Subcritical water-assisted fish gelatin hydrolysis for astaxanthin-loaded fish oil emulsion stability.

Though gelatin emulsifying properties have been intensively studied, how low-molecular-weight (LMW) fish gelatin affects astaxanthin (AST)-loaded fish oil emulsion stability remains elusive. In this study, subcritical water hydrolysis (SWH)-modified LMW fish gelatin (SWHG) was produced from 110 °C to 180 °C and used to enhance the AST steadiness in oil/water emulsions in the presence of an emulsifier, lecithin. In the prepared emulsions, the surface charge increased while droplet size decreased with the decrease in gelatin MW due to the reduced thickness of the adsorbed gelatin membrane. LMW gelatin and lecithin could form a firm-absorbed layer on the droplet surface by electrostatic interaction between amide groups of gelatin molecules and phosphate groups of lecithin, thus stabilizing the emulsions. SWHG improved the creaming stability of the emulsions and hindered the oxygen- and light-induced AST degradation for 11 months compared to high MW gelatin. Whereas, the control emulsion showed noticeable phase separation after two weeks of storage. These findings prove the advantage of the SWH approach and propose the use of SWHG in oil-in-water emulsions for AST stabilization.

In Vitro Biodegradation, Drug Absorption, and Physical Properties of Gelatin-Fucoidan Microspheres Made of Subcritical-Water-Modified Fish Gelatin.

This study aimed to prepare gelatin-fucoidan microspheres with enhanced doxorubicin binding efficiency and controllable biodegradation using fish gelatin combined with low molecular weight (LMW) gelatin and fucoidan at fixed ratios. The MW of gelatin was modified by subcritical water (SW), which is known as a safe solvent, at 120 °C, 140 °C, and 160 °C. In addition, gelatin-fucoidan microspheres were prepared using a solvent exchange technique. Our findings revealed that particle size decreased, the surface was rougher, the swelling ratio increased, and particle shape was irregular in microspheres composed of SW-modified gelatin. Doxorubicin binding efficiency was improved by fucoidan and SW-modified gelatin at 120 °C but not at 140 °C and 160 °C. Interestingly, an increase in in vitro enzymatic degradation was observed in the microspheres consisting of SW-modified fish gelatin, although the cross-linking degree between them was not significantly different. This is because LMW gelatin could form more cross-linked bonds, which might be weaker than the intramolecular bonds of gelatin molecules. Gelatin-fucoidan microspheres consisting of SW-modified fish gelatin with controlled biodegradation rates could be a candidate for a short-term transient embolization agent. In addition, SW would be a promising method to modify the MW of gelatin for medical applications.

Development of fish gelatin/carrageenan/zein bio-nanocomposite active-films incorporated with turmeric essential oil and their application in chicken meat preservation.

Gelatin/carrageenan (Ge/Car) active packaging films incorporated with turmeric essential oil (TEO) encapsulated in zein nanoparticles (ZNP) were developed. The efficacy of these active packaging films and their antimicrobial properties were also investigated to ensure their practical application. Three different types of nanocomposite films (Ge/Car, Ge/Car/TEO, and Ge/Car/ZNP) were prepared. The characterization of the films was elucidated using Fourier transform infrared (FTIR), X-ray diffraction analyses (XRD), and scanning electron microscope (SEM). Physicochemical and mechanical properties of the films were enhanced, owing to the application of TEO-containing nanocomposites. Supercritical-CO2 extracted TEO showed excellent biological activities, alongside GC-MS analysis identified that TEO contained 33 bioactive compounds where the major constituent was Zingiberene. ZNP proved an excellent carrier of TEO. The nanocomposite film sustainably released TEO, improving the shelf life of the chicken meat by reducing bacterial colonies from 3.08 log CFU/g to 2.81 log CFU/g after 14 days incubation against Salmonella enterica compared with 6.66 log CFU/g observed in the control film. The overall results of this study suggest that the nanocomposite active film is an excellent candidate for food packaging to ensure a better world.

Anticancer and Apoptotic Activity in Cervical Adenocarcinoma HeLa Using Crude Extract of Ganoderma applanatum.

