Valorization of Yellowfin Tuna Tails: From Proteolytic Enzyme Production to Gelatin and Antioxidant Hydrolysate Extraction.

This study investigates the valorization potential of yellowfin tuna (Thunnus albacares) tails to produce high-value commercial products. Firstly, the tuna tails were placed in a perforated stainless-steel cylinder, and hydraulic pressure was applied to separate the skin from the muscle in the tails. The extracted muscle was then utilized as a nitrogen source for the growth of the proteolytic enzyme producer Bacillus subtilis, while the skins were employed for gelatin extraction. The proteases from B. subtilis were partially purified and used to produce antioxidant peptides from the obtained gelatin. The gelatin formed a gel upon cooling, with gelling and melting temperatures of 16 °C and 22 °C, respectively, and a Bloom strength of approximately 160. Response Surface Methodology (RSM) was employed to determine the optimal hydrolysis conditions to achieve the highest antioxidant activity (35.96% measured as DPPH radical scavenging activity), which were 50 °C and 6.5 IU of enzyme. The findings emphasize the importance of an integrated approach to maximize the value of tuna by-products, promoting sustainability within the framework of a circular bioeconomy. Overall, these results contribute to the efficient utilization of tuna by-products, waste reduction, and enhanced economic viability of the tuna industry.

Optimization of fractionation with membranes of antioxidant enzymatic hydrolysate of Californian red worm (Eisenia fetida) protein.

Problem: Earthworm is a valuable source of biologically and pharmacologically active compounds, with applications in the treatment of various types of diseases; however, the main application they have been given is in the production of organic fertilizer. One of the alternatives for obtaining bioactive compounds is by means of enzymatic hydrolysis. Aim: This study proposes the optimization of the fractionation of the antioxidant enzymatic hydrolysate from Californian red worm (Eisenia fetida) protein. Methodology: For this purpose, the worms were separated and hydrolyzed using the enzyme Alcalase 2.4L for 4000s. The obtained hydrolysate was fractionated by means of a crossflow tangential ultrafiltration system, with a 3 kDa molecular weight cut-off ceramic membrane. A response surface design of the composite central factorial type was implemented to evaluate the effect of pH, transmembrane pressure, and flow factors on the response variables transmission, volume reduction factor (VRF) and permeate flow resistance. The transmissions focused on the antioxidant peptides, measured by three conventional methods such as TEAC, FRAP, ORAC, also known as TTEAC, TFRAP and TORAC, respectively. The evaluated resistances were the total resistance (Rtotal), fouling resistance (Rfouling), and gel resistance (Rgel). Result: The results showed that the three factors evaluated affect all the response variables either in their linear or quadratic terms or by some interaction. For each response variable, a mathematical model was obtained, with statistical significance and a non-significant lack of adjustment. The models obtained were used for a multi-objective optimization process in which transfers were maximized, and resistances were minimized. The efficiency of the optimum ultrafiltration process was 25 %. Conclusion: The neutral-alkaline pH is ideal for the ultrafiltration process of bioactive peptides, as it is where the highest transmissions of peptides with antioxidative capacity are found. Under optimal conditions, the 3 kDa membrane permeate was found to exhibit higher antioxidant capacity than the retentate and feed. Based on this, the fraction of less than 3 kDa emerges as a potential multifunctional ingredient, thanks to its antioxidant properties.

Pepsin immobilization on activated carbon and functionalized with glutaraldehyde and genipin for the synthesis of antioxidant peptides of goat casein.

In this article, the synthesis of antioxidant peptides in the enzymatic hydrolysis of caprine casein was analyzed at three different time points (60 min, 90 min, and 120 min) using immobilized pepsin on activated and modified carbon (AC, ACF, ACG 50, ACG 100). The immobilization assays revealed a reduction in the biocatalysts' activity compared to the free enzyme. Among the modified ones, ACG 50 exhibited greater activity and better efficiency for reuse cycles, with superior values after 60 min and 90 min. Peptide synthesis was observed under all studied conditions. Analyses (DPPH, ß-carotene/linoleic acid, FRAP) confirmed the antioxidant potential of the peptides generated by the immobilized enzyme. However, the immobilized enzyme in ACG 50 and ACG 100, combined with longer hydrolysis times, allowed the formation of peptides with an antioxidant capacity greater than or equivalent to those generated by the free enzyme, despite reduced enzymatic activity.

Antioxidant Potential of the Sweet Whey-Based Beverage Colada after the Digestive Process and Relationships with the Lipid and Protein Fractions.

Whey-based beverages could be an effective way of reusing a by-product of th cheese industry, mitigating environmental hazards and, at the same time, profiting a useful food with high nutritional and antioxidant properties. In this study, a traditional Ecuadorian beverage (Colada) was prepared combining sweet whey, Maracuyá and barley. Antioxidant properties before and after an in vitro digestion using the INFOGEST method were determined, and relationships with intestinal transformations of the lipid and protein fractions were analyzed. The digestive process had a positive effect on antioxidant properties based on increased values of ABTS and FRAP located in the bioaccessible fraction (BF), together with strong increments of total polyphenols. Moreover, pretreatment of Caco-2 cells with the BF of Colada significantly reduced ROS generation (p < 0.001) measured by the dichlorofluorescein assay. Substantial changes of the fatty acid profile occurred during digestion, such as a fall of saturated fatty acids and a rise of polyunsaturated. The protein profile, examined by SDS-PAGE and exclusion molecular chromatography in the BF, showed that the major part of the proteins were digested in the intestinal phase. Analysis of NanoLC-MS/MS revealed 18 antioxidant peptides originated from whey proteins, but also 16 peptides from barley with potential antioxidant properties. In conclusion, combining sweet whey with Maracuyá and barley constitutes an excellent nutritional beverage with a strong antioxidant potential.

