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Medical biotechnology serves as a foundational platform for identifying disease markers and developing specific diagnostic assays. Complementing this, biosensors and medical devices translate these biomarkers into quantifiable signals enabling real-time health assessment. Artificial intelligence (AI) augments this framework by processing vast datasets to recognize patterns and generate predictive insights thereby aiding clinicians in decision-making and streamlining personalized disease management. The integration of molecular-based companion diagnostics, biosensor technology, and medical AI forms the cornerstone of contemporary, data-driven precision medicine. This review explores the synergistic convergence of these fields specifically examining how AI-based approaches enhance molecular diagnostics and biosensor devices to optimize clinical outcomes. This interdisciplinary collaboration facilitates advancing precision medicine. Consequently, these discovery technologies profoundly impact disease prevention, early diagnosis, and targeted therapeutic strategies. Despite challenges such as data privacy and technology accessibility, this cross-disciplinary approach holds tremendous potential for transforming the future of precision healthcare.
Letter recognition is assumed to involve several levels of analysis, including coarse tuning for category and novelty and more fine tuning for specific features, related to letter orientation. We employed an oddball fast periodic visual stimulation (FPVS) paradigm with magnetoencephalography (Elekta VectorView, 306 sensors) to study neural discrimination responses in the source space. Using contrasts between native and foreign letters, digits, or inverted native letters, we aimed to isolate the neural responses to visual novelty, category, and orientation during character analysis. The study was conducted with a cohort of 25 adults. The response topography demonstrated bilateral organization, including language-related brain regions such as the ventral occipitotemporal cortex, inferior parietal cortex, and middle temporal areas. Comparing conditions, we revealed right lateralized parietal clusters, associated with novelty tuning, and left lateralized occipitotemporal clusters exhibiting higher activity for letters among digits discrimination, supporting the role of this area in letter processing. No distinct spatial patterns specific to orientation tuning were observed in comparison to novelty and category tuning. We propose that expertise-dependent orientation-specific tuning mechanisms may operate within an embedded neural framework that spatially overlaps with coarse tuning systems, but are characterized by specific spatiotemporal patterns.
Glycoprotein hormones, including follicle-stimulating hormone (FSH), luteinizing hormone (LH), and thyroid-stimulating hormone (TSH), play essential roles in the reproductive endocrinology of teleosts. However, research on these hormones in non-model species such as the Japanese eel (Anguilla japonica) has been constrained by the lack of specific immunological detection tools. Recombinant A. japonica FSH, LH, and TSH were expressed using a Bombyx mori-based silkworm-baculovirus system. Polyclonal antibodies were then generated using two distinct immunogen strategies: (i) chimeric constructs fusing β-subunits with a murine α-subunit, and (ii) synthetic peptides representing unique, non-conserved β-subunit epitopes. Both approaches induced antibody responses, but only peptide-based immunogens yielded polyclonal antisera with high specificity and sensitivity. These antibodies distinguished homologous glycoprotein hormones without cross-reactivity and retained target selectivity in mixtures simulating physiological serum conditions. Peptide-derived antibodies provide a robust and precise tool for the immunological discrimination of FSH, LH, and TSH in A. japonica. This strategy establishes a scalable framework for species-specific hormone assay development and offers translational potential for endocrine monitoring, reproductive management, and diagnostic applications in aquaculture biotechnology.
We investigated the effectiveness of hydrated deep eutectic solvents (DESs) to achieve tunable seaweed cellulose microstructures from Ulva lactuca. Cellulose was extracted via a sequential chemical protocol, then treated with 30% DESs (choline chloride (ChCl) or betaine as hydrogen bond acceptors and urea, citric acid, or oxalic acid as donors) combined with mechanical shearing. While most DESs combinations yielded spherical seaweed cellulose microparticles (dry diameter of 605-777 nm), the ChCl:urea formulation successfully produced seaweed cellulose microfibers (SCMFs). These SCMFs exhibited superior quality (dry diameter of 372 nm), and water dispersibility with a hydrodynamic diameter of 134 nm and a polydispersity index of 0.23. Crucially, the DES composition dictated cellulose structure: ChCl:urea-treated SCMF remained predominantly amorphous, whereas other DESs treatments increased microparticles crystallinity. Furthermore, ChCl:oxalic acid introduced carboxyl functional groups. Selecting appropriate hydrated DESs thus offers a sustainable biotechnology tool to tune cellulose morphology, crystallinity, and surface chemistry.
