Synthesis Of Antibiotics Research Articles

A rapid microwave approach for ‘one-pot’ synthesis of antibiotic conjugated silver nanoparticles with antimicrobial activity against multi-drug resistant bacterial pathogens

Deaths directly attributable to drug-resistant infections reached 1.27 million in 2019 and continue to rise. This escalating resistance to antibiotics has driven a resurgence in the exploration of ancient antimicrobials to develop efficacious alternatives. The modern field of nanomaterials is a promising area of research with silver nanoparticles performing well as antimicrobial agents due to their large surface area and multiple bacterial targets. In the current study antibiotic conjugated silver nanoparticles (3–35 nm) were synthesized using β-lactam antibiotic, ampicillin. The method of heating during synthesis either microwave (4 min) or convection (4 h) influenced the physical characteristics of the ampicillin coated silver nanoparticles, however both approaches produced nanomaterials with antimicrobial activity against a variety of multi-drug resistant (MDR) clinical isolates in physiologically relevant media (when present at <0.2–2.28 mg L−1 in defined media). Critically, the microwave method is five times faster than the traditional water bath method, allowing rapid synthesis of ampicillin-conjugated nanoparticles, which supports scale up processes for industry. We suggest that the combination of antibiotic and silver in these nanoparticles produces a synergistic effect that circumvents resistance mechanisms and has the potential to provide a new line of combinatorial agents able to treat multi-drug resistant infections.

Design, synthesis, and bioactivity investigation of novel cyclic lipopeptide antibiotics targeting top-priority multidrug-resistant gram-negative bacteria

ObjectivesPolymyxins are the last-line therapy for top-priority multidrug-resistant (MDR) gram-negative bacteria. However, polymyxin nephrotoxicity impedes its clinical application. This study aimed to design, synthesize, and identify a novel and promising polymyxin derivative with high efficacy and low toxicity. MethodsTo design polymyxin derivatives, we reduced the hydrophobicity of the two hydrophobic domains (fatty acyl chain and D-Phe6-L-Leu7) and modified the positive charged L-2,4-diaminobutyric acid (Dab) residues. Twenty-five derivatives were synthesized, and their antibacterial activities in vitro and renal cytotoxicities were determined. The nephrotoxicity and pharmacokinetic parameters of compound 12 were examined in rats. Antibacterial efficacy in vivo was evaluated using a mouse systemic infection model. Surface plasmon resonance analysis, compound 12-rifampicin combination therapy, and scanning electron microscopy were used to study the mechanism of action of compound 12. ResultsThis research found a new compound, identified as compound 12, which showed similar or increased antibacterial activity against all tested sensitive and carbapenem-resistant gram-negative bacteria. It exhibited reduced renal cytotoxicity and nephrotoxicity, a favorable pharmacokinetic profile, and maintained or improved antibacterial efficacy in vivo. Importantly, its anti-Pseudomonas aeruginosa activity significantly improved. Compound 12, when combined with rifampicin, enhanced the activity of rifampin against gram-negative bacteria. Compound 12 also showed a high affinity for lipopolysaccharide and disrupted cell membrane integrity. ConclusionReducing the hydrophobicity of the two domains reduced renal cytotoxicity and nephrotoxicity. Shortening the side chain of Dab3 by one carbon maintained or increased its antibacterial activity both in vitro and in vivo. Furthermore, only the length of the side chain of Dab9 could be shortened by one carbon among the Dab1,5 and Dab8,9 residues. The bactericidal effects of compound 12 were related to the disruption of cell membrane integrity. Compound 12 may be a promising candidate for combating sensitive and carbapenem-resistant gram-negative bacterial infections, especially Pseudomonas aeruginosa.

Heterologous Expression of Ketoreductase ChKRED20 Mutant in Pichia pastoris and Bioreductive Production of (R)-1, 3-Butanediol.

