. coelicolor. ICA of large transcriptomic datasets thus enlightens a new and fundamental understanding of transcriptional regulation of secondary metabolite synthesis along with interconnected metabolic processes in Streptomyces. [ABSTRACT FROM AUTHOR]

The thermo-solvent-tolerant lipase-producing actinomycete, Streptomyces sp. A3301, was utilized as a biocatalyst for poly (lactic acid) or PLA polymerization. The study aimed to optimize lipase immobilization conditions, characterize the immobilized lipase and apply it for PLA polymerization. The results showed using a sponge as the immobilizing matrix was the most effective method, achieving a maximum activity of 277 U/g of sponge. The optimal sponge size was determined to be 0.125 cm³ and pre-soaking the sponge in 0.1 M phosphate buffer at pH 7.0 for 24 h before use proved advantageous. Immobilization significantly enhanced the thermo-stability of the enzyme, with a relative activity ranging from 140 to 190% within the temperature range of 30 to 60 °C. In contrast, the crude lipase exhibited thermo-stability only within the 30 - 50 °C range. The immobilized lipase demonstrated stability under PLA polymerization conditions, which involved a reaction mixture containing toluene and lactic acid and performed at 60 °C for 8 h. The immobilized lipase maintained its activity under this condition for 5 h, retaining a relative activity of 230%, which was 1.2 times higher than the activity of the crude lipase. When the immobilized lipase was used in PLA polymerization, the resulting PLA product exhibited a molecular weight of 5,333 ± 0.02 Da, and the degree of polymerization was approximately 72. These findings underscore the potential of the immobilization technique to enhance lipase activity for PLA polymerization. [ABSTRACT FROM AUTHOR]

Actinobacterial species are mostly thought to be nonmotile. Recent studies have revealed the degenerate evolution of flagella in this phylum and different flagellar rod compositions from the classical model. Moreover, flagella-independent motility by various means has been reported in Streptomyces spp. and Mycobacterium spp., but the underlying mechanisms remain elusive. [ABSTRACT FROM AUTHOR]

Watermelon (Citrullus lanatus Thumb) is a significant vegetable crop globally and is prone to anthracnose disease caused by Colletotrichum spp. The use of chemical fungicides for managing crop diseases has adverse impacts on the environment and human health. Therefore, it is important to use biocontrol agents for disease management. In this study, streptomycetes were assessed as a potential biocontrol agent against anthracnose pathogen, Colletotrichum orbiculare WEC2. The potent Streptomyces sp. isolate WAB2 was isolated from healthy watermelon plants and showed remarkable antifungal efficacy against C. orbiculare WEC2. Rigorous assays confirmed its performance against C. orbiculare WEC2. Bioactive compounds produced by Streptomyces sp. isolate WAB2 were identified using Gas Chromatography–Mass Spectrometry (GC–MS). An SEM study indicated the parasitic action of Streptomyces sp. isolate WAB2 against the pathogen. The formulated talc-based product of Streptomyces sp. isolate WAB2 was effective when applied as a seed treatment + foliar spray against C. orbiculare WEC2 under pot culture conditions. These treatments reduced the disease incidence (31.81%) and exhibited a higher disease reduction (52.56%), compared to the untreated control. Therefore, treatment with talc-based Streptomyces sp. isolate WAB2 is a sustainable solution for anthracnose disease management, and it facilitates eco-friendly crop protection practices against the disease in watermelon. RESEARCH HIGHLIGHTS: Isolated streptomycete strains from healthy watermelon plants and screened for its efficacy against the pathogen, Colletotrichum orbiculare WEC2. Streptomyces sp. isolate WAB2 showed significant antifungal activity against the pathogen. Bio-active compounds produced by Streptomyces sp. isolate WAB2 was identified by Gas Chromatography–Mass Spectrometry (GC–MS). Using scanning electron microscopy (SEM), the parasitic activity of Streptomyces sp. isolate WAB2 against the pathogen was detected. Developed a talc-based formulation of Streptomyces sp. isolate WAB2 and tested it as a seed treatment plus foliar spray, effectively reducing the disease incidence. Graphical abstract illustrating the isolation and characterisation of Streptomyces sp. isolate WAB2 for the management of anthracnose pathogen (Colletotrichum orbiculare WEC2) in Watermelon. Antagonistic potential of Streptomyces sp. isolate WAB2 against Colletotrichum orbiculare: [ABSTRACT FROM AUTHOR]

Two undescribed frenolicins H and I (1 and 2) along with six previously described frenolicin analogues [frenolicins A (3), B (4), UCF76-B (5), E – G (6 – 8)] and two anthraquinones [3,8-dihydroxy-1-propylanthraquinone-2-carboxylic acid (9) and 3,8-dihydroxy-1-propylanthraquinone (10)] were isolated from a longkong bark eating caterpillar-derived Streptomyces sp. TBRC17107. The chemical structures were determined by NMR spectroscopic information and HRESIMS data. Frenolicins H (1) and I (2) showed weak cytotoxicity against malignant and non-malignant cells. Frenolicins A (3) and B (4) showed antimalarial activity against Plasmodium falciparum (IC50 17.4 and 1.37 μM), antibacterial activity against Bacillus cereus and Staphylococcus aureus (MIC 50.0 and 0.20 μg/mL). Only frenolicin B had anti-plant pathogenic fungal activity against Collectotrichum acutatum and Alternaria brassicicola with MIC values of MIC 1.56 and 6.25 μg/mL, respectively. Frenolicins A and G possessed anti-Mycobacterium tuberculosis with equal MICs of 25.0 μg/mL. [ABSTRACT FROM AUTHOR]

Iron homeostasis is vital for normal physiology, but in the majority of circumstances, like iron overload, this equilibrium is upset leading to free iron in the plasma. This condition with excess iron is known as hemochromatosis, which has been linked to many side effects, including cancer and liver cirrhosis. The current research aimed to investigate active molecules from Streptomyces sp. isolated from the extreme environment of Bahawalpur deserts. The strain was characterized using 16 S rRNA sequencing. Chemical analysis of the ethyl acetate cure extract revealed the presence of phenols, flavonoids, alkaloids, and tannins. Multiple ultraviolet (UV) active metabolites that were essential for the stated pharmacological activities were also demonstrated by thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC). Additionally, Gas chromatography/mass spectrometry (GC-MS) analysis revealed the primary constituents of the extract to compose of phenol and ester compounds. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay was used to assess the extract's antioxidant capacity, and the results showed a good half-maximal inhibitory concentration (IC50) value of 0.034 µg/mL in comparison to the positive control ascorbic acid's 0.12 µg/mL. In addition, iron chelation activity of extract showed significant chelation potential at 250 and 125 µg/mL, while 62.5 µg/mL showed only mild chelation of the ferrous ion using ethylene diamine tetra acetic acid (EDTA) as a positive control. Likewise, the extract's cytotoxicity was analyzed through 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay using varying concentrations of the extract and showed 51% cytotoxicity at 350 µg/mL and 65% inhibition of cell growth at 700 µg/mL, respectively. The bioactive compounds from Streptomyces sp. demonstrated strong antioxidant and iron chelating potentials and can prolong the cell survival in extreme environment. [ABSTRACT FROM AUTHOR]

