Search Results (1,414)

Some actinobacterial species have been reported to improve plant growth due to their roles as biostimulants and biological control agents. In this study, the effect of actinobacterial isolate 27, obtained from the rhizospheric soil of melon plants and identified as Streptomyces calvus, was evaluated on the growth of Arabidopsis thaliana and tomato plants. In Arabidopsis, in vitro assays showed that after seven days of interaction, isolate 27 increased fresh weight by 1.4-, 1.5-, and 2.3-fold and lateral root number by 1.7-, 1.3-, and 2.5-fold under physical contact and split-plate systems (MS and ISP2 media), respectively, compared with non-inoculated plants. An increased β-glucuronidase (GUS, encoded by the uidA gene) signal was observed in primary and lateral roots of the Arabidopsis DR5::uidA reporter line during both interaction types, suggesting the activation of auxin-responsive pathways. In addition, isolate 27 rescued the rhd6 (root hair defective 6) mutant phenotype, restoring root hair formation. Gas chromatography-mass spectrometry (GC-MS) analysis revealed that isolate 27 emitted volatile organic compounds (VOCs), including an alcohol and several sesquiterpenes, and that this profile changed during interaction with Arabidopsis plantlets. In soil-based pot assays, inoculation with isolate 27 significantly enhanced the development of Arabidopsis plants after 23 days, both when applied alone and in co-inoculation with Trichoderma atroviride. Furthermore, isolate 27 stimulated tomato plant growth, leading to significant increases in fresh and dry biomass, as well as shoot and root lengths after 28 days. Overall, these results demonstrate that S. calvus isolate 27 promotes plant growth and development through the production of bioactive compounds that modulate plant growth pathways, including hormonal responses, highlighting its potential as a bioinoculant for sustainable and productive agricultural systems. Full article

The olive tree (Olea europaea L.) is one of the oldest known cultivated trees worldwide and an iconic species within the Mediterranean Basin. This study evaluated the impact of three bacterial strains, Bacillus subtilis D3, Paenibacillus tundrae M4, and Streptomyces tricolor HM10, on the mortality of the following four mite pests: Oxycenus niloticus, Tegolophus hassani, Aceria olivi, and Tetranychus urticae. B. subtilis D3 confirmed the highest efficacy, causing 91.84–85.36% mortality in laboratory tests and 88.90–84.12% in field trials after five days. In addition, P. tundrae M4 ranked second, achieving 90.49–84.26% mortality in the lab and 87.87–83.81% in the field after one week. S. tricolor HM10 produced 80.06–74.09% mortality in laboratory assays and 76.73–73.36% under the field conditions. Effects on the predatory mites Agistemus exsertus and Amblyseius swirskii were minimal, with mortality ranging from 13.28 to 18.55% in the lab work and 12.46–16.74% in the field experiment. Genome analysis of strain HM10 revealed a biosynthetic gene cluster with predicted terpenes production. Terpenes can cause chemo-osmotic stress and broad membrane-disrupting capabilities. These results highlight the promise of microbial agents for sustainable mite management and provide a foundation for further optimization of bacterial biocontrol strategies. Full article

The increasing demand for sustainable plant protection products has intensified interest in microbial biocontrol agents (BCAs). This study aimed to evaluate the antifungal activity of selected Streptomyces, Bacillus, and Trichoderma asperellum strains against phytopathogenic fungi and to assess their potential as BCAs under in vitro conditions. The antifungal activity of ten Streptomyces strains was first evaluated against Botrytis cinerea, Colletotrichum salicis, Fusarium oxysporum, and F. graminearum using a dual-culture assay. All isolates exhibited antifungal activity, with Streptomyces venezuelae MSCL 350 showing the strongest inhibition. In addition, the antifungal activity of T. asperellum MSCL 309 and three Bacillus strains was assessed against twelve Fusarium spp. isolates obtained from oats. T. asperellum demonstrated broad-spectrum inhibition, with growth inhibition ranging from 44.6% to 78.4%, primarily due to soluble metabolites, while volatile compounds showed no significant effect. Among the other tested Bacillus strains, only Bacillus subtilis MSCL 1441 exhibited antifungal activity, inhibiting all tested isolates. These results demonstrate strong strain-dependent antifungal activity and highlight T. asperellum MSCL 309, S. venezuelae MSCL 350, and B. subtilis MSCL 1441 as promising candidates for the development of environmentally friendly biocontrol agents. Full article