(R)-1, 3-Butanediol (1, 3-BDO) is an important intermediate in the synthesis of aromatics, pheromones, insecticides, and beta-lactam antibiotics. The ChKRED20 is a robust NADH-dependent ketoreductase identified from Chryseobacterium sp. CA49. We obtained a ChKRED20 mutant (M12) through directed evolutionary screening of ChKRED20, the mutant with significantly improved activity to asymmetrically reduce 4-hydroxy-2-butanone (4H2B) to (R)-1, 3-BDO. So far, both ChKRED20 and its mutants have been expressed in intracellular in E. coli, the process of purification after intracellular expression is complicated, which leads to high cost. Here, we expressed M12 by constructing multicopy expression strains in P. pastoris, and the target protein yield was 302 mg/L in shake-flask fermentation and approximately 3.5 g/L in high-density fermentation. The recombinant M12 showed optimal enzyme activity at 30 °C and had high activity within a broad pH range of 6.0-8.0, and also showed high thermal stability. The recombinant M12 was further used for the reduction of 4H2B to (R)-1, 3-BDO, and 98.9% yield was achieved at 4540 mM 4H2B. The crude M12 enzyme extract was found to catalyze the bioreductive production of (R)-1, 3-BDO with excellent stereoselectivity (ee > 99%) and meet the production requirements. Our research shows that the M12 mutant can be used for the synthesis of (R)-1, 3-BDO, and the P. pastoris expression system is an ideal platform for the large-scale, low-cost preparation of ChKRED20 or its mutants, which may have applications in industrial settings.

Microbial synthesis of sedoheptulose from glucose by metabolically engineered Corynebacterium glutamicum

BackgroundSeven-carbon sugars, which rarely exist in nature, are the key constitutional unit of septacidin and hygromycin B in bacteria. These sugars exhibit a potential therapeutic effect for hypoglycaemia and cancer and serve as building blocks for the synthesis of C-glycosides and novel antibiotics. However, chemical and enzymatic approaches for the synthesis of seven-carbon sugars have faced challenges, such as complex reaction steps, low overall yields and high-cost feedstock, limiting their industrial-scale production.ResultsIn this work, we propose a strain engineering approach for synthesising sedoheptulose using glucose as sole feedstock. The gene pfkA encoding 6-phosphofructokinase in Corynebacterium glutamicum was inactivated to direct the carbon flux towards the pentose phosphate pathway in the cellular metabolic network. This genetic modification successfully enabled the synthesis of sedoheptulose from glucose. Additionally, we identified key enzymes responsible for product formation through transcriptome analysis, and their corresponding genes were overexpressed, resulting in a further 20% increase in sedoheptulose production.ConclusionWe achieved a sedoheptulose concentration of 24 g/L with a yield of 0.4 g/g glucose in a 1 L fermenter, marking the highest value up to date. The produced sedoheptulose could further function as feedstock for synthesising structural seven-carbon sugars through coupling with enzymatic isomerisation, epimerisation and reduction reactions.

Synthesis and Characterization of Nanosilica-Coated Antibiotics, TAP: A Comprehensive Study Utilizing XRD, EDS, FTIR, SEM, and TEM – Invitro Study

Abstract Aim: To synthesize nanosilica and investigate its properties, focusing on its interaction with antibiotics, particularly triple antibiotic paste (TAP). Various analytical techniques, including X-ray diffraction (XRD), energy-dispersive X-ray spectroscopy (EDS), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and transmission electron microscopy (TEM), were employed. Materials and Methods: The synthesis of nanosilica involved the meticulous combination of ammonia, ethanol, water, and tetraethyl orthosilicate in a sterile conical flask, followed by centrifugation and drying. Nanosilica was further explored by being combined with antibiotics, including doxycycline, Flagyl, and ciprofloxacin, to create novel antibiotic-coated variants. These combinations were subjected to various analytical techniques to assess their properties. Results: The FTIR spectra of silica nanoparticles revealed specific peaks associated with silicon-oxygen bonds and siloxane structures. Nanosilica coated with antibiotics exhibited distinctive peaks indicating the presence of organic components and Si-O-Si bonds, offering insights into their potential applications in drug delivery systems. SEM images displayed uniformly dispersed spherical particles, while TEM images and histograms provided valuable data about the size distribution of nanosilica, essential for applications like drug delivery. XRD analysis confirmed the crystalline nature of silica nanoparticles and offered insights into their crystallographic structure. Conclusion: This comprehensive study showed the synthesis and characterization of nanosilica and its antibiotic-coated variants, with a particular focus on TAP. The findings contribute to our understanding of their structural and compositional properties, providing potential avenues for applications in diverse scientific and technological fields, including drug delivery and nanotechnology.