The decline of new antibiotics and the emergence of multidrug resistance in pathogens necessitates a revisit of strategies used for lead compound discovery. This study proposes to induce the production of bioactive compounds with sub-lethal concentrations of silver nanoparticles (Ag-NPs). A total of Forty-two Actinobacteria isolates from four Saudi soil samples were grown with and without sub-lethal concentration of Ag-NPs (50 µg ml−1). The spent broth grown with Ag-NPs, or without Ag-NPs were screened for antimicrobial activity against four bacteria. Interestingly, out of 42 strains, broths of three strains grown with sub-lethal concentration of Ag-NPs exhibit antimicrobial activity against Staphylococcus aureus and Micrococcus luteus. Among these, two strains S4-4 and S4-21 identified as Streptomyces labedae and Streptomyces tirandamycinicus based on 16S rRNA gene sequence were selected for detailed study. The change in the secondary metabolites profile in the presence of Ag-NPs was evaluated using GC–MS and LC–MS analyses. Butanol extracts of spent broth grown with Ag-NPs exhibit strong antimicrobial activity against M. luteus and S. aureus. While the extracts of the controls with the same concentration of Ag-NPs do not show any activity. GC-analysis revealed a clear change in the secondary metabolite profile when grown with Ag-NPs. Similarly, the LC–MS patterns also differ significantly. Results of this study, strongly suggest that sub-lethal concentrations of Ag-NPs influence the production of secondary metabolites by Streptomyces. Besides, LC–MS results identified possible secondary metabolites, associated with oxidative stress and antimicrobial activities. This strategy can be used to possibly induce cryptic biosynthetic gene clusters for the discovery of new lead compounds. [ABSTRACT FROM AUTHOR]

During the course of development plants form tight interactions with microorganisms inhabiting their root zone. In turn, rhizosphere bacteria, in particular members of the phylum Actinomycetota, positively influence the host plant by increasing access to essential nutrients and controlling the pathogenic microorganism's population. Herein, we report the characterisation of the rhizosphere associated actinobacteria community of Phyllostachys viridiglaucescens growing in the Nikitsky Botanical Garden (Crimean Peninsula, Ukraine). The overall composition of the bacterial community was elucidated by 16S rRNA gene amplicon sequencing followed by isolation of culturable microorganisms with the focus on actinomycetes. The metagenomic approach revealed that the representatives of phylum Actinomycetota (57.1%), Pseudomonadota (20.0%), and Acidobacteriota (12.2%) were dominating in the studied microbiome with Ilumatobacter (phylum Actinomycetota) (13.1%) being the dominant genus. Furthermore, a total of 159 actinomycete isolates, belonging to eight genera of Streptomyces, Micromonospora, Nonomuraea, Arthrobacter, Actinomadura, Kribbella, Cellulosimicrobium, and Mumia, were recovered from P. viridiglaucescens rhizosphere. The isolated species were tested for antimicrobial activity. 64% of isolates were active against at least one bacterial test-culture and 7.5% against fungal test culture. In overall, the rhizosphere bacterial communities act as a great source of actinobacterial diversity with the high potential for production of new bioactive compounds. [ABSTRACT FROM AUTHOR]

Flavonoids are a large and important group of phytochemicals with a great variety of bioactivities. The addition of methyl groups during biosynthesis of flavonoids and other polyphenols enhances their bioactivities and increases their stability. In a previous study of our research group, we detected a novel flavonoid O-methyltransferase activity in Streptomyces albidoflavus J1074, which led to the heterologous biosynthesis of homohesperetin from hesperetin in feeding cultures. In this study, we identify the O-methyltransferase responsible for the generation of this methylated flavonoid through the construction of a knockout mutant of the gene XNR_0417, which was selected after a blast analysis using the sequence of a caffeic acid 3′-O-methyltransferase from Zea mays against the genome of S. albidoflavus J1074. This mutant strain, S. albidoflavus ∆XNR_0417, was no longer able to produce homohesperetin after hesperetin feeding. Subsequently, we carried out a genetic complementation of the mutant strain in order to confirm that the enzyme encoded by XNR_0417 is responsible for the observed O-methyltransferase activity. This new strain, S. albidoflavus SP43-XNR_0417, was able to produce not only homohesperetin from hesperetin, but also different mono-, di-, tri- and tetra-methylated derivatives on other flavanones, flavones and stilbenes, revealing a broad substrate flexibility. Additionally, in vitro experiments were conducted using the purified enzyme on the substrates previously tested in vivo, demonstrating doubtless the capability of XNR_0417 to generate various methylated derivatives. [ABSTRACT FROM AUTHOR]

Streptomyces genus produces a large number of antibiotics, which are always synthesized by specific biosynthetic gene clusters (BGCs). To resist autotoxicity, transporters encoded by genes located within BGC occasionally pump antibiotic along with transporter encoded by gene located outside BGC. Daunorubicin is an anthracycline antibiotic biosynthesized by Streptomyces species, playing a crucial role in the treatment of leukaemia. In existing studies, only one two‐component ATP‐binding cassette (ABC) transporter, encoded by drrA1‐drrB1 (abbreviated as drrAB1) and located within the daunorubicin BGC, has been proven to extrude daunorubicin. In this work, two other two‐component ABC transporters, encoded by drrAB2 and drrAB3 and located outside the cluster, were found to play the complementary role in daunorubicin efflux in S. coeruleorubidus. Disruption of three drrABs resulted in a 94% decrease in daunorubicin production. Furthermore, drrAB2 is regulated by the TetR family regulator DrrR1, responding to the intracellular accumulation of daunorubicin and suggesting its role in stress response and self‐resistance. Although the homologues of DrrAB1 are only found in three anthracycline BGCs, the homologues of DrrAB2 and DrrAB3 are spread in many Streptomyces strains which do not contain any known anthracycline BGC. This indicates that DrrAB2 and DrrAB3 may recognize and extrude a broader range of substrates besides daunorubicin, thus playing a more extensive role in cellular detoxification. [ABSTRACT FROM AUTHOR]

Antibiotic resistance is a major problem in the control of infectious diseases. To overcome this obstacle, a common approach is to search for novel antimicrobial drugs from natural sources, especially microorganisms such as Streptomyces. Streptomyces has long been demonstrated to produce bioactive secondary metabolites with antimicrobial activities. The objectives of this research were to isolate and characterize Streptomyces strains from the deciduous dipterocarp forest soils in the University of Phayao, and to evaluate their metabolites for antibacterial activities. The results show that the morphological and physiological characteristics of PA5.6 are consistent with those of the genus Streptomyces. Furthermore, a comparative analysis of the whole genome sequence verified that PA5.6 represents a novel species within the genus Streptomyces. The genome sequence analysis of Streptomyces sp. PA5.6 revealed a total length of 9,146,340 bp, a G + C content of 71.1 %, and an N50 scaffold of 320,345 bp. The antiSMASH analysis revealed putative secondary metabolite Biosynthetic Gene Clusters (BGCs) involved in the biosynthesis of antimicrobial metabolites, including terpene, Type I PKS, NRPS and lassopeptide gene clusters. Functional prediction of these gene clusters indicates that they are involved in the biosynthesis of several antimicrobial-associated metabolites, such as albaflavenone, monensin and citrulassin D. Streptomyces sp. PA5.6 culture supernatant and its crude ethyl acetate extract exhibited antibacterial activity against several drug-resistant, bacterial pathogens, including S. aureus, E. faecalis, E. coli, P. aeruginosa, A. baumannii, MRSA, VREF, ESBL-E. coli and CRPA. The GC-MS analysis of the crude ethyl acetate extracts revealed benzeneacetic acid, benzeneacetic acid, 4-hydroxy and benzeneacetamide as major active compounds metabolites. In conclusion, Streptomyces sp. PA5.6 exhibited promising potential for producing bioactive compounds against bacterial pathogens. [ABSTRACT FROM AUTHOR]