Cancer is currently one of the foremost health challenges and a leading cause of death worldwide. Cervical cancer is caused by cofactors, including oral contraceptive use, smoking, multiparity, and HIV infection. One of the major and considerable etiologies is the persistent infection of the oncogenic human papilloma virus. G. applanatum is a valuable medicinal mushroom that has been widely used as a folk medicine for the treatment and prevention of various diseases. In this study, we obtained crude extract from G. applanatum mushroom with a subcritical water extraction method; cell viability assay was carried out and the crude extract showed an antiproliferative effect in HeLa cells with IC50 of 1.55 ± 0.01 mg/mL; however, it did not show any sign of toxicity in HaCaT. Protein expression was detected by Western blot, stability of IκBα and downregulation of NFκB, IKKα, IKKß, p-NFκB-65(Ser 536) and p-IKKα/ß(Ser 176/180), suggesting loss of survival in a dose-dependent manner. RT-qPCR revealed RNA/mRNA expression; fold changes of gene expression in Apaf-1, caspase-3, cytochrome-c, caspase-9, Bax and Bak were increased, which implies apoptosis, and NFκB was decreased in a dose-dependent manner. DNA fragmentation was seen in the treatment groups as compared to the control group using gel electrophoresis. Identification and quantification of compounds were carried out by GC-MS and HPLC, respectively; 2(5H)furanone with IC50 of 1.99 ± 0.01 µg/mL could be the responsible anticancer compound. In conclusion, these findings suggest the potential use of the crude extract of G. applanatum as a natural source with anticancer activity against cervical cancer.

Subcritical Water Hydrolysis of Comb Pen Shell (Atrina pectinata) Edible Parts to Produce High-Value Amino Acid Products.

Artina pectinata (Comb pen shell, CPS) is a high-protein source that contains a variety of essential amino acids. Subcritical water hydrolysis (SWH) was used to recover amino acids from the posterior adductor muscle (PAM), anterior adductor muscle (ADM), and mantle. The temperatures ranged from 120 °C to 200 °C, and the pressure and time of hydrolysis were 3 MPa and 30 min, respectively. Further characterization of the hydrolysates was performed to ascertain amino acid profiles and biofunctional properties. The hydrolysates contained more free amino acids than the untreated samples. Antioxidant activity of treated samples increased as SW temperatures increased. At 200 °C, those inhibiting ACE had a maximum antihypertensive activity of 200 °C in 1% PAM, ADM, and mantle with 85.85 ± 0.67, 84.55 ± 0.18, and 82.15 ± 0.85%, respectively, compared to 97.57 ± 0.67% in 1% standard captopril. Perhaps the most significant finding was the predominance of taurine in the three parts following SW treatment at 120 °C. The hydrolysates may be of considerable interest for use in food or energy drinks. SWH demonstrates efficacy in recovering amino acids, particularly taurine, from edible parts of A. pectinata.

Lipid Indexes and Quality Evaluation of Omega-3 Rich Oil from the Waste of Japanese Spanish Mackerel Extracted by Supercritical CO2.

Japanese Spanish mackerel (JSM) (Scomberomorus niphonius) is a marine fish species containing health-beneficial polyunsaturated fatty acids (PUFAs). In the present study, the quality of JSM by-products oils extracted by supercritical CO2 (SC-CO2) and organic solvent extraction was compared in terms of physico-chemical properties of the oils. Eicosapentaenoic acid (EPA) is one of the important polyunsaturated fatty acids present in SC-CO2-extracted skin and muscle oil 5.81 ± 0.69% and 4.93 ± 0.06%, respectively. The amount of docosahexaenoic acid (DHA) in SC-CO2-extracted skin and muscle oil was 12.56 ± 0.38% and 15.01 ± 0.28%, respectively. EPA and DHA are considered as important PUFAs for the development of brain function and the prevention of coronary heart diseases. Extracted oils showed considerable antioxidant activity. In the obtained oils, atherogenic index (AI) values varied from 0.72 to 0.93 and thrombogenic index (TI) ranged from 0.75 to 0.92, which is considered an acceptable level. Fatty acid composition, bio potentiality, thermogravimetric, and vitamin D analysis showed that oils extracted from JSM by-products can be a good source of oil for application in food, pharmaceutical and cosmetic industries. Therefore, the present research revealed the potentiality of green valorisation of S. niphonius by-products as a possible sustainable approach for targeting the era of zero waste.

In vivo protective effect against ethanol metabolism and liver injury of oyster (Crassostrea Gigas) extracts obtained via subcritical water processing.

