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Alcohol-induced liver injury has become a leading risk for human health, however, effective strategies for the prevention or treatment are still lacking. Hence, the present study explored the potential of Musculus senhousei as a source of hepatoprotective peptides against alcoholic liver injury using in vitro, in vivo and in silico methods. Results indicated that Musculus senhousei peptides (MSP, extracted by simulated gastrointestinal digestion of cooked mussel) exhibited notable antioxidant (ABTS and DPPH assays) and alcohol dehydrogenase (ADH) stabilizing activity in vitro. The ingestion of MSP markedly alleviated alcohol-induced liver injury in mice, as indicated by the decrease of serum transaminases (AST and ALT). In line with in vitro assays, significantly increased hepatic ADH activity and activated antioxidative defense system (GSH, SOD, GSH-Px and CAT) were observed, whereas the oxidative stress (MDA) was decreased. Peptidomic analysis revealed over 6000 peptides with favorable amino acid compositions, and a total of 20 potentially novel peptides with bioactivity and bioavailability were excavated among 746 of the most influential peptides using an in silico strategy. Peptides (i.e. WLPMKL, WLWLPA, RLC and RCL) were further synthesized and validated in vitro to be bioactive. These findings suggest that Musculus senhousei can be an ideal source of bioactive peptides for the prevention of alcoholic liver injury., This work was supported by the Key-Area Research and Development Program of Guangdong Province (2022B0202030001 and 2021B0707060001), the National Natural Science Foundation of China (No. 32101896) and Self-innovation Research Funding Project of Hanjiang Laboratory (HJL202010B002).

Bile acids are involved in the modulation of various metabolic processes facilitating the biliary excretion of endogenous and exogenous cholesterol. The objective of this study was to determine the glycocholic acid binding capacity (BC) of chicken blood hydrolysates using an optimized RP-HPLC methodology. Samples were hydrolysed using a combination of five different enzymes. Alcalase and Protamex hydrolysates presented the highest BC, with mean values of 20.09% and 20.61%, respectively. Subsequently, both hydrolysates were ultrafiltered to obtain fractions >10 kDa, between 10 and 3 kDa, and 10 kDa. Finally, the protein fragments (MW > 10 kDa) potentially responsible for BC were identified by LC-MS/MS. The results confirmed the relation of BC with the molecular weight of the peptides generated, suggesting that certain protein fragments generated from chicken blood could contribute to a positive impact on health by interfering with cholesterol metabolism., Grant PID2020-119684RB-I00 funded by MCIN/AEI/10.13039/501100011033 is acknowledged. Grant GRISOLIAP/2020/021 de la Consellería de Innovació, Universitats, Ciència i Societat Digital de la Generalitat Valenciana (GCA) is also acknowledged. The proteomic analysis was performed in the proteomics facility of SCSIE University of Valencia belonging to Proteored.

The ultrasound-induced impacts on the peptide characteristics and taste of unsmoked bacon have been evaluated through the use of peptidomics and bioinformatics approaches. Furthermore, the effect of such ultrasound-induced changes on the main endogenous proteases responsible for peptide generation was also investigated. In fact, the activity of main endogenous proteases was significantly increased after ultrasonic treatment during the processing of unsmoked bacon, and contributed to an increased number and an enhanced LFQ intensity of peptides. Besides, such increased amount of peptides and LFQ intensity with up to 500 W ultrasonic treatment were beneficial for the taste improvement of the final products as shown by taste prediction analysis. Nevertheless, an excessive ultrasonic power like 750 W hindered protein hydrolysis and further exerted a negative effect on peptide generation. Therefore, ultrasound under controlled conditions could be considered as a promising way to improve the taste of unsmoked bacon., This work was financially supported by the earmarked fund for China Agriculture Research System (CARS-35) and the Kekedala City of the 4th Division Science and Technology Plan (2022GY03).

High cholesterolemia is a key risk factor for the development of cardiovascular diseases, which are the main cause of mortality in developed countries. Most therapies are focused on the modulation of its biosynthesis through 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoAR) inhibitors. In this sense, food-derived bioactive peptides might act as promising health alternatives through their ability to interact with crucial enzymes involved in metabolic pathways, avoiding the adverse effects of synthetic drugs. Dry-cured ham has been widely described as an important source of naturally-generated bioactive peptides exerting ACEI-inhibitory activity, antioxidant activity, and anti-inflammatory activity between others. Based on these findings, the aim of this work was to assess, for the first time, the in vitro inhibitory activity of HMG-CoAR exerted by dipeptides generated during the manufacturing of dry-cured ham, previously described with relevant roles on other bioactivities.The in vitro inhibitory activity of the dipeptides was assessed by measuring the substrate consumption rate of the 3-hydroxy-3-methylglutaryl CoA reductase in their presence, with the following pertinent calculations.Further research was carried out to estimate the possible interactions of the most bioactive dipeptides with the enzyme by performing in silico analysis consisting of molecular docking approaches.Main findings showed DA, DD, EE, ES, and LL dipeptides as main HMG-CoAR inhibitors. Additionally, computational analysis indicated statin-like interactions of the dipeptides with HMG-CoAR.This study reveals, for the first time, the hypocholesterolemic potential of dry-cured ham-derived dipeptides and, at the same time, converges in the same vein as many reports that experimentally argue the cardiovascular benefits of dry-cured ham consumption due to its bioactive peptide content.

Background: Evidence suggests that bioactive peptides reduce hypertension and affect certain metabolic pathways. Methods: Fifty-four volunteers with stage 1 prehypertension and/or hypercholesterolemia and/or basal glucose >100 mg/dL were recruited and randomized to pork dry-cured ham (n = 35) or cooked ham (placebo group; n = 19) for 28 days. After a wash-out period, meat products were changed for 28 additional days. Bioactive peptides composition and enzyme inhibitory activities of both products were characterized. Treatment comparisons for the main effects were made using a two (treatment) × two (times) repeated measures minus the effect of cooked ham (placebo). Results: 24 h mean systolic and diastolic pressures decreased up to 2.4 mmHg in the dry-cured ham period (treatment effect, p = 0.0382 y p = 0.0233, respectively) as well as the number of systolic pressure measures > 135 mmHg (treatment effect, p = 0.0070). Total cholesterol levels also decreased significantly after dry-cured ham intake (p = 0.049). No significant differences were observed between the two treatments for basal glucose, HOMA-IR index and insulin levels (p > 0.05). However, a significant rise of ghrelin levels was observed (treatment effect, p = 0.0350), while leptin plasma values slightly decreased (treatment effect, p = 0.0628). Conclusions: This study suggested the beneficial effects of regular dry-cured ham consumption on the improvement of systolic/diastolic blood pressures and facilitated the maintenance of metabolic pathways, which may be beneficial in the primary prevention of cardiovascular disease., This research was supported by grant RTC-2017-6500-1 funded by MCIN/AEI//10.13039/501100011033 and by ERDF a way of making Europe, and “Fundación Séneca de la Región de Murcia” through the project “Jovenes Líderes en Investigación“ (20646/JLI/18). AVS belongs to the “Programa de Doctorado en Ciencias de la Salud. Universidad Católica de Murcia (UCAM)”.