Methionine (Met), a sulfur-containing amino acid, is essential for the underlying biological processes in living organisms. In addition to its importance as a starting building block for peptide chain elongation in protein biosynthesis, Met is a direct precursor of S-adenosyl-L-methionine, an indispensable methyl donor molecule in primary and secondary metabolism. Streptomyces bacteria are well known to produce diverse secondary metabolites, but many strains lack canonical Met pathway genes for L-homocysteine, a direct precursor of Met in bacteria, plants, and archaea. Here, we report the identification of a novel gene (metM) responsible for the Met biosynthesis in Streptomyces strains and demonstrate the catalytic function of the gene product, MetM. We further identified the metO gene, a downstream gene of metM, and showed that it encodes a sulfur-carrier protein (SCP). In in vitro analysis, MetO was found to play an important role in a sulfur donor by forming a thiocarboxylated SCP. Together with MetO (thiocarboxylate), MetM directly converted O-phospho-L-homoserine to L-homocysteine. O-Phospho-L-homoserine is also known as an intermediate for threonine biosynthesis in bacteria and plants, and MetM shares sequence homology with threonine synthase. Our findings thus revealed that MetM seizes O-phospho-L-homoserine from the threonine biosynthetic pathway and uses it as an intermediate of the Met biosynthesis to generate the sulfur-containing amino acid. Importantly, this MetM/MetO pathway is highly conserved in Streptomyces bacteria and distributed in other bacteria and archaea. IMPORTANCE Methionine (Met) is a sulfur-containing proteinogenic amino acid. Moreover, Met is a direct precursor of S-adenosyl-L-methionine, an indispensable molecule for expanding the structural diversity of natural products. Because Met and its derivatives benefit humans, the knowledge of Met biosynthesis is important as a basis for improving their fermentation. Streptomyces bacteria are well known to produce diverse and valuable natural products, but many strains lack canonical Met pathway genes. Here, we identified a novel L-homocysteine synthase (MetM) in Streptomyces and demonstrated that it converts O-phospho-L-homoserine to L-homocysteine using a thiocarboxylated sulfur-carrier protein as a sulfur donor. Since the metM is distributed in other bacteria and archaea, our pioneering study contributes to understanding Met biosynthesis in these organisms. [ABSTRACT FROM AUTHOR]

The present study was conducted to select Streptomyces strains that exhibited characteristics of probiotics for application in aquaculture. Streptomyces were isolated from soil and sediment samples collected from various habitats such as riverside, shrimp ponds, pangasius ponds, mangrove forests, or estuaries along the coast of Vietnam. Two strains of Streptomyces (AG12.2 and ND10.1) were selected due to (1) their potent antimicrobial activity against common pathogens that cause serious diseases in shrimp and pangasius such as E. ictaluri, A. hydophila, V. harveyi, V. parahaemolyticus, and (2) their superior ability to produce several extracellular enzymes that strongly hydrolyze organic compounds such as starch, CMC, and skimmed milk, compared to other strains. AG12.2 and ND10.1 can tolerate salinity from 0 ppt to 5 ppt and pH from 3 to 8; however, their growth is most vigorous at salt concentrations and pH between 0 ppt to 2 ppt, and 7.2 to 8, respectively. In vivo evaluation on L. vannamei in this study shows that shrimp that were fed with diets containing AG12.2 and ND10.1 had WG (%) increasing by 1.59 times and 1.61 times respectively, DGW (g/day) increasing by 1.64 times, and FCR decreasing by 1.30 times and 1.35 times respectively. The shrimp treated with AG12.2 and ND10.1 were then challenged with V. parahaemolyticus, resulting in a survival rate increase of 3.9 times and 3.71 times, respectively. Phylogenetic analysis revealed that strains AG12.2 and ND10.1 belong to Streptomyces kunmingensis and Streptomyces angustmyceticus, respectively. The findings of the present study led to the conclusion that Streptomyces kunmingensis AG12.2 and Streptomyces angustmyceticus ND10.1 are excellent candidates for producing beneficial probiotics for aquaculture. [ABSTRACT FROM AUTHOR]

Angucyclines and angucyclinones represent a class of natural compounds that belong to the group of aromatic polyketides. They exhibit a wide array of biological properties, such as antimicrobial, antiviral, and cytotoxic. Their considerable therapeutic potential and diverse scaffolds have attracted many synthetic chemists to devise novel strategies to construct their intricate molecular architecture. Over 300 class members have been isolated from natural sources, mainly from bacterial strains of Streptomyces species. This review highlights recent advancements in their synthesis, such as oxidative cyclization, photooxidation, and metal‐catalyzed [4+2]‐cycloaddition, which has facilitated the efficient and practical total syntheses of various angucycline and angucyclinone natural products. [ABSTRACT FROM AUTHOR]

Istamycins (ISMs) are 2-deoxyfortamine-containing aminoglycoside antibiotics (AGAs) produced by Streptomyces tenjimariensis ATCC 31603 with broad-spectrum bactericidal activities against most of the clinically relevant pathogens. Therefore, this study aimed to statistically optimize the environmental conditions affecting ISMs production using the central composite design (CCD). Both the effect of culture media composition and incubation time and agitation rate were studied as one factor at the time (OFAT). The results showed that both the aminoglycoside production medium and the protoplast regeneration medium gave the highest specific productivity. Results also showed that 6 days incubation time and 200 rpm agitation were optimum for their production. A CCD quadratic model of 17 runs was employed to test three key variables: initial pH, incubation temperature, and concentration of calcium carbonate. A significant statistical model was obtained including, an initial pH of 6.38, incubation temperature of 30 ˚C, and 5.3% CaCO3 concentration. This model was verified experimentally in the lab and resulted in a 31-fold increase as compared to the unoptimized conditions and a threefold increase to that generated by using the optimized culture media. To our knowledge, this is the first report about studying environmental conditions affecting ISM production as OFAT and through CCD design of the response surface methodology (RSM) employed for statistical optimization. In conclusion, the CCD design is an effective tool for optimizing ISMs at the shake flask level. However, the optimized conditions generated using the CCD model in this study should be scaled up in a fermenter for industrial production of ISMs by S. tenjimariensis ATCC 31603 considering the studied environmental conditions that significantly influence the production proces. [ABSTRACT FROM AUTHOR]

Pigments are chemical compounds that impart color through mechanisms such as absorption, reflection, and refraction. While traditional natural pigments are derived from plant and insect tissues, microorganisms, including bacteria, yeasts, algae, and filamentous fungi, have emerged as promising sources for pigment production. In this study, we focused on pigment production by 20 Streptomyces isolates from our in-house actinobacteria strain collection, sourced from the Guaviare and Arauca Rivers in Colombia. The isolates were identified via 16S rRNA gene sequencing, and the bioactivities—including antioxidant, antibacterial, and cytotoxic properties—of their extracts obtained across four different culture media were assessed. Promising pigmented hydroalcoholic extracts demonstrating these bioactivities were further analyzed using LC-MS, leading to the annotation of a variety of pigment-related compounds. This study revealed that culture media significantly influenced both pigment production and bioactivity outcomes. Notably, anthraquinones, phenazines, and naphthoquinones were predominant pigment classes associated with cytotoxic and antimicrobial activities, while carotenoids were linked to antioxidant effects. For instance, S. murinus 4C171 produced various compounds exhibiting both cytotoxic and antioxidant activities. These findings highlighted a growth medium-dependent effect, as pigment production, coloration, and bioactivity outcomes were influenced by growth media. These results demonstrate the significant potential of Streptomyces isolates as sources of bioactive pigments for diverse applications. [ABSTRACT FROM AUTHOR]

