Targeting Bacterial Nanocellulose Properties through Tailored Downstream Techniques.

Bacterial nanocellulose (BNC) is a biodegradable polysaccharide with unique properties that make it an attractive material for various industrial applications. This study focuses on the strain Komagataeibacter medellinensis ID13488, a strain with the ability to produce high yields of BNC under acidic growth conditions and a promising candidate to use for industrial production of BNC. We conducted a comprehensive investigation into the effects of downstream treatments on the structural and mechanical characteristics of BNC. When compared to alkaline-treated BNC, autoclave-treated BNC exhibited around 78% superior flexibility in average, while it displayed nearly 40% lower stiffness on average. An SEM analysis revealed distinct surface characteristics, indicating differences in cellulose chain compaction. FTIR spectra demonstrated increased hydrogen bonding with prolonged interaction time with alkaline solutions. A thermal analysis showed enhanced thermal stability in alkaline-treated BNC, withstanding temperatures of nearly 300 °C before commencing degradation, compared to autoclaved BNC which starts degradation around 200 °C. These findings provide valuable insights for tailoring BNC properties for specific applications, particularly in industries requiring high purity and specific mechanical characteristics.

Exploring Microorganisms from Plastic-Polluted Sites: Unveiling Plastic Degradation and PHA Production Potential.

The exposure of microorganisms to conventional plastics is a relatively recent occurrence, affording limited time for evolutionary adaptation. As part of the EU-funded project BioICEP, this study delves into the plastic degradation potential of microorganisms isolated from sites with prolonged plastic pollution, such as plastic-polluted forests, biopolymer-contaminated soil, oil-contaminated soil, municipal landfill, but also a distinctive soil sample with plastic pieces buried three decades ago. Additionally, samples from Arthropoda species were investigated. In total, 150 strains were isolated and screened for the ability to use plastic-related substrates (Impranil dispersions, polyethylene terephthalate, terephthalic acid, and bis(2-hydroxyethyl) terephthalate). Twenty isolates selected based on their ability to grow on various substrates were identified as Streptomyces, Bacillus, Enterococcus, and Pseudomonas spp. Morphological features were recorded, and the 16S rRNA sequence was employed to construct a phylogenetic tree. Subsequent assessments unveiled that 5 out of the 20 strains displayed the capability to produce polyhydroxyalkanoates, utilizing pre-treated post-consumer PET samples. With Priestia sp. DG69 and Neobacillus sp. DG40 emerging as the most successful producers (4.14% and 3.34% of PHA, respectively), these strains are poised for further utilization in upcycling purposes, laying the foundation for the development of sustainable strategies for plastic waste management.

Triangular Silver Nanoplates as a Bioanalytical Tool: Potential COVID-19 Detection.

Nanotechnology offers new possibilities in molecular diagnostics, with nanoparticles gaining attention as biosensor upgrades. This study evaluates gold-coated silver nanoplates coated with PEG for enhanced protection, aiming to detect Spike protein with higher sensitivity, and emphasizes the importance of considering complex environments and appropriate controls for specific binding and accurate analysis. The sensitivity of antibody-coated PEGAuTSNPs as tools for immunoassays is demonstrated through fibronectin (Fn)- anti-fibronectin binding within an isolated extracellular matrix as a complex and native environment of Fn. Moreover, the optimal functionalization volume of Spike protein was determined (4 µg/mL of PEGAuTSNP). Anti-Spike was added to confirm binding, while the TJP1 protein was used as a negative control. The same experiment was used in the presence of horse serum to simulate a complex environment. According to Localized Surface Plasmon Resonance analysis and Dynamic Light Scattering size measurements, anti-Spike exhibited a stronger affinity for the nanoplates, causing TJP1 to be replaced by the antibody on the nanoplates' surface. Future research will involve exploring alternative complex environments, filtering larger molecules, and the optimization of immunoassay performance.

Enhanced Antimicrobial Activity of Biocompatible Bacterial Cellulose Films via Dual Synergistic Action of Curcumin and Triangular Silver Nanoplates.