BiVO4/sulfur-doped g-C3N4 nanocomposite as photocatalyst for degradation of ciprofloxacin under visible light irradiation

In recent years, the use of antibiotics has greatly increased, which has caused environmental pollution. We offer a photocatalyst based on BiVO4/sulfur-doped g-C3N4 nanocomposite, which was made by a one-step thermal condensation method from thiourea and bismuth vanadate to degradate ciprofloxacin (CIP). The BiVO4/sulfur-doped g-C3N4 photocatalyst was analyzed by certain techniques including XRD, FTIR, BET, FESEM, EDX, TEM, DRS, photoluminesence, and Mott-Schottky to determine various physicochemical properties. The photocatalytic degradation of CIP, as a model pollutant, under visible LED light irradiation was tested to investigate the catalytic performance of nanocomposite. Response Surface Methodology (RSM) was employed to optimize operational parameters, including photocatalyst dosage (50–150 mg), CIP concentration (10–30 mg/L), initial pH of the solution (3–11), and irradiation time (12–120 min). The BiVO4/sulfur-doped g-C3N4 nanocomposite, with a narrow bandgap of 2.15 eV, exhibited exceptional photocatalytic performance in degrading CIP. The optimal conditions for CIP removal were photocatalyst dosage of 120.9 mg, CIP concentration of 10 mg/L, and initial solution pH of 3, resulting in 93.5 % removal efficiency after 105 min of irradiation. The photocatalytic degradation kinetics of CIP followed a pseudo-first-order model with a rate constant (K) of −0.0221 L/min, indicating rapid degradation process. The photocatalyst maintained excellent performance after 5 reuses, with only a 3 % reduction in efficiency. XRD analysis confirmed the structural stability of the nanocomposite after reuse.

Large-scale production of cefazolin in a microreactor with a low impurity content and high yield

Cefazolin is crucial in the field of healthcare because of its efficacy against bacterial diseases, but trace amounts of polymer impurities produced during synthesis may cause severe allergic reactions. In this work, an efficient system based on a membrane dispersion microreactor was designed for scalable cefazolin production with a high yield and low impurity content. At a production scale of 200 kg, the yield could reach 90 %, and the polymer impurity content could reach <0.008 %. Density functional theory calculations were used to investigate the formation mechanisms of the polymer impurities. The results revealed that the activation energy of cefazolin ring-opening and substitution reactions decreased by 22.4 kJ/mol and 6.4 kJ/mol respectively under alkaline conditions, resulting in increased polymer content. Computational fluid dynamics simulation indicates that the organic base in the substrate can be rapidly dispersed in the membrane dispersion microreactor, thus avoiding excessive local base concentration. In conclusion, the development of a membrane dispersion microreactor system offers a scalable and efficient method for cefazolin production with significantly lower polymer impurity content, thereby addressing a critical challenge in antibiotic synthesis and ensuring safer and more reliable healthcare applications.

Uncoated gold nanoparticles create fewer and less localized defects in model prokaryotic than in model eukaryotic lipid membranes

The rise of the populations of antibiotic resistant bacteria represents an increasing threat to human health. In addition to the synthesis of new antibiotics, which is an extremely expensive and time-consuming process, one of the ways to combat bacterial infections is the use of gold nanoparticles (Au NPs) as the vehicles for targeted delivery of therapeutic drugs. Since such a strategy requires the investigation of the effect of Au NPs (with and without drugs) on both bacterial and human cells, we investigated how the presence of coating-free Au NPs affects the physicochemical properties of lipid membranes that model prokaryotic (PRO) and eukaryotic (EU) cells. PRO/EU systems prepared as multilamellar liposomes (MLVs) and hybrid structures (HSs) from 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-dipalmitoyl-sn-glycero-3-phosphatidylglycerol (DPPG)/1,2-dipalmitoyl-sn-glycero-3-phosphoserine (DPPS) in the absence (MLVs)/presence (HSs) of differently distributed Au NPs (sizes ∼20 nm) reported stabilization of the gel phase of PRO systems in comparison with EU one (DSC data of PRO/EU were Tm(MLVs) ≈ 41.8 °C/42.0 °C, Tm¯ (HSs) ≈ 43.1 °C/42.4 °C, whereas UV-Vis response Tm(MLVs) ≈ 41.5 °C/42.0 °C, Tm¯ (HSs) ≈ 42.9 °C/41.1 °C). Vibrational spectroscopic data unraveled a substantial impact of Au NPs on the non-polar part of lipid bilayers, emphasizing the increase of kink and gauche conformers of the hydrocarbon chain. By interpreting the latter as Au NPs-induced defects, which exert the greatest effect when Au NPs are found exclusively outside the lipid membrane, these findings suggested that Au NPs reduced the compactness of EU-based lipid bilayers much more than in analogous PRO systems. Since the uncoated Au NPs manifested adverse effects when applied as antimicrobials, the results obtained in this work contribute towards recognizing AuNP functionalization as a strategy in tuning and reversing this effect.