Hydrochar application to soil inevitably releases hydrochar-derived dissolved organic matter (HDOM), yet its specific impact on soil microbial communities, independent of the hydrochar solid matrix, remains poorly understood. This study investigated, for the first time, the dose-dependent effects of HDOM on bacterial communities in three distinct soil types (red, yellow-brown, and black soils). A concentration gradient, including undiluted stock solution and 10-, 100-, and 1000-fold dilutions with ultrapure water, was established to test for hormesis-like responses. High-throughput 16S rRNA gene sequencing revealed that HDOM induced profound, soil-specific shifts in bacterial community structure. The application of HDOM induced the emergence of numerous specific bacterial taxa, with unique ASVs reaching up to 15,372. However, no significant changes were observed in microbial community richness or evenness (alpha diversity). Drastic shifts in beta diversity were evident only in red soil and yellow-brown soil, and exclusively under the undiluted HDOM treatment. At the phylum level, HDOM application did not alter the dominant bacterial types (top 10 phyla); however, their relative abundances were jointly regulated by both HDOM dose and soil type. Significant HDOM-induced changes in key bacterial biomarkers were primarily detected in red soil (e.g., phylum Elusimicrobia, class Fimbriimonadia, and family Alicyclobacillaceae) and yellow-brown soil (e.g., phylum Proteobacteria, class Alphaproteobacteria, and family Rhizobiaceae), while in black soil, such changes were observed only under the undiluted HDOM treatment (e.g., species Streptomyces rochei). Predictive functional profiling suggested limited impact on major metabolic pathways, with soil type remaining the primary determinant. These findings demonstrate that HDOM exerts a direct, dose-dependent, and soil-specific influence on bacterial communities, providing key insights into the environmental behavior of hydrochar and guiding its safe application. Full article

The genus Streptomyces is a major driver of the soil microbial community. These filamentous, exospore-producing bacteria are copious producers of bioactive compounds that are not only used as antibiotics but also affect the soil microbial community in composition and activity. With an average of about 30 different bioactive compounds produced per species, the bacteria use complex regulatory mechanisms that respond to environmental as well as community signals. Understanding these mechanisms will be useful in harnessing the full potential of Streptomyces in biotechnology, e.g., to tackle the antibiotic resistance crisis. This includes the discovery of new antibiotics that are not produced under standard laboratory conditions, as well as being able to modulate the signaling cascades to produce other biotechnology products. As an example, the genus Streptomyces, as one of the few bacterial and archaeal taxa, produces cobalamin de novo through both the oxic and anoxic biosynthesis pathways. This feature adds to the importance of this genus for the soil microbial communities, as well as for applications in fermentation. Full article

Cannabinoids are high-value bioactive compounds whose sustainable production remains challenging, prompting interest in biocatalytic and microbial platforms as alternatives to plant extraction. In this study, we investigated the heterologous expression and functionality of two key cannabinoid-related enzymes in the photosynthetic microalga Chlamydomonas reinhardtii: the aromatic prenyltransferase, NphB_G286S/Y288A from Streptomyces sp., and the plant-derived cannabidiolic acid synthase (CBDAS) from Cannabis sativa. Codon-optimized genes were introduced into the nuclear genome of C. reinhardtii using several construct configurations and promoters, and stable transformants were generated and characterized for genomic integration, transcript accumulation, protein production, enzymatic activity, and cannabinoid-related metabolite formation. While NphB protein accumulation was achieved under the PSAD promoter control, CBDAS was not detected at the protein level under any condition tested. In vitro enzymatic assays using soluble algal protein extracts from NphB-expressing lines confirmed catalytic activity, yielding cannabigerolic acid (CBGA), reaching up to 633 ± 58 µg L⁻¹. However, no CBGA production was detected in vivo, despite substrate supplementation. These results indicate that, although bacterial prenyltransferase can be functionally expressed in C. reinhardtii, efficient metabolic conversion in vivo is limited by cellular and biochemical constraints, including substrate availability, intracellular compartmentalization, and potential competition with endogenous pathways. In contrast, the absence of detectable CBDAS highlights the challenges associated with expressing complex plant oxidocyclases in this photosynthetic host. Overall, this work provides mechanistic insights into enzyme compatibility and metabolic bottlenecks in microalgal systems and outlines key considerations for the future development of photosynthetic platforms for cannabinoid biocatalysis. Full article