Alcohol hydydrogenase (ADH) is a key enzyme that influences alcohol metabolism in vivo. Our previous study showed that chicken-derived tripeptide KPC could stabilize ADH in a dose-dependant manner. In this study, possible mechanism underlying how KPC could stabilize ADH was further investigated. Fluorescence quenching data showed that KPC induced a dynamic fluorescence quenching with a quenching rate constant value of 1.074 × 1010 M−1s−1, indicating a conformational change. Circular dichroism further suggested a possible transformation from α-helix (14.03%–13.90%) to random coil (31.98–33.04%) in ADH structure as affected by KPC (5 mmol/L). Molecular docking analysis predicted that KPC may bind to ADH through hydrophobic interaction and hydrogen bonds. Validation of ADH stabilizing activity of fragment peptides and amino acids from KPC implied the vital role of Cys residue to the bio-activity of KPC. These results indicated that KPC may stabilize ADH through peptide-enzyme interactions, and protect ADH against oxidative modification. Besides, a favorable absorption, distribution, metabolism, excretion, and toxicity (ADMET) profile of KPC was obtained using in silico assays. Overall, this study provided a comprehensive understanding on the interaction mechanism between KPC and ADH, and illustrated the potential applicability of KPC as drug or functional food ingredient., This work was supported by the National Natural Science Foundation of China (No. 32101896 and No. 31871746), Key-Area Research and Development Program of Guangdong Province (2021B0707060001), the Self-innovation Research Funding Project of Hanjiang Laboratory (HJL202010B002, HJL202101B001, HJL202101B004,and HJL202101B005), and the Chaozhou Science and Technology Project (2020PT01).

Chicken hydrolysates (CHs) have been reported to protect mice against alcoholic liver injury possibly through oxidative stress reduction. In this study, the antioxidant activity of CHs was studied. Results showed that CHs exhibited significant antioxidant activity (around 600 and 400 μM TEAC/g in ORAC and ABTS assay, respectively) and could resist simulated gastrointestinal digestion. A total of 22 peptides were identified after antioxidant activity-oriented isolation using size-exclusion chromatography and high-performance liquid chromatography. Further in silico analysis and the validation of antioxidant activity revealed that novel peptides (RWGG and YYCQ) exhibited strong antioxidant activity. The most active peptide YYCQ displayed a TEAC value of 3.54 and 4.28 μM TEAC/μM in ORAC and ABTS assay, respectively. These peptides could contribute to reduce oxidative stress and protect against alcohol-induced liver injury. However, further studies understanding the bioactivity of such peptides in vivo are necessary before further applying them as functional food ingredient., This work was supported by the National Natural Science Foundation of China (No. 32101896 and No. 31871746), Key-Area Research and Development Program of Guangdong Province (2021B0707060001), and Self-innovation Research Funding Project of Hanjiang Laboratory (HJL202010B002). Grant AGL2017-89381-R funded by MCIN/ AEI /10.13039/501100011033 by “ERDF A way of making Europe” is acknowledged and grant RYC-2016-19635 (L.M.) funded by MCIN/AEI /10.13039/501100011033 and “ESF Investing in your future” is also acknowledged.

Sweet dipeptide Ala-Ala (AA) could be one of the most abundant dipeptides naturally generated in meat products due to the high occurrence of its sequence all along endogenous proteins, which means it could play a key role in dry-cured ham flavour development and bioactivity. In this study, a mass spectrometry in tandem methodology was optimised to quantitate the dipeptide AA in dry-cured ham manufactured under different processing conditions. Dipeptide values reached 230 µg/g after 12-month of ripening, but statistically insignificant differences were observed with salt reduction. Regarding its antihypertensive activity, an IC50 value of 110.824 µM was determined from the in vitro inhibition of angiotensin-I converting (ACE-I) enzyme, and the oral administration of 1 mg of AA per kg of body weight showed a significant reduction of the systolic blood pressure (SBP) in spontaneous hypertensive rats (SHR). This study elucidates the importance of AA dipeptide and its potential role in cardiovascular health after dry-cured ham consumption., Grant AGL2017-89381-R funded by MCIN/ AEI /https://doi.org/10.13039/501100011033 by “ERDF A way of making Europe” is acknowledged. Grant PRE2018-083367 (A.H.) funded by MCIN/AEI /https://doi.org/10.13039/501100011033 and “ESF Investing in your future” and grant RYC-2016–19635 (L.M.) funded by MCIN/AEI /https://doi.org/10.13039/501100011033 and “ESF Investing in your future” are also acknowledged. Arihara funding KAKENHI (21H02348) from Japan Society for the Promotion of Science is also acknowledged.

We thank Dr. López-Moreno for the comment [...]

Large amounts of meat by- and co-products are generated during slaughtering and meat processing, and require rational management of these products for an ecological disposal. Efficient solutions are very important for sustainability and innovative developments create high added-value from meat by-products with the least environmental impact, handling and disposal costs, in its transition to bioeconomy. Some proteins have relevant technological uses for gelation, foaming and emulsification while protein hydrolyzates may contribute to a better digestibility and palatability. Protein hydrolysis generate added-value products such as bioactive peptides with relevant physiological effects of interest for applications in the food, pet food, pharmaceutical and cosmetics industry. Inedible fats are increasingly used as raw material for the generation of biodiesel. Other applications are focused on the development of new biodegradable plastics that can constitute an alternative to petroleum-based plastics. This manuscript presents the latest developments for adding value to meat by- and co-products and discusses opportunities for making meat production and processing more sustainable., Grant from AGL2017-89831-R from the Spanish Ministry of Economy, Industry and Competitivity and FEDER funds.

A systematic insight into the structure-function properties of small bioactive peptides is of great importance. Herein, peptidomics and computational methodology were adopted to investigate the stabilization patterns and building blocks of antioxidant peptides resulting from proteolysis during the ripening of Spanish dry-cured ham (9-24 months of processing). The results showed that native peptides underwent manufacture-induced steric/redox stress, while homogeneous/heterogeneous p-π/π-π interaction significantly improved the ABTS+ inhibition activity of hydrophobic peptides. However, for more hydrophilic peptides, the intrinsic π-interaction system (i.e., cation-π and π-π packing) substantially interfered with ABTS+/DPPH scavenging events when compared to the aromatic residues. Semi-quantitative peptidomics and molecular simulation/docking revealed that VFSSQGQSELILLQK and LCPSPDGLYL are two potential antioxidant peptides at the late ripening-drying. They had distinctive self-folding destinies following solvation owing to varying charged/hydrophobic properties of termini and hydrogen atom donor, allowing different flexibility of backbone and interactive surface towards free radicals ex situ followed by electron/proton transfer., Grant AGL2017-89381-R funded by MCIN/ AEI /10.13039/501100011033 and “ERDF A way of making Europe” and grant RYC-2016-19635 (L.M.) funded by MCIN/AEI /10.13039/501100011033 and “ESF Investing in your future” are also acknowledged. C.L. appreciated the financial support from China Scholarship Council (CSC), China through the State Scholarship Fund for Overseas Study (No. 201806760050).