This study evaluated real-world treatment patterns and 1-year outcomes in patients with neovascular age-related macular degeneration (nAMD) initiating faricimab. FARETINA-AMD was a retrospective study using data from the IRIS® Registry for patients diagnosed with nAMD initiating faricimab from February 2022 to March 2023. Included in the study were 2,025 treatment-naive patients (2,184 eyes) and 22,253 patients (26,851 eyes) previously treated with anti-vascular endothelial growth factor (anti-VEGF) therapy. Visual acuity improved by 2.0 ± 15.0 (mean ± SD) letters in treatment-naive eyes (P < .001) and was maintained in previously treated eyes at injection 7. Central subfield thickness (CST) improved by -53.1 ± 64.8 μm in treatment-naive and -28.5 ± 79.7 μm in previously treated eyes (both P < .0001); 78.4% and 66.8%, respectively, had achieved/maintained CST ≤ 280 μm at injection 7. Dosing frequency was reduced in the second 6 months (mean 2.4-3.2 injections) versus the first 6 months (4.0-4.2) of treatment. Outcomes among patients with nAMD receiving faricimab over 1-year follow-up support the real-world effectiveness and extended durability of treatment.
Bacillus subtilis is a key industrial microorganism widely used to produce enzymes, vitamins, antibiotics, and recombinant proteins. Its non-pathogenic nature, efficient protein secretion systems, and genetic tractability make it an ideal candidate for industrial biotechnology. Genome-scale metabolic modeling has emerged as a key systems biology approach to understand, explore and manipulate the metabolism of B. subtilis for improved strain performance. To date, the modeling efforts have resulted in fourteen published genome-scale metabolic reconstructions for B. subtilis; however, substantial opportunities for improvement remain, especially in integrating regulatory mechanisms for refining model accuracy. Furthermore, there is a growing trend toward modelling the cell as a whole system. This article is the first comprehensive and up-to-date review that highlights the genome sequencing, annotation efforts and the current state of genome-scale metabolic modeling for B. subtilis, emphasizing future directions.
Bacteria exhibit extraordinary evolutionary and ecological diversity. They range from dominant, well-characterized phyla to rare lineages that are known only through environmental sequencing. This chapter reviews four key bacterial phyla, including Pseudomonadota, Bacillota, Actinomycetota, and Bacteroidota. These phyla are widely distributed, metabolically versatile, and play a central role in ecosystem functioning and human health. We discuss unique phyla within the PVC superphylum (Planctomycetota, Verrucomicrobiota, Chlamydiota) for their unusual cell biology, compartmentalization, and host associations. We also highlight hyperthermophilic phyla, such as Thermotogota, Aquificota, and Thermodesulfobacteriota, that thrive in geothermal ecosystems and drive sulfur and carbon cycling. We consider less-cultivated lineages, including Deinococcota, Acidobacteriota, Nitrospirota, Fusobacteriota, Fibrobacterota, Synergistota, Deferribacterota, and Chrysiogenota, in terms of their ecological niches, metabolic specializations, and roles in biogeochemical cycles, symbiosis, and disease. Collectively, these examples demonstrate the remarkable metabolic flexibility and ecological impact of bacteria, ranging from host-associated commensals and pathogens to free-living autotrophs in extreme environments. Despite advances in genomics and cultivation-independent methods, vast portions of bacterial diversity remain uncultured and poorly understood. Continued exploration of both dominant phyla and rare lineages promises to refine bacterial taxonomy, expand our understanding of microbial evolution, and reveal novel metabolic pathways with implications for ecology, medicine, and biotechnology.
