Applied Biochemistry and Biotechnology最新文献

Copper-based nanozyme has shown the superior in the oxidase-like activities due to its electron transfer ability between the Cu(I) and Cu(II) sites during the catalytic reactions. Herein, a Cu(I)-MOF (Cu-Mel) was readily synthesized by a traditional hydrothermal process using the precursors of Cu⁺ and melamine, which was then used in the laccase-like catalytic reactions for the first time. Some means, such as X-ray diffraction (XRD), Scanning electron microscope (SEM), Fourier transform infrared spectroscopy (FT-IR), and X-ray photoelectron spectroscopy (XPS), were employed to character the microstructure of the Cu-Mel. The catalytic oxidation of the 4-aminoantipyrine (4-AP) and 2,4-dichlorophenol (2,4-DP) was adopted to evaluate the laccase-like catalytic ability of the resulting Cu-Mel. The catalytic conditions including the temperatures, the presence of alcohols, and the ionic concentrations were varied to optimize the laccase-like activities, based on that, the highest laccase-like catalytic activity is presented with a higher maximum reaction rate (V_max). The good storage stability is also presented by the Cu-Mel. The Cu-Mel was utilized in the degradation of Congo red, showing a good degradation efficiency. These findings facilitate the development of the laccase mimics and serve as a foundation for the design and applications of Cu-MOFs in the nanozyme realm.

Hepatocellular carcinoma (HCC) is a primary liver malignancy characterized by high morbidity and mortality. Recently, ferroptosis has been recognized as an important factor in regulating cell growth in HCC. However, the role of ferroptosis-related genes in HCC remains unclear. The SRP119173 dataset from the Sequence Read Archive database was used to screen differentially expressed genes (DEGs) related to ferroptosis. Meanwhile, weighted gene co-expression network analysis was conducted to identify the HCC-related gene modules in the TCGA-liver hepatocellular carcinoma (LIHC) cohort. Next, the candidate genes related to HCC progression and ferroptosis were identified by Venn analysis. Kaplan-Meier, multivariate COX regression, and CIBERSORT analyses were then performed. Our results found that the levels of PIF1 5'-to-3' DNA helicase (PIF1) were notably elevated in HCC tissues relative to normal tissues. Additionally, HCC patients with high PIF1 expression had worse overall survival outcomes than patients with low PIF1 expression. Additionally, the PIF1 gene could independently predict HCC patients' prognosis. Meanwhile, HCC patients with high PIF1 levels had a higher proportion of regulatory T cells (Tregs) and M0 macrophages, as well as higher expression of immune checkpoints such as PD-1 (PDCD1) and PD-L1 (CD274), compared with patients with low PIF1 levels. Our data suggested that a ferroptosis-related gene PIF1 may serve as a potential biomarker for predicting prognosis in HCC patients.

Cervical cancer is a common tumor in women and one of the common causes of cancer death in women. Due to the aggressive and non-specific nature of traditional chemotherapy, there is a growing need for new treatment modalities. Currently, tumor immunotherapy is increasingly garnering attention as a disruptive treatment approach. Therefore, we constructed CCTP-SmacN7, a delivery system capable of releasing active molecules in the tumor microenvironment. CCTP-SmacN7 can not only inhibit tumor proliferation and migration, but also induce tumors to produce large amounts of reactive oxygen species. The production of reactive oxygen species can activate tumors to release or expose damage-associated molecular patterns, promote DC cell maturation, and ultimately activate T cells. Here, we present an innovative targeted treatment approach for cervical cancer. While inducing tumor immunogenic cell death, this program can also improve the tumor microenvironment and initiate the tumor immune cycle.

In the context of agribusiness, the agricultural and livestock sectors generate a considerable quantity of waste on a daily basis. Solid-state fermentation (SSF) represents a potential alternative for mitigating the adverse effects of residue accumulation and for producing high-value products such as enzymes. Pleurotus pulmonarius is capable of producing a number of commercial enzymes, including amylases. Accordingly, the present study sought to produce, characterize, and apply amylases obtained from solid-state fermentation of cocoa and peach-palm waste by the fungus Pleurotus pulmonarius CCB19. The highest amylase production by P. pulmonarius was observed after 3 days of solid-state fermentation of the cocoa shells, with an activity of 83.90 U/gds. The physicochemical characterization of the crude amylase using the artificial neural network (ANN) revealed that the highest activity was observed at pH 9 and a temperature of 20 °C (120.7 U/gds). Furthermore, the amylase demonstrated stability in the majority of the tested conditions, maintaining up to 80% of its residual activity for up to 120 min of incubation. With regard to the impact of ions and reagents on enzymatic activity, a positive effect was observed in the presence of Co⁺ ions at concentrations of 1 and 5 mM, whereas Cu⁺ ions at 5 mM demonstrated an inhibitory effect. The addition of SDS and EDTA reagents did not affect the observed activity. Furthermore, the extract was tested in commercial detergent formulations and demonstrated enhanced compatibility (110%) and efficacy (270% with boiled detergent) in removing starch stains from fabrics with Ariel liquid detergent. In conclusion, amylase derived from the fungus Pleurotus pulmonarius CCB19 exhibited favorable properties that make it a suitable candidate for use as an additive in laundry detergent formulations.