This study focused on the isolation of actinobacteria capable of producing extracellular keratinase from keratin-rich residues, which led to the selection of an actinobacterial strain referenced as Streptomyces strain DZ 06 (ES41). The Plackett–Burman screening plan was used for the statistical optimization of the enzymatic production medium, leading to the identification of five key parameters that achieved a maximum activity of 180.1 U/mL. Further refinement using response surface methodology (RSM) with a Box–Behnken design enhanced enzyme production to approximately 458 U/mL. Model validation, based on the statistical predictions, demonstrated that optimal keratinase activity of 489.24 U/mL could be attained with 6.13 g/L of chicken feather meal, a pH of 6.25, incubation at 40.65 °C for 4.11 days, and an inoculum size of 3.98 × 107 spores/mL. The optimized culture conditions yielded a 21.67-fold increase in keratinase compared with the initial non-optimized standard conditions. The results show that this bacterium is an excellent candidate for industrial applications when optimal conditions are used to minimize the overall costs of the enzyme production process. [ABSTRACT FROM AUTHOR]

An investigation of ×19 soil samples collected under the auspices of the Australian citizen science initiative, Soils for Science, returned ×559 chemically dereplicated microbial isolates, of which ×54 exhibited noteworthy anthelmintic activity against either the heartworm Dirofilaria immitis microfilaria and/or the gastrointestinal parasite Haemonchus contortus L1–L3 larvae. Chemical (GNPS and UPLC-DAD) and cultivation (MATRIX) profiling prompted a detailed chemical investigation of Streptomyces sp. S4S-00196A10, which yielded new anthelmintic polyketide goondapyrones A–J (1–10), together with the known actinopyrones A (11) and C (12). Structures for 1–12 were assigned on the basis of detailed spectroscopic and chemical analysis, with preliminary structure activity relationship analysis revealing selected γ-pyrones >50-fold and >13-fold more potent than isomeric α-pyrones against D. immitis mf motility (e.g., EC50 0.05 μM for 1; EC50 2.7 μM for 5) and H. contortus L1–L3 larvae development (e.g., EC50 0.58 μM for 1; EC50 8.2 μM for 5), respectively. [ABSTRACT FROM AUTHOR]

Background/Objectives: Research-engaged academic institutions offer the opportunity to couple undergraduate education/citizen science projects with antimicrobial biodiscovery research. Several initiatives reflecting this ethos have been reported internationally (e.g., Small World, Tiny Earth, MicroMundo, Antibiotics Unearthed). These programs target soil habitats due to their high microbial diversity and promote initial screening with non-selective, nutrient media such as tryptic soy agar (TSA). However, evaluation of published outputs to date indicates that isolate recovery on TSA is consistently dominated by the genera Bacillus, Pseudomonas, and Paenibacillus. In this study, we evaluated the potential of soil extract agar to enhance soil isolate diversity and antibiosis induction outcomes in our undergraduate Antibiotics Unearthed research program. Methods: We comparatively screened 229 isolates from woodland and garden soil samples on both tryptic soy agar (TSA) and soil extract agar (SEA) for antimicrobial activity against a panel of clinically relevant microbial pathogens. Results: On one or both media, 15 isolates were found to produce zones of clearing against respective pathogens. 16S rRNA gene sequencing linked the isolates with three genera: Streptomyces (7), Paenibacillus (6), and Pseudomonas (2). Six of the Streptomyces isolates and one Pseudomonas demonstrated antimicrobial activity when screened on SEA, with no activity on TSA. Furthermore, incorporation of the known secondary metabolite inducer N acetyl-glucosamine (20 mM) into SEA media altered the pathogen inhibition profiles of 14 isolates and resulted in broad-spectrum activity of one Streptomyces isolate, not observed on SEA alone. In conclusion, SEA was found to expand the diversity of culturable isolates from soil and specifically enhanced the recovery of members of the genus Streptomyces. SEA was also found to be a superior media for antibiosis induction among Streptomyces isolates when compared to TSA. It was noted that Paenibacillus isolates' antibiosis induction demonstrated a strain-specific response with respect to the growth media used. Conclusions: The authors propose SEA inclusion of in soil screening protocols as a cost-effective, complementary strategy to greatly enhance outcomes in undergraduate/citizen science-engaged antimicrobial biodiscovery initiatives. [ABSTRACT FROM AUTHOR]

Metribuzin, a widely used triazine herbicide, persists in agricultural soils and poses significant environmental pollution threats globally. The aim of this study was to investigate the biodegradation of metribuzin by actinobacterial strains in vitro at different environmental conditions. From an initial screen of 12 actinobacterial strains, four bacteria exhibited robust growth in the presence of the metribuzin as the sole carbon source at 50 mg/L concentration. The optimization of metribuzin biodegradation under different conditions (pH, temperature and inoculum size) using a spectrophotometric method revealed that maximum degradation of metribuzin occurred at a pH of 7.2, a temperature 30 °C, and at an inoculum volume of 4%. Subsequent GC-MS validation confirmed the remarkable biodegradation capabilities of the actinobacterial isolates, where the strain C1 showed the highest rate of metribuzin degradation of 83.12%. Detailed phylogenetic identified the active strains as Streptomyces toxytricini (CH), Streptomyces stelliscabiei (B2), and two Streptomyces heliomycini (C1, C3). Structural analysis by ATR-FTIR spectroscopy confirmed the extensive biotransformation of the herbicide molecule. Our findings highlight the immense untapped potential of soil actinobacteria, particularly the Streptomyces heliomycini C1 strain, as versatile bioremediation agents for removing persistent agrochemical pollutants. [ABSTRACT FROM AUTHOR]

Background/Objectives: Recently, antimicrobial-resistant pathogens and cancers have emerged as serious global health problems, highlighting the immediate need for novel therapeutics. Consequently, we aimed to isolate and characterize endophytic Streptomyces strains from the rhizospheres of the Himalayan region of Nepal and identify specialized metabolites with antibacterial, antifungal, and cytotoxic potential. Methods: To isolate Streptomyces sp., we collected two soil samples and cultured them on an ISP4 medium after pretreatment. We isolated and identified the strains PY108 and PY109 using a combination of morphological observations and 16S rRNA gene sequencing. Results: The BLAST results showed that PY108 and PY109 resembled Streptomyces hundungensis PSB170 and Streptomyces sp. Ed-065 with 99.28% and 99.36% nucleotide similarity, respectively. Antibacterial assays of ethyl acetate (EA) extracts from both isolates PY108 and PY109 in a tryptic soy broth (TSB) medium were conducted against four pathogenic bacteria. They showed significant antibacterial potential against Staphylococcus aureus and Klebsiella pneumoniae. Similarly, these extracts exhibited moderate antifungal activities against Saccharomyces cerevisiae and Aspergillus niger. Cytotoxicity assays on cervical cancer cells (HeLa) and breast cancer cells (MCF-7) revealed significant potential for both extracts. LC-MS/MS profiling of the EA extracts identified 27 specialized metabolites, including diketopiperazine derivatives, aureolic acid derivatives such as chromomycin A, and lipopeptide derivatives. In comparison, GC-MS analysis detected 34 metabolites, including actinomycin D and γ-sitosterol. Furthermore, a global natural product social molecular networking (GNPS)-based molecular networking analysis dereplicated 24 metabolites in both extracts. Conclusions: These findings underscore the potential of endophytic Streptomyces sp. PY108 and PY109 to develop new therapeutics in the future. [ABSTRACT FROM AUTHOR]

