Synergistic effects of starch and carrageenan from Kappaphycus alvarezii in composite film formation: Physicochemical and degradable properties.

Rising concerns around plastic pollution from single-use plastic (SUPs), especially food packaging, have driven interest in sustainable alternatives. As such, algae biomass has gained attention for bioplastic production due to algae's rapid growth and abundant polysaccharides. This research focuses on extracting carrageenan from Kappaphycus alvarezii, extensively cultivated in Sabah, Malaysia, and utilizing it in combination with starch and glycerol to develop algae-based films. The physicochemical properties and degradation rate of these films were evaluated, revealing that the addition of carrageenan enhanced overall thermal stability meanwhile increasing water solubility, water content but reducing the degradation rate and swelling degree. This is primarily due to the crystalline structures of carrageenan, which provide a more rigid arrangement compared to the network of starch polymers. However, the incorporation of starch into the blends has enhanced the elongation and surface morphology, resulting in more balanced properties. Overall, these carrageenan films displayed impressive thermal, mechanical, and biodegradability characteristics, establishing their viability as substitutes for conventional plastics.

Potentilla tormentilla Extract Loaded Gel: Formulation, In Vivo and In Silico Evaluation of Anti-Inflammatory Properties.

The objective of the study was to develop a novel topical gel by mixing Potentilla tormentilla ethanolic extract, thermosensitive poloxamer 407, and carbomer 940 and evaluating its stability and rheological behavior. The irritation potential of the gel was evaluated in accordance with the Organization for Economic Cooperation and Development Guidelines 404. The potential anti-inflammatory effects of the developed gel were evaluated in vivo in rats using the carrageenan-induced paw edema test. Moreover, the in silico binding affinity for chlorogenic and ellagic acid, as dominant components in the extract, against cyclooxygenase (COX) 1 and 2 was also determined. Our findings suggest that the gel containing Potentilla tormentilla extract remained stable throughout the observation period, exhibited pseudoplastic behavior, and caused no irritation in rats, thus being considered safe for topical treatment. Additionally, the developed gel showed the capability to reduce rat paw edema, which highlights significant anti-inflammatory potential. In silico analysis revealed that chlorogenic and ellagic acid exhibited a reduced binding affinity against COX-1 but had a similar inhibitory effect on COX-2 as flurbiprofen, which was confirmed by molecular dynamics results. The study proposes the possible application of Potentilla tormentilla ethanolic extract gel for the alleviation of localized inflammatory diseases; however, future clinical evaluation is required.

Formulation of novel low-sodium salts using potassium salts and dietary polysaccharide.

Potassium citrate (KC) and potassium lactate (KL) are considered as salt replacers due to their saltiness, processing advantages, and health benefits. However, the obvious bitter taste associated with these compounds has limited their use in salt substitutes. Despite this challenge, little attention has been paid to improving their sensory properties. This study provided evidence that dietary polysaccharide carrageenan can effectively mask the bitterness of KC and KL by specifically binding K+ and forming double helix chains. A highly accurate prediction model was then established for the saltiness and bitterness of low-sodium salts using mixture design principles. Three low-sodium salt formulas containing different potassium salts (KC, KL, KCl), NaCl, and carrageenan were created based on the prediction model. These formulas exhibited favorable saltiness potencies (>0.85) without any noticeable odor, preserving the sensory characteristics of high-sodium food products like seasoning powder while significantly reducing their sodium content. This research provides a promising approach for the food industry to formulate alternative low-sodium products with substantially reduced sodium content, potentially contributing to decreased salt intake.

Investigation of the Aggregation of Aß Peptide (1-40) in the Presence of κ-Carrageenan-Stabilised Liposomes Loaded with Homotaurine.

The kinetics of amyloid aggregation was studied indirectly by monitoring the changes in the polydispersity of mixed dispersion of amyloid ß peptide (1-40) and composite liposomes. The liposomes were prepared from the 1,2-dioleoyl-sn-glicero-3-phoshocholine (DOPC) phospholipid and stabilised by the electrostatic adsorption of κ-carrageenan. The produced homotaurine-loaded and unloaded liposomes had a highly negative electrokinetic potential and remarkable stability in phosphate buffer (pH 4 and 7.4). For the first time, the appearance and evolution of the aggregation of Aß were presented through the variation in the standard percentile readings (D10, D50, and D90) obtained from the particle size distribution analysis. The kinetic experiments indicated the appearance of the first aggregates almost 30 min after mixing the liposomes and peptide solution. It was observed that by adding unloaded liposomes, the size of 90% of the particles in the dispersion (D90) increased. In contrast, the addition of homotaurine-loaded liposomes had almost minimal impact on the size of the fractions of larger particles during the kinetic experiments. Despite the specific bioactivity of homotaurine in the presence of natural cell membranes, this study reported an additional inhibitory effect of the compound on the amyloid peptide aggregation due to the charge effects and 'molecular crowding'.