Background: Dry-cured ham is one of the most valued food products by Mediterranean consumers. In this sense, the appropriate development of its different production stages is essential to ensure the quality requirements. For this reason, non-invasive technologies have gained popularity and have been reported as useful not only to ensure the food safety of different products, but also to monitor fundamental stages in the production process, such as the salting stage, to analyze the content of different compounds without sample losses, and to correct possible defects in the final product. Scope and approach: This work has been focused on summarizing the studies that describe and have successfully applied these techniques, as well as on mentioning other technologies with potential use in dry-cured ham manufacture which have not been studied enough. Finally, the potential next steps to improve and optimize the process, as well as the suitability of creating new products with added value based on the new quality standards, have also been evaluated. Key findings and conclusions: Innovative non-invasive technologies such as high pressure (HP), ultrasound (US), pulsed electric fields (PEF), microwaves, irradiation, etc. can be used as promising tools to effectively control salting and curing stages as well as for checking defects of the final product and/or ensuring food safety. HP and US are useful tools for the determination of salt and fat content, and for monitoring the salting process. Moreover, HP enhances salty taste perception, which makes it a useful tool to reduce salt addition. Both, HP and US, can correct texture defects. In addition, NIRS allows predicting the state of the meat to remove those pieces that could result in defective products. Moreover, RAMAN or MRI are able to detect anomalous textures at the end of the process. Microwaves could be useful for the online estimation of salt, water and fat contents easily with portable equipment. Finally, data mining, that allows to make predictions based on an immense data file, is the most promising discovery in recent years for detecting defects or classifying products according to sensory attributes., This research was supported by Grant RTA 2013-00030-CO3-03 from INIA (Spain). Acknowledgements to INIA for granting Cristina Pérez Santaescolástica with a predoctoral scholarship (grant number CPD2015-0212). José M. Lorenzo is member of the MARCARNE network, funded by CYTED (ref. 116RT0503).

Foods and their industry by-products constitute very good sources of bioactive peptides, which can be naturally generated during processing but are also extensively produced through enzymatic hydrolysis, microbial fermentation, and even during gastrointestinal digestion in the human body [...]

Recently, the use of bioactive compounds in improving human health has received more attention. The aim of the present study was to hydrolyze orange seed proteins using pepsin enzyme to obtain bioactive peptides as well as to study the stability of such activity after simulated gastrointestinal digestion conditions. The method was optimized using different enzyme concentrations from 1% to 3%, hydrolysis times between 2 and 5 h, and an optimal temperature of 33 °C. Biological activities including α-glucosidase inhibition, α-amylase inhibition, Angiotensin I-Converting Enzyme (ACEI) inhibition, ferric reducing antioxidant power, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity were evaluated. According to the results, a significant higher value of the biological activity (p < 0.05) was observed using an enzyme ratio of 0.03 E/S and hydrolysis time of 3.5 h. After size-exclusion chromatography separation, fractions 45–49 and 50–54 showed the highest biological roles such as antioxidant, ACEI inhibitory, and hypoglycemic. Fractions with the highest biological activity were purified using RP-HPLC and analyzed using nano-liquid chromatography and mass spectrometry. The results obtained after simulated gastrointestinal digestion indicated that peptide fractions obtained after chromatographic separation significantly maintain their activity., This research was funded by grant AGL2017-89381-R, and FEDER funds from the Spanish Ministry of Economy, Industry, and Competitiveness and the Ramón y Cajal postdoctoral contract by L.M. are acknowledged.

Meat and meat products constitute an important source of nutrients and play vital roles for growth, maintenance and repair of the body. In addition to the high quality of proteins, meat is also regarded as a major resource to produce bioactive peptides. Meat processing industry also produces by-products such as bones, blood and viscera, which could be further used for the production of bioactive compounds. In the physiological analysis, meat bioactive peptides have been reported to exert antioxidant, anti-hypertensive, anti-inflammatory, anti-microbial and antitumoral activities, which endow nutritional and functional value of meat. With the objective to exert the functional effect, the bioavailability should also be considered due to the degradation by digestion enzymes and the absorption process in intestinal mucosa. In this chapter, the general source, the enzymatic hydrolysis, the physiological effects as well as the bioavailability of bioactive peptides in meat are discussed.

Microenvironmental factors may influence the antioxidant efficacy of food-derived peptides. This study evaluated the in vitro antioxidant properties of peptides released during the ripening (9 to 24 months of processing) of Spanish dry-cured ham (Biceps femoris muscle) assisted by spectral-chromatographic methodologies. Results indicated that 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical-cation (ABTS●+) quenching capacity of peptide extracts significantly increased (P < 0.05) until 24 months whereas peroxyl radical (ROO●) scavenging activity increased slowly and remained with non-significant change (P > 0.05) between 15 and 24 months. However, both ferric reducing antioxidant power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl radical (DPPH●) scavenging ability significantly decreased (P < 0.05) at 24 months. Additionally, morphological traits of peptide extracts suggested that a prolonged ripening enabled the formation/reconstruction of intra-/inter-molecular interactions in dispersion medium. Meta-analysis of chromatographic and spectral fingerprinting proved that the non-conjugated/π-conjugated oligomers mediated by aromatic moieties probably differentiated DPPH● and ABTS●+ antioxidant performance of peptides, showing a potentially altered solvent polarization process., Grant AGL2017-89381-R and FEDER funds from the Spanish Ministry of Economy, Industry and Competitiveness, and Ramón y Cajal postdoctoral contract by L.M. are acknowledged. C.L. appreciated the financial support from China Scholarship Council (CSC) through the State Scholarship Fund for Overseas Study (No. 201806760050).

© 2020 by the authors., In this study, orange seed proteins were hydrolyzed by Alcalase enzyme at different enzyme concentrations 1–3% (v/w) and hydrolysis times (2–5 h), to obtain bioactive peptides showing antioxidant, Angiotensin-converting enzyme (ACE) -inhibitory, and hypoglycemic activities. The highest biological activities (p < 0.05) were achieved by using a hydrolysis time of 5 h and an enzyme concentration of 2%. Orange seed protein hydrolysate (OSPH) was prepared under these conditions, and peptides were isolated and purified by using size-exclusion chromatography and high-performance liquid chromatography, respectively. The fractions that showed the highest biological activities were analyzed by mass spectrometry in tandem, and a total of 63 peptide sequences were found. Moreover, the effect of simulated gastrointestinal digestion on the bioactivity of the fractions was studied, and the novel peptide sequences generated were also identified. Overall, despite there being some differences in the profile of peptide sequences obtained, the main results showed non-significant differences in the analyzed bioactivities after simulated gastrointestinal digestion., Grant AGL2017-89381-R and FEDER funds from the Spanish Ministry of Economy, Industry and Competitiveness, and Ramón y Cajal postdoctoral contract by L.M. are acknowledged. This work was also supported by Intramural project from CSIC number 201870E006.