Insect cell-baculovirus expression vector systems (IC-BEVS) are valuable tools in pharmaceutical bioprocesses for producing complex proteins like immunogenic virus-like particles. In this context, standardization is key to guarantee consistency in process yield, productivity, and product quality attributes. The study investigates the nine-month stability of the main biotechnological input used in IC-BEVS, the recombinant baculovirus vectors produced in chemically defined medium. Viral titer (pfu/mL), zeta potential (mV), and mean hydrodynamic particle size (μm), were employed to assess viral inactivation under eight combinations of generation and storage conditions. First-order, Weibull, and biphasic models were applied to describe viral decay. The critical parameters (factors) analyzed were the size of the heterologous gene inserted (719 and 1621 bp), storage temperature (- 80 and 1.5 °C), and infection time used for baculovirus batch generation (48 or 72 h post-infection). They were mainly explored according to a two-level factorial design (23). The primary quality attribute evaluated in this study was the one-log10 decay time of the viral titer (td), which exhibited an overall mean value of 80 days across eight batches. The biphasic model best fits the dispersion of the viral titer data collected over the assessed time in all considered combinations of factors and was employed to find significant factors over td values. Gene size was the only factor with a statistically significant effect on viral titer decay; additionally, the study indicates the occurrence of particle aggregation over the course of the analysis.
The growing demand for biopharmaceuticals has increased the need for efficient and cost-effective recombinant protein production platforms. Transcriptional gene silencing after random genomic integration of expression vectors remains a major limitation in mammalian expression systems and often reduces protein yield. Incorporation of ubiquitous chromatin opening elements (UCOEs) into expression cassettes has emerged as a promising strategy to mitigate position effects and enhance transgene expression. This study examined the effect of a UCOE-containing expression system on erythropoietin (EPO) production in Chinese hamster ovary (CHO) DG44 cells. A codon-optimized EPO expression cassette was introduced into CHO DG44 cells using either a conventional pOptiVEC vector and a UCOE-containing vector following random genomic integration. Recombinant EPO expression was assessed at the transcriptional and protein levels using reverse transcription quantitative polymerase chain reaction (RT-qPCR), western blotting, and enzyme-linked immunosorbent assay (ELISA). The UCOE-containing cell pool exhibited a significant enhancement in recombinant EPO expression, with approximately a 3.8-fold increase in mRNA levels and a sevenfold increase in secreted protein levels compared to the non-UCOE control cell pool. These results show that incorporating a UCOE reduces position-dependent gene silencing and increases recombinant mRNA and protein expression in CHO DG44 cell pools. This strategy supports improved efficiency during the early stages of cell line development for recombinant protein production.
P. aeruginosa is an opportunistic pathogen that causes various nosocomial infections. The ability of P. aeruginosa to form biofilms is one of the main factors contributing to its pathogenicity. Due to biofilm formation, bacteria get embedded in it and is able to withstand extreme environmental conditions like chemicals, UV, temperature, pH, salinity, and antibiotics. Biofilm formation is an important virulence factor associated with quorum sensing (QS), which is a cell-to-cell communication system that is influenced by cell density. P. aeruginosa is a notorious pathogen that is known to cause severe complications in patients suffering from cystic fibrosis and immuno-compromised individuals in hospital setting as a result of biofilm formation. We might limit P. aeruginosa infection pathogenesis and biofilm formation if we can disrupt the signalling molecules involved in QS. In this study, we suggested that already-approved medications by the FDA could be employed as anti-quorum and anti-biofilm agents and potentially be helpful in curing P. aeruginosa associated infections. The anti-quorum and anti-biofilm properties of FDA-approved medications have been investigated here. We have also performed RT-PCR analysis and molecular docking experiments to assess the mechanism of action of these drugs. We have found that all four drugs have anti QS activity. Out of these four drugs Flurbiprofen was found to be more effective. RT data confirms that these drugs have significantly downregulated all four QS gene and therefore are able to inhibit the P. aeruginosa virulence factors. This study significantly opens up the new horizons for the development of novel therapeutics against P. aeruginosa and infections associated with it.