Chitosan-carrageenan microbeads containing nano-encapsulated curcumin: Nano-in-micro hydrogels as alternative-therapeutics for resistant pathogens associated with chronic wounds.

Antimicrobial resistance is an issue of global relevance for the treatment of chronic wound infections. In this study, nano-in-micro hydrogels (microbeads) of chitosan and κ-carrageenan (CCMBs) containing curcumin-loaded rhamnosomes (Cur-R) were developed. The potential of Cur-R-CCMBs for improving the antibacterial activity and sustained release of curcumin was evaluated. Curcumin-loaded rhamnosomes (rhamnolipids functionalized liposomes) had a mean particle size of 116 ± 7 nm and a surface-charge of -24.5 ± 9.4 mV. The encapsulation efficiency of curcumin increased from 42.83 % ± 0.69 % in Cur-R to 95.24 % ± 3.61 % respectively after their embedding in CCMBs. SEM revealed smooth surface morphology of Cur-R-CCMBs. FTIR spectroscopy confirmed the presence of weak electrostatic and hydrophobic interactions among curcumin, rhamnosomes, and microbeads. Cur-R-CCMBs had demonstrated significant antibacterial activity against multi-drug resistant chronic wound pathogens including Staphylococcus aureus and Pseudomonas aeruginosa. Cur-R-CCMBs also exhibited significantly higher anti-oxidant (76.85 % ± 2.12 %) and anti-inflammatory activity (91.94 % ± 0.41 %) as well as hemocompatibility (4.024 % ± 0.59 %) as compared to pristine microbeads. In vivo infection model of mice revealed significant reduction in the viable bacterial count of S. aureus (∼2.5 log CFU/mL) and P. aeruginosa (∼2 log CFU/mL) for Cur-R-CCMBs after 5 days. Therefore, nano-in-micro hydrogels can improve the overall efficacy of hydrophobic antimicrobials to develop effective alternative-therapeutics against resistant-pathogens associated with chronic wound infections.

Exploring the aroma profile and biomedical applications of Scutellaria nuristanica Rech. F.: A new insight as a natural remedy.

BACKGROUND: The Scutellaria genus has promising therapeutic capabilities as an aromatherapy. Based on that and local practices of S. nuristanica Rech. F. The essential oil was studied for the first time for its diverse biomedical applications. PURPOSE: This study aimed to evaluate and validate their therapeutic capabilities by screening the essential oil ingredients and examining their antimicrobial, antioxidant, carbonic anhydrase, and antidiabetic using further In silico assessment and In vivo anti-inflammatory and analgesic capabilities to devise novel sources as natural remedies alternative to the synthetic drugs. METHODS: Essential oil was obtained through hydrodistillation, and the constituents were profiled using GC-MS. The antimicrobial assessment was conducted using an agar well diffusion assay. Free radical scavenging capabilities were determined by employing DPPH and ABTS assay. The carbonic anhydrase-II was examined using colorimetric assay, while the antidiabetic significance was performed using α-Glucosidase assay. The anti-inflammatory significance was examined through carrageenan-induced paw edema, and the analgesic features of the essential oil were determined using an acetic acid-induced writhing assay. RESULTS: Fifty constituents were detected in S. nuristanica essential oil (SNEO), contributing 95.93 % of the total EO, with the predominant constituents being 24-norursa-3,12-diene (10.12 %), 3-oxomanoyl oxide (9.94 %), methyl 7-abieten-18-oate (8.85 %). SNEO presented significance resistance against the Gram-positive bacterial strains (GPBSs), Bacillus atrophaeus and Bacillus subtilis, as compared to the Salmonella typhi and Klebsiella pneumoniae, Gram-negative bacterial strains (GNBSs) as well as two fungal strains Aspergillus parasiticus and Aspergillus niger associated with their respective standards. Considerable free radical scavenging capacity was observed in DPPH compared to the ABTS assay when correlated with ascorbic acid. In addition, when equated with their standards, SNEO offered considerable in vitro carbonic anhydrase II and antidiabetic capabilities. Additionally, the antidiabetic behavior of the 9 dominant compounds of SNEO was tested via In silico techniques, such as molecular docking, which assisted in the assessment of the significance of binding contacts of protein with each chemical compound and pharmacokinetic evaluations to examine the drug-like characteristics. Molecular dynamic simulations at 100 ns and binding free energy evaluations such as PBSA and GBSA models explain the molecular mechanics and stability of molecular complexes. It was also observed that SNEO depicted substantial anti-inflammatory and analgesic capabilities. CONCLUSION: Hence, it was concluded that the SNEO comprises bioactive ingredients with biomedical significance, such as anti-microbial, antioxidant, CA-II, antidiabetic, anti-inflammatory, and analgesic agents. The computational validation also depicted that SNEO could be a potent source for the discovery of anti-diabetic drugs.