Predictive Functional Profiling Reveals Putative Metabolic Capacities of Bacterial Communities in Drinking Water Resources and Distribution Supply in Mega Manila, Philippines

Assessing bacterial communities across water resources is crucial for understanding ecological dynamics and improving water quality management. This study examines the functional profiles of bacterial communities in drinking water resources in Mega Manila, Philippines, including Laguna Lake tributaries, pre-treatment plant sites, groundwater sources, and post-treatment plant sites. Using eDNA sequencing, flux balance analysis, and taxonomy-to-phenotype mapping, we identified metabolic pathways involved in nutrient metabolism, pollutant degradation, antibio- tic synthesis, and nutrient cycling. Despite site variations, there are shared metabolic pathways, suggesting the influence of common ecological factors. Site-specific differences in pathways like ascorbate, aldarate, and phenylalanine metabolism indicate localized environmental adaptations. Antibiotic synthesis pathways, such as streptomycin and polyketide sugar unit biosynthesis, were detected across sites. Bacterial communities in raw and pre-treatment water showed potential for pollutant degradation such as for endocrine-disrupting chemicals. High levels of ammonia-oxidizing and sulfate-reducing bacteria in pre- and post-treatment water suggest active nitrogen removal and pH neutralization, indicating a need to reassess existing water treatment approaches. This study underscores the adaptability of bacterial communities to environmental factors, as well as the importance of considering their functional profiles in assessing drinking water quality resources in urban areas.

Chemical Predictive Modelling and Natural Product-based Divergent Synthesis - Design of Type B PPAPs with Nanomolar Activities against MRSA.

In response to the pressing global challenge of antibiotic resistance, time efficient design and synthesis of novel antibiotics are of immense need. Polycyclic polyprenylated acylphloroglucinols (PPAP) were previously reported to effectively combat a range of gram-positive bacteria. Although the exact mode of action is still not clear, we conceptualized a late-stage divergent synthesis approach to expand our natural product-based PPAP library by 30 additional entities to perform SAR studies against methicillin-resistant Staphylococcus aureus (MRSA). Although at this point only data from cellular assays are available and understanding of molecular drug-target interactions are lacking, the experimental data were used to generate 3D-QSAR models via an artificial intelligence training and to identify a common pharmacophore model. The experimentally validated QSAR model enabled the estimation of anti-MRSA activities of a virtual compound library consisting of more than 100,000 in-silico generated B PPAPs, out of which the 20 most promising candidates were synthesized. These novel PPAPs revealed significantly improved cellular activities against MRSA with growth inhibition down to concentrations less than 1 μm.

Bdelloid rotifers deploy horizontally acquired biosynthetic genes against a fungal pathogen

Coevolutionary antagonism generates relentless selection that can favour genetic exchange, including transfer of antibiotic synthesis and resistance genes among bacteria, and sexual recombination of disease resistance alleles in eukaryotes. We report an unusual link between biological conflict and DNA transfer in bdelloid rotifers, microscopic animals whose genomes show elevated levels of horizontal gene transfer from non-metazoan taxa. When rotifers were challenged with a fungal pathogen, horizontally acquired genes were over twice as likely to be upregulated as other genes — a stronger enrichment than observed for abiotic stressors. Among hundreds of upregulated genes, the most markedly overrepresented were clusters resembling bacterial polyketide and nonribosomal peptide synthetases that produce antibiotics. Upregulation of these clusters in a pathogen-resistant rotifer species was nearly ten times stronger than in a susceptible species. By acquiring, domesticating, and expressing non-metazoan biosynthetic pathways, bdelloids may have evolved to resist natural enemies using antimicrobial mechanisms absent from other animals.