Bacillus subtilis strain AF2 was obtained from the Mediterranean Sea and its identification was determined by its phylogenetic analysis of 16S rRNA sequences, as well as their morphological, physiological, and biochemical characteristics. The isolate yielded EPSS4 at a concentration of 7.4 g/l. EPSS4 constituted a significant proportion of the EPS recovered through purification using DEAE-Cellulose. The sample consisted of sulfate (24.71%) and uronic acid (18.55%). The fraction's viscosity was measured to be 1.1 mm2/sec, and the total hexose amine content was measured to be 9.06%. The monosaccharide composition of this fraction consists of glucose, fructose, xylose, and glucuronic acid in a molar ratio of 1.0:0.5:1.0:1.0, respectively. The toxicity studies on EPSS4 demonstrated its safety at 5 g/kg doses. EPSS4 underwent evaluation for its antioxidant, anticancer, and anti-inflammatory properties. The results indicated that the antioxidant activity percentages were as follows: 85.17 ± 0.77% for DPPH, 79.63 ± 1.05% for ABTS, 52.20 ± 0.67% for Fe2+ ion chelation ability, 75.80 ± 0.28% for Lipid peroxidation Inhibition capacity, 84.66 ± 1.21% for O2- radicals scavenging capacity, and 55.09 ± 0.34% for NO scavenging capacity. The IC50 value of EPSS4 for HepG-2 is 238.05 µg/ml. A-549 has determined the IC50 values to be 102.71 µg/ml. The concentration of HCT-116 was measured to be 328.01 µg/ml, while the IC50 value for the MCF-7 cell line was determined to be 204.77 µg/ml. The IC50 values for the HEP-2 and PC-3 cell lines were 118.95 and 128.94 µg/ml, respectively. The anti-inflammatory activity of EPSS4 was assessed using various methods, including measuring its inhibitory effects on lipoxygenase (LOX) and cyclooxygenase (COX2). At a dosage of 100 µg/ml, EPSS4 had a LOX inhibitory activity of 80.54 ± 1.09% and a COX2 inhibitory activity of 84.70 ± 1.05%. Due to its antioxidant capabilities, the EPS produced from Bacillus subtilis strain AF2 is a promising option for application as a potent antioxidant agent in the healthcare system. So, from observed activities demonstrate that it has the potential to be employed as an anticancer, antioxidant and non-steroidal anti-inflammatory substance via improving adaptive immune responses.
The increasing incidence of breast cancer is leading researchers to investigate new treatment approaches. Targeted therapy approaches are particularly attractive because they minimize the detrimental effects of therapeutic agents on healthy tissues and cells by focusing on tumor sites. This study focuses on synthesizing mPEG-modified triblock copolymers as carrier materials for drug delivery applications, enabling the efficient encapsulation of DOX, and evaluating the cytotoxic effects of the resulting nanocarriers on breast cancer cell lines. In this study, mPEG-poly(butylene adipate)-mPEG and mPEG-poly(ethylene adipate)-mPEG triblock copolymers were synthesized by a step-growth polycondensation polymerization method. Firstly, poly(butylene adipate) (pBAd) and poly(ethylene adipate) (pEAd) were synthesized to form the body of the triblock copolymer, and their chemical structures were characterized using Fourier transform infrared (FT-IR) and 1H NMR spectroscopy. The end-group analysis method was applied to determine the average molecular weights of the pBAd and pEAd polymers before their modification with mPEG-500. The nanocarriers produced by the double emulsion method were analyzed using the dynamic light scattering (DLS) method, while encapsulation efficiency and the DOX release profile were measured using a spectrofluorometer. The antiproliferative effects and cellular uptake capacities of the resulting nanocarriers were subsequently examined in MCF-7 and MDA-MB-231 cells. The cytotoxicity of DBANP and DEANP nanocarriers was lower than that of free DOX, demonstrating that encapsulation reduces drug-associated toxicity and may enhance safety. These findings suggest that the nanocarrier systems developed in this study show strong potential as promising candidates for breast cancer therapy.