Chitinases have vast applications due to their chitin degrading potential, as chitin is the main component of the cell-wall and exoskeleton of fungi and insects, respectively. In the late 20th century, wide research was conducted to explore their antifungal and insecticidal potential. The unavailability of complete sequences of genes, and less enzyme production from natural sources have restricted their applications at the industrial scale. In the present day, the availability of gene sequences may help to accomplish the long-awaited targets. In previous studies, Streptomyces sp. strain 130 was studied for chitinase production, where ten chitinase-producing genes were observed. In the current study, chitinases from strain 130 were validated using proteomics, where eight chitinases were found. Out of all chitinases, Chi65_1884 was selected for protein structure prediction, molecular docking and cloning of the chitinase-producing gene. Protein structure prediction was performed using homology modeling (Modeller 9.21). Molecular docking was conducted using Autodock 4.2 and MGL tools to examine bond interactions. The binding efficiency of Chi65_1884 with ligand was found to be -6.3 kcal/mol and the inhibition constant was 24.18 μM. Based on the molecular docking results, the chi65 gene was cloned into the vector pET32a and transformed into E. coli DH5α. Subsequent transformation into E.coli BL21 was facilitated for protein expression. The size of the expressed protein was observed to be ~85kDa (65kDa chitinase and ~20kDa carrier proteins of the vector). In future studies, chitinase produced by the cloned gene may tested against a range of pathogenic fungi to check its antifungal potential. [ABSTRACT FROM AUTHOR]

Trialeurodes vaporariorum Westwood poses a significant threat to vegetable and ornamental crops in temperate zones, resulting in notable reductions in yield and substantial economic burdens. In order to find compounds with high insecticidal activity against T. vaporariorum, five compounds were isolated and identified from the crude extract of Streptomyces sp. SA61. These include three new polyketides, named strekingmycins F–H (1–3); one new diterpenoid, named phenalinolactone CD8 (4); and one known compound, strekingmycin A (5). Their structures were analyzed using high-resolution electrospray ionization mass spectrometry and one-dimensional and two-dimensional nuclear magnetic resonance spectroscopy data and by comparing them with previously reported data. The insecticidal activities of compounds 1–5 against T. vaporariorum were evaluated. Among them, compound 5 exhibited the highest insecticidal activity, with an LC50 of 6.949 mg/L against T. vaporariorum at 72 h using the leaf-dip method. Lower insecticidal activities were found in compounds 1–4, with LC50 values of 22.817, 19.150, 16.981 and 41.501 mg/L, respectively. These data indicate that strekingmycin could be a potential candidate for a novel insecticide to control T. vaporariorum. [ABSTRACT FROM AUTHOR]

The Streptomyces sp. is considered the vast reservoir of bioactive natural products belonging to different classes like polyketides, terpenoids, lanthipeptides, and non-ribosomal peptides to name a few. The ubiquitous distribution of the genus makes them capable of producing distinct compounds. Many of those compounds contain a unique γ-pyrone with various chemical structures and exhibit different bioactivities. One such class, nitrophenyl-γ-pyrone, constitutes different bioactive compounds isolated from Streptomyces sp. from different sources ranging from soil to marine environments. In addition, such compounds have antinematodal, cytotoxicity activities, and inhibition of adipogenesis. These compounds include aureothin (3), spectinabilin (7), and their derivatives. Moreover, there are other compounds like actinopyrones (11–16), benwamycins (22–23), and peucemycin and its derivatives (24–26) that also have antibacterial and anticancer activities. The other group classified as anthra-γ-pyrone has various bioactive natural products. For instance, tetrahydroanthra-γ-pyrone, shellmycin A-D (27–30) possess antibacterial as well as anticancer activities. In addition, the pluramycin family compounds belonging to anthra-γ-pyrone group also possess cytotoxic activity, for instance, kidamycin (31), rubiflavin, and their derivatives (33–37). Xanthones are another important group of natural products that also contain γ-pyrone ring producing different bioactivities. Albofungin (42) and its derivatives (43–46) belong to subgroup polycyclic tetrahydro xanthones that possess antibacterial, anticancer, and antibiofilm, antimacrofouling activities. Similarly, other compounds, belonging to this subgroup, exhibit different bioactivities like antifungal, antimalarial, and antibacterial activities and block transient receptor potential vanilloid 1 (TRPV1). These compounds include cervinomycins (48–55), citreamycins (56–57), sattahipmycin (59), and chrexanthomycins (60–63). This review gives succinct information on the γ-pyrone containing natural products isolated from Streptomyces sp. focusing on their structure and bioactivities. Key points: • The Streptomyces sp. is the producer of various bioactive natural products including the one with γ-pyrone ring. • These γ-pyrone compounds are structurally different and possess different bioactivities. • The Streptomyces has the potential to produce such compounds and the reservoir of these compounds is expected to increase in the future. [ABSTRACT FROM AUTHOR]

Potato common scab (PCS), a significant potato disease, negatively impacts tuber quality. The present study was conducted to isolate and characterize pathogenic Streptomyces species associated with PCS disease in Türkiye based on their morphological, physiological and molecular characteristics. Field‐grown potatoes that exhibited scab lesions were collected from four provinces in 2020–2021, and 200 bacterial isolates were obtained from netted, superficial and deep‐pitted common scab lesions. Pathogenicity assays, including in vitro tuber slice and in planta radish seedling bioassays, identified 150 pathogenic isolates. Morphological and physiological characterization of 92 selected isolates revealed several Streptomyces species that exhibited diverse mycelium colours, sporulation patterns and pigmentation. Molecular analysis using 16S rRNA sequencing, species‐specific primers, PCR‐RFLP of the 16S‐23S (ITS) region with Hpy99I restriction enzyme, and multilocus sequence analysis (MLSA) based on atpD, recA, rpoB and trpB genes revealed that S. scabiei was the dominant species, followed by S. europaeiscabiei, S. caniscabiei, S. bottropensis, S. stelliscabiei and S. turgidiscabies. PCR analysis revealed the presence of the thaxtomin synthetase genes (txtAB) in all tested samples, while the necrogenic protein (nec1) and tomatinase (tomA)‐encoding genes were absent from three and two isolates, respectively. Phylogenetic analysis of 32 representative isolates conducted with sequences from 16S rRNA, species‐specific PCR and MLSA confirmed their morphological identification and clustered them with reference strains. This study contributes to the understanding of distribution of Streptomyces species associated with PCS, and to our knowledge, is the first molecular confirmation of S. caniscabiei and S. turgidiscabies causing potato scab in Türkiye. [ABSTRACT FROM AUTHOR]

The glycoside hydrolase family 3 (GH3) β-glucosidases from filamentous fungi are crucial industrial enzymes facilitating the complete degradation of lignocellulose, by converting cello-oligosaccharides and cellobiose into glucose. Understanding the diverse domain organization is essential for elucidating their biological roles and potential biotechnological applications. This research delves into the variability of domain organization within GH3 β-glucosidases. Two distinct configurations were identified in fungal GH3 β-glucosidases, one comprising solely the GH3 catalytic domain, and another incorporating the GH3 domain with a C-terminal fibronectin type III (Fn3) domain. Notably, Streptomyces filamentous bacteria showcased a separate clade of GH3 proteins linking the GH3 domain to a carbohydrate binding module from family 2 (CBM2). As a first step to be able to explore the role of accessory domains in β-glucosidase activity, a screening system utilizing the well-characterised Aspergillus niger β-glucosidase gene (bglA) in bglA deletion mutant host was developed. Based on this screening system, reintroducing the native GH3-Fn3 gene successfully expressed the gene allowing detection of the protein using different enzymatic assays. Further investigation into the role of the accessory domains in GH3 family proteins, including those from Streptomyces, will be required to design improved chimeric β-glucosidases enzymes for industrial application. [ABSTRACT FROM AUTHOR]