Curcumin and triangular silver nanoplates (TSNP)-incorporated bacterial cellulose (BC) films present an ideal antimicrobial material for biomedical applications as they afford a complete set of requirements, including a broad range of long-lasting potency and superior efficacy antimicrobial activity, combined with low toxicity. Here, BC was produced by Komagataeibacter medellinensis ID13488 strain in the presence of curcumin in the production medium (2 and 10%). TSNP were incorporated in the produced BC/curcumin films using ex situ method (21.34 ppm) and the antimicrobial activity was evaluated against Escherichia coli ATCC95922 and Staphylococcus aureus ATCC25923 bacterial strains. Biological activity of these natural products was assessed in cytotoxicity assay against lung fibroblasts and in vivo using Caenorhabditis elegans and Danio rerio as model organisms. Derived films have shown excellent antimicrobial performance with growth inhibition up to 67% for E. coli and 95% for S. aureus. In a highly positive synergistic interaction, BC films with 10% curcumin and incorporated TSNP have shown reduced toxicity with 80% MRC5 cells survival rate. It was shown that only 100% concentrations of film extracts induce low toxicity effect on model organisms' development. The combined and synergistic advanced anti-infective functionalities of the curcumin and TSNP incorporated in BC have a high potential for development for application within the clinical setting.

Streptomyces sp. BV410: Interspecies cross-talk for staurosporine production.

AIMS: Sequencing and genome analysis of two co-isolated streptomycetes, named BV410-1 and BV410-10, and the effect of their co-cultivation on the staurosporine production. METHODS AND RESULTS: Identification of two strains through genome sequencing and their separation using different growth media was conducted. Sequence analysis revealed that the genome of BV410-1 was 9.5 Mb, whilst that of BV410-10 was 7.1 Mb. AntiSMASH analysis identified 28 biosynthetic gene clusters (BGCs) from BV410-1, including that responsible for staurosporine biosynthesis, whilst 20 BGCs were identified from BV410-10. The addition of cell-free supernatant from BV410-10 monoculture to BV410-1 fermentations improved the staurosporine yield from 8.35 mg L-1 up to 15.85 mg L-1 , whilst BV410-10 monoculture ethyl acetate extract did not have the same effect. Also, there was no improvement in staurosporine production when artificial mixed cultures were created using three different BV410-1 and BV410-10 spore ratios. CONCLUSIONS: The growth of BV410-10 was inhibited when the two strains were grown together on agar plates. Culture supernatants of BV410-10 showed potential to stimulate staurosporine production in BV410-1, but overall co-cultivation attempts did not restore the previously reported yield of staurosporine produced by the original mixed isolate. SIGNIFICANCE AND IMPACT OF STUDY: This work confirmed complex relations between streptomycetes in soil that are difficult to recreate under the laboratory conditions. Also, mining of streptomycetes genomes that mainly produce known bioactive compounds could still be the fruitful approach in search for novel bioactive molecules.

Macro and Micro Routes to High Performance Bioplastics: Bioplastic Biodegradability and Mechanical and Barrier Properties.

On a score sheet for plastics, bioplastics have a medium score for combined mechanical performance and a high score for biodegradability with respect to counterpart petroleum-based plastics. Analysis quickly confirms that endeavours to increase the mechanical performance score for bioplastics would be far more achievable than delivering adequate biodegradability for the recalcitrant plastics, while preserving their impressive mechanical performances. Key architectural features of both bioplastics and petroleum-based plastics, namely, molecular weight (Mw) and crystallinity, which underpin mechanical performance, typically have an inversely dependent relationship with biodegradability. In the case of bioplastics, both macro and micro strategies with dual positive correlation on mechanical and biodegradability performance, are available to address this dilemma. Regarding the macro approach, processing using selected fillers, plasticisers and compatibilisers have been shown to enhance both targeted mechanical properties and biodegradability within bioplastics. Whereas, regarding the micro approach, a whole host of bio and chemical synthetic routes are uniquely available, to produce improved bioplastics. In this review, the main characteristics of bioplastics in terms of mechanical and barrier performances, as well as biodegradability, have been assessed-identifying both macro and micro routes promoting favourable bioplastics' production, processability and performance.

Antimicrobial Active Bioplastics Using Triangular Silver Nanoplate Integrated Polycaprolactone and Polylactic Acid Films.