Carrot (Daucus carota L.), a key crop in the Apiaceae family, adapts to environmental changes through its endogenous circadian clock, which coordinates physiological processes over a 24-h cycle. As a central physiological activity of carrot growth, photosynthesis, including parameters such as photosynthetic efficiency and stomatal conductance, is influenced by environmental factors. Among them, photoperiod is one of the main regulatory factors for development and chlorophyll synthesis. In this study, we investigated the effects of three different light cycles (16 h of light/8 h of darkness, 16L/8D, 12 h of light/12 h of darkness, 12L/12D, and 8 h of light/16 h of darkness, 8L/16D) on the expression of circadian rhythm genes in carrot. The core clock genes, DcLNK1, DcLNK3, DcRVEa, DcRVEb, and DcGI exhibited distinct transcriptional responses to these light cycles. Three different photoperiod treatments led to changes in the expression profiles of circadian rhythm related genes, photosynthetic parameters, and stomatal activity in carrot. The results elucidated a potential molecular regulatory network linking photoperiod perception to physiological pathway in carrot, providing potential insight into the intrinsic mechanisms of photo-entrainment within its circadian system.
Adeno-associated virus (AAV), a 4.7 kb single-stranded DNA virus, is widely used as a gene therapy vector, but its limited packaging capacity poses challenges for delivering large genes, always resulting in truncation during packaging. Quantifying truncation is difficult because both strands of the plasmid can be packaged from the 3' end. In this article, we aim to first produce single-polarity AAV and then explore its truncation pattern. To address this, we modified one of the ITRs and created an oversized self-complementary AAV, which functions as a single-polarity vector to some extent. Using this modified backbone, we generated a series of oversized single-polarity AAV (spAAV) vectors of varying lengths and sequences, analyzing DNA truncation patterns via quantitative PCR (qPCR). The results show that as the distance from the 3'-ITR increased, less DNA was detected. At 3000 bp from the 3'-ITR, 70% of the genomic DNA remained; this dropped to 50% at 4000 bp, 20% at 4500 bp, and almost none beyond 5000 bp. Additionally, reporter gene expression significantly decreased when the expression cassette extended to 4.5 kb compared to 2.7 kb under identical in vitro conditions. Our results show that DNA will be truncated far earlier before 4.5 kb during the packaging of very large genomes. This study provides important insights into the truncation patterns of AAV genomes, which is crucial for optimizing AAV vector design in gene therapy.
Great mass of tea tree seeds (TTS) are naturally rotted away as agricultural waste because there is no suitable technology used for extracting oil from TTS. The fermentation method is a new process and that can simultaneously recover oil, starch and saponins from TTS. In this study, a key bacterial strain named JJZ21 was isolated from fermented TTS milk (ground TTS and water) by the serial dilution method and identified as Lactiplantibacillus plantarum strain through molecular analyses. The production of cellulase, pectinase, amylase and protease of JJZ21 were visualized on agar plates containing specific enzyme substrates. Compared with the control, addition treatment with JJZ21 dramatically improved the activities of cellulase, pectinase, amylase and protease, reduced content of soluble sugar, protein and dry matter and pH in the TTS milk, shortened the fermentation time and increased the yield of TTS oil. Meanwhile, the addition of JJZ21 had no significant effect on quality of TTS oil. However, the yield of TTS oil decreased with excessive fermentation. Response surface methodology was used to evaluate the optimum conditions for fermentation process to obtain the maximum oil yield.
