BMC Biotechnology最新文献

Background: Algae are microscopic or macroscopic organisms that can photosynthesize and are found in both freshwater and saltwater ecosystems and are considered one of the cornerstones of life. In our study, the biological activities of extracts produced under the extract conditions that provided the highest biological activity of Ulva lactuca L. were determined.

Methods: Two different methods, Response Surface Method (RSM) and Artificial Neural Network-Genetic Algorithm (ANN-GA) integration were used for optimization. In this context, the optimum extract conditions were determined as 54.940 ˚C temperature, 6.513 h, 42.109 ethanol/water ratio according to the RSM method and 56.286 ˚C temperature, 7.2793 h, 36.8625 ethanol/water ratio according to ANN-GA. The antioxidant activities of the optimized extracts were evaluated by Rel Assay kits, DPPH and FRAP methods. Anticholinesterase activities were determined by testing against acetylcholinesterase and butyrylcholinesterase enzymes. In addition, antiproliferative effects were examined in A549 lung cancer cell line and phenolic compound contents were analyzed by LC-MS/MS.

Results: As a result of the analyzes, it was seen that the extract obtained from the conditions recommended by ANN-GA exhibited higher activities. Optimized extracts of U. lactuca were found to have potential in terms of antioxidant activities. The highest total antioxidant value was determined as 6.272 ± 0.024 mmol/L. In addition, it was determined that extracts of U. lactuca produced under optimum conditions showed strong cytotoxic effects against A549 lung cancer cell line. In addition, gallic acid, 4-hydroxybenzoic acid, caffeic acid, vanillic acid, syringic acid, quercetin and kaempferol were found in the extracts of the algae produced under optimum conditions. The highest detected compound was caffeic acid. In addition to all these properties, it was seen that U. lactuca has anticholinesterase potential in our study.

Conclusion: In conclusion, optimized extracts of U. lactuca attract attention with their antioxidant and cytotoxic activities as well as anticholinesterase potential and can be considered as a potential source for cancer treatment and management of neurological diseases.

This study investigates the potential of Tetradesmus obliquus for lipid accumulation under heavy metal stress and evaluates it's aviability for biodiesel production. We surveyed how different concentrations of heavy metals, including manganese (Mn), cobalt (Co), and zinc (Zn), influence the carbohydrate & protein, lipid yield, and fatty acid profiles of T. obliquus cultures. Our results demonstrated that while lipid content increased under heavy metal stress, the extent of accumulation was highly dependent on metal type and concentration. Notably, the algal culture treated with 0.04 mM Co²⁺ showed the highest lipid accumulation. Treatment with 0.3 mM Zn²⁺ resulted in the highest proportion of saturated fatty acids (SFA). The Relative Enrichment Efficiency Coefficient (REEC) analysis demonstrated that 0.04 mM and 0.07 mM Co²⁺ led to the highest lipid and carbohydrate content stimulation. Additionally, GC-MS analysis revealed increased monounsaturated fatty acids (MUFA) under several metal stress conditions. The study demonstrated that exposure to specific concentrations of heavy metals can significantly enhance lipid accumulation and alter the fatty acid profiles of T. obliquus, which are crucial for improving biodiesel quality. The implications of these findings suggest that heavy metal-induced stress could be a feasible approach to enhancing lipid accumulation for sustainable biodiesel production, and T.obliquus is a promising candidate for future biodiesel production.

Background: Candida species are commensal fungi that can become opportunistic pathogens under specific host and environmental conditions. The emergence of multidrug-resistant Candida strains poses a significant challenge. Nanotechnology represents a cutting-edge field offering precise and targeted delivery systems for combating fungal infections, leveraging the unique properties of plant-derived bioactive compounds. This investigation employed a biogenic approach utilizing polyherbal leaf extracts from Citrus limon and Citrus medica, known for their abundant Citral content.

Results: Citrus sp. extracts were used to synthesize Citrusfusion silver nanoparticles (CitAgNPs) through a green synthesis method. Characterization of CitAgNPs was carried out using advanced analytical methods ensuring the quality, uniformity, size, and charge. The synthesized CitAgNPs exhibited non toxic effect when tested on Vigna radiata and Danio rerio, highlighting their potential for sustainability and safe therapeutic use. Antifungal assays demonstrated the potent efficacy of CitAgNPs in various Candida strains, with low MIC and MFC. CitAgNPs exhibited remarkable biofilm inhibition capabilities and elucidated specific mechanisms of action in Candida species, surpassing the performance of fluconazole.