Antibacterial activity and biosynthetic potential of Streptomyces sp. PBR19, isolated from forest rhizosphere soil of Assam.

An Actinomycetia isolate, designated as PBR19, was derived from the rhizosphere soil of Pobitora Wildlife Sanctuary (PWS), Assam, India. The isolate, identified as Streptomyces sp., shares a sequence similarity of 93.96% with its nearest type strain, Streptomyces atrovirens. This finding indicates the potential classification of PBR19 as a new taxon within the Actinomycetota phylum. PBR19 displayed notable antibacterial action against some ESKAPE pathogens. The ethyl acetate extract of PBR19 (EtAc-PBR19) showed the lowest minimum inhibitory concentration (MIC) of ≥ 0.195µg/mL against Acinetobacter baumannii ATCC BAA-1705. A lower MIC indicates higher potency against the tested pathogen. Scanning electron microscope (SEM) findings revealed significant changes in the cytoplasmic membrane structure of the pathogen. This suggests that the antibacterial activity may be linked to the disruption of the microbial membrane. The predominant chemical compound detected in the EtAc-PBR19 was identified as phenol, 3,5-bis(1,1-dimethylethyl), comprising 48.59% of the area percentage. Additionally, PBR19 was found to contain the type II polyketide synthases (PKS type II) gene associated with antibiotic synthesis. The predicted gene product of PKSII was identified as the macrolide antibiotic Megalomicin A. The taxonomic distinctiveness, potent antibacterial effects, and the presence of a gene associated with antibiotic synthesis suggest that PBR19 could be a valuable candidate for further exploration in drug development and synthetic biology. The study contributes to the broader understanding of microbial diversity and the potential for discovering bioactive compounds in less-explored environments.

Micro-nano H2 bubbles enhanced hydrodehalogenation of 3-chloro-4-fluoroaniline: Mass transfer and action mechanism

3-chloro-4-fluoraniline (FCA) is an important intermediate for the synthesis of antibiotics, herbicides and insecticides, and has significant environmental health hazards. Catalytic hydrogenation technology is widely used in pretreatment of halogenated organics due to its simple process and excellent performance. However, compared with the research of high activity hydrogenation catalyst, the research of efficient utilization of hydrogen source under mild conditions is not sufficient. In this work, micro-nano H2 bubbles are produced in situ by electrolytic water and active metal replacement, and their apparent properties are studied. The result show that the H2 bubbles have a size distribution in the range of 150–900 nm, which can rapidly reduce the REDOX potential of the water and maintain it in a hydrogen-rich state for a long time. Under the action of Pd/C catalyst, atomic hydrogen (H•) produced by dissociative adsorption can sequentially hydrogenate FCA to aniline. The H• utilization ratios of the above two hydrogen supply pathways reach 6.20% and 4.94% respectively, and H2 consumption is reduced by tens of times (≥50 → ≈1.0 mL/min). The research provides technical support for the efficient removal of halogenated refractory pollutants in water and the development of hydrogen economy.

Metagenomic and metabolomic analysis showing the adverse risk–benefit trade-off of the ketogenic diet

BackgroundKetogenic diets are increasingly popular for addressing obesity, but their impacts on the gut microbiota and metabolome remain unclear. This paper aimed to investigate how a ketogenic diet affects intestinal microorganisms and metabolites in obesity.MethodsMale mice were provided with one of the following dietary regimens: normal chow, high-fat diet, ketogenic diet, or high-fat diet converted to ketogenic diet. Body weight and fat mass were measured weekly using high-precision electronic balances and minispec body composition analyzers. Metagenomics and non-targeted metabolomics data were used to analyze differences in intestinal contents.ResultsObese mice on the ketogenic diet exhibited notable improvements in weight and body fat. However, these were accompanied by a significant decrease in intestinal microbial diversity, as well as an increase in Firmicutes abundance and a 247% increase in the Firmicutes/Bacteroidetes ratio. The ketogenic diet also altered multiple metabolic pathways in the gut, including glucose, lipid, energy, carbohydrate, amino acid, ketone body, butanoate, and methane pathways, as well as bacterial secretion and colonization pathways. These changes were associated with increased intestinal inflammation and dysbiosis in obese mice. Furthermore, the ketogenic diet enhanced the secretion of bile and the synthesis of aminoglycoside antibiotics in obese mice, which may impair the gut microbiota and be associated with intestinal inflammation and immunity.ConclusionsThe study suggest that the ketogenic diet had an unfavorable risk–benefit trade-off and may compromise metabolic homeostasis in obese mice.