Application of purple basil leaf anthocyanins-loaded alginate-carrageenan emulgel beads in gelatin-based jelly candies.

Jelly candies could be considered promising food matrices for the delivery of bioactive compounds such as anthocyanins (ACNs). In this study, gelatin-based jelly candies were fortified with free ACNs-rich purple basil leaf extract (PBLE) and PBLE-loaded emulgel beads. The interaction between free ACNs and gelatin in the jelly matrix resulted in a lower release of ACNs in the mouth (8.27 %) and gastric stage (74.44 %) compared to those of free extract (24.92 and 86.13 %), as well as some protection in the intestinal stage. The release of ACNs from the jellies enriched with PBLE-loaded emulgel beads only began in the intestinal stage and reached 66.34-70.75 % of the initial load. Compared to those of PBLE-loaded emulgel beads, the introduction of beads within the jelly matrix increased the in vitro release of ACNs. The jelly samples enriched with PBLE-loaded emulgel beads yielded higher hardness, adhesiveness, flexibility, and chewiness compared to samples with free PBLE. While the color and appearance of jellies fortified with encapsulated PBLE were impaired, they got the highest sensory acceptance scores due to the masking of the bitter taste of the free extract. ACNs-loaded emulgel beads could be a viable method for fortifying jelly candies.

ShSPI Inhibits Thrombosis Formation and Ischemic Stroke In Vivo.

Thrombotic diseases, emerging as a global public health hazard with high mortality and disability rates, pose a significant threat to human health and longevity. Although current antithrombotic therapies are effective in treating these conditions, they often carry a substantial risk of bleeding, highlighting the urgent need for safer therapeutic alternatives. Recent evidence has increasingly pointed to a connection between elastase activity and thrombosis. In the current study, we investigated the antithrombotic effects of ShSPI, an elastase inhibitor peptide derived from the venom of Scolopendra hainanum. Results showed that ShSPI significantly attenuated carrageenan-induced thrombosis in vivo. Furthermore, ShSPI effectively inhibited the carrageenan-induced decrease in serum superoxide dismutase (SOD) activity and increase in prothrombin time, fibrinogen level, and endothelial nitric oxide synthase (eNOS) activity. In addition, ShSPI reduced intracerebral thrombosis and improved functional outcomes following ischemic stroke in a transient middle cerebral artery occlusion (tMCAO) mouse model. Collectively, these findings suggest that ShSPI is a promising candidate for the development of novel thrombotic therapies.

Fabrication of smart nanogel based on carrageenan and green coffee extract as a long-term antifouling agent to improve biofilm prevention in food production.