Actinomycetes are among the richest sources of bioactive secondary metabolites in biotechnology, owing to their remarkable metabolic diversity. Although the genus Streptomyces has been extensively explored and has yielded many clinically important antibiotics, rare actinomycetes remain comparatively underinvestigated. In this study, Kitasatospora sp. SeTe27, isolated from uncontaminated soil in Sicily (Italy), was investigated for its antibacterial activity and fermentation-driven enhancement of secondary metabolite production. The strain inhibited Staphylococcus aureus ATCC 25923, prompting physiological and genomic analyses. Spore conditioning was evaluated in four media (R5A, GYM, TSB, and YEME) to enhance antibiotic production. Conditioned cultures exhibited markedly increased antibacterial activity in TSB and YEME, moderate production in R5A, and no detectable activity in GYM. Whole-genome sequencing revealed an 8.5 Mb genome (73.5% GC) containing 48 biosynthetic gene clusters (BGCs), including NRPS, PKS, terpene, and hybrid pathways. Several clusters showed high similarity to known antibiotic-associated BGCs, such as clifednamide- and phenazine-related pathways, while numerous orphan clusters indicated significant unexplored biosynthetic potential. These findings identify Kitasatospora sp. SeTe27 as a promising antimicrobial producer and demonstrate that spore conditioning in complex media is an effective strategy to enhance antibiotic production in rare actinomycetes. Full article

Maize (Zea mays L.) cultivation is severely affected by Fusarium verticillioides, a highly adaptable systemic pathogen that causes serious yield losses, reduces grain quality, and produces toxic fumonisin, posing significant health risks to humans and livestock. A biological control approach to combating it was investigated. Streptomyces sp. BPTC-684 showed strong inhibitory activity (53.11%) against F. verticillioides BNGO-16, isolated from a diseased tissue sample. Based on physiological and biochemical characteristics, 16S rRNA gene sequencing, average nucleotide identity, and digital DNA–DNA hybridization, strain BPTC-684 is considered a candidate new species belonging to the genus Streptomyces. In silico analysis of Streptomyces sp. BPTC-684 showed that it expresses diverse biosynthetic gene clusters encoding potential bioactive compounds, notably antibiotics (kinamycin, antimycin, fuelimycins A-C, hangtaimycin, and deoxyhangtaimycin) and siderophores (desferrioxamines B and E). In addition, plant growth-promoting behaviors, such as indole-3-acetic acid production; phosphate solubilization; and the production of extracellular lytic enzymes that degrade cellulose, chitin, proteins, amylose, and xylan, were also discovered in Streptomyces sp. BPTC-684. The pot experiments demonstrated that plant height, fresh weight, and dry root weight were increased in strain BPTC-684 by 37.88%, 132.50%, and 223.81%, respectively, compared to F. verticillioides BNGO-16 on the 15th day of infection. These findings suggest that Streptomyces sp. BPTC-684 is a promising biological control agent for inhibiting fungal diseases and promoting maize growth. Full article