Isolation and evaluation of quinoa (Chenopodium quinoa Willd.) protein fractions. A nutritional and bio-functional approach to the globulin fraction.

This study evaluated the solubility profiles of quinoa grain proteins and applied a complete process for the isolation of its main protein fractions, namely: albumins, globulins, prolamins and glutelins, which corresponded to 26.96%, 41.3%, 1.7% and 23.16% of the total protein content, respectively. When these fractions were digested with pepsin followed by pancreatin, the degrees of hydrolysis achieved varied between 26.62% (for unheated globulin fraction) and 38.97% (for unheated glutelin), with casein reached 33.73% hydrolysis. After heating, the globulin hydrolysis degree increased to 34.7%, not significantly differing from casein. These results reflect its good susceptibility to hydrolysis by digestive enzymes, and this observation is reinforced with assays with pepsin, trypsin and chymotrypsin tested separately. Globulins, the largest protein fraction, showed promising results in additional assays regarding the amino acid profile, with limitation only for lysine in relation to the FAO standard, and the potential for releasing bioactive peptides after digestion. Although pepsin-digested globulin inhibited only 5% of ACE activity under the conditions tested, after 24h with the addition of pancreatin, the inhibition was 100%. Antioxidant activity (DPPH assay) also indicated very similar results, when hydrolysis with pepsin was inefficient in releasing antioxidant peptides, while hydrolysis by pancreatin led to 35 times greater results.

Antarctic fungus proteases generate bioactive peptides from caseinate.

The extracellular serine protease produced by Acremonium sp. L1-4B isolated from the Antarctic continent, was purified and used for the proteolysis of bovine and caprine sodium caseinate. Protein hydrolysates were evaluated in vitro to determine their antioxidant and antihypertensive potential, and later characterized by mass spectrometry. Bovine and caprine hydrolysates produced over 24 h showed a higher content of copper chelation (25.8 and 31.2% respectively), also at this time the ABTS+• scavenging was 65.2% (bovine sample) and 67.5% (caprine sample), and bovine caseinate hydrolysate (8 h) exhibited higher iron chelation capacity (43.1%). Statistically (p < 0.05), caprine caseinate hydrolysates showed relatively higher antioxidant potential in this study. All hydrolysates showed antihypertensive potential; however peptides released from caprine caseinate after 8 h of hydrolysis were able to inhibit 75% of angiotensin-converting enzyme (ACE) activity. Nano-ESI-Q-TOF-MS/MS analysis prospected a total of 23 different peptide sequences in the bovine hydrolysate fraction, originated from the αS1- and ß-casein chain, whilst in caprine hydrolysate, 31 sequences were detected, all from ß-casein. The low molecular weight bovine and caprine hydrolysates obtained in this research have the potential to act in the prevention of disorders caused by oxidative reactions and in the regulation of blood pressure. These findings support the development of new functional food and nutraceutical formulations.

Changes in digestibility of proteins from chickpeas (Cicer arietinum L.) germinated in presence of selenium and antioxidant capacity of hydrolysates.

Germination in the presence of selenium (Se) is an alternative to increase the healthy properties of seeds. This study aimed to compare the Se accumulation in different protein fractions from germinated chickpea (Cicer arietinum L.) and the effect on digestibility and cellular antioxidant activity (CAA) of protein hydrolysates. Chickpeas were germinated during four days after soaking with sodium selenite (0, 1, or 2 mg/100 g seeds). Total protein (TP) and glutelin (Glu), albumin (Alb) and globulin (Glo) fractions were digested and ultrafiltrated through a 10 kDa membrane. Se accumulated in the order of Glu > Alb > Glo. Ultrafiltrated Glu hydrolysate of four days germinated chickpeas treated with 2 mg Na2SeO3/100 g increased CAA (51.47%), demonstrating the potential health benefits of selenization. The intensity of vicilin bands (34-37 kDa) increased from the second to the fourth day compared with the control samples. Glo digestibility was higher in selenized chickpea sprouts.

Potential antioxidant peptides produced from whey hydrolysis with an immobilized aspartic protease from Salpichroa origanifolia fruits.

An aspartic protease from Salpichroa origanifolia fruits was successfully immobilized onto an activated support of glutaraldehyde agarose. The immobilized enzyme presented higher thermal stability than the free enzyme from 40°C to 50°C and high reusability, retaining 54% of the initial activity after ten cycles of the process. Whey protein concentrates (WPC) were hydrolyzed with both free and immobilized enzyme, reaching a similar degree of hydrolysis of approximately 6-8% after 20h. In addition, the immobilized derivate hydrolyzed α-lactalbumin protein with a higher affinity than ß-lactoglobulin. The hydrolysate was ultra-filtrated, and the fractions were evaluated for antioxidant activities with the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity method. The fraction containing peptides with a molecular mass below 3kDa demonstrated a strong radical quenching effect (IC50: 0.48mg/ml). These results suggest that hydrolyzed WPC could be considered as a promising source of natural food antioxidants for the development of functional food.