The present study aimed to investigate the effect of cooking and simulated gastrointestinal digestion on the antioxidant and alcohol dehydrogenase (ADH) stabilization activity of peptides extracted from chicken breast muscle. Results showed that cooking would not affect peptide bioactivity, whereas further digestion using gastrointestinal enzymes could lead to significant changes, producing an increase in ORAC (112.5 to 682.0 uM TE/g) and ABTS radical scavenging activities (164.0 to 848.9 uM TE/g), whereas a decrease in DPPH radical scavenging (from 36.1% to 4.4%), ferric-reducing power (OD 700 from 0.50 to 0.15) and ADH stabilization activities (from 44.1% to 20.5%) was observed. The peptidomic analysis resulted in the identification and relative quantitation of 777 peptides from 76 different parent proteins and evidenced that peptides derived from titin and collagen were mainly responsible for the differences detected in the peptide profile. The decrease of DPPH radical scavenging, ferric reducing power, and ADH stabilization activity may result from the release of inactive peptides containing oxidized residues, mainly from collagen, leading to the loss of efficacy of active sequences. The results confirmed the importance of collagen derived peptides on the antioxidant and ADH stabilization activity observed in chicken breast as well as the negative impact of oxidation on the bioactivity of generated peptides after simulated gastrointestinal digestion. Nevertheless, further work would be needed to confirm the peptide sequences responsible for the observed bioactivity., This work was supported by Grant AGL2017-89381-R and FEDER funds from the Spanish Ministry of Economy, Industry and Competitiveness, and the National Natural Science Foundation of China (No. 31701539 and No.31871746), and the Natural Science Foundation of Guangdong Province (No. 2017A030313127). Ramón y Cajal postdoctoral contract by LM and the China Scholarship Council (CSC) research program for C. X. are also acknowledged.

Dry-cured hams are a good source of bioactive peptides, whose stability to gastrointestinal (GI) digestion determines their bioaccessibility and thus their bioavailability to exert beneficial effects. The aim of this study was to evaluate the effect of in vitro GI digestion of dry-cured hams on the peptide profiles and their antioxidant activity. Results showed that the antioxidant activity decreased in the digested samples when measured by DPPH radical scavenging activity and ferric-reducing antioxidant power methods, but increased when assayed by ABTS radical scavenging assay. Chromatographic and mass spectrometry analysis revealed changes in the peptide profiles and evidenced the degradation of proteins, mainly titin, by the digestive enzymes. The study of the peptide profiles before and after digestion of dry-cured hams would help to extend the knowledge about the influence of peptide characteristics on their antioxidant activity and resistance to hydrolysis by digestive enzymes, allowing a better understanding of the bioaccessibility of the bioactive peptides generated in dry-cured hams., Grant AGL2017-89381-R and FEDER funds from the Spanish Ministry of Economy, Industry and Competitiveness are acknowledged.

Nitrite and nitrate have been traditionally used for the preservation of meat products because of the effective antimicrobial action of nitrite against Clostridium botulinum, the outgrowth of its spores as well as other bacteria. However, the use of nitrite and nitrate has been questioned in last half century due to the possible generation of N-nitrosamines through reaction of nitrite with secondary amines. Nitrite replacement strategies began in the 70s addressing these issues and instigated searches for natural alternatives to nitrate and nitrite, or for natural sources of nitrite and nitrate such as vegetable extracts. These alternatives have been considered by producers and consumers as an attractive practice even though they may also have some risks. This manuscript reviews and discusses the chemistry, safety, and regulatory considerations in the use of nitrite and nitrate from natural origin for the preservation of meat products.

Peptidyl post-translational modifications (PTMs) could influence the final quality of processed meat. In this study, the peptide oxidative phenomena in Spanish dry-cured ham (Biceps femoris muscle) was evaluated at different ripening times (9, 12, 15, 18 and 24 months of processing) evidencing interactions amongst the lipid and protein oxidation, major peptidyl PTMs and the release of free amino acids (FAAs). Results showed that 12 months of processing enabled the most abundant protein-bound carbonyls, while TBARS value was significantly favored (p < 0.001) by ripening. However, FAAs were still intensively generated during overall ripening. Peptidomics and chemometrics further revealed that proteolysis mostly hampered the oxidized peptides rather than the deamidated ones during ripening. Myosin light chain (MYL1 and MYL3) showed high oxidative susceptibility owing to peptidyl methionine and proline oxidation as well as acetaldehyde adduct formation on lysine or histidine residues., Grant AGL2017-89381-R and FEDER funds from the Spanish Ministry of Economy, Industry and Competitiveness, and Ramón y Cajal postdoctoral contract by L.M. are acknowledged. C.L. appreciated the financial support from China Scholarship Council (CSC) through the State Scholarship Fund for Overseas Study (No. 201806760050). The peptidomic analysis was performed in the proteomics facility of SCSIE University of Valencia. This proteomics laboratory is a member of Proteored, PRB3 and is supported by grant PT17/0019, of the PE I + D + i 2013-2016, funded by ISCIII and ERDF

Peptides and free amino acids are naturally generated in dry-cured ham as a consequence of proteolysis phenomenon exerted by muscle peptidases. The generation of bioactive peptides in different types of dry-cured ham produced in Spain, Italy and China is reviewed in this manuscript. Major muscle proteins are extensively hydrolysed firstly by endogenous endo-peptidases followed by the successive action of exo-peptidases, mainly, tri- and di-peptidylpeptidases, aminopeptidases and carboxypeptidases. Such proteolysis is very intense and consists of the generation of large amounts of free amino acids and a good number of peptides with different sequences and lengths, some of them exerting relevant bioactivities like angiotensin converting enzyme inhibitory activity, antioxidant activity, di-peptidylpeptidase IV inhibitory activity among other and in vivo antihypertensive, hypoglycemic or anti-inflammatory activity. This manuscript reviews the recent findings showing that dry-cured ham constitutes a good source of natural bioactive peptides that have potential benefit for human health., The research leading to these results received funding from Grant AGL2017-89831-R from the Spanish Ministry of Economy, Industry and Competitivity and FEDER funds The Ramón y Cajal postdoctoral contract to LM is also acknowledged.

There is a wide range of chemicals that may be present in meat as residues either during processing or during cooking and household management. Analytical technologies are constantly being developed and improved, allowing the correct detection and determination of such compounds and contributing to a better meat safety. Some toxic compounds that may be present in meat or meat products are N-nitrosamines as a consequence of nitrite use, polycyclic aromatic hydrocarbons generated from smoking, oxidation of lipids and proteins, and, finally, other substances resulting from the raw materials used in animal production like veterinary drug residues or environmental contaminants. All these are harmful compounds and the available detection analyses are described in this chapter.

Small peptides such as dipeptides contribute to a great extent to the characteristic taste of dry-cured hams. In this study, hydrophilic interaction liquid chromatography (HILIC) combined to tandem mass spectrometry was used to separate, identify, and quantify seven dipeptides in dry-cured hams sampled at different processing times (6, 12, 18, and 24 months). Results showed an increased concentration of dipeptides DA, DG, EE, ES, and EV with the length of processing, obtaining values up to 23 μg/g of dry-cured ham, which suggests an intense action of muscle enzymes dipeptidyl peptidases during the process. The dipeptide VG significantly decreased from 7 to 4 μg/g of dry-cured ham as the processing increased from 6 to 24 months, whereas the dipeptide PA showed low values between 380 and 550 ng/g of dry-cured ham at all the sampling times. Additionally, in silico analyses reported the sensory characteristics of the studied dipeptides, mostly giving bitter and umami taste, and predicted their allergenicity, toxicity, and physicochemical properties. These results could be useful for further studies related to the pleasant taste of dry-cured hams., Grant AGL2017-89381-R and FEDER funds from the Spanish Ministry of Economy, Industry and Competitiveness are acknowledged.