Effects of the pleiotropic regulator DasR on lincomycin production in Streptomyces lincolnensis

The lincoamide antibiotic lincomycin, derived from Streptomyces lincolnensis, is widely used for the treatment of infections caused by gram-positive bacteria. As a common global regulatory factor of GntR family, DasR usually exists as a regulatory factor that negatively regulates antibiotic synthesis in Streptomyces. However, the regulatory effect of DasR on lincomycin biosynthesis in S. lincolnensis has not been thoroughly investigated. The present study demonstrates that DasR functions as a positive regulator of lincomycin biosynthesis in S. lincolnensis, and its overexpression strain OdasR exhibits a remarkable 7.97-fold increase in lincomycin production compared to the wild-type strain. The effects of DasR overexpression could be attenuated by the addition of GlcNAc in the medium in S. lincolnensis. Combined with transcriptome sequencing and RT-qPCR results, it was found that most structural genes in GlcNAc metabolism and central carbon metabolism were up-regulated, but the lincomycin biosynthetic gene cluster (lmb) were down-regulated after dasR knock-out. However, DasR binding were detected with the DasR responsive elements (dre) of genes involved in GlcNAc metabolism pathway through electrophoretic mobility shift assay, while they were not observed in the lmb. These findings will provide novel insights for the genetic manipulation of S. lincolnensis to enhance lincomycin production.Key points• DasR is a positive regulator that promotes lincomycin synthesis and does not affect spore production• DasR promotes lincomycin production through indirect regulation• DasR correlates with nutrient perception in S. lincolnensis

Improving the enzymatic activity and stability of N-carbamoyl hydrolase using deep learning approach

BackgroundOptically active D-amino acids are widely used as intermediates in the synthesis of antibiotics, insecticides, and peptide hormones. Currently, the two-enzyme cascade reaction is the most efficient way to produce D-amino acids using enzymes DHdt and DCase, but DCase is susceptible to heat inactivation. Here, to enhance the enzymatic activity and thermal stability of DCase, a rational design software “Feitian” was developed based on kcat prediction using the deep learning approach.ResultsAccording to empirical design and prediction of “Feitian” software, six single-point mutants with high kcat value were selected and successfully constructed by site-directed mutagenesis. Out of six, three mutants (Q4C, T212S, and A302C) showed higher enzymatic activity than the wild-type. Furthermore, the combined triple-point mutant DCase-M3 (Q4C/T212S/A302C) exhibited a 4.25-fold increase in activity (29.77 ± 4.52 U) and a 2.25-fold increase in thermal stability as compared to the wild-type, respectively. Through the whole-cell reaction, the high titer of D-HPG (2.57 ± 0.43 mM) was produced by the mutant Q4C/T212S/A302C, which was about 2.04-fold of the wild-type. Molecular dynamics simulation results showed that DCase-M3 significantly enhances the rigidity of the catalytic site and thus increases the activity of DCase-M3.ConclusionsIn this study, an efficient rational design software “Feitian” was successfully developed with a prediction accuracy of about 50% in enzymatic activity. A triple-point mutant DCase-M3 (Q4C/T212S/A302C) with enhanced enzymatic activity and thermostability was successfully obtained, which could be applied to the development of a fully enzymatic process for the industrial production of D-HPG.

A smooth vetch (Vicia villosa var.) strain endogenous to the broad-spectrum antagonist Bacillus siamensis JSZ06 alleviates banana wilt disease.