We have created a novel synthetic biology expression system allowing easy refactoring of biosynthetic gene clusters (BGCs) as monocistronic transcriptional units. The system is based on a set of plasmids containing a strong kasOp* promoter, RBS and terminators. It allows the cloning of biosynthetic genes into transcriptional units kasOp *-gene(s)-terminator flanked by several rare restriction cloning sites that can be sequentially combined into the artificial BGC in three compatible Streptomyces integration vectors. They allow a simultaneous integration of these BGCs at three different attB sites in the Streptomyces chromosome. The system was validated with biosynthetic genes from two known BGCs for aromatic polyketides landomycin and mithramycin. • A new synthetic biology-based system for investigating antibiotic biosynthesis. • Efficient rabelomycin production using the initial landomycin biosynthetic genes. • Efficient 4-DMPC production using the initial mithramycin biosynthetic genes. [ABSTRACT FROM AUTHOR]

The influence of talc microparticles on metabolism and morphology of S. rimosus at various initial organic nitrogen concentrations was investigated. The shake flask cultivations were conducted in the media with yeast extract (nitrogen source) concentration equal to 1 g YE L− 1 and 20 g YE L− 1. Two talc microparticle concentrations of 5 g TALC L− 1 and 10 g TALC L− 1 were tested in microparticle-enhanced cultivation (MPEC) runs. A high nitrogen concentration of 20 g YE L− 1 promoted the development of small agglomerates (pellets) of projected area lower than 105 µm2 and dispersed pseudohyphae. A low nitrogen concentration of 1 g YE L− 1 led to the limitation of S. rimosus growth and, in consequence, the development of the smaller number of large pseudohyphal agglomerates (pellets) of projected area higher than 105 µm2 compared to the culture containing a high amount of nitrogen source. In both cases talc microparticles were embedded into pellets and caused the decrease in their sizes. The lower amount of talc (5 g TALC L− 1) usually caused the weaker effect on S. rimosus morphology and metabolite production than the higher one. This correlation between the microparticles effect on morphology and metabolism of S. rimosus was especially noticeable in the biosynthesis of oxytetracycline, 2-acetyl-2-dicarboxamide oxytetracycline (ADOTC) and spinoxazine A. Compared to the control run, in MPEC their levels increased 4-fold, 5-fold and 1.6-fold respectively. The addition of talc also improved the production of 2-methylthio-cis-zeatin, lorneic acid J and milbemycin A3. [ABSTRACT FROM AUTHOR]

Background: Abamectin (ABA) is considered a powerful insecticidal and anthelmintic agent. It is an intracellular product of Streptomyces avermitilis; is synthesized through complicated pathways and can then be extracted from mycelial by methanol extraction. ABA serves as a biological control substance against the root-knot nematode Meloidogyne incognita. This investigation is intended to reach a new strain of S. avermitilis capable of producing ABA effectively. Results: Among the sixty actinobacterial isolates, Streptomyces St.53 isolate was chosen for its superior nematicidal effectiveness. The mycelial-methanol extract of isolate St.53 exhibited a maximum in vitro mortality of 100% in one day. In the greenhouse experiment, the mycelial-methanol extract demonstrated, for the second-stage juveniles (J2s), 75.69% nematode reduction and 0.84 reproduction rate (Rr) while for the second-stage juveniles (J2s), the culture suspension demonstrated 75.38% nematode reduction and 0.80 reproduction rate (Rr). Molecular identification for St.53 was performed using 16 S rRNA gene analysis and recorded in NCBI Genbank as S. avermitilis MICNEMA2022 with accession number (OP108264.1). LC-MS was utilized to detect and identify abamectin in extracts while HPLC analysis was carried out for quantitative determination. Both abamectin B1a and abamectin B1b were produced and detected at retention times of 4.572 and 3.890 min respectively. Conclusion: Streptomyces avermitilis MICNEMA2022 proved to be an effective source for producing abamectin as a biorational agent for integrated nematode management. [ABSTRACT FROM AUTHOR]

Recent advances in whole genome sequencing have revealed an immense microbial potential for the production of therapeutic small molecules, even from well‐known producers. To access this potential, we subjected prominent antimicrobial producers to alternative antiproliferative assays using persistent cancer cell lines. Described herein is our discovery of hirocidins, novel secondary metabolites from Streptomyces hiroshimensis with antiproliferative activities against colon and persistent breast cancer cells. Hirocidin A is an unusual nine‐membered carbocyclic maleimide and hirocidins B and C are relatives with an unprecedented, bridged azamacrocyclic backbone. Mode of action studies show that hirocidins trigger mitochondrion‐dependent apoptosis by inducing expression of the key apoptotic effector caspase‐9. The discovery of new cytotoxins contributes to scaffold diversification in anticancer drug discovery and the reported modes of action and concise total synthetic route for variant A set the stage for unraveling specific targets and biochemical interactions of the hirocidins. [ABSTRACT FROM AUTHOR]

Lasso peptides, ribosomally synthesized and post-translationally modified peptides, are primarily produced by bacteria and some archaea. Streptomyces lasso peptides have been known for their antimicrobial, anticancer, and antiviral properties. However, understanding their role in the morphology and production of secondary metabolites remains limited. We identified a previously unknown lasso peptide gene cluster in the genome of Streptomyces sp. L06. This gene cluster (LASS) produces two distinct lasso peptides, morphosin-1 and − 2. Notably, morphosin-2 is a member of a new subfamily of lasso peptides, with BGCs exhibiting a similar structure. When LASS was expressed in different Streptomyces hosts, it led to exciting phenotypic changes, including the absence of spores and damage in aerial mycelium development. In one of the hosts, LASS even triggered antibiotic formation. These findings open up a world of possibilities, suggesting the potential role of morphosins in shaping Streptomyces' morphological and biochemical development. [ABSTRACT FROM AUTHOR]

Fusarium head blight (FHB) of wheat, mainly caused by Fusarium graminearum, leads to severe economic losses worldwide. Effective management measures for controlling FHB are not available due to a lack of resistant cultivars. Currently, the utilization of biological control is a promising approach that can be used to help manage FHB. Previous studies have confirmed that Streptomyces pratensis S10 harbors excellent inhibitory effects on F. graminearum. However, there is no information regarding whether invasive hyphae of F. graminearum are inhibited by S10. Thus, we investigated the effects of S10 on F. graminearum strain PH-1 hypha extension, toxisome formation, and TRI5 gene expression on wheat plants via microscopic observation. The results showed that S10 effectively inhibited the spread of F. graminearum hyphae along the rachis, restricting the infection of neighboring florets via the phloem. In the presence of S10, the hyphal growth is impeded by the formation of dense cell wall thickenings in the rachis internode surrounding the F. graminearum infection site, avoiding cell plasmolysis and collapse. We further demonstrated that S10 largely prevented cell-to-cell invasion of fungal hyphae inside wheat coleoptiles using a constitutively green fluorescence protein-expressing F. graminearum strain, PH-1. Importantly, S. pratensis S10 inhibited toxisome formation and TRI5 gene expression in wheat plants during infection. Collectively, these findings indicate that S. pratensis S10 prevents the spread of F. graminearum invasive hyphae via the rachis. [ABSTRACT FROM AUTHOR]