Azo dye was used to prepare a new series of complexes with chlorides of rhodium (Rh⁺³), ruthenium (Ru⁺³), and corona (Au⁺³). The prepared materials were subjected to infrared, ultraviolet-visible, and mass spectrometry, as well as thermogravimetric analysis, differential calorimetry, and elemental analysis. Conductivity, magnetic susceptibility, metal content, and chlorine content of the complexes were also measured. The complexes prepared from the dye were used to determine their ability to inhibit free radicals by measuring their antioxidant capacity using DPPH as a free radical and ascorbic acid as a standard substance and then determining the IC₅₀ value. The ability to inhibit free radicals of the complexes varied according to the IC₅₀ value and its comparison with ascorbic acid. The gold complex gave the highest ability to inhibit free radicals compared to the rest of the complexes, and this was the case and it was. The results are as follows (ascorbic acid >[Au(L)Cl]>[Rh(L)(H₂O)₂Cl]>[Ru(L)(H₂O)₂ Cl]. The effectiveness of corona and rhodium complexes as anticancer agents has been studied for specific types of breast cancer. Five different concentrations at a wavelength of 570 nm were used. From this, the average percentage of cell survival was calculated. It has been found that the highest concentration of 600 µg/ml increases the inhibition of cancer cells. The gold complex exhibited the highest inhibition, providing the strongest anticancer activity and the most effective inhibition of free radicals.

Diabetes affects approximately 422 million people worldwide, leading to 1.5 million deaths annually and causing severe complications such as kidney failure, neuropathy, and cardiovascular disease. Aldose reductase (AR), a key enzyme in the polyol pathway, is an important therapeutic target for managing these complications. The high cost, severe side effects, and rising drug resistance in traditional diabetes treatments underscore the urgent need for novel AR-targeting antidiabetic agents. Ficus benjamina used in traditional medicine demonstrates promising potential for diabetes management. This study investigated the antidiabetic potential of F. benjamina phytocompounds targeting AR receptor employing a structure-based drug design approach to identify potential antidiabetic drug agents. Using molecular docking, ADMET analysis, molecular dynamics (MD) simulation, MM/GBSA, MM/PBSA, and DFT calculations, we identified three promising lead compounds: adenocarpine (- 9.2 kcal/mol), marmesin (- 8.8 kcal/mol), and lycocernuine (- 8.4 kcal/mol). These compounds presented favorable pharmacokinetic, pharmacodynamic, and toxicity profiles, with a 500-ns MD simulation confirming their stability, supported by PCA and Gibbs FEL analysis. MM/GBSA study identified adenocarpine (- 72.53 kcal/mol) as the best compound, outperforming marmesin (- 70 kcal/mol) and lycocernuine (- 61.95 kcal/mol). DFT analysis revealed that adenocarpine exhibited the highest molecular reactivity (3.914 eV), while lycocernuine demonstrated the greatest kinetic stability (6.377 eV). Marmesin and lycocernuine showed increased reactivity upon transitioning from the free states (4.441 eV and 6.377 eV, respectively) to the bound states (4.359 eV and 6.231 eV, respectively). These results could lead to the development of adenocarpine, marmesin, and lycocernuine as novel drug candidates for diabetes, warranting further in vitro and in vivo validation.

γ-Glutamylcysteine (γ-EC) can increase intracellular glutathione (GSH) levels, which may prevent and alleviate age-related disorders and chronic diseases caused by oxidative damage. However, the commercial availability of γ-EC remains limited owing to its complex chemical synthesis from glutamate and cysteine. In this study, we have developed the method of the effective conversion of GSH to γ-EC to achieve the optimal reaction conditions for repeated batch production and potential application in industrial γ-EC production using the phytochelatin synthase-like enzyme NsPCS. For repeated batch conversion reactions, the optimal temperature was determined at 25 °C, where γ-EC showed good stability compared with that at 37 °C, leading to higher overall productivity. Cellulose sponges and microcrystalline cellulose (MCC) showed superior mechanical strength as immobilization carriers and greater stability and productivity than other materials. The total amounts of γ-EC obtained by NsPCS immobilized on the cellulose sponge and MCC were 305 mg and 291 mg, respectively, in a 5 mL reaction over five repeated batch reactions. These simple production processes are easily reproduced, and their high volumetric efficiency is promising for the industrial production of stable and low-cost γ-EC.