Fusarium wilt, caused by Fusarium oxysporum f. sp. cubense Tropical Race 4 (Foc TR4), poses a significant threat to banana production globally, thereby necessitating effective biocontrol methods to manage this devastating disease. This study investigates the potential of Bacillus siamensis strain JSZ06, isolated from smooth vetch, as a biocontrol agent against Foc TR4. To this end, we conducted a series of in vitro and in vivo experiments to evaluate the antifungal activity of strain JSZ06 and its crude extracts. Additionally, genomic analyses were performed to identify antibiotic synthesis genes, while metabolomic profiling was conducted to characterize bioactive compounds. The results demonstrated that strain JSZ06 exhibited strong inhibitory activity against Foc TR4, significantly reducing mycelial growth and spore germination. Moreover, scanning and transmission electron microscopy revealed substantial ultrastructural damage to Foc TR4 mycelia treated with JSZ06 extracts. Genomic analysis identified several antibiotic synthesis genes, and metabolomic profiling revealed numerous antifungal metabolites. Furthermore, in pot trials, the application of JSZ06 fermentation broth significantly enhanced banana plant growth and reduced disease severity, achieving biocontrol efficiencies of 76.71% and 79.25% for leaves and pseudostems, respectively. In conclusion, Bacillus siamensis JSZ06 is a promising biocontrol agent against Fusarium wilt in bananas, with its dual action of direct antifungal activity and plant growth promotion underscoring its potential for integrated disease management strategies.

Application of enzymes in drug discovery research: A Review

Enzymes have effectively substituted many conventional chemical catalysts-reliant pharmaceutical drug manufacturing procedures. From this v-prospect, the use of enzymes in drug discovery research is quite encouraging. Numerous enzymes have been demonstrated to perform detoxification at the cellular level, for instance, the antioxidant enzymes, catalase, and superoxide dismutase (SOD). These enzymes work simultaneously to neutralize oxygen-free radical species. The enzyme-mediated pharmaceutical procedures include the synthesis of diverse semi-artificial antibiotics, dynamic enantiomers of medicines through kinetic determination, production of enantiomerically unadulterated types of amino acids, and others. Enzymes are being used to treat cancer and infectious disorders where antibiotics are no longer effective because of antibiotic resistance. Thus, the present article aims to review the diverse use of enzymes in the drug industries and discuss the features of enzymes that make them suitable drug candidates.

Comparative Evaluation of the Effectiveness of Biocatalytic Synthesis and Antibacterial Activity of Known Antibiotics and “Chimeric” Cephalosporin Compounds

Comparative Evaluation of the Effectiveness of Biocatalytic Synthesis and Antibacterial Activity of Known Antibiotics and “Chimeric” Cephalosporin Compounds

ANALYSIS OF BIOSYNTHETIC GENE CLUSTERS OF BACILLUS VELEZENSIS ONU 553 IN SILICO

The aim of the work was to analyse biosynthetic gene clusters (BGC) of Bacillus velezensis ONU 553 based on bioinformatics approach. Methods. Identification of species was processed with tools of TYGS server; EzBioCloud was used to calculate ANI. Analysis of biosynthetic gene, bacteriocin, and antibiotic resistance gene clusters using antiSMASH, Bagel4, respectively. The results. It is shown that the results of identification, phylogenetic analysis and DNA-DNA hybridization (DDH) carried out in silico proved that the strain Bacillus velezensis ONU 553 belongs to the operational group B. amyloliquefaciens (OGBa). Sequences identified as possible phages and CpG-islands were found in the genome of our strain. 12 biosynthetic gene clusters (BGC) were identified using antiSMASH. One new cluster capable of synthesizing a new metabolite was identified (region 11). The presence of two clusters of bacteriocins in the genome of Bacillus velezensis ONU 553, which are assigned to uberolysin/carnocyclin and the antimicrobial peptide LCI based on the identification of the core gene, is shown. Conclusions. The preliminary identification of the Bacillus velezensis ONU 553 strain as a representative of the Bacillus velezensis strain of the B. amyloliquefaciens group (OGBa) was confirmed. The presence of gene clusters of secondary metabolites responsible for the synthesis of surfactins, polyene antibiotics, antimicrobial peptides, macrolide antibiotics and bacteriocins was shown. The obtained results indicate that the Bacillus velezensis ONU 553 strain is promising for use in the field of "Blue Biotechnology" for the development of new drugs with antimicrobial and antifungal activity.