Applied Biochemistry and Biotechnology最新文献

As a novel protein post-translational modification, lysine succinylation is widely involved in metabolism regulation. To describe succinylated lysine's physiological functions and distribution patterns in Saccharopolyspora erythraea, a large and global protein succinylome was identified in a hypersuccinylated strain E3ΔsucC, using high-resolution 4D label-free mass spectrometry. Bioinformatic analysis was conducted to examine the succinylated proteins further in this study. The results showed that succinylated proteins were identified to be predominantly involved in protein synthesis, central carbon and nitrogen metabolism, and secondary metabolism. The process of lysine succinylation was found intricately regulated by a delicate interplay of factors, such as the relative abundance of lysine within the protein, the strategic positioning of polar amino acids flanking the succinylated sites, and the degree to which lysine residues are exposed to the solvent, thereby shaping the landscape of post-translational modifications. This systematic analysis has represented the global analysis of lysine succinylation in S. erythraea and has provided an important resource for exploring the function and regulation of lysine succinylation in S. erythraea and likely in all actinomycetes.

Edible mushrooms have been used as sustainable sources of proteases of industrial interest. The aim of this research was to investigate the influence of different culture media on mycelial growth and the potential of an Amazonian mushroom species, Auricularia fuscosuccinea DPUA 1624, in the biosynthesis of bovine milk coagulant enzymes. The species was cultivated on Sabouraud agar, malt, glucose, and peptone agar, malt extract agar, and glucose and peptone agar, supplemented with yeast extract for mycelial development. Enzyme biosynthesis was evaluated by submerged fermentation. Subsequently, the cultures were incubated at 28 °C for 8 days. Proteolytic and coagulant activities were determined using 1% azocasein solution and milk powder as substrates, respectively. In the results of radial growth speed of A. fuscosuccinea, the values were significant in the GYP and SAB + YE culture media. However, GYP agar favored the growth and mycelial vigor of A. fuscosuccinea; therefore, this medium was selected to obtain inoculum in the tests. In submerged fermentation, the MGYP medium favored the synthesis of proteases for A. fuscosuccinea and synthesized coagulant proteases in 100% of the media, in which significant activity was observed in SAB + YE. The significant production of coagulant proteases of A. fuscosuccinea was obtained under the following conditions: inoculum size 10%, 8 days of fermentation period, and 8 days of inoculum age. The results indicate that A. fuscosuccinea DPUA 1624 has potential for use in industrial manufacturing, especially in dairy products.

Glucan 1,4-alpha-maltohydrolase (3.2.1.133, GMH) is an important biocatalyst in the baking industry, which could delay the retrogradation of bread and improve its cold-storage durability. In the present study, a newly cloned Thgmh was characterized and secreted by Pichia pastoris (Komagataella pastoris). After computationally assisted rational design that promotes peptide folding, the maltogenic activity in supernatant was enhanced 1.6-fold in comparison with the base strain. The signal leading sequence screening and the gene dosage increment further improved secretion by approximately 6.4-fold. The purified rationally designed ThGMHs exhibited maximal activity against soluble starch at pH 7.0 and 60 ℃, and maltose is the main catalytic product. In a 5-L bioreactor, conventional fed-batch fermentation resulted in 6130 U mL^-1 extracellular maltogenic activity. Therefore, a promising strain for GMH production was developed, which provides a useful reference for the secretory production of other industrial enzymes.

The worldwide obesity prevalence is increasing, affecting around 4 million individuals annually. This research critically evaluated the anti-obesity efficacy of the Korean mudflat halophyte herb Suaeda japonica (Suaeda japonica Makino). In the obese mice model, the administration of 200 mg/kg b.w. of S. japonica extract (SJE) significantly mitigated obesity by modulating body and organ weight, food efficiency ratio, energy expenditure, multiple blood chemistry parameters, lipid accumulation, adipose tissue hypertrophy, and various gene expressions associated with lipogenesis and thermogenesis. The significant obesity control (80%) of the aforementioned concentration of SJE treatment in mice mimics the plant-derived commercial anti-obesity drug Garcinia cambogia (Garcinia gummi-gutta) (80%, 245 mg/kg) b.w. Since SJE has not been extensively studied for obesity management, this study demonstrated that it might influence physiological, biochemical, and molecular pathways to combat obesity and related metabolic illnesses.

