Applied Biochemistry and Biotechnology最新文献

In this study, the interaction of melatonin (ME) and its metabolites 5-Methoxytryptamine (MT) and N-Acetylserotonin (NAS) with fish sperm DNA (dsDNA) was investigated using various methods such as spectrophotometry, spectrofluorimetric and viscosity measurements. The results obtained demonstrated that these molecules exhibited a significant binding interaction with dsDNA. The intercalation binding mode was confirmed by competitive binding studies with ethidium bromide and rhodamine B, UV–Vis spectrophotometry and viscosity measurements. The molecules were also evaluated for their cholinesterase (AChE and BChE) inhibition properties in vitro. Compared to the reference drug donepezil, the molecules exhibited higher inhibitory activity. They showed high AChE inhibitory activity with IC₅₀ values of 2.87 ± 0.0958–17.48 ± 0.0785 µM and BChE inhibitory activity with IC₅₀ values of 3.12 ± 1.74–22.48 ± 1.58 µM. Especially NAS molecule shows dual inhibitory effect on AChE and BChE with IC₅₀ values of 2.87 ± 0.0958 and 3.12 ± 1.74 μM, respectively. In addition to cholinesterase inhibitory activity, the molecules showed good antioxidant activity and metal chelating capacity. Antioxidant activity was determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) methods and it was found that the molecules showed various levels of antioxidant activity. These findings not only reveal the molecular interactions of ME, MT, and NAS, but also highlight their therapeutic potential, providing a basis for future drug development studies targeting oxidative stress and cholinergic dysfunction.

In this study, a research design was implemented to investigate the potential of PPy-COOH/Ni micromotors for the degradation of oils as water pollutants. The study's objective was to determine the micromotors' capacity for oil removal. For this purpose, PPy-COOH/Ni micromotors were prepared by template-directed electrodeposition method, and lipase enzyme (from Porcine Pancreas) was covalently bound to these micromotors. The micromotors were characterized by SEM, EDX, and FTIR techniques. From the SEM photographs, it was found that the synthesized micromotors were cylindrical, about 5 μm in diameter and 15 μm in size. In addition, from the FTIR spectra, the binding of lipase enzyme to the micromotors was shown by amide I, II, and III bonds at 1640 cm⁻¹, 1540 cm⁻¹, and 1240 cm⁻¹. The present study has determined that the optimal pH level for lipase-attached micromotors is 8.0. Furthermore, the optimal temperature has been ascertained to be 45°C. In addition, the results of this study indicate that lipase-attached micromotors exhibit superior thermal and storage stability in comparison to free lipase. When the reusability of the immobilised lipase was examined, the immobilised lipase showed 73.5% activity even after five uses. The lipase-attached micromotors were tested on tributyrin triglyceride and were found to degrade 90% of the initial amount of tributyrin within 90 min. The findings obtained are consistent with existing literature on the subject, and these lipase-attached micromotors have the potential to be utilized in environmental applications, such as oil breakdown.

In recent years, using modern technologies, researchers have harnessed the potential of yeast species for various industrial uses, such as the bioproduction of biopharmaceuticals, food additives, industrial biocatalysts, and biofuels. To improve the efficiency and potential of yeast species for industrial uses, genetic modification is carried out. Various genome engineering techniques, including Cre-loxP, homing endonucleases, zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9), have been employed by different research groups for the genetic manipulation of yeast species. Among different genome engineering techniques, CRISPR/Cas9 has become popular because of its precise editing at targeted loci with increased efficiency. The ease of use, effectiveness, and adaptability of CRISPR/Cas9 make multiplexing possible for simultaneously targeting multiple genes, which was earlier very challenging through traditional methods. Moreover, the ability to perform marker-free editing is the significant advantage offered by CRISPR/Cas9. This review focuses on the applications of the CRISPR/Cas9 system in both conventional and non-conventional yeast species. Further, we discussed the advancements of CRISPR/Cas9, including the regulation of gene transcription-activation/repression and other genome engineering aspects. Additionally, innovations in CRISPR/Cas9, such as cloning-free CRISPR/Cas9 assembly, CRISPR-targeted in vivo editing (ACtive), CRISPR/Cas9-induced gene conversion, and selective ploidy ablation (CRI-SPA) are also discussed for enhancing the potential applications of CRISPR/Cas9 in diverse yeast species.