This study investigates the extract of the bioactive compounds from green coffee extract (GCE) and the loading of two different concentrations of GCE (1% and 2%) onto carrageenan nanogels (CAR NGs) to compare their antibacterial and antibiofilm effects with unloaded nanogels (NGs). The bioactive compounds of GCE were characterized using GC-MS analysis. The GCE1 and GCE2 were successfully deposited onto the surface of CAR NGs. The antibacterial and antibiofilm potential of prepared NGs were conducted against some foodborne pathogens (E. coli O157, Salmonella enterica, Staphylococcus aureus, and Listeria monocytogenes). The results of GC-MS analysis indicated that there were identified 16 bioactive compounds in GCE, including caffeine (36.27%), Dodemorph (9.04%), and D-Glycero-d-ido-heptose (2.44%), contributing to its antimicrobial properties. The antibacterial coatings demonstrated a notable antimicrobial effect, showing zone of inhibition (ZOI) diameters of up to 37 mm for GCE2 loaded CAR NGs. The minimum inhibitory concentration (MIC) values for GCE2 loaded CAR NGs were 80 ppm for E. coli O157, and 120 ppm for S. enterica, S. aureus, and L. monocytogenes, achieving complete bacterial inactivation within 10-15 min of exposure. Both GCE1 and GCE2 loaded CAR NGs significantly reduced biofilm cell densities on stainless steel (SS) materials for E. coli O157, S. enterica, S. aureus, and L. monocytogenes, with reductions ranging from 60% to 95%. Specifically, biofilm densities were reduced by up to 95% for E. coli O157, 89% for S. enterica, 85% for S. aureus, and 80% for L. monocytogenes. Results of the toxicity evaluation indicated that the NGs were non-toxic and biocompatible, with predicted EC50 values proved their biocompatibility and safety. These results recommended that GCE loaded CAR NGs are promising as natural antimicrobial agents for enhancing food safety and extending shelf life. Further, the study concluded that incorporating GCE into CAR NGs is an effective strategy for developing sustainable antimicrobial coatings for the food industry and manufacturing.

Synthesis, characterization and biocompatibility of hybrid hydrogels based on alginate, κ-carrageenan, and chitosan filled with montmorillonite clay.

New hybrid hydrogel composites based on a mixture of natural polysaccharides (sodium alginate, κ-carrageenan, and chitosan) filled with the clay mineral of natural origin, montmorillonite (MMT), were studied. The structure of intercalated/flocculated MMT distribution in the interpenetrating network of polysaccharide matrix was characterized using FTIR, X-ray diffraction, and SEM techniques. Swelling kinetics was investigated using the weight analysis, whereas the phase transition of water in the composition of hybrid hydrogels, by DSC method. Their biosafety was estimated using the Nelyubov method, germination test on cress (L. sativum) seeds, and metabolic fingerprinting of microbial communities and dehydrogenase assay. The obtained results indicated promising water-retaining properties of the synthesized materials. The hydrogels had a good sorption affinity for cadmium (Cd) ions confining bioavailability of the selected toxic heavy metal. They were safe for soil microorganisms and did not generate metabolic stress for them. Moreover, they did not reduce the viability of pea seeds. Thus, the development of biosafe hybrid hydrogel composites with a comprehensive, good effect on the environment could be considered as successful.

Physicochemical properties and in vitro digestive behavior of astaxanthin loaded Pickering emulsion gel regulated by konjac glucomannan and κ-carrageenan.

This study aimed to elaborate the combination effect of polysaccharides on physicochemical properties and in vitro digestive behavior of astaxanthin (AST)-loaded Pickering emulsion gel. AST-loaded Pickering emulsion gel was prepared by heating Pickering emulsion with konjac glucomannan (KGM) and κ-carrageenan (CRG). The microstructure revealed that adding the two polysaccharides resulted in Pickering emulsion forming a network structure. It exhibited a denser and more uniform network structure, enhancing its mechanical properties four times and increasing its water-holding capacity by 20 %. In vitro digestion experiments demonstrated that the release of free fatty acids from the Pickering emulsion gel (4.25 %) was notably lower than that from conventional Pickering emulsion (17.19 %), whereas AST bioaccessibility was remarkably low at 0.003 %. It provided a feasible strategy to regulate the bioaccessibility in Pickering emulsion, which has theoretical significance to guide the current eutrophic diet people.

Novel analgesic peptide derived from Cinobufacini injection suppressing inflammation and pain via ERK1/2/COX-2 pathway.

Inflammatory pain is a chronic pain caused by peripheral tissue inflammation, seriously impacting the patient's life quality. Cinobufacini injection, as a traditional Chinese medicine injection preparation, shows excellent efficacy in anti-inflammatory and analgesic treatment in patients with advanced tumors. In this study, a novel analgesic peptide CI5 with anti-inflammatory and analgesic bio-functions that naturally presents in Cinobufacini injection and its regulatory mechanism are reported. Our results showed that the administration of CI5 significantly relieved the pain of mice in the acetic acid twisting analgesic model and formalin inflammatory pain model. Furthermore, CI5 effectively reduced the inflammatory cytokines (IL-6, TNF-α and IL-1ß) and inflammatory mediator (PGE2) expressions, and prevented the carrageenan-induced paw edema in mice. Further LC-MS/MS results showed the anti-inflammatory and analgesic bio-functions of CI5 depended on its interaction with the Rac-2 protein upstream of ERK1/2 and the inflammatory signaling pathway (ERK1/2/COX-2 axis). In summary, CI5, as a novel natural candidate identified from Cinobufacini injection, showed substantial clinical promise for inflammatory pain treatments.