This study investigated the valorization of chicken feet, an underutilized poultry by-product, through enzymatic hydrolysis to obtain a protein hydrolysate with improved functional properties. Enzymatic treatment was carried out using Enzy-Mix U100 and collagenase from Streptomyces lavendulae, with cheese whey applied as a process medium. The resulting protein hydrolysate contained 59.1% protein and was characterized by high levels of glycine (31.64 g/100 g protein), hydroxyproline (10.91 g/100 g protein), and alanine (10.58 g/100 g protein). The hydrolysate exhibited strong techno-functional performance, with a water-binding capacity of 580%, an emulsifying activity index of 166 m²/g, and an emulsion stability index of 31 min. Microstructural analysis revealed irregular porous particles typical of freeze-dried protein hydrolysates, reflecting structural modification of collagen during enzymatic treatment. Mineral analysis showed notable levels of sodium (463.1 mg/100 g) and magnesium (351.8 mg/100 g). Microbiological evaluation demonstrated high sanitary quality, with a total viable count of 100 CFU/g and absence of coliforms, Escherichia coli, yeasts, and molds in 1 g of product. The technological process reduced the characteristic odor of chicken feet while maintaining a light color and good dispersibility. These findings confirm the potential of enzymatic hydrolysis as a sustainable strategy for converting poultry by-products into safe, value-added functional protein ingredients for food applications. Full article

Genome sequencing has revealed that microorganisms have the potential to produce many more natural products than previously thought; the challenge is to establish efficient ways to “wake up” those “sleeping” biosynthetic pathways or genes, which are undoubtedly expressed in nature under specific conditions that are not normally reproduced in the laboratory. To activate these cryptic natural products, co-cultivation of cross-kingdom microorganisms between Candida albicans and Streptomyces longshengensis was performed in this study. A novel peak generated through co-culture was isolated and analyzed by a high-performance liquid chromatograph (HPLC), and its chemical structure was further determined by using mass spectrum (MS) and nuclear magnetic resonance (NMR) analyses. Bioassays of antimicrobial and antitumor activities were performed, and heterologous expression in Escherichia coli was attempted. The chemical structure was identified as tryptophol, and the bioassays revealed that tryptophol showed antitumor activity with IC₅₀ values of 154.5, 144.3, 122.6, and 110.7 μg/mL against A549, MC38, HepG2, and MCF-7 cells, respectively. As a valuable compound, tryptophol was also heterologously expressed in E. coli C41 to address the drawbacks of chemical synthesis. These findings combine co-cultivation with genetic engineering for tryptophol biosynthesis, expanding its antitumor application and laying a foundation for its industrial and sustainable production. Full article

Members of the genus Streptomyces possess large genomes, a vast and largely unexplored metabolic potential, and a distinctive life cycle characterized by pronounced morphological differentiation. However, despite extensive molecular, genomic, and microbiological research, the functions of many genes in this genus remain poorly characterized. In this study, 929 complete Streptomyces genomes were analyzed. From the predicted proteomes of these genomes, proteins conserved in at least 75% of strains and lacking annotation in the KEGG GENES database were identified and clustered. To expand the annotation, synteny and co-occurrence analyses were performed between these unannotated proteins and annotated genes. A total of 330 conserved clusters were identified; 284 out of 330 clusters contain proteins encoded by genes that are syntenic with those associated with transcriptional regulation, fatty acid metabolism, two-component signaling systems, and morphological development. Additional clusters included metalloproteins and enzymes such as dehydrogenases, suggesting a wide functional spectrum. The conserved yet uncharacterized proteins identified in this analysis represent promising targets for future research, both for elucidating the molecular biology of Streptomyces and for expanding the range of secondary metabolites produced by these ecologically and industrially significant microorganisms Full article

Milbemycin D is a promising 16-membered macrolide insecticide with reported superior efficacy, but its commercial development has been hindered by extremely low natural yields. This study aimed to construct a high-yielding microbial platform for milbemycin D production using combinatorial biosynthesis and advanced genome editing. An optimized CRISPR/Cas9-AcrIIA4 system was employed to seamlessly replace the aveA3 polyketide synthase (PKS) gene in the ivermectin B1b-producing strain Streptomyces avermitilis HU501 with the heterologous milA3 PKS from S. bingchenggensis. The engineered strain was validated genetically and metabolically, followed by high-throughput screening and fermentation optimization in various media. The biosynthesized compound was structurally confirmed by spectroscopy. Bioactivity was evaluated against Bursaphelenchus xylophilus, Hyphantria cunea, and Plutella xylostella. The engineered strain S. avermitilis HU501-M successfully shifted its major product to milbemycin D, reaching a final titer of 679.03 mg/L. Bioassays revealed that milbemycin D exhibited significantly enhanced potency, with LC₅₀ values 8–24% lower than those of milbemycin A3/A4. This work demonstrates an efficient CRISPR/Cas9-mediated PKS replacement strategy to achieve the high-yield production of milbemycin D, offering a promising microbial source and a generalizable framework for engineering complex polyketide pathways. This proof-of-concept establishes a foundation for future process development toward potential commercial application. Full article