Creatinine clearance is used to reflect the glomerular filtration rate to assess kidney function. Creatinine degradation-related enzymes have been used for creatinine detection in clinical medicine. The mixture of spores encapsulating either creatininase or creatinase or sarcosine oxidase could mediate a three-step reaction to produce hydrogen peroxide from creatinine. In this study, to achieve consecutive and efficient creatinine detection, degradation enzymes creatininase, creatinase, and sarcosine oxidase were co-encapsulated in a single Saccharomyces cerevisiae spore. The co-encapsulation spores performed high specific activity and enzymatic properties and converted creatinine to H₂O₂, which was 160% higher than the mixture of spores that individually expressed these three enzymes. The detection condition of co-encapsulation was optimized for the store at room temperature and resistance to environmental stresses. The S. cerevisiae spores can co-encapsulate enzyme families and catalyze consecutive reactions in the spore wall, having potential application prospects.

JAK1, a key regulator of multiple oncogenic pathways, is a sought-out target, and its expression in immune cells and tumour-infiltrating lymphocytes (TILs) is associated with a favorable prognosis in breast cancer. JAK1 activates IL-6 via ERBB2 receptor tyrosine kinase signalling and promotes metastatic cancer and STAT3 activation in breast cancer cells. Hence, targeting JAK1 in breast cancer is being explored as a potential therapeutic strategy. A comprehensive in silico approach was utilised in this study to identify selective JAK1 inhibitors from the Life chemicals database. First, we utilised an anticancer focussed library and performed molecular docking to screen against JAK1 protein. The top 10 compounds from docking were taken for cross-docking, to assess the selectivity towards JAK1 target. Lipinski's RO5 was checked for eliminating the compounds that violate rules. Toxicity, biological activity and reactivity for the identified best compounds were predicted by Protox-II server, PASS server and cDFT analysis respectively. MD simulations were carried out to examine the stability and dynamic behaviour of the top leads, including the long-term stability of the ligand-receptor complex and any conformational changes. Lastly, the MM/PBSA method was used to determine the binding free energy of the protein-ligand complex. Our in silico approach has yielded a promising set of compounds F2638-0133, F3408-0020 and F5833-7435 with the potential to selectively target JAK1, a critical player in breast cancer progression. The docking, simulation and MM/PBSA results were compared with standard drug abrocitinib. Identified compounds exhibit favorable binding interactions, electronic properties and robust stability profiles compared to standard drug, making them promising leads for further experimental validation.

Multi-targeted therapies are gaining attention in the management of multifactorial diseases due to their poly pharmacology, enhanced potency and reduced toxicity. Metabolic disorders like Type 2 diabetes mellitus (T2DM) and obesity necessitate multi-targeted therapy to improve insulin sensitivity, regulate glucose homeostasis and support weight loss. Medicinal plants rich in bioactive compounds exhibit multi-targetted action with minimal side effects. In the current study, Cocculus hirsutus methanol extract (CME) and its hydromethanolic fraction (HMF) were investigated for their multi-target potential. Significant inhibition of Dipeptidyl peptidase IV (DPP-IV), a key enzyme in glucose metabolism was observed due to CME (54%) and HMF (70%) at 10 µg/ml and 1 µg/ml respectively. Protein Tyrosine Phosphatase 1B (PTP-1B), involved in the regulation of insulin signalling, was also inhibited by CME (67%) and HMF (73%) at 10 µg/ml concentration. An increase in glucose uptake was observed due to CME (62% and 65%) and HMF (63% and 68%) in 3T3-L1 adipocytes and L6 myotubes at 100 ng/ml. Further, investigation of HMF showed a decrease in lipid accumulation by 63% at 1 µg/ml in 3T3-L1 cells. Interestingly, HMF improved insulin sensitivity by upregulating GLUT4 expression (p < 0.05) via the PI3K/AKT pathway in both 3T3-L1 adipocytes and L6 myotubes. An inhibition in lipid accumulation was also observed by suppression of Peroxisome proliferator-activated receptor γ (PPARγ) (p < 0.05), a key regulator of adipogenesis in 3T3-L1 adipocytes. Gas chromatography-mass spectrometry analysis of the HMF showed the major component to be 3-methylmannoside (26.52%).

