Salt-cured Atlantic cod skin: a sustainable source of acid-soluble type I collagen

Collagen is the most abundant protein in the animal kingdom. Industrial collagen is mainly bovine and porcine origin. However, due to religious beliefs, allergic issues, and infectious diseases, alternative sources of collagen as marine are gaining increasing interest. In this work, the acid-soluble collagen (ASC) were extracted from salt-cured Atlantic cod (Gadus morhua) skin and characterized. The extraction yield was about 2.0%, equivalent to the extraction yield reported for other fish skins. The electrophoretic pattern showed the typical type I structure (α, β and γ chains). UV-VIS and FTIR absorbance spectra suggested a very pure ASC with an intact triple helical structure. The integrity and the adequate porosity required for different applications were then confirmed by electron micrograph. Our findings allow us to say that, for the first time, we extracted acid-soluble type I collagen from salt-cured Atlantic cod skin, with characteristics suitable for application in various fields, such as biomedical.info:eu-repo/semantics/publishedVersio

Non-ionic aqueous micellar extraction of trypsin inhibitors and isoflavones from soybean meal: process optimization

Simultaneous extraction of trypsin inhibitors and soy isoflavones from soybean meal was investigated using the non-destructive phytochemical extraction process, namely aqueous micellar system. The ethoxylated aliphatic alcohols Genapol X-080, Tergitol 15-S-7, and Tergitol 15-S-9, all non-toxic and biodegradable surfactants, were assessed as potential extractants. A Box-Behnken multifactorial design with the application of the Derringer desirability was used to determine the conditions that maximized the trypsin inhibitors and isoflavone extraction while minimizing the protein extraction. The optimum condition of 5% m/m of surfactant in 50 mM aqueous sodium citrate solution pH 4.5, at 45 °C for 45 min, was established for the three surfactants. The novel methodology would allow the extraction of the main soybean antinutritional factors, trypsin inhibitors, and the valuable isoflavones, preserving the nutritional quality of the treated material. This represents a sustainable alternative methodology for industrial purposes due to its low cost, biodegradability, non-toxicity, and easy scaling up.info:eu-repo/semantics/publishedVersio

Applications in medicine: joint health

Rheumatic and joint diseases are among the most widespread pathologies associated with pain and cause a great impact in society across the globe. Given the continuing rise in life expectancy, their prevalence is destined to grow. Osteoarthritis, a degenerative joint disease, is on its way to becoming the fourth leading cause of disability worldwide by 2020. Accompanying osteoarthritis is rheumatoid arthritis, which is a chronic systemic disease that often causes pain and deformity. The joint function involves several processes and understanding the mechanism of action of the joint disease process (proliferation, degradation, regeneration, and infection) is important for the diagnostic, prophylaxis, and treatment. These mechanisms are associated with active peptides that are involved in almost all physiological processes, including cell differentiation, proliferation, inflammatory processes, and immune regulation. Therefore the study of bioactive peptides as biological markers and therapeutic targets for the diagnosis, prevention, and therapy of joint diseases has a very important research ahead. In this context, the present chapter will highlight the importance of bioactive peptides in joint health, underline peptides natural sources, their mechanisms of action, and their potential applications on joint diseases.info:eu-repo/semantics/acceptedVersio

Peptide fraction identification by SE-HPLC and LC-MS/MS analysis of the body mucus from Portugal coastal fish Halobatrachus didactylus

The mucus covers the fish's body, working as a protective barrier. Besides physical protection, mucus provides molecules that protect the fish from pathogens damaging. 1,2These include antimicrobial peptides secreted in the mucus, which play an essential role in defense against microbial pathogens since these belong to the innate immune system2,3. In this study, two adult Halobatrachus didactylus individuals were captured from the wild in Sesimbra. Then, mucus collection was performed by scraping the dorsal-lateral body of the fish with a sponge. Our objective was the identification of new peptides with bioactive potential in mucus samples by chromatography analysis. Size exclusion high-performance liquid chromatography (SE-HPLC) analysis performed on mucus samples from the two individuals revealed a similar profile with an intense highlight peak which resulted in a distribution of about 775 Dalton. With interest in that peak, the two mucus samples were pooled for fractionation by SEC. The resulting fraction was analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify the most probable peptide sequences. Identification from databases did not provide reliable results, indicating a lack of information on the matrix analyzed. We resorted to de novo sequencing with good results using PEAKS Studio software. Five identified peptides were selected according to their bioactivities predicted in silico. Furthermore, the five identified peptides were synthesized, and the molecular size was validated by SE-HPLC analysis. Overall, this chromatographic approach enabled the identification of promising peptides, which bioactivities will be evaluated in vitro in future work.info:eu-repo/semantics/publishedVersio

First bioactive characterization of the skin mucus from Portugal coastal fish Halobatrachus didactylus

info:eu-repo/semantics/publishedVersio

Bioactive potential and chromatographic characterization of body mucus from Portugal coastal fish halobatrachus didactylus