Acute myocardial infarction (AMI) triggers systemic biochemical responses, including dynamic changes in the phosphorylation status of circulating proteins. However, the phosphoproteomic profile of serum in the context of AMI remains insufficiently characterized. This study aimed to investigate serum phosphoproteomic alterations associated with AMI and to explore potential correlations with markers of cardiac injury. A comparative phosphoproteomic analysis was performed on serum samples obtained from eight patients with AMI and pooled healthy control samples. High-abundance serum proteins were depleted, and phosphopeptides were enriched using TiO2 phosphopeptide enrichment kit. Samples were analyzed by liquid chromatography-tandem mass spectrometry using a Q Exactive HF-X Orbitrap mass spectrometer. Data were searched against the Homo sapiens database using Sequest HT with a 1% false discovery rate and were quantified by label-free quantification using Proteome Discoverer version 2.4. A total of 46 phosphoproteins were confidently identified, revealing distinct phosphorylation profiles between AMI and control samples. Increased phosphorylation levels were observed for solute carrier family 12 member 5, apolipoprotein L1, the low-molecular-weight isoform of kininogen-1, and osteopontin in AMI serum. Conversely, phosphorylated inter-alpha-trypsin inhibitor heavy chain H2, antithrombin III, histidine-rich glycoprotein, peroxiredoxin-4, GTPase ERas, and the 26S proteasome non-ATPase regulatory subunit 1 were reduced or undetectable. A strong negative correlation was found between apolipoprotein L1 phosphorylation and cardiac troponin I concentrations (r = -0.91; p = 0.0016). These findings demonstrate that serum phosphoproteomics can provide valuable insights into the molecular events associated with AMI. The inverse relationship between apolipoprotein L1 phosphorylation and cardiac troponin I levels suggests that phosphoproteomic profiling may aid in understanding myocardial injury mechanisms.
Hemp (Cannabis sativa L.) is one of the oldest cultivated plants in human history. It is a valuable plant that has economic value for all industrial branches. The hemp plant has fibers of the highest quality. Accordingly, research aiming to enhance the fiber yield of hemp has significant agronomic and industrial relevance. To this end, three soilless media treatments were applied to the modified Hoagland solution. The effects of N: P:K/255:45:267 (A1), A1 + 10 mg/L silicon (A2), and A1 + 1 mg/L indole butyric acid (A3) on plant growth and fiber yield were investigated in Narlı hemp (Cannabis sativa L.), a registered local variety (Türkiye), in deep water culture and soil media (A4). The analysis determined that the soilless medium was more productive than the soil medium in terms of root fresh-dry weight, stem fresh-dry weight, stem length, stem thickness, leaf length and width, relative water content (RWC), photosynthetic pigment content, fiber content, and total cellulose content. According to these results, the lowest values were obtained in the soilless medium in terms of malondialdehyde (MDA) and leaf secondary metabolite (cannabidiol: CBD, tetrahydrocannabinol: THC, and cannabinol: CBN) contents, which indicate stress in the plant. CBD was measured %0.0213, %0.0134, %0.0395, %0.0326, THC was measured %0.002, %0.0017, %0.0032, %0.0033, CBN was measured %0.0003, %0.0002, %0.0002, %0.0009 (A1, A2, A3, A4 respectively). Considering all the parameters studied, the hemp plant grown in the deep water culture medium containing Si yielded the best results among the treatments. The fiber content was 29.12% in A2, 24.02% in A1, 19.99% in A4, and 13.74% in A3. The cellulose content was determined to range from 61% to 82% on average in the treatments tested. The highest total cellulose content of 82% was obtained from treatment A2. The current study revealed that silicon (Si) treatment significantly increased fiber and cellulose contents. Incorporating such treatments into production models is regarded as a cost-effective and efficient strategy.