A wide variety of dry-fermented sausages are produced in European countries, where are considered valued traditional products. An intense proteolysis takes place during the processing of dry-fermented sausages due to the combined action of muscle and microbial peptidases, generating large amounts of peptides and free amino acids. These compounds participate in the development of the characteristic flavor of dry-fermented products, but some peptides can also exert certain bioactivities such as antioxidant and ACE inhibitory activities. This study has evaluated the changes in peptide profile and amino acid contents of three European dry-fermented sausages produced in Spain, Italy and Belgium, proving the intense degradation of proteins, mainly myofibrillar, and the generation of high amounts of different size peptides and free amino acids. The changes observed between the profiles of European sausages could be due to differences in product formulation, processing conditions and starter cultures used, which influence the activity of enzymes, both from muscle and bacterial origin. On the other hand, the bioactivity profile of each type of dry-fermented sausage was evaluated through the measurement of the ACE inhibitory and antioxidant activities in water-soluble peptide extracts fractionated by size-exclusion chromatography. Spanish and Belgian dry-fermented sausages showed values of ACE inhibition around 85%, whereas Belgian samples presented the highest DPPH radical-scavenging activity and ferric reducing power capacity. These results evidence the potential of Spanish, Italian and Belgian dry-fermented sausages as natural sources of bioactive peptides, giving an added-value to these traditional products., Authors acknowledge European Union FP7 under Grant Agreement 312090 (BACCHUS). This publication reflects only the authors views and the Community is not liable for any use made of the information contained therein. Grant AGL2014-57367-R and FEDER funds from the Spanish Ministry of Economy, Industry and Competitiveness are acknowledged. Juan de la Cierva de Incorporación postdoctoral contract to LM is also acknowledged. The proteomic analysis was performed in the proteomics facility of SCSIE University of Valencia that belongs to ProteoRed, PRB2-ISCIII, (IPT13/0001 - ISCIII-SGEFI/FEDER).

Oxidative modifications of proteins and peptides can negatively affect nutritional, sensory and quality characteristics of dry-cured hams. The aim of this study was to use a peptidomics strategy for the identification and relative quantification of oxidised peptides in two different muscles, the external Semimembranosus (SM) and the internal Biceps femoris (BF), from dry-cured hams of 12 months of curing. The analysis by nanoliquid chromatography-tandem mass spectrometry identified peptides showing oxidation at different amino acid residues such as methionine, cysteine, histidine, tryptophan, and phenylalanine. The highest percentages of oxidised peptides identified in SM and BF muscles were from pyruvate kinase protein (16.7 and 28.6%, respectively). The relative quantitation of oxidised peptides using mass spectrometry in tandem label-free methodology evidenced significant differences between muscles, with myosin and troponin fragments as main responsible peptides for the clustering of data. The existing differences between BF and SM peptides were confirmed using MALDI imaging mass spectrometry technique. The obtained results suggest that differences in physicochemical characteristics such as water and salt contents between SM and BF muscles could affect the quantity of peptide oxidation. This peptidomics approach has allowed to analyse differences in peptide oxidation between dry-cured ham muscles as well as to improve the knowledge about the oxidative processes that occur in dry-cured hams., Grant AGL2014-57367-R from MINECO and FEDER funds are acknowledged. Ramón y Cajal 2016 postdoctoral contract to LM is also acknowledged. The proteomic analysis was performed in the proteomics facility of SCSIE University of Valencia that belongs to ProteoRed, PRB2-ISCIII, (IPT13/0001 - ISCIII-SGEFI/FEDER).

This study aims to examine the effects of non-hydrolyzed octopus (Octopus vulgaris) muscle proteins (NHOPs) and their hydrolysates (OPHs) on alloxan induced diabetes in Wistar rats (AIDR). Animals were allocated into seven groups of six rats each: control group (C), diabetic group (D) and diabetic rats treated with acarbose (D + Acar), non-hydrolyzed octopus proteins (D + NHOPs) and octopus proteins hydrolysates (D + OPHs) groups. The diabetic rats presented a significant increase in glycemic status such as α-amylase activity (in plasma, pancreas and intestine), hepatic glycogen, blood glucose and glycated hemoglobin (HbA1c) levels, as well as a significant decrease in the levels of plasma insulin and total hemoglobin compared to control group. In addition, plasma and liver contents in total cholesterol, triglycerides and LDL-cholesterol significantly increased in AIDR compared to control group. However, the daily administration of OPHs for 30 days improved the glucose tolerance test, the glycemic status of diabetic rats and corrected the lipid profiles. Further, a significant increase in the activities of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and gamma-glutamyl transpeptidase as well as in the level of plasma bilirubin on diabetic status was observed, indicating considerable hepatocellular injury. OPHs treatment was found to attenuate the increased activities of the plasma enzymes produced by diabetes and caused a subsequent recovery towards normalization compared to the control group. By contrast, the NHOPs treatment was found to increase the glucose metabolic disorders in AIDR. These beneficial effects of OPHs were confirmed by histological findings in the hepatic and pancreatic tissues of diabetic treated rats. Indeed, they avoid lipid accumulation in the hepatocytes and protect the pancreatic β-cells from degeneration. Our results thus suggest that OPHs may be helpful in the preventing from diabetic complications by reversing hepatotoxicity., This work was funded by the Ministry of Higher Education and Scientific Research, Tunisia. The authors wish to express their sincere gratitude to Pr Najiba ZEGHAL, professor in the Sfax Faculty of sciences for her comments that greatly improved the manuscript.

Food-derived bioactive peptides are promising components for the prevention and treatment of cardiovascular diseases including hypertension. Recently, there has been an increased knowledge about the variety of complex and interrelated mechanisms for blood pressure regulation as well as the potential targets for peptides to exert their antihypertensive effects. Empiric and bioinformatics studies have provided large amounts of data regarding characteristics, structure–activity relationships and bioavailability of antihypertensive peptides through the use of in vitro assays, cell cultures, and animal studies. However, the scarce number of robust clinical trials to prove their efficacy in humans is the main reason of the limited use of antihypertensive peptides as functional foods for promoting health. Further research is needed to overcome the challenges for the application of food-derived antihypertensive peptides as functional ingredients with health benefits in the human body., Grant AGL2014-57367-R and FEDER funds from the Spanish Ministry of Economy, Industry and Competitiveness are acknowledged. Juan de la Cierva de Incorporación postdoctoral contract to LM is also acknowledged.

Smooth-hound viscera hydrolysates (SHVHs) were prepared by treatment with Neutrase (SHVH-N) and Purafect (SHVH-P). Hydrolysates were then separated according to their molecular weight, using the ultra-filtration membrane system, into 5 fractions (≥50, 50-5, 5-3, 3-1 and ≤1kDa). Fractions showed different amino acid compositions and angiotensin I-converting enzyme (ACE) inhibitory potentials. The SHVH-P-FV (≤1kDa) and SHVH-N-FIV (3-1kDa) fractions showed the best ACE-inhibitory activities with IC50 values of 53.31 and 75.05µg/ml, respectively. According to their high ACE-inhibitory potential, FIV and FV were fractionated by RP-HPLC and then analyzed by LC-MS/MS to identify peptide sequences. A systematic peptidomic study resulted in the identification of numerous novel sequences. Furthermore, in silico data, based on the molecular docking simulation, showed that GPAGPRGPAG, AVVPPSDKM, TTMYPGIA, and VKPLPQSG could bind ACE active site with low interaction scores. Indeed, they share hydrogen bonds and Van der Waals and electrostatic interactions with ACE catalytic pockets.