Spinal cord injury (SCI) is one of the devastating neurological disorders that leads to a loss of motor and sensory functions. Long non-coding RNA small nucleolar RNA host gene 6 (lncRNA SNHG6) plays a crucial role in inflammatory regulation across various diseases. This study investigates the role of SNHG6 in SCI development and its underlying regulatory mechanisms. Two experimental models were established: an in vitro model using LPS-challenged (100 ng/mL) mouse microglia BV2 cells and an in vivo model employing controlled spinal cord impact in mice. SNHG6, miR-182-5p, and NEUROD4 expression levels were quantified through RT-qPCR and Western blot. Functional and histological assessments were performed using the Basso mouse scale (BMS) and Nissl staining, respectively. Putative binding sites between SNHG6 and miR-182-5p, as well as between miR-182-5p and NEUROD4, were predicted using the ENCORI/starBase platform. These molecular interactions were validated through dual-luciferase reporter assays and RNA pull-down experiments, with further confirmation by qRT-PCR and Western blot analyses. Both LPS-stimulated BV2 cells and spinal cord tissues from SCI mice exhibited elevated SNHG6 expression. Downregulation of SNHG6 enhanced LPS-induced polarization of BV2 cells from M1-type to M2-type, significantly modulated the expression of pro-inflammatory factors (TNF-α, IL-1β, and IL-6) and anti-inflammatory factors (TGF-β, IL-10, and IL-13), and reduced injury severity in SCI mice. Our mechanistic studies revealed that SNHG6 functions as a molecular sponge for miR-182-5p to regulate NEUROD4 expression. This study demonstrates that SNHG6 knockdown promotes microglial M2-type polarization and alleviates inflammatory responses through modulation of the miR-182-5p/NEUROD4 axis, suggesting SNHG6 as a potential therapeutic target for SCI treatment.

Circular RNAs (circRNAs), along with their pathogenic property in non-small cell lung cancer (NSCLC), require comprehensive analyses and explanations. The study is established with the purpose to elucidate the potential molecular mechanism of circATP9A in NSCLC. CircATP9A and microRNA (miR)-582-3p were evaluated by real-time quantitative polymerase chain reaction, and ribosomal protein large P0 (RPLP0), cleaved caspase-3, cleaved Ki-67, epithelial-to-mesenchymal transition (EMT)-associated proteins (N-cadherin and E-cadherin), and core proteins of the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) pathway were by Western blot. The processes of proliferation, apoptosis, migration, and invasion were measured by cell counting kit-8, 5-ethynyl-2'deoxyuridine, flow cytometry, and Transwell. Gene interaction was verified by RNA immunoprecipitation and dual luciferase reporter assay. CircATP9A and RPLP0 were abnormally highly expressed in both NSCLC tissues and cell lines, while miR-582-3p was abnormally low. Knockdown of circATP9A reduced NSCLC proliferation, invasion migration, and EMT and promoted apoptosis. This was further validated in nude mouse xenograft experiments. The inhibitory effect of knockdown of circATP9A on NSCLC was reversed by knockdown of miR-582-3p. In addition, the promoting effect of overexpression of circATP9A on NSCLC was reversed by knockdown of RPLP0. Mechanistically, circATP9A acted as a competitive endogenous RNA, sequestering miR-582-3p away from its target, which in turn modulated the expression of RPLP0. CircATP9A activated the miR-582-3p/RPLP0 axis by regulating the PI3K/Akt pathway in NSCLC cells. CircATP9A stimulates NSCLC progression via miR-582-3p/RPLP0 axis and PI3K/AKT cascade activation.

New methodologies have been evaluated for validating analytical characterization with artificial neural networks (ANNs). Compared to previous machine learning models, these provide more accurate and automated results with high testing accuracy. The Schiff base ruthenium complexes used in the proposed study were synthesized using 4-aminoantipyrine derivatives. 4-Aminoantipyrine is a biologically active pharmacophore. The geometry of the complexes was confirmed by IR, electronic, and magnetic measurements. XRD analysis pointed out the nanocrystalline behavior of the chelates. The molecular structures have been optimized using DFT calculations. The ruthenium complexes are one of the main chemotherapeutic agents in anticancer therapy over platinum drugs due to a wide range of peculiarities. Complexes exhibit octahedral geometry as confirmed by magnetic measurements exhibiting more biological activity. The complexes are redox active depicting high biological potency. The Ru chelates also display high photocatalytic efficiency. The chelates also adhere to Lipinski's rule of five as evidenced from mole inspiration calculations. Among the chelates, RuL³ exhibits high anticancer potency suggesting a valuable candidate for the treatment of cancer. RuL⁵ has high antibacterial efficiency, and RuL⁴ complex possesses high antifungal activity. The chelates may serve as potential antimicrobial agents.