Basal stem rot (BSR), a devastating disease caused by Ganoderma boninense (synonym G. orbiforme) significantly affects the oil palm industry, particularly in Indonesia and Malaysia. Current control measures for BSR are limited, prompting the need for environmentally sustainable biocontrol alternatives. In this study, the antagonistic potential of medicinal mushrooms against G. orbiforme to mitigate BSR in oil palm were evaluated. Among the tested species, G. lucidum exhibited the highest percentage inhibition of radial growth (PIRG) at 60.96%, followed by G. neojaponicum (50.37%) and Lignosus rhinoceros (13.50%). The biocontrol efficacy of G. lucidum was further assessed using a modified air-lift bioreactor (MALB) system, demonstrating that G. lucidum is non-pathogenic to oil palm and enhanced plant growth by 41.67% in terms of dry weight. In contrast, G. orbiforme inoculation resulted in typical symptoms such as leaf necrosis, yellowing and browning, Cercospora leaf spots and extensive hyphal mass growth, thereby confirming the pathogenicity of this species in oil palm. Infected plants experienced a significant reduction in dry weight by 115.31%. Furthermore, inoculating G. lucidum into plants either before or after inoculation with G. orbiforme positively impacted plant growth, with more pronounced effects when G. lucidum was introduced first, followed by the pathogenic G. orbiforme as tested in MALB system. These findings suggest that G. lucidum could be effectively utilized as a biocontrol agent to control BSR.
Immobilization of lipase enzyme is a promising approach towards cost-effective production of biodiesel to protect the enzyme from denaturation during the transesterification process. In the present study, hydrogen titanate nanotubes (HTNTs), synthesized by simple hydrothermal method, was used to immobilize lipase from Aspergillus niger via physical adsorption with the following ratios of HTNTs and lipase; 1 T: 1 L, 1 T: 0.75 L, 1 T: 0.5 L, and 1 T: 0.25 L. All prepared samples were characterized by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and scanning electron microscopy (SEM). The catalytic activities of free and immobilized lipases were evaluated for biodiesel production using sunflower oil, methanol to oil molar ratio of 4: 1 at 40 °C for 90 min. The biodiesel yields (0% water content, 0.877 g/cm3 density, and 0.041 mg acid value) were 79.2 ± 0.01, 82.3 ± 0.9, 79.1 ± 1.2, 81.97 ± 1.4, 78 ± 0.1 percent for 1 T: 1 L, 1 T: 0.75 L, 1 T: 0.5 L, 1 T: 0.25 L, and free lipase (1 L), respectively. Compared with the control (1 L), immobilizing lipase using (HTNTs) enabled a 50- 75% reduction in lipase required quantity while maintaining or even increasing biodiesel production levels. The work establishes a promising method for lipase immobilization in biodiesel production that can be evaluated for large-scale application in further studies.
The present study aimed to valorize rainbow trout (Oncorhynchus mykiss) processing waste through enzymatic hydrolysis using Alcalase and Flavourzyme, to produce protein hydrolysates with enhanced biofunctional properties. Hydrolysates were evaluated for their chemical composition, antimicrobial activity, antioxidant capacity, and molecular effects on Pseudomonas aeruginosa. Biochemical analysis revealed that enzymatic treatment significantly increased protein content and reduced moisture levels compared to crude extracts. Antimicrobial assays showed that while crude proteins exhibited no inhibitory effect, hydrolysates particularly those derived from Alcalase demonstrated clear inhibition zones against P. aeruginosa. Quantitative PCR confirmed microbial suppression by showing delayed amplification curves in treated samples. Antioxidant activity, measured via IC₅₀ values, indicated that Alcalase-generated hydrolysates possessed superior radical-scavenging capacity. The observed bioactivity was closely linked to peptide size distribution and enzyme specificity. These findings suggest that rainbow trout waste hydrolysates are promising candidates for use as natural antimicrobial and antioxidant agents in food, pharmaceutical, and environmental applications. Further studies are recommended to isolate and characterize the active peptides and assess their efficacy in real-world systems.