In the current study, gelatin was extracted from black-barred halfbeak ( Hemiramphus far ) skin by successive alkaline and acid treatments and then hydrolyzed with Purafect ® . The black-barred halfbeak gelatin (BG) and its hydrolysate (BGH) were characterized and compared to the commercial bovine gelatin (CG). Samples were evaluated for their antioxidant, antibacterial and angiotensin I-converting enzyme (ACE) inhibitory activities. Results obtained using size exclusion chromatography showed that BG contained lower level of high molecular weight proteins, compared to CG. In addition, the amino acids composition revealed that BG contained lower level of imino acids (Pro+Hpx), compared to CG. These differences reflect the variations observed in the gel strength and gelling and melting temperatures of both skin gelatins. Furthermore, high similarities were observed between CG and BG in terms of their Fourier transform infrared (FTIR) spectra, while their amino acid compositions were quite different. BGH, with a degree of hydrolysis of 12.5%, showed high antioxidant potential that was assessed by the scavenging activity, reducing power, oxygen radical absorbance capacity, β-carotene bleaching protection and lipid per-oxidation inhibition assays. In addition, BGH sample exhibited antibacterial activity against different Gram+ and Gram- bacteria. The ACE-inhibitory activity was also investigated. BGH showed an inhibitory effect of 80.76%, while BG inhibited the ACE only by 36.51% at 1 mg/ml. Thus, black-barred halfbeak gelatin represents a promising source of antioxidant, ACE-inhibitory and antimicrobial peptides that might prevent humans from several diseases.

A nitrate reductase immobilized system with an oxygen electrode has been developed and optimized to determine the nitrate content in dry-cured ham. The obtained amperometric signal was recorded at 5 s in the immobilized nitrate reductase sensor and the reaction rates (slope) were related to the nitrate content. A linear relationship between the reduction nitrate rate by action of nitrate reductase and the nitrate concentration was found within the range 10–70 μM (R2 = 0.9761). The immobilized enzyme showed a high specificity and sensibility and was stable enough to allow the reutilization of the membranes up to eight times without loss of activity. This reduces the cost of the analysis as well as the necessary equipment that is cheap and the short average measurement time for each sample. The analysis of nitrate in dry-cured ham samples with the sensor and by HPLC revealed very good agreement (R2 = 0.971). The use of this sensor may constitute an interesting and valid alternative for the quantification of nitrate in dry-cured ham., Funded by the Spanish Ministry of Economy, Industry and Competitiveness, AGL2014-57367-R and FEDER funds. Felipe Jadán enjoyed a scholarship from Secretaria de Educación Superior, Ciencia, Tecnología e Innovación (SENESCYT), Ecuador.

Ageing is widely used in the meat industry to improve tenderness mainly as a result of the breakdown of muscular proteins through the action of endopeptidases during storage time. In addition, meat contains a large pool of other proteolytic enzymes, mainly exopeptidases, which cut from the outer edges of proteins and polypeptides generating a vast array of peptides and free amino acids. Some of these peptides could potentially exert bioactivities of interest for human health. In this study, ACE-inhibitory and antioxidant activities during meat ageing in chilled-storage at 4 °C for a period of time of 0, 1, 2, 3 and 4 weeks have been determined. Beef loin steak samples were analysed before and after cooking, and gastrointestinal digestion was simulated in order to study its effect on the bioactivity. Control and cooked samples showed an increase in the ACE-inhibitory activity from the third week of chilled storage whereas non-significant differences in the antioxidant activity between control and cooked samples were detected during the studied time. After gastrointestinal digestion of samples, there was a significant increment of the ACE-inhibitory and antioxidant activities in comparison with control and cooked samples at all the studied times. As a main conclusion, cooking does not significantly influence the bioactivity detected whereas the in vitro gastrointestinal digestion produces a significant increase in the ACE-inhibitory and antioxidant activities from the first week, probably due to the intense generation of small peptides as a result of the action of gastrointestinal enzymes.

The in vivo wound-healing potential of gels and films based on cuttlefish skin gelatin (CSG) incorporated with aqueous henna extract (AHE) (at 50 and 500 μg/ml) was studied. As wound healing is related to reactive oxygen species (ROS), this study was first conducted to evaluate the antioxidant activity of AHE by DPPH radical-scavenging, β-carotene-linoleate bleaching, and reducing power assays. The major volatile compounds of AHE were lawsone, dimethyl sulfoxide, dimethyl sulfide, eugenol and maltol. The finding of the study showed that the incorporation of AHE into gelatin gels and films enhanced the antioxidant activity in a dose-dependent manner. In the wound-healing study, topical application of gelatin hydrogels or films enriched by AHE, on the wound site in a rat model, enhanced significantly wound healing activity and helped to prevent from inflammation damage, when compared to the control and CICAFLORA®-treated groups. Further, all preparations were found to improve the antioxidant status of treated rats as was evidenced by increased antioxidant enzymatic activities and reduced malondialdehyde (MDA) content. The increase in the activities of catalase (CAT), superoxide dismutase (SOD) and gluthatione peroxydase (GPx) in wound tissues revealed that some bioactive compounds in AHE are able to enhance the healing of wounds by reducing the damage caused by ROS. Wound-healing activity was confirmed by histopathology studies. The obtained results showed that the addition of AHE to the gelatin matrix could improve the wound healing activity., This work was funded by the Ministry of Higher Education and Scientific Research, Tunisia. Emerging Research Group Grant from Generalitat Valenciana in Spain (GV/2015/138) and JAEDOC-CSIC postdoctoral contract of L.M. cofounded by the European Social Found are acknowledged.

Organophosphate pesticides are frequently used to eliminate or prevent insects in poultry. However, their residues may continue in meat after slaughtering. In this study, proteomics and peptidomics approaches were used to evaluate their oxidative impact on myosin and chicken breast proteins under in vitro conditions. Myosin protein was exposed to diazinon and chlorpyrifos showing an increase in its oxidation by increasing times, especially with chlorpyrifos. Then, chicken breast was contaminated with chlorpyrifos to evaluate carbonylation and the effect of simulated gastrointestinal digestion. Proteins were isolated using size-exclusion-chromatography and identified by mass spectrometry in tandem. Myosin, β-enolase, CK-M-type and actin were identified as main proteins susceptible to oxidation. Also, oxidised peptides obtained before and after simulated gastrointestinal digestion were identified. Collagen peptides the most susceptible to oxidation. These results suggest that the presence of chlorpyrifos residues on meat could have a negative effect on its final quality and nutritional value., Grant AGL2017-89381-R and FEDER funds from the Spanish Ministry of Economy, Industry and Competitiveness, and Ramón y Cajal postdoctoral contract by L.M. are acknowledged. Also COLCIENCIAS and University of Cartagena for financial support (2019 and 2020) to the project Code 1107-711-50102 are acknowledged. J.M.L. thanks to Colciencias for the scholarship No 647-2014.