Acyl-CoA carboxylase (YCC) complexes generate essential starter and extender units for fatty acid and polyketide biosynthesis in Actinobacteria. In Streptomyces coelicolor, two tri-subunit YCC complexes, acetyl-CoA carboxylase (ACC) and propionyl-CoA carboxylase (PCC), have been characterized. However, comparative genomic analyses indicate that β/ε subunits are more diversified than currently appreciated. Here, we identify a previously unrecognized β/ε pair, AccB2 and AccE2, and demonstrate that they assemble with the canonical α subunit to form a functional YCC complex. Both genes are transcribed in vivo, and co-immunoprecipitation (Co-IP) reveals association with AccA1 and AccA2, with AccE2 showing stronger relative association with AccA1-containing pull-downs. In vitro reconstitution confirms carboxylation activity toward acetyl-CoA, propionyl-CoA, and butyryl-CoA, which is strongly dependent on AccE2. These findings expand the YCC repertoire in S. coelicolor and support a modular assembly model in which alternative β/ε combinations contribute to functional diversification of YCC complexes. Full article

Chemical investigation of the marine sponge-derived Streptomyces xiamenensis 1310KO-148 afforded six polycyclic tetramate macrolactams (PTMs), including three known compounds (1–3) and three previously undescribed chlorohydrin-containing analogues, chlokamycins B–D (4–6). Their planar structures were elucidated by extensive analysis of 1D and 2D NMR spectra and HR-ESIMS data, while the relative configurations were assigned using NOESY correlations. The absolute configurations were further confirmed by electronic circular dichroism (ECD) calculations. Compounds 3–6 exhibited significant cytotoxic activity against 14 human cancer cell lines (GI₅₀ = 2.68–24.92 μM) and antibacterial activity against Staphylococcus aureus (MIC = 16.00–32.00 μg/mL) and Micrococcus luteus (MIC = 4.00–32.00 μg/mL) among six tested bacterial strains. Full article

L-glutamate (L-Glu) is the primary excitatory neurotransmitter in the mammalian central nervous system. Developing a real-time monitoring system is essential to understanding the onset and progression of related conditions. However, the absence of an L-Glu dehydrogenase that is insensitive to oxygen limits the development of oxygen-independent electrochemical enzymatic sensors. Additionally, the most commonly used L-Glu-specific oxidase requires site-specific proteolytic post-translational modifications in specific host microorganisms, which makes protein engineering difficult. To address these issues, L-Glu oxidase derived from Streptomyces mobaraensis (SmEOx), which does not require post-translational modifications, was engineered to function as a dehydrogenase. Residues crucial for the oxidative half reaction with oxygen in SmEOx were identified, and mutagenesis studies were conducted. Mutant SmEOx variants with suppressed oxidase activity and improved dye-mediated dehydrogenase activity compared to the wild-type enzyme were successfully obtained. The ratio of dehydrogenase activity to oxidase activity (Dh/Ox) increased ~2900-fold in mutant M117I and ~6700-fold in mutant M117F/K400N compared to wild-type recombinant SmEOx. The resulting virtually L-Glu dehydrogenases (vEDHs) were modified with a redox mediator and evaluated using transient open-circuit potential (OCP)-based L-Glu measurements. As a result, the vEDH (M117F/K400N mutant)-immobilized electrode enabled electrochemical L-Glu detection under ambient oxygen without the need for an external electron mediator, unlike the wild-type enzyme. The created vEDH, together with the OCP sensor developed using it, paves the way for future development of miniaturized, real-time L-Glu monitoring systems with high temporal and spatial resolution. Full article