Applied Biochemistry and Biotechnology最新文献

The current study examines the anti-dengue and antibacterial potential and in silico drug development of the andrographolide against the pathogens Staphylococcus aureus (1JIJ) and Pseudomonas aeruginosa (6MVN) and the dengue viral protein NS5. The phytochemical analysis identified the presence of flavonoids, alkaloids, saponins, tannins, phenols, glycosides, and steroids in the Andrographis paniculata methanolic leaf extract. Based on the GC-MS analysis, andrographolide was identified as the primary chemical constituent of the medicinal plant A. paniculata and the compound with the largest peak area among other compounds. The agar well diffusion method was employed to observe the antibacterial potential of the methanolic leaf extract against S. aureus and P. aeruginosa. The maximal inhibition zones were 15.6 ± 0.34 mm and 11.7 ± 0.42 mm, respectively. Additionally, andrographolide was docked with the NS5 viral protein and bacterial proteins, including S. aureus TyrRS (PDB: 1JIJ) and P. aeruginosa LasR PDB (ID: 6MVN), resulting in a docking score of - 20.7384 kJ/mol, - 15.0969 kJ/mol, and - 11.1171 kJ/mol, respectively. In summary, our molecular docking experiments with the identified andrographolide compound demonstrated its potential as a drug with anti-dengue viral and antibacterial properties.

In this study, adding mulberry leaf flavonoids (MLFs) during the liquid-state fermentation of Monascus purpureus significantly improved the production of Monascus pigments (MPs). Compared with the control group, the red, yellow, and orange pigment levels increased by 1.69, 1.4, and 1.29 times, respectively. Metabolomic analysis suggested adding rutin, the primary component of MLFs, induced changes in 32 significantly different metabolites. These changes included a significant increase in (S)-2,3,4,5-tetrahydropyridine-2-carboxylate, D-4'-phosphopantothenate, cis-4-hydroxy-D-proline, and L-isoleucine. Enriched Kyoto Encyclopedia of Genes and Genomics (KEGG) metabolic pathways revealed valine, leucine, and isoleucine biosynthesis, and phenylalanine metabolism improvements. These improvements help to activate the pigment synthesis pathway and promote pigment synthesis. These findings offer crucial insights into the impact of MLFs on Monascus purpureus metabolic pathways and suggest potential methods for increasing MP production.

This study explores the green synthesis of silver (Ag-NPs), copper (Cu-NPs), and silver-copper bimetallic nanoparticles (Ag-Cu Bimetallic NPs) using the leaf extract of Peganum harmala, a sustainable and renewable plant source. This eco-friendly synthesis method avoids hazardous chemicals and minimizes waste, advancing the principles of green nanotechnology. Characterization demonstrated distinct properties; Ag-NPs, Cu-NPs, and Ag-Cu bimetallic NPs exhibited surface plasmon resonance peaks at 425 nm, 555 nm, and 525 nm, respectively, and crystallite sizes of 21.42 nm, 21.40 nm, and 26.29 nm as confirmed by X-ray diffraction. Morphological analysis revealed spherical Ag-NPs, flake-like Cu-NPs, and a mix of cubic and spherical Ag-Cu NPs. Catalytic activity tests showed Ag-Cu bimetallic NPs achieved 95% degradation of methylene blue dye, highlighting their potential in environmental remediation. Antibacterial assays demonstrated that Ag-NPs exhibited the highest inhibition zones against Bacillus subtilis (27.0 mm) and Escherichia coli (27.67 mm), while Cu-NPs were effective against Klebsiella pneumoniae at higher concentrations. Ag-NPs also exhibited significant antifungal activity, particularly against Aspergillus niger (15.00 mm). Antioxidant assays revealed Ag-Cu bimetallic NPs displayed the highest free radical scavenging capacity (59.18%), followed by Ag-NPs (35.41%). Cytotoxicity tests indicated Ag-Cu bimetallic NPs had an IC₅₀ value of 6.13 µg/mL, reflecting high cytotoxicity, whereas Ag-NPs demonstrated better biocompatibility. Hemolysis assays showed P. harmala extract had the highest hemolytic activity (IC₅₀ = 4.49 µg/mL). This study provides a scalable, sustainable method for synthesizing multifunctional nanoparticles with promising applications in environmental remediation, antimicrobial resistance management, and biomedicine. Future studies should focus on scaling up this sustainable synthesis method, investigating the biological interactions of these nanoparticles, and optimizing their dosages to reduce potential environmental and health risks.

