BMC Biotechnology最新文献

Background: The numerous hydroxyl groups in tannic acid (TA) and its tendency to form hydrogen bonds with proteins and carbohydrates are key factors contributing to its utility. TA-based enzyme immobilization supports provide flexible platforms for enzyme immobilization. chia gum (CG) can also trap significant amounts of water between its chains, preventing water loss from the gum and inhibiting enzyme leaching.

Results: In this study, TA, a natural linker molecule, was used to immobilize horseradish peroxidase (HRP) on chia gum (CG), serving as the natural support. On the created TA-CG support, HRP was successfully immobilized, with the maximum immobilization recovery (80%) observed at pH 7.0, 9 mg TA, and 20 U of the enzyme. Using a scanning electron microscope (SEM) and Fourier transform infrared (FTIR) technology, the chemical functional groups and morphological characteristics of the synthesized CG-TA-HRP were illustrated. After ten reuses, CG-TA-HRP demonstrated good reusability, with 65% retention. By comparing CG-TA-HRP to soluble HRP, the former exhibited better thermal stability up to 40 °C, a higher temperature optimum at 50 °C, and a pH optimum at 6.0. According to the K_m data, CG-TA-HRP exhibited a lower affinity for hydrogen peroxide (H₂O₂) and guaiacol and a higher oxidizing affinity to certain phenolic substrates. The prepared CG-TA-HRP demonstrated greater resistance to heavy metals, isopropanol, urea, and Triton X-100. During the 6-hour incubation period, soluble HRP removed 44% of the methyl orange, while immobilized HRP decolorized 78% of the dye.

Conclusions: The TA-linker molecule and chia gum are considered environmentally safe components in the production of CG-TA-HRP, making this an easy and eco-friendly method for enzyme immobilization.

Background: The Arctic environment, characterized by extreme conditions, hosts a largely untapped reservoir of fungal communities that have adapted to these harsh conditions by producing specialized bioactive secondary metabolites. Among these, Acremonium species also remain underexplored, despite their potential to produce structurally diverse and biologically active secondary metabolites. This is largely due to difficulties in sampling in remote Arctic regions and limited research focus on fungi from such environments. This study aimed to use an integrated workflow combining metabolomics, chemometrics, and bioactivity screening to prioritize Acremonium strains for the identification of bioactive secondary metabolites. We applied this workflow to investigate six Acremonium strains associated with driftwood from the Arctic: A. ellipsoideum (F1, F2), A. synnematoferum (F3, F4, F5), and A. multiramosum (F6), aiming to identify cytotoxic secondary metabolites.

Results: The integrated metabolomics and chemometrics approach revealed unique chemical fingerprints for A. ellipsoideum (F1) and A. synnematoferum (F5) among the six strains. By further combining bioactivity screening results, strain F5 was prioritized for further detailed study. Five compounds were isolated from F5 and structurally elucidated as cyclic depsipeptides: destruxin-A4 chlorohydrin (1), trichomide D (2), destruxin-A5 (3), homodestruxin (4), and homodestcardin (5). All compounds exhibited cytotoxic effects against the tested cell lines but did not exhibit activity in the targeted bioassays against the kinase FLT3 or the phosphatase PTP1B. Cytotoxic destruxin-type compounds likely play a significant ecological role, as they have been reported to exhibit entomopathogenic, nematocidal, and phytotoxic effects.

Conclusion: The identification of five known cytotoxic destruxin-type depsipeptides from A. synnematoferum as a new source expands the chemical diversity known for this genus and underscores their potential for the development of cytotoxic agents. These findings highlight not only the value of Arctic fungi as a reservoir of bioactive compounds but also the necessity of deciphering the ecological roles of cytotoxic metabolites produced by these organisms as they adapt to extreme environments. Furthermore, this study highlights the effectiveness of multi-informative-driven strain prioritization in uncovering bioactive metabolites from new fungal sources, emphasizing the significance of exploring Arctic fungal diversity for its potential to enhance chemical diversity, contribute to drug development, and broaden our understanding of ecological roles.