Traditionally, Bidens pilosa L. is an edible herb utilized for various ailments. The study accomplished a complete analysis of B. pilosa extract including UPLC/T-TOF-MS/MS, GC-MS, and in vitro antiproliferative activity, in addition to molecular docking on kinase and aldose reductase enzymes. From GC-MS analysis, the percentage of identified unsaturated fatty acids (FAs) (11.38%) was greater than saturated FAs (8.69%), while the sterols percent (39.92%) was higher than the hydrocarbons percent (6.6%). Oleic and palmitic acids are the major FAs (9.48% and 6.14%, respectively). Phytochemical profile uncovered the presence of quercetin, kaempferol, myricetin, and isorhamnetin aglycones and/or glycoside derivatives alongside apigenin, acacetin, and luteolin derivatives. B. pilosa extract suppressed cell proliferation in a concentration-dependent manner against SNB-19 and SK-MEL-5 cell lines (IC₅₀ 1.66 ± 0.06 and 4.04 ± 0.14 mg/mL, respectively). These potentials aligned with the molecular docking results on aldose reductase and kinase enzymes with promising binding affinities (- 5.3 to - 8.89 kcal mol^-1). B. pilosa metabolites were found as kinases and aldose reductase inhibitors, which rationalize their antiproliferative activity. Unfortunately, toxicity assessments were not performed to assess the safety of B. pilosa extract. Assessment of the therapeutic efficiency via in vivo and clinical studies is required.

In this present investigation, plant-mediated synthesis of titanium oxide (TiO₂) nanoparticles was synthesized from seagrass (Thalassia hemprichi) using the hot plate combustion method (HPCM). Synthesized TiO₂ nanoparticles optical, functional, structural, and morphology properties were analyzed by UV-visible spectroscopy, Fourier transform infrared spectroscopy (FT-IR), powder X-ray diffraction (XRD), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDX). SEM analysis confirmed the spherical shape of the TiO₂ nanoparticles were observed in various sizes, viz., 50 nm and 78 nm. The XRD analysis revealed that TiO₂ nanoparticles have a body-centred cubic structure without a secondary phase. Green synthesized TiO₂ nanoparticle applications were studied against the antimicrobial, antioxidant, anticancer, and photocatalytic activity. The pathogenic bacterial strains, including Staphylococcus epidermidis, Staphylococcus aureus, Klebsiella pneumonia, and Pseudomonas aeruginosa, were tested against TiO₂ nanoparticles; the maximum level of activity was seen at a concentration of 50 µg/mL. The antioxidant assays were performed against TiO₂ nanoparticles, and inhibitory concentration values (IC₅₀) were 40.28 μg/mL of the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, 52.04 µg/mL of the acrylamide tertiary butyl sulfonic acid (ATBS) assay, and 16.91 µg/mL of the metal chelating assay. The anticancer activity was analyzed against MCF-7 cancer cells using TiO₂ nanoparticles, and the IC₅₀ value showed 64.14 µg/mL concentration. An eco-friendly and convenient method was formulated for the production of titanium oxide nanoparticles utilizing seagrass extract. The potential employment of TiO₂ involves water treatment, biomedicine, biosensors, and nanotechnology.

The present study investigated the genomic and functional potential of Burkholderia contaminans PB_AQ24, a bacterial strain isolated from the municipal solid waste dumpsite, for boosting the growth of Dendrocalamus strictus (Male bamboo) seedlings. The isolated strain exhibited high potency for metal solubilization and ACC (1-Aminocyclopropane-1-carboxylate) deaminase activity. Its genome harbored diverse genes responsible for nitrogen and phosphorus utilization (trpABCDES, iaaH, acdS, pstABCS, phoAUD, pqqABCDE, kdpABC, gln, and nirBD) and also an abundance of heavy metal tolerant genes (ftsH, hptX, iscX-fdx-hscAB-iscAUR, mgtA, corA, and copC). Seeds priming experiments carried out in heavy metal contaminated soils (such as waste dumpsite soil (WDS), fly ash dumpsite soil (FAS) and natural garden soil (NGS control)) augmented with Burkholderia contaminans sp. PB_AQ24 showed 85% sustenance of seedlings in WDS and 80% in FAS. The study thus highlighted the potential features in isolated bacterial strain Burkholderia sp. PB_AQ24 (NCBI accession no. JAQOUG000000000), which could boost the growth of bamboo seedlings in heavy metal contaminated soils and may be applied for restoration and rejuvenation of contaminated sites.