Conclusion: This study underscores the immense potential of nanotechnology-driven approaches harnessing Citrus leaf extract for synthesizing highly effective antifungal nanoparticles. The fusion of biogenic nanoparticles with Citrus bioactive compounds presents a sustainable strategy for addressing the escalating challenge of azole-resistant Candida infections. The research outcomes suggest that CitAgNPs have promising applications in inhibiting Candida biofilms, offering potential solutions for infections caused by diaper rashes and onychomycosis, providing safe and effective alternatives to antifungal therapies.

Background: Antibiotic resistance is a growing global threat due to antibiotic overuse and limited treatment options. Multidrug-resistant bacteria, like Staphylococcus aureus and Escherichia coli, increase infection complexity and mortality. This study explores nanocomposite films of ZIF-8 nanoparticles and Doxycycline (Dox) to enhance antibacterial efficacy. In this study, nanocomposite films composed of chitosan (CS) and polyethylene glycol (PEG), incorporating zeolitic imidazolate framework-8 (ZIF-8) nanoparticles and DOX, were developed. These films were characterized by their morphological, mechanical, antibacterial, and drug-release properties. Antibacterial efficacy was evaluated using disk diffusion, broth microdilution, and checkerboard assay methods to determine MICs and potential synergistic effects.

Results: The nanocomposite films demonstrated flexibility, semi-transparency, and a yellowish-brown hue, with films containing ZIF-8 nanoparticles being thicker (79 ± 0.2 μm) than those without (54 ± 0.5 μm). The tensile strength was enhanced with the incorporation of ZIF-8, peaking at 53.12 MPa for the CS-PEG-G-10% DOX-4% ZIF-8 film. XRD analysis confirmed the crystallinity of the ZIF-8 and DOX, with distinct peaks observed for each material. The drug release studies revealed an initial burst followed by sustained release, with higher release rates in acidic environments compared to neutral and alkaline media. The CS-PEG-G-10% DOX-4% ZIF-8 nanocomposite film demonstrated significantly higher antibacterial activity, achieving the lowest MIC values, particularly against S. aureus (22.5 mm inhibition zone) compared to E. coli (14 mm inhibition zone). Additionally, a notable synergistic effect was observed between CS-PEG-G-10% DOX and CS-PEG-G-10% DOX, with FICI values below 0.5.

Conclusions: The CS-PEG-G-10% DOX-4% ZIF-8 nanocomposite exhibits enhanced antibacterial efficacy and optimal properties, positioning it as a strong candidate for developing effective treatments against multidrug-resistant pathogens.

Background: Spinal muscular atrophy (SMA) is a devastating neuromuscular condition resulting from the loss of the survival motor neuron 1 (SMN1) gene. Precise genetic testing has become essential after the authorization of several potent medications. To achieve this objective, the use of dried blood spot (DBS) has assured convenient and extensive testing from a distance. Nevertheless, developing countries such as Indonesia sometimes lack access to standard filter papers like FTA or Guthrie cards for DBS processing. Here, we aim to develop a cellulose-based card as an alternative filter paper for DBS preparation suitable for the genetic testing of SMA including but not limited to a direct polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and multiplex allele-specific amplification (multi-ASA).

Results: An in-house paper was developed from a 180 gsm cellulose-based paper and was used for DBS preparation. The performance of dried blood spotted on the cellulose-based card (DBSc) was compared to pure genomic DNA (gDNA) isolate and dried blood spotted on FTA cards (DBSf) for genetic testing. The results of the genetic testing of our cellulose-based card were completely matched with those of gDNA and DBSf in both direct PCR-RFLP and Multi-ASA to separate SMN1 from SMN2. In addition, after three months of storing, the DBSc continued to exhibit a clear result, suggesting its high stability for DNA storage.

Conclusion: Our cellulose-based card has the potential to be used for DBS carrier and for further genetic testing using PCR. Our findings can assist physicians in sending DBS samples from SMA suspicion cases to genetic testing centers, thereby preventing diagnosis delay or misdiagnosis.