Streptomyces aureofaciens widely produces the antibiotic tetracycline and many other compounds during fermentation. The compounds have yet to be known for their antibacterial potential. This work aims to determine new antibiotics or other possible antibacterial compounds produced by marine S.aureofaceiens A3 through an in silico molecular docking method. The ethyl acetate (EA) extracts from fermented marine S. aureofaciens A3 in ISP4 medium enriched with seawater components showed strong antibacterial activity. The antibacterial activity of EA extracts during 6-12 days of fermentation was carried out by the Kirby-Bauer method and the compounds of EA extracts were analyzed by GC/MS. Compounds identified by GC/MS were ligands for an in silico molecular docking study against four target proteins (DNA gyrase, topoisomerase IV, PBP 1a, and DHFR) of pathogenic bacteria. The drug-likeness of selected chemicals as antibacterial agents was assessed using Lipinski's Rule of Five. The results showed the prospective compounds as a narrow-spectrum antibacterial, including 3,5-di-tert-Butyl-4-hydroxyphenylpropionic acid against PBP 1a and Benzenepropanoic acid, and 3,5-bis (1,1-dimethyl ethyl)-4-hydroxy-, methyl esters against DHFR. Substances with broad-spectrum antibacterial activity, such as 3-Acetylphenanthrene and 3-(p-Ethoxyphenyl)-5-(O-tolyloxymethyl)-2-oxazolidone, against multitarget DNA gyrase B and DHFR, 7,9-Di-tert-butyl-1-oxaspiro (4,5) Deca-6,9-diene-2,8-dione against PBP1a and DHFR, and isobenzofuro [5,6-b] benzofuran-8-carboxylic acid, 1,3-dihydro-7,10-dimethoxy-9-methyl-1-oxo-, methyl ester against DNA gyrase B, PBP 1a, and DHFR. On the 12th day of fermentation, two compounds were identified: isobenzofuro[5,6-b] benzofuran-8-carboxylic acid, 1,3-dihydro-7,10-dimethoxy-9-methyl-1-oxo-, methyl ester, and 3-(p-Ethoxyphenyl)-5-(O-tolyl oxy methyl)-2-oxazolidone. This is the first report that these two compounds, known as potential drugs like antibiotics through in silico molecular docking, were first produced by Streptomyces species. [ABSTRACT FROM AUTHOR]

Fusarium‐induced wilt significantly affects the cultivation and yield of pigeon peas. This warrants sustainable disease management while promoting plant growth. The present study investigated the biopotential of coinoculation of Streptomyces pseudogriseolus S‐9 and Rhizophagus irregularis for plant growth promotion and mitigation of the impact of Fusarium wilt on pigeon pea over three seasons at pot and field levels. Pigeon pea plants were subjected to Fusarium wilt stress and treated with different inoculation strategies, including single and combined applications of S. pseudogriseolus S‐9 and R. irregularis. Plant growth parameters and yields were assessed to evaluate the efficacy of the coinoculation. In the pot experiment, T‐6 treatment resulted in the longest root (62.56 ± 0.01 cm) and shoot (70.24 ± 0.01 cm) lengths compared to the application of commercial biofungicide T‐8 (Trichoderma). This treatment also significantly influenced the yield of potted plants. It resulted in the highest fresh root weight (62.27 ± 0.01 g), fresh shoot weight (70.24 ± 0.02 g), maximum root (55.25 ± 0.01 g) and shoot dry weights (52.25 ± 0.01 g). In the field experiment, pigeon pea plants treated with the bioinoculant also demonstrated a substantial increase (р ≤ 0.05) in total grain yield, the weight of 100 grains, and the number of filled grains compared to the control group in all experimental seasons. In vitro, antagonism assay of compatibility of mycorrhizae and bacteria showed good activity using powder formulation. Thus, the consortium application inspired the broad application of Streptomyces and Trichoderma as effective bioinoculants for wilt management and yield improvement in pigeon peas. [ABSTRACT FROM AUTHOR]

Natural products play a crucial role in drug development, addressing the escalating microbial resistance to antibiotics and the treatment of emerging diseases. Progress in genome sequencing techniques, coupled with the development of bioinformatics tools and the exploration of uncharted habitats, has highlighted the biosynthetic potential of actinomycetes. By in silico screening for diazo-related gene genomes from twelve Streptomyces strains isolated from Attini leaf-cutting ants, the new crx biosynthetic gene cluster (BGC) was identified in Streptomyces sp. CS057. This cluster, highly conserved in several Streptomyces strains, contains genes related to diazo group formation and genes for the biosynthesis of 3,4-AHBA. By overexpressing the LuxR-like regulatory gene crxR1, we were able to activate the crx cluster, which encodes the biosynthesis of three 3,4-AHBA-derived compounds that we named crexazones (CRXs). The chemical structure of crexazones (CRXs) was determined by LC-DAD-HRMS-based dereplication and NMR spectroscopic analyses and was found to correspond to two known compounds, 3-acetamido-4-hydroxybenzoic acid (CRX1) and the phenoxazinone texazone (CRX3), and a novel 3,4-AHBA-containing compound herein designated as CRX2. Experimental proof linking the crx BGC to their encoded compounds was achieved by generating mutants in selected crx genes. [ABSTRACT FROM AUTHOR]

Streptomyces offer a wealth of naturally occurring compounds with diverse structures, many of which possess significant pharmaceutical values. However, new product exploration and increased yield of specific compounds in Streptomyces have been technically challenging due to their slow growth rate, complex culture conditions and intricate genetic backgrounds. In this study, we screened dozens of Streptomyces strains inhabiting in a plant rhizosphere for fast-growing candidates, and further employed CRISPR/Cas-based engineering techniques for stepwise refinement of a particular strain, Streptomyces sp. A-14 that harbors a 7.47 Mb genome. After strategic removal of nonessential genomic regions and most gene clusters, we reduced its genome size to 6.13 Mb, while preserving its growth rate to the greatest extent. We further demonstrated that cleaner metabolic background of this engineered strain was well suited for the expression and characterization of heterologous gene clusters, including the biosynthetic pathways of actinorhodin and polycyclic tetramate macrolactams. Moreover, this streamlined genome is anticipated to facilitate directing the metabolic flux towards the production of desired compounds and increasing their yields. [ABSTRACT FROM AUTHOR]

Mollemycin A (MOMA) is a unique glyco-hexadepsipeptide-polyketide that was isolated from a Streptomyces sp. derived from the Australian marine environment. MOMA exhibits remarkable inhibitory activity against both drug-sensitive and multidrug-resistant malaria parasites. Optimizing MOMA through structural modifications or product enhancements is necessary for the development of effective analogues. However, modifying MOMA using chemical approaches is challenging, and the production titer of MOMA in the wild-type strain is low. This study identified and characterized the biosynthetic gene cluster of MOMA for the first time, proposed its complex biosynthetic pathway, and achieved an effective two-pronged enhancement of MOMA production. The fermentation medium was optimized to increase the yield of MOMA from 0.9 mg L−1 to 1.3 mg L− 1, a 44% boost. Additionally, a synergistic mutant strain was developed by deleting the momB3 gene and overexpressing momB2, resulting in a 2.6-fold increase from 1.3 mg L− 1 to 3.4 mg L− 1 . These findings pave the way for investigating the biosynthetic mechanism of MOMA, creating opportunities to produce a wide range of MOMA analogues, and developing an efficient strain for the sustainable and economical production of MOMA and its analogues. [ABSTRACT FROM AUTHOR]