The body mucus in fish provides a stable physical or chemical barrier against invading pathogens. In mucus, antimicrobial peptides are secreted as a response to immunestimulation. Studies have shown antimicrobial activity against multidrug­resistant pathogens and low toxicity to eukaryotic cells. Previously, body mucus from five captive Halobatrachus didactylus individuals was collected. We aim to explore the mucus studying molecules with bioactive potential. Size exclusion high­performance liquid chromatography (SE­HPLC) analyses were performed on the five body mucus samples showing a similar molecular size distribution with a maximum peak of ca. 800 Da. These five mucus samples were pooled to assess the following bioactivities: antioxidant (ABTS and ORAC), antimicrobial (minimal inhibitory concentration), andcytotoxicity (Caco­2 and HaCaT human cell lines). The protein content in the mucus, determined by the bicinchoninic acid methodology, was 16836 ± 1020 µg BSA/mL. The antioxidant activity resulted in 268 ± 11 µmol TE/g mucus protein for ABTS and 306 ± 11 µmol TE/g mucus protein for ORAC. The antibacterial activity was assessed against five pathogenic bacteria: Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella enterica, Staphylococcus aureus, within minimal inhibitory concentrations of 421 to 105 µg mucus protein/mL. Moreover, the mucus showed non­cytotoxic for Caco­2 cells in concentrations between 196 to 25 µg mucus protein/mL, while it showed cytotoxicity for HaCaT cells. In the future, liquid chromatography­tandem mass spectrometry (LC­MS/MS) analysis will be performed to determine the molecules behind these bioactivities, namely antimicrobial peptides.info:eu-repo/semantics/publishedVersio

Bioactive potential and chromatographic characterization of body mucus from Portugal coastal fish halobatrachus didactylus

The body mucus in fish provides a stable physical or chemical barrier against invading pathogens. In mucus, antimicrobial peptides are secreted as a response to immune stimulation. Studies have shown antimicrobial activity against multidrug-resistant pathogens and low toxicity to eukaryotic cells. Previously, body mucus from five captive Halobatrachus didactylus individuals was collected. We aim to explore the mucus studying molecules with bioactive potential. Size exclusion high­performance liquid chromatography (SE­HPLC) analyses were performed on the five body mucus samples showing a similar molecular size distribution with a maximum peak of ca. 800 Da. These five mucus samples were pooled to assess the following bioactivities: antioxidant (ABTS and ORAC), antimicrobial (minimal inhibitory concentration), and cytotoxicity (Caco­2 and HaCaT human cell lines). The protein content in the mucus, determined by the bicinchoninic acid methodology, was 16836 ± 1020 µg BSA/mL. The antioxidant activity resulted in 268 ± 11 µmol TE/g mucus protein for ABTS and 306 ± 11 µmol TE/g mucus protein for ORAC. The antibacterial activity was assessed against five pathogenic bacteria: Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella enterica, Staphylococcus aureus, within minimal inhibitory concentrations of 421 to 105 µg mucus protein/mL. Moreover, the mucus showed non­cytotoxic for Caco­2 cells in concentrations between 196 to 25 µg mucus protein/mL, while it showed cytotoxicity for HaCaT cells. In the future, liquid chromatography­tandem mass spectrometry (LC­MS/MS) analysis will be performed to determine the molecules behind these bioactivities, namely antimicrobial peptides.info:eu-repo/semantics/publishedVersio

Methods for the collection of fish mucus: a systematic review

The aquatic environment holds a vast source of organisms that provide numerous opportunities to bioprospect new molecules. Notably, fish are producers of an epidermal mucus that offers protection against pathogens, making it a promising source of bioactive molecules. This source of molecules, however, has yet to be thoroughly explored, and particularly, optimization of methods for collection and study is needed. This review concentrates on the methods of mucus collection employed to secure high-quality samples, enabling the extraction and characterization of molecules with bioactive potential. A comprehensive search was conducted, and publications were selected based on the following criteria: (i) the mucus has been collected from the external body of the fish, not involving dissection or damage; (ii) mucus crude extracts have undergone a chemical or genetic characterization; (iii) mucus was used in bioactivity assays (e.g., antimicrobial or immune-related). Scraping, bagging, and absorption are the primary methods for collecting fish mucus. They were assessed based on fish handling, sample volume, and processing, including anesthesia and starvation. Scraping with a soft tool, such as cotton balls or sponges, proved most effective and minimized contamination, dilution, and injury risk. This review aids future studies of mucus composition and properties.info:eu-repo/semantics/acceptedVersio

Thermosonication applied to kiwi peel: a healthy source of nutrients

info:eu-repo/semantics/publishedVersio

Thermosonication applied to kiwi peel: impact on nutritional and microbiological indicators

The peels of many fruits are rich sources of nutrients, although they are not commonly consumed. If they are properly decontaminated, they can be used as healthy food ingredients reducing food waste. The objective was to apply thermosonication processes to kiwi peel and evaluate the impact on Listeria innocua survival (a non-pathogenic surrogate of L. monocytogenes) and key nutrients and quality indicators: proteins, fibers, minerals (Ca, K, Mg, Na, and P), chlorophylls, and phenolic contents. Kiwi peels were artificially inoculated with L. innocua and thermal and thermosonication treatments were performed at 55 °C and 60 °C for 30 and 15 min maximum, respectively. Bacteria were enumerated through treatment time, and quality indicators were assessed before and at the end of treatments. A Weibull model with a decimal reduction time (D-value) was successfully used in L. innocua survival data fits. Results showed that coupling temperature to ultrasound had a synergistic effect on bacteria inactivation with significant decreases in D-values. Thermosonication at 60 °C was the most effective in terms of protein, fiber, chlorophylls, and phenolics retention. Minerals were not significantly affected by all treatments. Applying thermosonication to kiwi peel was more effective for decontamination than thermal treatments at the same temperature while allowing the retention of healthy compounds.info:eu-repo/semantics/publishedVersio