An innovative antimicrobial technology for plastic surfaces is presented. We report the synthesis and scale-up of triangular silver nanoplates (TSNPs) and their integration into polycaprolactone (PCL) and polylactic acid (PLA) polymers through a solvent-casting technique. The TSNPs have a high geometric aspect ratio and strong local surface plasmon resonance (LSPR) response, which provides an effective tool for monitoring their integrity during processing and integration with the biodegradable plastics. An aqueous-based seed-mediated chemical method was used to synthesize the TSNPs, and characterisation was carried out using TEM and UV (Ultraviolet)-VIS (Visible) spectroscopy to measure LSPR profiles. The UV-VIS spectra of silver seeds and TSNPs exhibited characteristic peaks at 395 and 600 nm respectively. Synthesized TSNPs were coated with thiol-terminated polyethylene glycol (SH-PEG) and transferred into chloroform in order to effect compatibility with PCL and PLA. TSNP/PCL and TSNP/PLA composite films were prepared by solvent casting. The morphological structure, thermal, mechanical, and antimicrobial properties of the TSNP-incorporated composite films were evaluated. Results showed the TSNP-treated films had a rougher surface than the bare films. Insignificant changes in the thermal properties of TSNP-treated films compared to bare ones were also observed, which indicated the thermal stability of the composite films. The tensile strength and antimicrobial properties of the composite films were increased after TSNP incorporation. TSNP/PCL and TSNP/PLA films exhibited improved antimicrobial activity against Escherichia coli and Staphylococcus aureus with antimicrobial effect (AE) values ranging between 0.10 and 0.35. The obtained results and demonstrated TSNP production scalability validate the TSNP treated PCL and PLA films as a composite material with desirable antimicrobial effect for wide-ranging surface applications.

Production of Polyhydroxybutyrate (PHB) and Factors Impacting Its Chemical and Mechanical Characteristics.

Plastic pollution is fueling the grave environmental threats currently facing humans, the animal kingdom, and the planet. The pursuit of renewable resourced biodegradable materials commenced in the 1970s with the need for carbon neutral fully sustainable products driving important progress in recent years. The development of bioplastic materials is highlighted as imperative to the solutions to our global environment challenges and to the restoration of the wellbeing of our planet. Bio-based plastics are becoming increasingly sustainable and are expected to substitute fossil-based plastics. Bioplastics currently include both, nondegradable and biodegradable compositions, depending on factors including the origins of production and post-use management and conditions. Among the most promising materials being developed and evaluated is polyhydroxybutyrate (PHB), a microbial bioprocessed polyester belonging to the polyhydroxyalkanoate (PHA) family. This biocompatible and non-toxic polymer is biosynthesized and accumulated by a number of specialized bacterial strains. The favorable mechanical properties and amenability to biodegradation when exposed to certain active biological environments, earmark PHB as a high potential replacement for petrochemical based polymers such as ubiquitous high density polyethylene (HDPE). To date, high production costs, minimal yields, production technology complexities, and difficulties relating to downstream processing are limiting factors for its progression and expansion in the marketplace. This review examines the chemical, mechanical, thermal, and crystalline characteristics of PHB, as well as various fermentation processing factors which influence the properties of PHB materials.

Pepper Bacterial Spot Control by Bacillus velezensis: Bioprocess Solution.

Pepper bacterial spot is one of the most severe plant diseases in terms of infection persistence and economic losses when it comes to fresh pepper fruits used in nutrition and industrial processing. In this study, Bacillus velezensis IP22 isolated from fresh cheese was used as a biocontrol agent of pepper bacterial spot, whose main causal agent is the cosmopolitan pathogen Xanthomonas euvesicatoria. After optimization of the cultivation medium composition aimed at maximizing of the antimicrobial activity against X. euvesicatoria and validation of the optimized medium at the scale of a laboratory bioreactor, in planta tests were performed. The results have showed significant suppression of bacterial spot symptoms in pepper plants by the produced biocontrol agent, as well as reduction of disease spreading on the healthy (uninoculated) pepper leaves. Furthermore, HPLC-MS (high pressure liquid chromatography-mass spectrometry) analysis was employed to examine antimicrobial metabolites produced by B. velezensis IP22, where lipopeptides were found with similar m/z values compared to lipopeptides from fengycin and locillomycin families. The bioprocess solution developed at the laboratory scale investigated in this study represents a promising strategy for production of pepper bacterial spot biocontrol agent based on B. velezensis IP22, a food isolate with a great perspective for application in plant protection.

Dinuclear silver(i) complexes with a pyridine-based macrocyclic type of ligand as antimicrobial agents against clinically relevant species: the influence of the counteranion on the structure diversification of the complexes.