Active pharmaceutical ingredients (APIs) have poor efficacy and low bloodstream and target site concentrations due to their limited water solubility. Pregabalin is essential product to control pain and inflammation. We have synthesized novel pregabalin derivative called gabsali ((S, E)-3-(((2-hydroxybenzylidene)amino)methyl)-5-methylhexanoic acid. We have synthesized it by using salicylaldehyde to functionalize pregabalin (PG). The research was divided into two parts. The first part of the process was making a new kind of pregabalin called gabsali (GS). Second, MSNs were created using the sol-gel technique and then the synthesized GS was added to them. This study introduces a unique drug delivery method that improves bioavailability, stability, and anti-inflammatory activity by incorporating chemically modified pregabalin (GS) into engineered MSNs. This strategy has not been documented in the literature before. Zeta sizer, scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), BET analysis, and high-performance liquid chromatography (HPLC) were some of the modern tools used to determine particle size, shape, drug release, and compatibility. Under all tested conditions, the inclusion compound showed a considerable improvement in dissolving rate relative to the crystalline medicine in in vitro dissolving assays. The produced MSNs had a particle size of about 500 nm and were nano-sized, spherical. The FTIR spectroscopy was used to examine the chemical GS. The medicine was successfully added into blank MSNs, resulting in a decrease in both their specific surface area (602.5 ± 0.7 m²/g) and pore width (5.9 nm). In order to identify the potential benefits of GS, this study evaluated a preclinical model of inflammation that was created using ovalbumin (OVA). In a comparison to pregabalin, its anti-inflammatory activity was tested at doses of 50, 75, and 100 mg/kg. Using reverse transcription-polymerase chain reaction (RT-PCR), the levels of inflammatory mediators (cytokines IL-2 and IL-6) in the blood were measured and discovered to be significantly reduced. Cell viability was evaluated using the MTT assay, which showed a higher IC50 compared to pregabalin, the unmodified medicine. According to in vivo pharmacokinetic studies, MSN-GS significantly increased bioavailability compared to the pure drug. Using MSN-GS, the aforementioned results clearly show biocompatibility, improved in-vivo bioavailability, and satisfactory in-vitro performance.

Background: Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-based gene editing has become a promising approach for enhancing traits in aquaculture species. Nevertheless, traditional CRISPR-Cas systems encounter challenges, including significant off-target effects and strict protospacer adjacent motif (PAM) requirements, which constrain their use in crustaceans such as Penaeus monodon. To address these limitations, this research has developed PmMAD7, a codon-optimized CRISPR system specifically designed for P. monodon, which incorporates nuclear localization signals to improve editing efficiency and precision.

Results: This research successfully synthesized and delivered PmMAD7 mRNA and crRNAs targeting the ECH1 and AQP4 genes into the hemocytes of P. monodon. Quantitative PCR analysis demonstrated that PmMAD7 achieved significant gene silencing, reducing the expression levels of ECH1 and AQP4 by 81.5% and 78.33%, respectively. Next-generation sequencing confirmed targeted insertions and deletions at the gene loci, with knockout efficiencies of 14.81% for ECH1 and 20.57% for AQP4, which were significantly higher than those obtained with LbCas12a (7.14% and 12.43%, respectively). Furthermore, functional analysis indicated that ECH1 knockout resulted in increased cell volume and mortality, while AQP4 knockout led to decreased cell volume and reduced viability. These specific results highlight the first successful demonstration of MAD7-based genome editing in shrimp. The broader PAM compatibility and enhanced editing efficiency of PmMAD7 provide a versatile platform for gene editing in shrimp.

Conclusion: PmMAD7 constitutes an enhanced CRISPR editing tool specifically designed for P. monodon, exhibiting superior precision, expanded PAM compatibility, and enhanced editing efficacy relative to conventional Cas12a systems. These results lay the groundwork for the advancement of gene editing applications in crustaceans and contribute to sustainable genetic improvements in aquaculture.