Till now, limited information was available on salt tolerance chickpea genotypes. Therefore, in comparison to CSG 8962 (check for salinity tolerance), an experiment on nine chickpea genotypes with different background (BG 1103, DCP 92-3, S7, ICCV 10, BG 256, KWR 108, JG 16, K 850, and ICC 4463) was conducted under medium salt stress of EC_iw ~ 6 dS m^-1 and high salt stress of 9 dS m^-1 to evaluate their salt tolerance potential. Different morphological, physiological, biochemical, and molecular traits were studied to characterize these genotypes. It was also noted that growth of all the genotypes was affected by salinity, but more reduction was shown by the genotypes BG 256, DCP 92-3, and ICC 4463. Irrigation water loaded with salts disrupted the water relations as displayed by the reducing values of RWC, water potential, and osmotic potential. Chlorophyll content, when compared with control, reduced in the range of 7.06 to 28.93% at moderate salinity level (EC_iw ~ 6 dS m^-1) and 23.71 to 55.83% at higher salinity level (EC_iw ~ 9 dS m^-1). S7, ICCV 10, KWR 108, and CSG 8962 (salt-tolerant check) maintained optimum gas exchange traits, i.e., photosynthetic rate, stomatal conductance, and transpiration rate with increasing salinity and osmoregulatory compounds, imino acid proline, and total soluble sugars were also higher in these genotypes. Na⁺/K⁺ ratio at control was 0.084 and it enhanced with increasing salinity and noted mean genotypic values of 0.399 and 0.758 at moderate and higher salinity levels, respectively. Antioxidative defense mechanism was quite active in the genotypes (S7, ICCV 10, KWR 108, and check CSG 8962) because higher values of antioxidative enzymes and low increment in the content of hydrogen peroxide and malondialdehyde were noted in these genotypes. Based on the results, genotypes with salinity contrasting response (KWR 108 as tolerant and ICC 4463 as sensitive) were selected, and gene expression studies were conducted along with CSG 8962 (the check). It was found that KWR 108 showed higher expression of Δ1-pyrroline-5-carboxylate synthetase (P5CS), pyrroline-5-carboxylate reductase (P5CR), Na⁺/H⁺ antiporter (NHX1), and sodium transporter HKT1 and downregulation of proline dehydrogenase gene than the genotype CSG 8962 (salt-tolerant check). So, it was concluded that genotypes, i.e., S7, KWR 108, and ICCV 10, maintained higher physiological and biochemical efficiency in terms of lower ψ_w, ψ_s, and membrane stability, higher RWC, photosynthetic rate, and osmolyte accumulation as well as antioxidative enzyme activities in comparison to the salt-tolerant check used in the study. Further, these results were validated through gene expression studies which revealed similar results that categorized these genotypes to be salt tolerant.

Antibiotics, the full-stop of invasive bacteria, have been used in clinical setups from unthreatening fever to massive challenging therapies. Constant dependency on medication upsurges the evasion of microbes from antibiotics contemporarily along with ecological footprint. Thus, the infested pathogen became resilient to antibiotics, disguised as multidrug-resistant bacteria (MDR), pandrug-resistant bacteria (PDR), and extensively drug-resistant bacteria (XDR). The etymology of genetic modifications and horizontal gene transfer played an external influence on the arising resurgence. Also, intrinsic parameters, such as antibiotic efflux pumps and the formation of biofilms, encouraged intense resistance to antibiotic drugs. This aggravated resistance in microbes builds up resistome in the environment due to selective pressure; thereby drastic devastation of people suffering from disastrous diseases is mournful. Since novelite approaches for broad-spectrum antibiotics against drug resistance microbes are grueling challenges in these crucial times. This scientific study has come up with neoteric methodologies to elude immediate consequences and health hazards. Inculcating ancestral treatment towards pharmacognosy as adjuvants to the prevailing hi-fi nanotechnology, phage and algal therapy, genome mining, and bioinformatics databases are the optimizing inventions for actual and prospective living.