New dinuclear silver(i) complexes with N,N',N'',N'''-tetrakis(2-pyridylmethyl)-1,4,8,11-tetraazacyclotetradecane (tpmc), [Ag2(NO3)(tpmc)]NO3·1.7H2O (1), [Ag2(CF3SO3)2(tpmc)] (2), and [Ag2(tpmc)](BF4)2 (3) were synthesized and characterized by NMR (1H and 13C), IR and UV-Vis spectroscopy, cyclic voltammetry and molar conductivity measurements. The molecular structures of the complexes were determined by single-crystal X-ray diffraction analysis. The spectroscopic and crystallographic data showed that the structure of the complexes strongly depends on the nature of the counteranion of silver(i) salt used for their synthesis. The antimicrobial activity of complexes 1-3 was examined against Gram-positive and Gram-negative bacteria and different species of unicellular fungus Candida spp. The ability of these complexes to inhibit the formation of Candida biofilms and to eradicate the already formed biofilms was tested in the standard microtiter plate-based assay. In addition, a bioelectrochemical testing of the antimicrobial activity of complex 1 against early biofilm was also performed. The obtained results indicated that complexes 1-3 showed increased activity toward Gram-negative bacteria and Candida spp. and could inhibit the formation of biofilms. In most cases, these complexes had positive selectivity indices and showed similar or even better activity with respect to the clinically used silver(i) sulfadiazine (AgSD). The values of the binding constants for complexes 1-3 to bovine serum albumin (BSA) were found to be high enough to indicate their binding to this biomolecule, but not so high as to prevent their release upon arrival at the target site. Moreover, the positive values of partition coefficients for these complexes indicated their ability to be transported through the cell membrane. Once inside the cell, complexes 1-3 could induce the formation of the reactive oxygen species (ROS) in C. albicans cells and/or interact with DNA. Taken together, silver(i) complexes with the tpmc ligand could be considered as novel antimicrobial compounds with favourable pharmacological properties, being safer than AgSD.

Zinc(II) complexes with aromatic nitrogen-containing heterocycles as antifungal agents: Synergistic activity with clinically used drug nystatin.

Three novel Zn(II) complexes, [ZnCl2(qz)2] (1), [ZnCl2(1,5-naph)]n (2) and [ZnCl2(4,7-phen)2] (3), where qz is quinazoline, 1,5-naph is 1,5-naphthyridine and 4,7-phen is 4,7-phenanthroline, were synthesized by the reactions of ZnCl2 and the corresponding N-heterocyclic ligand in 1:2 molar ratio in ethanol at ambient temperature. The characterization of these complexes was done by NMR, IR and UV-Vis spectroscopy, and their crystal structures were determined by single-crystal X-ray diffraction analysis. Complexes 1 and 3 are mononuclear species, in which Zn(II) ion is tetrahedrally coordinated by two nitrogen atoms belonging to two qz or 4,7-phen ligands, respectively, and by two chloride anions, while complex 2 is a 1D coordination polymer that contains 1,5-naph as bridging ligand between two metal ions. In agar disc-diffusion assay, complexes 1-3 manifested good inhibitory activity against two investigated Candida strains (C. albicans and C. parapsilosis), while not inducing toxic effects on the healthy human fibroblast cell line (MRC-5). This activity was not fungicidal, as revealed by the broth microdilution assay, however complex 3 showed the ability to modulate Candida hyphae formation, which is an important process during infection and showed significant synergistic effect with clinically used antifungal polyene nystatin.

Streptomyces sp. BV410 isolate from chamomile rhizosphere soil efficiently produces staurosporine with antifungal and antiangiogenic properties.

Applying a bioactivity-guided isolation approach, staurosporine was separated and identified as the active principle in the culture extract of the new isolate Streptomyces sp. BV410 collected from the chamomile rhizosphere. The biotechnological production of staurosporine by strain BV410 was optimized to yield 56 mg/L after 14 days of incubation in soy flour-glucose-starch-mannitol-based fermentation medium (JS). The addition of FeSO4 significantly improved the staurosporine yield by 30%, while the addition of ZnSO4 significantly reduced staurosporine yield by 62% in comparison with the starting conditions. Although staurosporine was first isolated in 1977 from Lentzea albida (now Streptomyces staurosporeus) and its potent kinase inhibitory effect has been established, here, the biological activity of this natural product was assessed in depth in vivo using a selection of transgenic zebrafish (Danio rerio) models, including Tg(fli1:EGFP) with green fluorescent protein-labeled endothelial cells allowing visualization and monitoring of blood vessels. This confirmed a remarkable antiangiogenic activity of the compound at doses of 1 ng/ml (2.14 nmol/L) which is below doses inducing toxic effects (45 ng/ml; 75 nmol/L). A new, efficient producing strain of commercially significant staurosporine has been described along with optimized fermentation conditions, which may lead to optimization of the staurosporine scaffold and its wider applicability.

New polynuclear 1,5-naphthyridine-silver(I) complexes as potential antimicrobial agents: The key role of the nature of donor coordinated to the metal center.