The present study assesses hepatoprotective effects of raw oyster lyophilized powder (OP) and subcritical water treated oyster powder (SOP) on D-galactosamine (D-GalN)-induced toxicity and acute ethanol intoxication in mice. High-performance liquid chromatography analysis revealed that four phenolic compounds and glucose were identified from the SOP. The levels of aspartate aminotransferase, alanine aminotransferase, and malondialdehyde were considerably lower for the oyster extracts and the levels of glutathione, γ-glutamylcysteinesynthetase, glutathione S-transferase, and glutathione reductase were higher in the D-GalN induced mice compared with those in the controls. Histology analysis suggested that SOP can protect against and heal the D-GalN toxified liver. For the acute ethanol intoxication study, the enzymatic activity of acetaldehyde dehydrogenase and SOP's alcohol dehydrogenase appeared better than that of OP. Overall, SOP may protect the liver from acute ethanol intoxication and D-GalN persuaded hepatitis. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10068-021-00941-9.

In vitro characterization of bioactive compounds extracted from sea urchin (Stomopneustes variolaris) using green and conventional techniques.

This study investigated the in vitro bioactivities of extracts obtained from viscera, spines, shells, and gonads of Stomopneustes variolaris using subcritical water extraction (SWE) at 110 °C, 150 °C, 190 °C, and 230 °C and Soxhlet extraction. The highest amounts of phenolics (22.68 ± 0.05 mg GAE/g), flavonoids (27.11 ± 0.10 mg RE/g), and proteins (40.25 ± 0.84 mg BSA/g) were recorded from gonads at 230 °C, whereas maximum sugar content (23.38 ± 1.30 mg glucose/g) was in viscera at 150 °C. Gonads at 230 °C exhibited the highest DPPH activity (78.68 ± 0.18%), whereas viscera at 150 °C exhibited the highest ABTS+ (98.92 ± 1.27%) and protein denaturation inhibition activity (37.13 ± 9.94%). Viscera at 110 °C claimed the highest amylase inhibition (42.46 ± 0.83%), and spines at 150 °C had the highest anticancer activity (IC50 = 767.47 µg/mL). SWE achieved superior results in bioactive compound recovery and detected higher levels of bioactivities (p < 0.05). Results suggest processing sea urchin extracts via SWE has potential application to the food and pharmaceutical industries.

Amino Acid Profiles and Biopotentiality of Hydrolysates Obtained from Comb Penshell (Atrina pectinata) Viscera Using Subcritical Water Hydrolysis.

The recovery of amino acids and other important bioactive compounds from the comb penshell (Atrina pectinata) using subcritical water hydrolysis was performed. A wide range of extraction temperatures from 140 to 290 °C was used to evaluate the release of proteins and amino acids. The amount of crude protein was the highest (36.14 ± 1.39 mg bovine serum albumin/g) at 200 °C, whereas a further increase in temperature showed the degradation of the crude protein content. The highest amount of amino acids (74.80 mg/g) was at 230 °C, indicating that the temperature range of 170-230 °C is suitable for the extraction of protein-rich compounds using subcritical water hydrolysis. Molecular weights of the peptides obtained from comb penshell viscera decreased with the increasing temperature. SDS-PAGE revealed that the molecular weight of peptides present in the hydrolysates above the 200 °C extraction temperature was ≤ 1000 Da. Radical scavenging activities were analyzed to evaluate the antioxidant activities of the hydrolysates. A. pectinata hydrolysates also showed a particularly good antihypertensive activity, proving that this raw material can be an effective source of amino acids and marine bioactive peptides.

Influences of Molecular Weights on Physicochemical and Biological Properties of Collagen-Alginate Scaffolds.

For tissue engineering applications, biodegradable scaffolds containing high molecular weights (MW) of collagen and sodium alginate have been developed and characterized. However, the properties of low MW collagen-based scaffolds have not been studied in previous research. This work examined the distinctive properties of low MW collagen-based scaffolds with alginate unmodified and modified by subcritical water. Besides, we developed a facile method to cross-link water-soluble scaffolds using glutaraldehyde in an aqueous ethanol solution. The prepared cross-linked scaffolds showed good structural properties with high porosity (~93%) and high cross-linking degree (50-60%). Compared with collagen (6000 Da)-based scaffolds, collagen (25,000 Da)-based scaffolds exhibited higher stability against collagenase degradation and lower weight loss in phosphate buffer pH 7.4. Collagen (25,000 Da)-based scaffolds with modified alginate tended to improve antioxidant capacity compared with scaffolds containing unmodified alginate. Interestingly, in vitro coagulant activity assay demonstrated that collagen (25,000 Da)-based scaffolds with modified alginate (C25-A63 and C25-A21) significantly reduced the clotting time of human plasma compared with scaffolds consisting of unmodified alginate. Although some further investigations need to be done, collagen (25,000 Da)-based scaffolds with modified alginate should be considered as a potential candidate for tissue engineering applications.