Food waste is an increasing cause of concern in India. Its management through composting plays a vital role in managing the biodegradable fraction of municipal solid waste. However, the existing composting process has many challenges, such as the lack of optimum microenvironment and microbiome knowledge, which limits efficient outcomes. Therefore, the present study aims to bridge the gap by applying metagenomics to study microbial community dynamicity during different stages of composting. The bacterial community analysis showed that genus Marionobacter (9.4%) and Halomonas (7.4%) were prevalent during the mesophilic stage, whereas the Bacillus (12.2%) and Cellulomonas (0.1%) were prevalent during the thermophilic and maturation stage of composting. The functional profiling of metagenome indicated the abundance of genes involved in degradation of polymeric compounds such as carbohydrates, lipids, and proteins. The relative abundance of arginine and proline metabolisms increased during the thermophilic stage. Whereas the relative abundance of genes involved in fatty acid, tryptophan, galactose, and propanoate metabolisms declined. Similarly, the CAZyme tool predicted that the genes encoding for glycoside hydrolase (GH) families were higher during the mesophilic and thermophilic stages of composting. These enzymes play an important role in degradation of complex polysaccharides such as cellulose and hemicellulose. The data obtained from the present study could be utilized for the optimization and improving the composting process.

Osteoporosis (OP) is a genetic disorder characterized by an imbalance between osteoblast-mediated bone formation and osteoclast-induced bone resorption. However, the underlying gene-related mechanisms of its pathogenesis remain to be fully elucidated. Aberrantly expressed neuralized E3 ubiquitin-protein ligase 3 (NEURL3), which is related to osteoclastic differentiation, was identified through the analysis of the microarray profile GSE176265. Bone marrow-derived macrophages (BMMs) were isolated from the femurs and tibias of C57BL/6 J mice and treated with 30 ng/mL macrophage-colony-stimulating factor (M-CSF) and 100 ng/mL receptor activator of nuclear factor-kappa B ligand (RANKL) to induce osteoclastic differentiation, thereby mimicking OP in vitro. To model OP in vivo, ovariectomy (OVX)-induced bone loss was performed in mice. High expression levels of NEURL3 were confirmed in clinical samples, OP model cells, and OP model mice using quantitative real-time polymerase chain reaction (qRT-PCR). The impact of NEURL3 on osteoclastic differentiation was assessed by evaluating cell viability and the expression levels of osteoclastogenesis-related marker genes. Additionally, bone loss in mice was quantified using micro-computed tomography before and after NEURL3 inhibition. Mechanistically, the effects of NEURL3 on osteogenic differentiation were investigated by determining the protein levels of osteogenic markers via Western blotting. NEURL3 was markedly overexpressed in serum samples collected from patients with OP, OVX-induced OP mouse models, and induced osteoclasts. Inhibition of NEURL3 leads to a 20% decrease in BMM survival rate and a reduction in the number of tartrate-resistant acid phosphatase (TRAP) positive cells, which is a characteristic of mature osteoclasts. Furthermore, the expression levels of osteoclastogenesis-related marker genes were reduced by 50%. In vivo studies revealed that suppressing NEURL3 resulted in a 38% improvement in trabecular bone volume (BV/TV) and a 28% increase in bone mineral density (BMD) in the OVX-induced OP mice. Mechanistically, NEURL3 promoted osteoclast differentiation by increasing the ubiquitination levels of BMP7. Inhibition of BMP7 reversed the effects of NEURL3 on osteoclast differentiation in BMMs. Suppression of NEURL3 inhibits osteoclast differentiation of BMMs in vitro and alleviates bone loss in vivo. The underlying mechanism may involve NEURL3-induced ubiquitination of BMP7. Collectively, the downregulation of NEURL3 represents a promising therapeutic strategy for suppressing osteoclast differentiation and treating OP.