Biocide physically cross-linked hydrogels based on carrageenan and guanidinium polyampholytes for wound healing applications.

Over last years, hydrogels based on natural polymers have attracted considerable interest as materials for wound healing. Herein, hydrogel films based on kappa-carrageenan and guanidinium polyampholytes were prepared by the in situ physical cross-linking with potassium chloride and borax, respectively. The polyampholytes were obtained by a free radical copolymerization of 2,2-diallyl-1,1,3,3-tetraethylguanidinium chloride and unsaturated acids. To characterize the composite films, NMR, FTIR, SEM, TGA, XRD, element analysis and tensile test were used. Ampicillin was incorporated into the hydrogels to enhance wound healing potential. The healing-related characteristics, including swelling ratio, drug release and antimicrobial activity, were assessed. The equilibrium swelling ratios were in the range of 3.9-6.5 depending on the polyampholyte composition. According to the in vitro ampicillin release studies, 30-43 % of ampicillin was released from the hydrogels after 5 h at 37 °C and pH 7.4, with drug release being temperature and pH dependent. The ampicillin-loaded films showed a remarkable antimicrobial effect. The inhibition sizes for Escherichia coli and Staphylococcus aureus were 1.10-1.85 and 1.95-2.60 cm, respectively. Although the bi-polymeric hydrogels were thoroughly characterized, with the in vitro study of their biocidal effects carried out in this work, the in vivo drug release assessment needs to be further explored.

Amine-Rich Carbon Dots Synthesized from Kappa-Carrageenan and l-Lysine as a Dual Probe for Detection of Folic Acid and Tumor-Targeted Delivery of Therapeutics.

Strategically designed, heteroatom-rich surface functionalized blue fluorescent carbon dots (CDs) were synthesized for high-throughput detection of folic acid (vitamin B9). The highly stable CDs could particularly detect vitamin B9 in the presence of 35 analytes, even up to 40 nM of the vitamin. The versatile CDs were found to have a high affinity for folic acid in wastewater, folic acid tablets, and food samples enriched with folic acid. The hemocompatibility of the CDs was also studied by using a hemolysis assay, confirming the CDs to be nontoxic to human blood samples up to 400 µg/mL. The CDs were then covalently conjugated to biotin, which possesses receptors that are overexpressed in tumor cells. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide dye) assay and confocal bioimaging studies proved the biotin-modified CDs (CDBT) were remarkably nontoxic in healthy cell lines (HEK-293) and highly target-specific toward tumor cells (HeLa), including triple-negative breast cancer cells (MDA-MB-231). The cytotoxicity assay of 5-fluorouracil encapsulated CDs (CDBTFu) showed the IC50 value to be 81 µM in HeLa cells and 185 µM in MDA-MB-231 cells, respectively, and significantly higher in HEK-293 cells (over 300 µM), owing to high specificity toward tumor cells.

Alaska pollock surimi-based meat analogs by high-moisture extrusion: Effect of konjac glucomannan/curdlan/carrageenan/sodium alginate on fibrous structure formation.

This study investigated the effects of the addition of konjac glucomannan (KGM), curdlan (CD), carrageenan (CA), and sodium alginate (SA) on fibrous structure formation in surimi-based meat analogs to livestock meat. Meat analogs were prepared using high-moisture extrusion with Alaskan pollock surimi and soy protein isolate at a ratio of 7:3 (w/w). The meat analogs samples were labeled as SSP. Macrostructure observation showed that the best fibrous structure was obtained in SSP containing 2% SA. Mesostructure and microstructure observations revealed that 2% CD, CA or SA promoted the formation of a less tight three-dimensional network structure, which contributed to the formation of fiber filaments. Increased ß-sheet structure content, ordered degree, fractal dimension and thermal stability were observed in SSP with the three colloids. Moreover, fibrous texture was closely associated with the thermal stability and fractal dimension. This study has provided useful information for colloid application in surimi-based meat analogs.

A novel biodegradable film based on chicken gelatin and κ-carrageenan cross-linked with oxidized phenolic compounds.