Green extraction of polyphenolic-polysaccharide conjugates from Pseuderanthemum palatiferum (Nees) Radlk.: Chemical profile and anticoagulant activity.

In this study, pressurized liquid extraction (PLE) of polyphenolic-polysaccharide (PP) from Pseuderanthemum palatiferum (Nees) Radlk. leaves was carried out and compared with a conventional technique using 0.1 M sodium hydroxide. The extracts were purified according to the method reported previously to obtain PP conjugates which were further studied about chemical profiles and anticoagulant activity. Fourier-transform infrared spectroscopy (FTIR), UV-Vis, nuclear magnetic resonance (NMR), gel permeation chromatography (GPC), and spectrophotometry analysis were used to characterize the selected PP conjugates. The results showed that PP conjugates comprised of carbohydrate, phenolic, and protein constituents with the yield ranged from 2.76% to 14.34%. Seven mono sugars containing in all conjugates were determined using high-performance liquid chromatography (HPLC), namely, arabinose, fucose, galactose, glucose, mannose, rhamnose, and xylose. PP conjugates obtained from PLE at 150 °C (PP-PLE5) exhibited better anticoagulant activity than those found at 200 °C and comparable to that of the conventional technique. On gel permeation chromatography, PP-PLE5 showed a broad molecular mass from 6 to 642 kDa. From the obtained results, PLE can be used as a green effective technique for the recovery of PP conjugate from P. palatiferum leaves.

Extraction of Bioactive Compounds from Pseuderanthemum palatiferum (Nees) Radlk. Using Subcritical Water and Conventional Solvents: A Comparison Study.

The present study investigated the recovery of antioxidative compounds obtained from Pseuderanthemum palatiferum (Nees) Radlk. and their biological activities using subcritical water in comparison with hot water, Soxhlet, and methanol extraction. In this study, high-performance liquid chromatography (HPLC) and three commonly used assays, that is, 2,2-diphenyl-1-picrylhydrazyl assays, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), and ferric reducing antioxidant power, were employed to measure the number of valuable compounds and their antioxidant capacities, respectively. Additionally, the antimicrobial ability of the various extracts against seven pathogenic bacteria strains was assessed. The results demonstrated that the extracts obtained from subcritical water extraction (SWE) contain the highest amount of saponin (33.82 ± 2.08 mg diosgenin/g), phenolic (34.87 ± 0.29 mg CE/g), and protein (104.66 ± 7.63 mg BSA/g), which are obtained at 170 °C, 190 °C, and 230 °C, respectively. The highest amounts of total sugar and flavonoids (211.73 ± 1.58 mg glucose/g and 20.71 ± 0.42 mg RE/g, respectively) were obtained at lower temperature (130 °C and 130 °C, respectively), and these values were higher than those obtained from hot water, Soxhlet, and methanol extraction. HPLC analysis indicated that the highest amount of apigenin (3.46 ± 0.03 mg/g) and kaempferol (2.43 ± 0.03 mg/g) were obtained by SWE at 170 °C and 190 °C, respectively. Furthermore, the extracts from SWE exhibited higher antimicrobial activity against five of the seven pathogenic bacterial strains tested compared with those obtained from conventional extraction methods. Therefore, subcritical water could be utilized as a cost-effective and green solvent to extract valuable compounds from P. palatiferum (Nees) Radlk. leaf. PRACTICAL APPLICATION: Subcritical water was able to recover more bioactive compounds from the sample, such as phenolics, flavonoids, saponin, protein, and polysaccharides, than conventional solvents. The mixture of polyphenolic-polysaccharide-protein conjugates could be used in further steps, isolation, and purification, and applied to functional food.

Deep eutectic solvent-based extraction and fabrication of chitin films from crustacean waste.

In this study, chitin was exclusively extracted from shrimp shells (Marsupenaeus japonicas) through a green solvent called deep eutectic solvent (DES), and various types of DES were utilized to extract chitin. The physicochemical properties of the obtained chitin were compared with the conventional method. A high purity of chitin was obtained while using DES-8 (choline chloride-malonic acid) with a yield of 19.41% ±â€¯1.35%, and purity was confirmed using 13C nuclear magnetic resonance. The DES-produced chitin was utilized to produce chitin films and was compared with standard chitin films. The obtained films were characterized by SEM, AFM, TGA, DSC, FTIR, mechanical properties, moisture sorption, swelling behavior, and biodegradation. The DES film showed similar properties to the standard film, while the mechanical properties, swelling behavior, and biodegradation of the DES chitin films proved to be similar to standard chitin film. These chitin films can be used as wound healing resources.