The tree bean (Parkia timoriana (DC). Merr) is an underutilized legume and is abundantly found in Southeast Asia. It is valued for its nutritious pods and cultivated for food and timber. Despite of the presence of several nutrients, the regulatory networks involved in secondary metabolite biosynthesis in the tree bean remain largely unexplored. Recent studies have highlighted that consumption of its pods provides numerous health benefits, including antioxidant, α-glucosidase inhibitory, antibacterial, antidiabetic, and insecticidal activities. To elucidate the biosynthesis of specific metabolites in this plant, a comparative metabolite and transcriptomic analysis of the leaf and root tissues of P. timoriana was carried out. The study revealed that P. timoriana leaf and root tissues contain varying levels of phenolics, flavonoids, and terpenoids. ¹H nuclear magnetic resonance (¹H NMR) analysis identified 16 significant metabolites in the leaf and root tissues, including sugars, amino acids, and organic acids. L-dihydroxyphenylalanine (L-DOPA), an amino acid derivative and precursor to dopamine, was detected for the first time in the seeds. Additionally, the presence of pinitol in P. timoriana was also confirmed. De novo RNA-sequence analysis identified differentially expressed genes (DEGs) in both the tissues. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis identified pathways associated with shikimate pathway, such as phenylpropanoid and flavonoid biosynthesis. MapMan pathway analysis revealed a high number of transcripts related to phenylalanine, tryptophan, tyrosine, and condensed tannin biosynthesis. The research conducted identified secondary metabolites in P. timoriana, and their probable biosynthetic pathway which can be used for medicinal and nutritional purposes.

Enzymatic oligomerization of flavonoids enables the synthesis of biomolecules with different structures and improved physicochemical and biological properties and can therefore broaden their application in industry. In this study, the influence of the key reaction parameters temperature, solvent, substrate, and enzyme concentrations on the synthesis of esculin oligomers was investigated. The reaction was optimized using response surface methodology (RSM) in order to obtain the highest products' concentration and specific products' yield (per mass of enzyme). Mass spectrometry revealed that oligomers with a degree of polymerization of up to 4 were synthesized in which dimers were the most abundant, while the NMR analysis of the esculin dimer product showed that C8-C8 link between two esculin units was formed. Maximum products' concentration was obtained at 60 °C, in 14% (v/v) methanol, 7 mg/mL of esculin, and 54.6 U/L of laccase after 7 h, while the optimal conditions for specific products' yield differed in the aspect of optimal laccase concentration which was 19 U/L for this output. Synthesized esculin oligomers exhibited higher iron chelating and cupric reducing antioxidant capacities and similar or even superior free radical scavenging activity compared to monomeric esculin. Moreover, the mixture of synthesized esculin oligomers has shown a promising potential to be used as a skin prebiotics, suggesting novel applications in skincare industries.

Microplastic pollution has emerged as a new global concern because of its ubiquitous and persistent nature. Due to the rising use of plastics and discharge of plastic waste into coastal water bodies from point and non-point sources, the occurrence of microplastics along coastal ecosystems has become very prevalent. The current study is the first of its kind to evaluate the presence of microplastics in the surface water of river estuary along the coast of Odisha. Six GPS-fixed locations were used to collect the surface water samples from the Budhabalanga river estuary in Chandipur, Odisha, India. The samples were then subjected to further investigation to determine the types of microplastics present. The average microplastic abundance, according to our findings, ranged from 9.33 ± 2.11 items L^-1 to 28.50 ± 2.77 items L^-1. Microplastics come in a variety of colours and shapes, but the most prevalent kind is fibre-shaped and black in colour. The pollution load index of the sampling area was calculated to be 4.25 which is categorized under ecological risk level I. FE-SEM images clearly showed the topology of microplastics and ATR-FTIR analysis confirmed the presence of polyethylene, polypropylene, polyvinyl chloride (PVC), nylon, polycarbonate (PC), ethylene vinyl acetate (EVA) and polystyrene (PS) at sampling stations. Our investigation provides useful information that helps to reduce the ecological risk in habitats connected with contaminated sites, including both aquatic and terrestrial habitats.