In the battle against clinical infections particularly the resistant pathogens, the creation of new antimicrobial drugs is essential. This study focuses on synthesis and characterization of Lipase-CuO nanoparticle conjugates in order to investigate their antibacterial efficiency. Lipase enzyme and CuO nanoparticles were synthesized biologically by specific selected fungal strains. Statistical optimization of lipase enzyme was done using a Plackett-Burman design giving two enhancement models for lipase production with increasing in productivity up to 143.43% (2800 U/ml). Copper oxide (CuO) nanoparticles were characterized using visual indication of greenish color formation, UV-vis spectrum analysis which revealed a strong peak at 300 nm. Also, CuO nanoparticles appeared as distinct, well-dispersed spherical particles with average size of 71.035 nm using TEM, while conjugate appears as large protein molecules linked to the nanoparticles. Also, using techniques like energy dispersive X-ray (EDAX) the resultant conjugates formation was confirmed as the elemental analysis approved its formation. The antimicrobial activity of Lipase-CuO nanoparticles conjugates was tested against a range of clinical pathogens. The results demonstrated a significant increase in antimicrobial potency compared to both CuO nanoparticles and lipase alone particularly against E. coli strain NRC B-3703 with remarkable increase of 373.6% and 75% followed by S. aureus with increase of 50 and 42.8%compared to that of individual CuO nanoparticles and lipase enzyme, respectively. These findings suggest that Lipase-CuO nanoparticle conjugates hold great promise as a novel antimicrobial strategy, offering a potential solution to combat bacterial infections, especially those caused by multidrug-resistant strains. The study highlights the importance of nanotechnology in enhancing the efficacy of traditional antimicrobial agents and opens new avenues for targeted antimicrobial therapies.

Background: Proteases are essential in various industries due to their unique substrate specificities and robustness in different operational conditions. Bacillus strains consist of a genotype favorable for rapid growth whilst secreting enzymes extracellularly, thereby simplifying recombinant protease production. Despite the widespread use of batch and fed-batch fermentations for their ease and robustness, these cultivation types are often marred by significant energy requirements and prolonged downtimes. The switch towards continuous cultivation methods promises reduced carbon footprints and improved equipment efficiency. Yet, research focusing on Bacillus strains is limited, therefore we aimed to establish a continuous cultivation as a competitive alternative to fed-batch.

Results: Therefore, this study aimed to explore the potential of chemostat cultivations for producing a protease from Bacillus licheniformis utilizing a derepressed induction system, and comparing specific productivities and space-time yields to fed-batch cultivations. The continuous cultivations were described in a hybrid model, considering the effect of productivity as function of the applied dilution rate as well as the generation time. The workflow of this study demonstrates that screenings in a fed-batch mode and chemostat cultivations conducted at the same growth rate, result in different specific productivities for derepressible systems.

Conclusion: The results of this study highlight that the feeding rate's impact on specific productivity varies significantly between fed-batch and chemostat cultivations. These differences suggest that fed-batch screenings may not be adequate for developing a continuous process using a derepressed promoter system in B. licheniformis. Although the space-time yield of fed-batch cultivations has not been surpassed by stable continuous operations-achieving only a third of the highest space-time yield observed in fed-batch-valuable mechanistic insights have been gained. This knowledge could facilitate the transition towards a more sustainable mode of cultivation for industrial protease production.

Background: Tissue engineering for bone regeneration aims to heal severe bone injuries. This study aimed to prepare and assess the early osteogenic differentiation effects of a gelatin/calcium phosphate- Punica granatum nanocomposite scaffold on stem cells from human exfoliated deciduous (SHED) and human dental pulp stem cells (HDPSCs).

Methods: The electrospinning method was used to prepare a gelatin/calcium phosphate nanocomposite scaffold containing pomegranate (Punica granatum) extract. The physicochemical properties of the scaffold were evaluated. The effect of the scaffold on the selected cells was done by the cell viability evaluation. A special alkaline phosphatase (ALP) kit was utilized to investigate the early osteogenic differentiation effects of the prepared scaffold on HDPSCs and SHED.

Results: The results showed that the scaffold had uniformly accumulated in the networked form. Besides, the prepared scaffold did not have beads (structural defects). No new interactions were observed in the spectroscopic spectra of the scaffold and these peaks showed the successful formation of the fibrous nanocomposite as well. Furthermore, cell viability percentage was significantly higher for the scaffold compared with the control group (cells without any material) for both HDPSCs and SHED. Early osteogenic differentiation results specified that the ALP activity was significantly higher for the scaffold compared with the control group (cells without any material) for both HDPSCs and SHED.

Conclusion: The appropriate physicochemical assay and cellular results (cell viability and early osteogenic differentiation) for the prepared fibrous nanocomposite showed that the use of this nanocomposite can be considered in the construction of various scaffolds in bone and dental tissue engineering.