New polynuclear silver(I) complexes with 1,5-naphthyridine (1,5-naph), [Ag(NO3)(1,5-naph)]n (Ag1), [Ag(CF3COO)(1,5-naph)]n (Ag2) and [Ag(CF3SO3)(1,5-naph)]n (Ag3) were synthesized by the reaction of the corresponding silver(I) salt and 1,5-naph in ethanol at room temperature. These complexes were characterized by NMR, IR and UV-Vis spectroscopy, while their crystal structures were determined by single-crystal X-ray diffraction analysis. In all these complexes, 1,5-naph acts as a bridging ligand between two Ag(I) ions, while the remaining coordination sites are occupied by oxygen atom(s) of the corresponding anion. The antimicrobial efficiency of these silver(I) complexes was evaluated against the broad panel of Gram-positive and Gram-negative bacteria and fungi. The complexes showed good to moderate antibacterial activity with the minimal inhibitory concentration (MIC) values being in the range 2.5-100 µg/mL (6.5-333.3 µM), while their antifungal activity against the investigated Candida spp. was significantly higher (MIC = 0.78-6.25 µg/mL; 2.6-20.8 µM). Moreover, complexes Ag1 and Ag2 effectively inhibited C. albicans biofilms formation, while Ag1 was also shown to inhibit the formation of mixed C. albicans/Pseudomonas aeruginosa biofilms. Toxicological evaluations on zebrafish (Danio rerio) embryos revealed that all silver(I) complexes could be applied as antifungal agents, whereas Ag3 had the best therapeutic potential showing both the lowest MIC values against the tested Candida strains and the non-toxic in vivo response in the zebrafish embryos at these doses.

Antifungal potential of bacterial rhizosphere isolates associated with three ethno-medicinal plants (poppy, chamomile, and nettle)

The objective of the present study was to isolate Actinobacteria, preferably Streptomyces spp. from the rhizosphere soils of three ethno-medicinal plants collected in Serbia (Papaver rhoeas, Matricaria chamomilla, and Urtica dioica) and to screen their antifungal activity against Candida spp. Overall, 103 sporulating isolates were collected from rhizosphere soil samples and determined as Streptomyces spp. Two different media and two extraction procedures were used to facilitate identification of antifungals. Overall, 412 crude cell extracts were tested against Candida albicans using disk diffusion assays, with 42% (43/103) of the strains showing the ability to produce antifungal agents. Also, extracts inhibited growth of important human pathogens: Candida krusei, Candida parapsilosis, and Candida glabrata. Based on the established degree and range of antifungal activity, nine isolates, confirmed as streptomycetes by 16S rRNA sequencing, were selected for further testing. Their ability to inhibit Candida growth in liquid culture, to inhibit biofilm formation, and to disperse pre-formed biofilms was assessed with active concentrations from 8 to 250 μg/mL. High-performance liquid chromatographic profiles of extracts derived from selected strains were recorded, revealing moderate metabolic diversity. Our results proved that rhizosphere soil of ethno-medicinal plants is a prolific source of streptomycetes, producers of potentially new antifungal compounds

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Antifungal potential of bacterial rhizosphere isolates associated with three ethno-medicinal plants (poppy, chamomile, and nettle).

The objective of the present study was to isolate Actinobacteria, preferably Streptomyces spp. from the rhizosphere soils of three ethno-medicinal plants collected in Serbia (Papaver rhoeas, Matricaria chamomilla, and Urtica dioica) and to screen their antifungal activity against Candida spp. Overall, 103 sporulating isolates were collected from rhizosphere soil samples and determined as Streptomyces spp. Two different media and two extraction procedures were used to facilitate identification of antifungals. Overall, 412 crude cell extracts were tested against Candida albicans using disk diffusion assays, with 42% (43/103) of the strains showing the ability to produce antifungal agents. Also, extracts inhibited growth of important human pathogens: Candida krusei, Candida parapsilosis, and Candida glabrata. Based on the established degree and range of antifungal activity, nine isolates, confirmed as streptomycetes by 16S rRNA sequencing, were selected for further testing. Their ability to inhibit Candida growth in liquid culture, to inhibit biofilm formation, and to disperse pre-formed biofilms was assessed with active concentrations from 8 to 250 µg/mL. High-performance liquid chromatographic profiles of extracts derived from selected strains were recorded, revealing moderate metabolic diversity. Our results proved that rhizosphere soil of ethno-medicinal plants is a prolific source of streptomycetes, producers of potentially new antifungal compounds.