This manuscript is reviewing the presence of substances of safety concern that may be generated in different types of meat products. Such substances include N-nitrosamines that can be generated under certain conditions when using nitrite as preservative, the polycyclic aromatic hydrocarbons generated as a consequence of particular smoking processes, heterocyclic aromatic amines generated under particular cooking conditions, compounds released from the oxidation of lipids and proteins, Maillard reaction products like acrylamide and carboxymethyl lysine, and amines that can be generated and accumulated in fermented meats. These hazardous compounds, their mechanisms of generation, risks for health and ways of preventing its presence in meat products are briefly described in this review., Grants AGL2017-89831-R, AGL2015-64673-R and FEDER funds from the Spanish Ministry of Economy, Industry and Competitiveness are acknowledged. Ramón y Cajal postdoctoral contract to LM is also acknowledged.

The worldwide consumption of high-protein food has notoriously increased in recent years. Meat industry generates substantial quantities of protein-rich raw material, which are often discarded as low-value by-products. However, several bioactive compounds can be isolated from these products giving an added value to them. In addition to conventional extraction methods, emerging technologies, including high-pressure processing (HPP), ultrasound (US), pulsed electric fields (PEF) can be used for peptide isolation from meat by-products allowing to maintain the functional properties of these compounds. Antioxidant and antimicrobial activities are between the properties associated with peptides, what would enable their introduction in foods as ingredient and preservative. This review is focused to gather accurate information about the entire extraction process, from the source used until the final peptides obtained. In this sequence are included the pretreatment of the by-product, the extraction procedure, the fractionation and purification, as well as the assay used for the determination of their antioxidant and antimicrobial activities., This research was supported by Grant RTA 2017-00024-CO4-04 from INIA (Spain) and Grant AGL2017-89381-R from Spanish Ministry of Economy, Industry and Competitiveness and Fondo Europeo de Desarrollo Regional (FEDER). Acknowledgments to INIA for granting Paula Borrajo with a predoctoral scholarship (grant number CPD2016-0030). LM was supported by Ramón y Cajal postdoctoral contract. José M. Lorenzo is member of the HealthyMeat network, funded by CYTED (ref. 119RT0568). Thanks to GAIN (Axencia Galega de Innovación) for supporting this research (grant number IN607A2019/01).

Myofibrillar proteins are extensively degraded by muscle endo- and exopeptidases during the ageing of meat and the processing of meat products. One of the most studied products is dry-cured ham. This degradation implies changes in the product in terms of texture (mainly due to calpains and cathepsins endopeptidases) and flavour (due to the action of exopeptidases) and defines its final quality. During the last decade, naturally generated peptides from the myofibrillar proteins titin, myosin light chain, troponin T, LIM domain-binding protein 3, and actin have been identified using peptidomic approaches, also showing the potential to act as bioactives in the human body when ingested. In this study, tandem mass spectrometry has been used for the identification of the peptides generated during the proteolysis of myosin heavy chain protein after 9 months of Spanish dry-cured ham processing. The size, sequence, and properties of some of the peptides showed their potential to act as bioactives., Grant AGL2014-57367-R and FEDER funds from the Spanish Ministry of Economy, Industry and Competitiveness are acknowledged. Ramón y Cajal postdoctoral contract to LM is also acknowledged.

The present study aims to characterize black-barred (Hemiramphus far) skin gelatin (BG) gel enriched or not with its hydrolysate (BGH) in terms of their rheological and micro-structural properties, as well as to evaluate their effect on smooth hound fillet quality during chilled storage. The results of the rheological properties showed that peptides addition influenced elastic and loss modulus as observed from temperature sweep. Based on the microstructure, BG gel had a fine network with small voids that were filled by the presence of BGH peptides. On the other hand, when used as coating agents for smooth hound fillets, BG and BG-BGH showed that, in comparison to control group, fish spoilage was significantly delayed in coated samples. In fact, after 8 days of storage, samples coated with BG and BG+BGH showed the lowest weight loss levels, and preserved their initial water activity (a) values. In addition, fillet coating was able to significantly reduce TVB-N content and oxidation process, as reflected by the low malondialdehyde and carbonyl contents. Furthermore, compared to the control group, all the treatments were found to reduce the degree of microbial deterioration of the fillets, leading to low the free amino acids and nucleotides contents in wrapped fish samples. Therefore, gelatin coating helps to prevent fish fillets from deterioration and oxidation processes., This work was funded and financially supported by the Ministry of Higher Education and Scientific Research, Tunisia (Grant no. 18PJEC06-03).

[EN] The interest in using food byproducts as a source of bioactive peptides has increased significantly in the recent years. The goal of this work was to determine the presence and stability of peptides showing angiotensin I-converting enzyme (ACE-I), endothelin-converting enzyme (ECE), dipeptidyl peptidase-IV (DPP-IV), and platelet-activating factor-acetylhydrolase (PAF-AH) inhibitory activity derived from dry-cured ham bones, which could exert cardiovascular health benefits. ACE-I and DPP-IV inhibitory peptides were stable against heating typically used in Mediterranean household cooking methods and also to in vitro digestion. PAF-AH inhibitory activity significantly increased following simulated gastrointestinal digestion whereas ECE inhibitory significantly decreased (P < 0.05). The mass spectrometry analysis revealed a notable degradation of hemoglobin-derived peptides after simulated digestion, and the release of a large number of dipeptides that may have contributed to the observed bioactivities. These results suggest that natural peptides from Spanish dry-cured ham bones could contribute to a positive impact on cardiovascular health., This study was funded by the Emerging Research Group Grant from Generalitat Valenciana in Spain (GV/2015/138). A Ramon y Cajal postdoctoral contract to L.M. is acknowledged. Proteomic analysis was performed in the proteomics facility of SCSIE University of Valencia that belongs to ProteoRed, PRB2-ISCIII, supported by grant PT13/0001.

The effect of chicken hydrolysates (CHs) on alcohol dehydrogenase (ADH) stability was investigated, together with further bioactivity-oriented isolation and identification of CHs. A total of 82 peptides were identified using mass spectrometry in tandem after consecutive separation by size-exclusion chromatography and high-performance liquid chromatography. The identified peptides were then subjected to in silico gastrointestinal digestion and 154 peptides were generated. The potential bioactivity, safety and applicability of the peptides were assessed using multiple predictive programs. A total of 21 among the 154 peptides were predicted to be potentially active with applicability. Four peptides (DPQYPPGPPAF, QKPVL, KPC, and APGH) obtained after in silico digestion were synthesized and validated their activity. Results showed that DPQYPPGPPAF, KPC, and APGH could stabilize ADH in a dose-dependent manner. This study further indicated that chicken hydrolysate could be a novel functional food ingredient in facilitating alcohol metabolism and protection against alcoholic liver injury., This work was supported by the National Natural Science Foundation of China (Nos. 31701539 and 31871746) and the Natural Science Foundation of Guangdong Province (No. 2017A030313127). Grant AGL2017-89381-R and FEDER funds from the Spanish Ministry of Economy, Industry and Competitiveness, and Ramón y Cajal postdoctoral contract by L.M. are acknowledged. We also thank the China Scholarship Council (CSC) research program for providing funding for C.X.