Natural and renewable polymers are gradually replacing petroleum-based plastics, mostly as a result of environmental concerns. Moreover, upcycling industrial food waste into new added-value products is a creative approach that is crucial for cleaner and more sustainable manufacturing. The aim of this study was to obtain an environmentally friendly biodegradable film using a combination of k-carrageenan (KCAR) and chicken gelatin (CGEL), which obtained from poultry by-products. The effects of varying concentrations of KCAR (0-2%) on the physical, permeability, textural, thermal, and microstructural properties of CGEL/KCAR composite films were evaluated. The findings demonstrated that an increase in KCAR enhanced the lightness and opacity levels of the films. Water vapor permeability (WVP) values reduced as the KCAR concentration increased. The lowest WVP value (0.0012 g.mm/h.m2.kpa) was seen in the treatment with 2% KCAR. Tensile strength (TS) values increased with increasing KCAR. The films' thermal stability was increased by the addition of KCAR. Microstructure assessments revealed a more compact and smooth structure in the KCAR-containing treatments, indicating improvements in WVP, thermal stability, and TS. Compared to the commercial cattle gelatin film, the CGEL film had higher TS and lower water solubility (WS). Overall, this study showed that the physical, mechanical, barrier and thermal and microstructural qualities of gelatin-based films may be enhanced by combining CGEL and KCAR to create an effective biodegradable film. Moreover, the comparison study between commercial cattle and chicken gelatin films revealed that cross-linked chicken gelatin films would be a suitable alternative for bovine gelatin films in the production of biodegradable film.

Structural Insights into the Substrate Recognition and Catalytic Mechanism of a GH16 ßκ-Carrageenase from Wenyingzhuangia fucanilytica.

Understanding the substrate specificity of carrageenases has long been of interest in biotechnology applications. So far, the structural basis of the ßκ-carrageenase that hydrolyzes furcellaran, a major hybrid carrageenan, remains unclear. Here, the crystal structure of Cgbk16A_Wf, as a representative of the ßκ-carrageenase from GH16_13, was determined, and the structural characteristics of this subfamily were elucidated for the first time. The substrate binding mode was clarified through a structure analysis of the hexasaccharide-bound complex and molecular docking. The binding pocket involves a conserved catalytic motif and several specific residues associated with substrate recognition. Functions of residues R88, E290, and E184 were validated through site-directed mutagenesis. Comparing ßκ-carrageenase with κ-carrageenase, we proposed that their different substrate specificities are partly due to the distinct conformations of subsite -1. This research offers a comprehensive understanding of the recognition mechanism of carrageenases and provides valuable theoretical support for enzyme modification and carrageenan oligosaccharide preparation.

Synthesis and characterization of polysaccharide-cryogel and its application to the electrochemical detection of DNA.

The main goal of our study is to demonstrate the applicability of the PPy-cryogel-modified electrodes for electrochemical detection of DNA. First, a polysaccharide-based cryogel was synthesized. This cryogel was then used as a template for chemical polypyrrole synthesis. This prepared polysaccharide-based conductive cryogel was used for electrochemical biosensing on DNA. Carrageenan (CG) and sodium alginate (SA) polysaccharides, which stand out as biocompatible materials, were used in cryogel synthesis. Electron transfer was accelerated by polypyrrole (PPy) synthesized in cryogel networks. A 2B pencil graphite electrode with a diameter of 2.00 mm was used as a working electrode. The prepared polysaccharide solution was dropped onto a working electrode as a support material to improve the immobilization capacity of biomolecules and frozen to complete the cryogelation step. PPy synthesis was performed on the electrodes whose cryogelation process was completed. In addition, the structures of cryogels synthesized on the electrode surface were characterized by thermogravimetric analysis (TGA), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). Surface characterization of the modified electrodes was performed by energy-dispersive X-ray spectroscopy (EDX) analysis. Electrochemical determination of fish sperm DNA (fsDNA) was performed using a PPy-cryogel-modified electrode. The use of a porous 3D cryogel intermediate material enhanced the signal by providing a large surface area for the synthesis of PPy and increasing the biomolecule immobilization capacity. The detection limit was 0.98 µg mL-1 in the fsDNA concentration range 2.5-20 µg mL-1. The sensitivity of the DNA biosensor was estimated to 14.8 µA mM-1 cm-2. The stability of the biosensor under certain storage conditions was examined and observed to remain 66.95% up to 45 days.