Biotechnology Journal最新文献

Magnesium (Mg) is necessary for plants as a structural element or an enzyme cofactor. It also plays a significant role in the light and dark responses of photosynthesis. Moringa plants are considered one of the most important medicinal plants that humans need for nutrition. The aim of the study is to provide Mg to plants in an easy and simple way. Therefore, it is widely available in human food due to its nutritional importance. In this experiment, the treatment was done on the Moringa plant as, one of the medicinal plants that is increasingly used in fresh form. The experiment was conducted at the research farm of the National Research Center. Magnesium oxide has been extracted and prepared in a natural way that is safe for humans, plants, and soil. Trichoderma viride filtrate was used as a chelating agent in an environmentally friendly process to create magnesium oxide nanoparticle (MgO-NPs). FTIR, TEM, and SEM-EDX were utilized to analyze and characterize the synthesized. Plants were treated with different doses to determine the optimal concentration for the plant. The effect of treatment on plants was to improve vegetation such, as a number of leaves (232.15), leaf area (273.97 cm²), and fresh weight (3693.17 g). It also increased dry matter in the plant (171.8 g), chemical properties, and elements.

Pleuromutilin, a tricyclic diterpene compound with significant inhibitory activity against gram-positive bacteria and mycoplasmas, serves as a precursor for various veterinary and human medicines. Previous efforts have primarily focused on strain screening and fermentation process optimization to enhance pleuromutilin production in native pleuromutilin-producing strains, with the absence of genetic engineering strategies. In this study, we performed whole-genome sequencing of the pleuromutilin-producing strain Clitopilus passeckerianus T6 to identify the biosynthetic genes related to pleuromutilin production. Transcriptomic and metabolomic data were collected during the fermentation of C. passeckerianus T6, and gene transcription and metabolite accumulation in the pleuromutilin biosynthetic pathway were analyzed to identify the rate-limiting steps in pleuromutilin biosynthesis. Overexpression of the key genes ple-ggpps and ple-cyc increased pleuromutilin production by 50%, achieving a titer of 6.9 g/L. This study is the first to employ metabolic engineering to enhance pleuromutilin production in a native strain, providing a strategy for efficient pleuromutilin production.

Monkeypox (mpox), a zoonotic disease, has rapidly spread globally, prompting the WHO to declare it a public health emergency. The long incubation period, early symptoms resembling respiratory infections, and diagnostic challenges hinder timely epidemic control and accurate clinical diagnosis. The monkeypox virus (MPXV) encodes the A29L protein, which binds to cellular heparan sulfate to facilitate infection and serves as a target for treatment and diagnostics. Thus, developing effective diagnostic tools and treatments is critically important. In this study, we expressed and purified Escherichia coli-derived A29L protein, which was used for chicken immunization to generate specific polyclonal IgY antibodies. The results demonstrated a successful elicitation of a humoral immune response. Subsequently, two single-chain variable fragments (scFv) antibody libraries were constructed using phage display technology, comprising 2.6 × 10⁸ and 3.8 × 10⁸ transformants. After bio-panning, phage-based ELISA indicated the enrichment of specific clones. Three scFv-expressing clones, including cA29LS1, cA29LS5, and cA29LS13, were selected from 13 randomly chosen clones and classified based on nucleotide sequence analysis. Their binding activities were evaluated through ELISA and Western blot, followed by purification for affinity determination via competitive ELISA. Among the selected clones, cA29LS5 demonstrated the highest binding affinity (1.3 × 10⁻⁶ M), followed by cA29LS1 (5.3 × 10⁻⁶ M). Additionally, both IgY and all three clones demonstrated binding activity to cell-derived and commercially purchased A29L proteins, as confirmed by Western blot and ELISA. Overall, these findings suggested that the IgY and scFv antibodies developed hold promise as potential diagnostic and therapeutic agents against MPXV infections.

The full approval of two SARS-CoV-2 mRNA vaccines, Comirnaty and Spikevax, has greatly accelerated the development of numerous mRNA vaccine candidates targeting infectious diseases and cancer. mRNA vaccines provide a rapid, safe, and versatile manufacturing process while eliciting strong humoral and cellular immune responses, making them particularly beneficial for addressing emerging pandemics. Recent advancements in modified nucleotides and lipid nanoparticle delivery systems have further emphasized the potential of this vaccine platform. Despite these transformative opportunities, significant improvements are needed to enhance vaccine efficacy, stability, and immunogenicity.

This review outlines the fundamentals of mRNA vaccine design, the manufacturing process, and administration strategies, along with various optimization approaches. It also offers a comprehensive overview of the mRNA vaccine candidates developed since the onset of the COVID-19 pandemic, the challenges posed by emerging SARS-CoV-2 variants, and current strategies to address these variants. Finally, we discuss the potential of broad-spectrum and combined mRNA vaccines and examine the challenges and future prospects of the mRNA vaccine platform.

Nicotinamide mononucleotide (NMN), a precursor of nicotinamide adenine dinucleotide (NAD), provides a direct method for maintaining NAD levels, which may alleviate aging and metabolic disorders. However, the enzymatic conversion of NMN in cascade reactions is limited by intermediate product inhibition, and quantitative insights into these limitations remain scarce. Here, an efficient multienzyme cascade system was developed by quantifying intermediate inhibition, which synthesizes NMN from D-ribose in three tandem reactions with an Adenosine Triphosphate (ATP) regeneration system and pyrophosphatase (PPase). A critical Adenosine Diphosphate (ADP) concentration of 0.5 mM was determined, which inhibits phosphoribosyl pyrophosphate synthetase (Prs) at 0.08 µM. The incorporation of an ATP regeneration system and PPase markedly increased the NMN yield to 81.3%. The intermediate phosphoribosyl pyrophosphate (PRPP) hydrolysis rate was measured at 3 µM/min. The highly active nicotinamide phosphoribosyltransferase (Nampt) could compete with PRPP hydrolysis, thereby increasing the yield of NMN. This research facilitates large-scale, efficient NMN manufacturing.