As a sustainable energy source and potential petroleum-free fuel, microalgae biomass is gaining popularity. The varied metabolic activities of microalgae species allow the extraction of bioproducts for animal feed, food, nutraceuticals, cosmetics, and biopharmaceuticals. Concentrating and measuring microalgal biomass biochemicals requires a lot of raw materials due to the low biomass-to-liquid ratio. This makes sustainable energy equilibrium in microalgae processing difficult. Thus, microalgae collection is a difficult field that needs more research. This paper presents the latest chemical/physical flocculation, bioflocculation, and electrical-based microalgal biomass harvesting methods. Physical flocculation needs energy and equipment, which may make microalgal biomass collection expensive. However, chemical flocculation, which uses inorganic and organic flocculants, is becoming more successful. This review also discusses biomolecule separation using low-liquefying ionic liquids or salts. This approach allows varied biorefinery theories to value microalgal biomass while maintaining biomolecule structural integrity and activity. The article uses comprehensive methods to examine scale up, industrial application, life cycle analysis problems, and future research prospects in microalgae collection for sustainable biofuel generation.

Septic encephalopathy (SE), a severe complication of systemic inflammatory response syndrome, continues to lack effective therapeutic options due to its complex pathophysiology. Ginsenoside Rc (GRc), a principal bioactive component in several medicinal herbs, has demonstrated neuroprotective and anti-tumor properties. However, its potential role in the pathogenesis of SE and the identification of its molecular targets remain unclear. Rho-associated kinase (ROCK) has been implicated in the onset and progression of sepsis, although its precise mechanism of action is not fully understood. In this study, we demonstrated that GRc significantly alleviated cognitive impairment in a murine model of SE, as assessed by the Morris water maze and open-field tests. To explore the ROCK-mediated mechanisms underlying SE and the anti-septic effects of GRc, we examined changes in ROCK2 protein expression in LPS-induced SE mice and identified NF-κBp65 as a downstream phosphorylation target. GRc treatment effectively reduced ROCK2 expression and suppressed the secretion of inflammatory cytokines. These findings elucidate the pathological role of the ROCK2/NF-κBp65 signaling axis in SE and provide compelling evidence that GRc mitigates SE, at least in part, by directly binding to ROCK2 and inhibiting NF-κBp65 phosphorylation.

Polyunsaturated fatty acids (PUFAs) are essential dietary lipids, vital for health and cellular integrity, with growing applications in biofuel, food, and pharmaceuticals. In this sense, this work investigated the production of single cell oil (SCO) from newly isolated Mortierellaceae strains from Antarctica. These strains were identified through ITS region sequencing and one of them (Linnemannia sp. ACF038) was selected for scaling up studies. The study of the physiological characterization of the selected strain was made by assessing various parameters including pH, temperature, nitrogen sources, and concentrations through solid and flask cultures. Based on these results, it was possible to scale up and optimize the culture conditions for SCO production using bench-scale bioreactors with submerged cultures. This process involved the use of two different culture media and an in-depth kinetic study to identify the most effective strategy for SCO production. The study also examined the volumetric oxygen transfer coefficient and oxygen consumption rate. The results showed that the culture medium with high initial glucose content and sodium nitrate significantly increased lipid production, reaching 65% of lipid content. Linoleic acid (LA) comprised 42% of the lipids, indicating strong potential for large-scale LA production. Linnemannia sp. ACF038 proved to be a promising candidate for sustainable industrial LA production, meeting the rising demand for PUFAs.