Pork, one of the most consumed meats worldwide, has been facing major challenges regarding its low sensory quality and unhealthy image of fat. This chapter addresses current feeding strategies to ameliorate pork sensory attributes and nutritional quality by increasing intramuscular fat deposition and improving fatty acid composition, respectively. Dietary protein reduction, alone or combined with some components, contributes to satisfy consumer requirements and enhances the competitiveness of the meat industry with higher pork quality and lower production costs. In addition, feeding sources of n-3 polyunsaturated fatty acids to pigs, mainly from marine origin (rich in eicosapentaenoic and docosahexaenoic acids), increases their content in pork, thus improving the health value of its fatty acid profile. In the near future, the inclusion of microalgae and seaweeds in feed represents a promising approach for the maintenance and development of the livestock sector, as an environmental friendly alternative to balance food and feed industries.

Peptides with a similar antioxidant and ACE-inhibitory activity of royal jelly (RJ) generated from Alcalase hydrolysated pollen (AHP) were predicted by Response Surface Methodology (RSM). The model equations were proposed according to the effects of time and enzyme concentration on the antioxidant and ACE-inhibitory activity. The optimum values for Alcalase concentration and hydrolysis time were 1.5% and 4 h, respectively. Later, AHP was prepared and deproteinised to be further analysed using size-exclusion chromatography (SEC). After SEC separation, fractions with the highest activity of ACE-inhibitory, DPPH radical scavenging and ferric-reducing power were purified by RP-HPLC. The highest ACE-inhibitory and DPPH scavenging activity of fractions was found 100% and 66.61%, respectively. The most active fractions were analysed by nano-liquid chromatography and mass spectrometry in tandem (nLC-MS/MS) and a total of 195 peptide sequences were identified. The origins of all peptides were herbal proteins and certain coincidences with previously described bioactive sequences were discussed., This work was funded by the Ministry of Science, Research and Technology of Iran and the Grant AGL2014-57367-R and FEDER funds from the Spanish Ministry of Economy, Industry and Competitiveness. Ramón y Cajal postdoctoral contract to LM is acknowledged. LC-MS/MS analysis was carried out by in the SCSIE University of Valencia Proteomics Unit (Spain), a member of ISCIII Proteo Red Proteomics Platform.

In the present study, response surface method was used to optimize hydrolysis condition to generate potential bioactive peptides from pollen protein using pepsin (pepsin hydrolysated pollen—PHP) and trypsin (trypsin hydrolysated pollen—THP). Then PHP and THP prepared under optimized conditions were analyzed by size-exclusion chromatography. The fractions possessing the maximum ACE-inhibitory, DPPH radical scavenging, and ferric-reducing power were further purified by RP-HPLC. A heterogeneous composition of hydrophobic and hydrophilic peptides in both fractions was obtained. Finally, peptide sequences in active fractions of PHP and THP were identified by mass spectrometry in tandem. All the identified peptides had herbal protein origins. These were 6–21 amino acids in length, and Glycine and Alanine were two main hydrophobic amino acids present in their sequences. The results proved that using controlled enzymatic hydrolysis of pollen protein is possible to generate bioactive peptides with high ACE-inhibitory and antioxidant activity in final product. Practical applications: Pollen is well-known as an interesting protein source. Compared to other types of hydrolysis, enzymatic hydrolysis of vegetable proteins has few or no undesirable side reactions or products. In this study, controlled enzymatic hydrolysis of pollen protein was applied as a suitable method to produce bioactive peptide. The results proved that using controlled enzymatic hydrolysis of pollen protein is possible to generate bioactive peptides with high ACE-inhibitory and antioxidant activity in final product. This product can be used as functional and health promoting ingredient in different food formulations., This work was financially supported by the Ministry of Science, Research and Technology of Iran, Grant AGL2014‐57367‐R and FEDER funds from the Spanish Ministry of Economy, Industry, and Competitiveness. Ramón y Cajal postdoctoral contract of LM is also acknowledged. MS/MS analysis was conducted in SCSIE University of Valencia Proteomics Unit (Spain) which is registered in ISCIII ProteoRed Proteomics Platform.

Proteolytic activity and physico-chemical characteristics were studied for Norwegian dry-cured ham at four different times of processing: raw hams, post-salted hams (3 months of processing), hams selected in the middle of the production (12 months of processing) and hams at the end of the processing (24 months). Cathepsin H activity decreased until negligible values after 3 months of processing, whereas cathepsins B and B + L were inactive at 12 months. AAP was the most active aminopeptidase whereas RAP and MAP were active just during the first 12 months of processing. Proteolysis index reached a value of 4.56 ± 1.03 % with non-significant differences between 12 and 24 months of ripening. Peptide identification by LC-MS/MS was done and two peptides (GVEEPPKGHKGNKK and QAISNNKDQGSY) showing a linear response with the time of processing were found. Unfreezable water content and glass transition temperature were investigated using differential scanning calorimetry (DSC) technique with non-significant differences in the temperature of glass transition for 12 and 24 months of processing., The work was supported by the Research Council of Norway (project 225262/E40 – DryMeat)

This study evaluates the effect of dietary selenium (Se) supplementation source (organic, Se-enriched yeast; SY vs. inorganic, sodium selenite; SS), dose (0.2: L vs. 0.4: H mg/kg) and the combination of Se and vitamin E (VITE + SS) for 26 days on drip loss, TBARS, colour changes, myofibrillar protein pattern and proteolysis in pork. The lowest water losses were observed in the SY-H group when compared to the others. SY-H and VITE + SS groups presented lower myofibrillar protein hydrolysis/oxidation. VITE + SS supplementation also resulted in higher PRO, TRP and PHE content at days 2 and 7, whereas the SY group showed increased GLY and CAR and tended to have higher TAU and ANS at day 2. The myofibrillar fragmentation index was not modified by the dietary treatment; however, at day 8, it tended to be higher in groups supplemented with SeY and VITE + SS. The results of the present study might indicate a possible relation between muscle proteolysis and water loss., This research was supported by projects CDTI IDI-20111017 and S2013/ABI-2913-MEDGAN-CAM.

Spanish dry-cured ham is a high quality product whose economic value is mainly given by its curing time. An intense proteolysis takes place throughout the dry-cured processing, which results in the generation of a high amount of peptides and free amino acids responsible for the final quality of dry-cured hams. In this work, a peptidomics approach has been used to study the evolution of peptides throughout the ham dry-curing process, identifying and quantifying the generated peptides in order to define potential quality biomarkers. For this purpose, dry-cured ham extracts at different processing times (0, 2, 3.5, 5, 6.5 and 9 months) were fractionated by size-exclusion chromatography and analysed by nanoliquid chromatography coupled to tandem mass spectrometry. Differences obtained in the relative quantification of peptides by using a label-free methodology were useful to establish differences between processing times, being peptides generated by the degradation of myosin light chain 1 protein mainly responsible for the observed differences during the last stages of curing. In particular, APAPAPAPPKEEKI and PAPAPAPAPAPAPAPPKE, exclusively identified at 9 months of curing, would be potential markers to control the time of curing and thus the final quality of dry-cured hams., The FPI Scholarship BES-2011-046096 from MINECO (Spain) to M.G. and Grants AGL2013-47169-R from MINECO and FEDER funds are acknowledged. The research also received funding from the European Union Seventh Framework Programme FP7-PEOPLE-2013-CIG under Grant Agreement 614281 (HIGHVALFOOD). The JAEDOC-CSIC postdoctoral contract to L.M. co-funded by the European Social Fund is also acknowledged.