Type I CRISPR-Cas systems are widely distributed, found in over 40% of bacteria and 80% of archaea. Among genome-sequenced actinomycetes (particularly Streptomyces spp.), 45.54% possess type I CRISPR-Cas systems. In comparison to widely used CRISPR systems like Cas9 or Cas12a, these endogenous CRISPRCas systems have significant advantages, including better compatibility, wide distribution, and ease of operation (since no exogenous Cas gene delivery is needed). Furthermore, type I CRISPR-Cas systems can simultaneously edit and regulate genes by adjusting the crRNA spacer length. Meanwhile, most actinomycetes are recalcitrant to genetic manipulation, hindering the discovery and engineering of natural products (NPs). The endogenous type I CRISPR-Cas systems in actinomycetes may offer a promising alternative to overcome these barriers. This review summarizes the challenges and recent advances in CRISPR-based genome engineering technologies for actinomycetes. It also presents and discusses how to establish and develop genome editing tools based on type I CRISPR-Cas systems in actinomycetes, with the aim of their future application in gene editing and the discovery of NPs in actinomycetes. [ABSTRACT FROM AUTHOR]

To investigate the biocatalytic potential of Amazonian actinomycetes for monoterpenes biotransformation. To carry out the present study, eleven actinomycetes of the genus Streptomyces isolated from inga-cipó (Inga edulis Mart.) rhizospheres were tested for their ability to bioconvert the substrates R-(+)-limonene, S-(-)-limonene, 1S-(-)-α-pinene, and (-)–β–pinene as sole carbon and energy source. According to gas chromatography-mass spectrometry analysis, three strains, LabMicra B270, LaBMicrA B310, and LaBMicrA B314, were able to biotransform 1S-(-)-α-pinene after 96 h of growth. However, Streptomyces LaBMicrA B270 was the most promising since it converted after only 72 h all the 1S-(-)-α-pinene mainly into cis-verbenol (74.9±1.24%) and verbenone (18.2±1.20%), compounds that have important biological activities and great industrial interest as additives in foods and cosmetics. These findings can stimulate the development of natural aromas using naturally abundant monoterpenes, ratify the potential of microorganisms from almost unexplored niches such as the Amazonian rhizosphere, and reinforce the importance of preserving those niches. [ABSTRACT FROM AUTHOR]

Nocardioform placentitis is a poorly understood disease of equine late gestation. The presence of nocardioform, filamentous branching gram-positive bacteria, has been linked to the disease, with Crossiella equi , Amycolatopsis spp., and Streptomyces spp. being the most frequently identified bacteria. However, these bacteria are not found in all clinical cases in addition to being isolated from healthy, normal postpartum placentas. To better understand this form of placentitis, we analyzed the microbial composition in the equine placenta (chorioallantois) of both healthy postpartum (control; n = 11) and nocardioform-affected samples (n = 22) using 16S rDNA sequencing. We found a lower Shannon index in nocardioform samples, a higher Chao1 index in nocardioform samples, and a difference in beta diversity between control and nocardioform samples (p < 0.05), suggesting the presence of dysbiosis during the disease. In the majority of the NP samples (77 %), one of the following genera— Amycolatopsis , Crossiella , Lentzea , an unidentified member of the Pseudonocardiaceae family, Mycobacterium, or Enterococcus —represented over 70 % of the relative abundance. Overall, the data suggest that a broader spectrum of potential opportunistic pathogens could be involved in nocardioform placentitis, extending beyond the traditionally recognized bacteria, resulting in a similar histomorphological profile. • Non-nocardioform opportunistic pathogens may be associated with placentitis. • Nocardioform placentitis is associated with dysbiotic changes. • Nocardioform placentitis exhibits a unique microbial diversity distinct from control samples. [ABSTRACT FROM AUTHOR]

Microbial‐derived natural products remain a major source of structurally diverse bioactive compounds and chemical scaffolds that have the potential as new therapeutics to target drug‐resistant pathogens and cancers. In particular, genome mining has revealed the vast number of cryptic or low‐yield biosynthetic gene clusters in the genus Streptomyces. However, low natural product yields—improvements to which have been hindered by the lack of high throughput methods—have slowed the discovery and development of many potential therapeutics. Here, we describe our efforts to improve yields of landomycins—angucycline family polyketides under investigation as cancer therapeutics—by a genetically modified Streptomyces cyanogenus 136. After simplifying the extraction process from S. cyanogenus cultures, we identified a wavelength at which the major landomycin products are absorbed in culture extracts, which we used to systematically explore culture medium compositions to improve total landomycin titers. Through correlational analysis, we simplified the culture optimization process by identifying an alternative wavelength at which culture supernatants absorb yet is representative of total landomycin titers. Using the subsequently improved sample throughput, we explored landomycin production during the culturing process to further increase landomycin yield and reduce culture time. Testing the antimicrobial activity of the isolated landomycins, we report broad inhibition of Gram‐positive bacteria, inhibition of fungi by landomycinone, and broad landomycin resistance by Gram‐negative bacteria that is likely mediated by the exclusion of landomycins by the bacterial membrane. Finally, the anticancer activity of the isolated landomycins against A549 lung carcinoma cells agrees with previous reports on other cell lines that glycan chain length correlates with activity. Given the prevalence of natural products produced by Streptomyces, as well as the light‐absorbing moieties common to bioactive natural products and their metabolic precursors, our method is relevant to improving the yields of other natural products of interest. [ABSTRACT FROM AUTHOR]

Chemical examination of an actinomycete strain Streptomyces malaysiensis DSM 4137 derived from a soil sample derived isolate Streptomyces sp. DSM 3816, yielded a new C2-asymmetric elaiophylin derivative efomycine U (1) and a known analogue halichoblelide D (2). These structures were unambiguously elucidated on the basis of extensive NMR spectroscopic and mass spectrometric analyses. All compounds isolated were subjected to antimicrobial, cytotoxic and immnosuppressive activities evaluation. [ABSTRACT FROM AUTHOR]

The increasing prevalence of antibiotic-resistant bacteria has prompted a global search for new antibiotics. Streptomycetaceae is one of the major families that produce antibiotics for medicinal use. Researchers have investigated the genus Streptomyces to identify new species that could produce potent antibacterial substances. This investigation was conducted to isolate and characterize Streptomyces strains with antibacterial potential from different soil samples in the Al-Karak Governorate, Jordan. Soil samples were obtained from caves, home gardens, greenhouses, and agricultural farmlands in the Governorate. Streptomyces were isolated and identified from the various soil samples. The antibacterial activity of the Streptomyces isolates was examined against a group of test bacteria using primary (modified cross-streak method) and secondary (agar diffusion test) screening methods. The isolate with the highest antimicrobial activity was identified based on morphological, biochemical, and molecular characterization. The results indicated that twenty-four Streptomyces strains were isolated from the different soil samples, all showing antimicrobial potential against at least two of the test bacteria. One of the isolates, the C8 strain had the highest antimicrobial potential in the primary screening against all test bacteria and displayed activity in the secondary screening against four types of test bacterial strains. The sequence analysis of the 16S rDNA gene showed that isolate C8 was 98.7% similar to Streptomyces pratensiss. The present study's findings suggested that Streptomyces pratensiss strain C8 from cave soil could produce antibacterial compounds, perhaps supporting the search for naturally occurring bioactive medications. [ABSTRACT FROM AUTHOR]