Vagus nerve stimulation (VNS) has demonstrated cardioprotective effects in a variety of cardiovascular diseases, including cardiac ischemia and reperfusion (IR) injury. However, the mechanisms responsible for these effects have not been completely understood. The present work aimed to uncover the potential mechanisms through which VNS confers protection against cardiac IR injury. Rats subjected to cardiac IR injury received electrical VNS through the right cervical vagus nerve. This intervention led to a notable reduction in cardiac dysfunction and injury, as well as decreased cardiac apoptosis, oxidative stress, and inflammation. Moreover, VNS treatment improved mitochondrial biogenesis by upregulating estrogen-related receptor α (ERRα), nuclear respiratory factor 1 (NRF-1), and transcriptional factor A mitochondrial (TFAM). In addition, VNS treatment not only increased the copy number of mitochondrial DNA (mtDNA) and the content of adenosine triphosphate (ATP), but also effectively reduced mitochondrial damage. VNS also upregulated the expression of silent information regulator 1 (SIRT1) and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) in IR-injured hearts. Inhibition of either SIRT1 or PGC-1α significantly reversed the effects of VNS on mitochondrial biogenesis and abolished its cardioprotective benefits. Notably, VNS increased the level of acetylcholine (ACh) in IR-injured hearts. Administration of atropine, a muscarinic ACh receptor (mAChR) antagonist, counteracted the effects of VNS on the SIRT1/PGC-1α pathway, mitochondrial biogenesis, and the associated cardioprotective outcomes. These findings suggest that VNS protects against cardiac I/R injury by enhancing mitochondrial biogenesis. This beneficial effect of VNS on mitochondrial biogenesis is attributed to activation of the SIRT1/PGC-1α pathway through the ACh/mAChR axis. Therefore, this research offers fresh perspectives on the mechanisms underlying the cardioprotective effects of VNS.

Viruses are minuscule entities that cannot survive independently without a Living host. Pathogenic viruses pose a significant threat to global health, resulting annually in the deaths of thousands of people. Recent studies indicate that medicinal plants may serve as an effective source of sustainable natural antiviral agents. Saudi Arabia exhibits diverse climates, resulting in a variety of ecosystems and environmental setups with a diverse array of plant species. Diversified flora thriving in various climatic conditions produces a wide spectrum of phytochemical constituents, many of which possess antiviral and other pharmacological properties. Saudi Arabia, home to approximately 2285 distinct plant species, renders an advantageous situation for the development of novel chemopreventive and therapeutic agents for combating infectious diseases. This review seeks to secure information regarding the antiviral properties of medicinal plants and their constituent phytochemicals from Saudi Arabia. The review provides a Listing of the phytoconstituents responsible for antiviral effects, their mechanistic workings, and pathways associated with the activity. Data mined from PubMed, Scopus, ScienceDirect, Google Scholar, EMBASE, and ChemSpider are included. The findings indicate that under 5% of these plant species have been investigated for their antiviral potential, thereby highlighting a significant gap in the information source. Investigating the antiviral potential of these unexplored plant species, in all probability, may result in the identification of novel compounds for the treatment of viral infections.

Gastric cancer (GC) is a malignant tumor originating from the epithelial cells of the gastric mucosa. The 5-methylcytosine (m⁵C) modification refers to the addition of a methyl group to the fifth carbon atom of cytosine in RNA molecules. This study aimed to investigate the role of NOL1/NOP2/SUN domain (NSUN)6 in GC and its underlying molecular mechanisms. Human gastric mucosa cells and gastric cancer cells were used for in vitro experiments. m⁵C level was quantified using dot blot analysis. Cell viability and proliferation were evaluated via cell counting kit-8 and colony formation assays. Apoptosis rates were analyzed by flow cytometry. Autophagy-related protein expression was detected through Western blot analysis. The interaction between NSUN6 and CCAAT/Enhancer Binding Protein Zeta (CEBPZ) was validated by RNA immunoprecipitation. Results demonstrated that NSUN6 functioned as an oncogene in GC. Furthermore, NSUN6 inhibition suppressed GC cell proliferation while promoting apoptosis and autophagy. CEBPZ was identified as a target gene of NSUN6 in GC through bioinformatic analysis. Mechanistically, NSUN6 enhanced CEBPZ mRNA stability via m⁵C methylation. Subsequent rescue experiments revealed that CEBPZ overexpression increased cell proliferation and reduced apoptosis and autophagy in GC. Additionally, NSUN6-mediated m⁵C methylation of CEBPZ suppressed autophagy by activating the p53/mTOR pathway. In conclusion, NSUN6 promoted GC progression by stabilizing CEBPZ mRNA in an m⁵C-dependent manner. However, further in vivo and clinical studies are warranted to validate these findings and explore their translational potential.