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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.

This study reports the effect of thermal pretreatment and the use of different commercial proteolytic enzymes (Protamex, Flavourzyme, Protana prime, and Alcalase) on the free amino acid content (FAA), peptide profile, and antioxidant, antidiabetic, antihypertensive, and anti-inflammatory potential (DPPH, FRAP, and ABTS assay, DPP-IV, ACE-I, and NEP inhibitory activities) of dry-cured ham bone hydrolyzates. The effect of in vitro digestion was also determined. Thermal pretreatment significantly increased the degree of hydrolysis, the FAA, and the DPP-IV and ACE-I inhibitory activities. The type of peptidase used was the most significant factor influencing antioxidant activity and neprilysin inhibitory activity. Protana prime hydrolyzates failed to inhibit DPP-IV and neprilysin enzymes and had low values of ACE-I inhibitory activity. After in vitro digestion, bioactivities kept constant in most cases or even increased in ACE-I inhibitory activity. Therefore, hydrolyzates from dry-cured ham bones could serve as a potential source of functional food ingredients for health benefits.

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 aimed to determine the effect of sealing process at different internal temperatures on the cooking loss, lipid oxidation, textural, and sensory properties, as well as the protein profile of cooked meat. For this aim, four different groups were formed. The control group was not sealed, while the other three groups were sealed until their internal temperature reached 20, 30, and 40 °C. Then, all samples were baked until their internal endpoint temperature reached 71 °C to ensure food safety. The lowest cooking loss (11.57%) was detected in the control group, and increased sealing core temperature resulted in an increase in cooking loss. The sealing did not show a significant effect (P > 0.05) on the TBARS value. On the other hand, myofibrillar and sarcoplasmic protein profile were influenced by sealing. Myofibrillar protein profile showed that protein denaturation rate was higher in groups sealed at the internal temperature of 30 and 40 °C. The sealing process affected only the hardness value among the textural properties examined. With the sealing process, the hardness value of the meat significantly (P < 0.05) reduced and the lowest hardness value was determined in the sample sealed at the internal temperature of 20 °C. Sealing positively affected the overall acceptability score of the samples, and the samples sealed until their internal temperatures reached 30 and 40 °C had higher overall acceptability scores.

The utilization of animal bones as a protein source could be used as a sustainable pathway for the production of bioactive compounds. In this study, bones were pretreated with pepsin enzyme (PEP) and then sequentially hydrolyzed with Alcalase (PA) and Alcalase, as well as Protana prime (PAPP). The degree of hydrolysis, antioxidant activity, and DPP-IV inhibitory activity were measured. All three hydrolysates showed antioxidant and DPP-IV inhibitory activity; however, the highest result in both bioactivities was obtained with the PAPP hydrolysate. The obtained free amino acid content was 54.62, 88.12, and 668.46 mg/100 mL of hydrolyzed in PEP, PA, and PAPP, respectively. Pepsin pretreatment did not significantly affect the degree of hydrolysis; however, it is suggested that it promoted the cleavage of certain bonds for subsequent protease action. Accordingly, a total of 550 peptides were identified in PEP hydrolysate, 1087 in PA hydrolysate, and 1124 in PAPP hydrolysate using an LC-MS/MS approach. Pepsin pretreatment could be an effective method in the utilization of bone sources for the production of antioxidant and hypoglycemic peptides.

A number of bioactive peptides with health benefits have been reported to be generated in meat through the proteolysis phenomena taking place during postmortem aging or further processing. Bioactive peptides consist of short sequences, less than 20 amino acid residues, that are inactive while in the parent protein. These sequences may be activated when they are released from the parent protein by peptidases. Such peptides are released through the hydrolysis of major muscle proteins by muscle endopeptidases, like calpains and cathepsins, and resulting protein fragments and polypeptides are further hydrolyzed by muscle exopeptidases, mainly, tri-and di-peptidylpeptidases, aminopeptidases, and carboxypep-tidases. As a result, a variety of peptides with different sequences and lengths and large amounts of free amino acids are generated. The highest amounts of bioactive peptides, and their major health benefits, have been reported in dry-cured ham as a result of the intense proteolysis experienced during the long processing time under mild conditions that favor the action of muscle peptidases. The generated peptides must be bioaccesible and bioavailable to exert their physiological benefit. This means that they must be resistant to gastrointestinal digestion and be absorbed through the intestinal barrier. Some bioactive peptides may be generated through hydrolysis during gastrointestinal digestion. This review is focused on the generation of bioactive peptides in meat and processed meats, on the most relevant bioactivities exerted by such peptides (such as inhibition of angiotensin I–converting enzyme and dipeptidyl peptidase IV as well as antioxidant and anti-inflam-matory activity), and on their reported benefits to consumers’ health.

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.

The defatted viscera from smooth hound were the raw material used to produce four protein hydrolysates using three different microbial proteases (Neutrase, Esperase and Purafect) and the endogenous enzymes, named VPH-N, VPH-E, VPH-P and VPH-EE, respectively. Hydrolysates showed different degrees of hydrolysis (DH) depending on the enzyme used, where the VPH-P had the highest DH (30%). The amino acids (AA) characterization showed that Gly was the prominent AA (≥ 22%), followed by Ala, Glx, Lys, Asx, and Pro. In addition, the assessment of the antioxidant, ACE-inhibitory and antimicrobial activities revealed the high potentiality of the Purafect-hydrolysate. Therefore, it was selected to be fractionated by ultra-filtration according to their molecular weight (MW). Data showed that FIV (MW < 5 kDa) exhibited the best antioxidant and ACE inhibition potentials. The differences detected in the bio-activities may be attributed, not only to the peptides size, but also to their sequences and hydrophobic AA contents. The antibacterial activity showed that the best inhibition values were recorded with FIV, particularly against S. typhi. Thus, the present results demonstrated the high potential of low MW peptides from viscera hydrolysate to be used as a promising natural source of bioactive agents in functional food formulations. Graphical Abstract: [Figure not available: see fulltext.].

Heterocyclic aromatic amines (HAAs) are detrimental substances can develop during the high-temperature cooking of protein-rich foods, such as meat. They are potent mutagens and carcinogens linked to an increased risk of various cancers. HAAs have complex structures with nitrogen-containing aromatic rings and are formed through chemical reactions between amino acids, creatin(in)e, and sugars during cooking. The formation of HAAs is influenced by various factors, such as food type, cooking temperature, time, cooking method, and technique. HAAs exert their toxicity through mechanisms like DNA adduct formation, oxidative stress, and inflammation. The research on HAAs is important for public health and food safety, leading to risk assessment and management strategies. It has also led to innovative approaches for reducing HAAs formation during cooking and minimizing related health risks. Understanding HAAs' chemistry and formation is crucial for developing effective ways to prevent their occurrence and protect human health. The current review presents an overview about HAAs, their formation pathways, and the factors influencing their formation. Additionally, it reviews their adverse health effects, occurrence, and the analytical methods used for measuring them.

Lawries’ Meat Science, Ninth Edition continues to be a classic reference in the meat world. It has been used by numerous generations of meat professionals since its first edition in 1966. The new edition brings four new chapters and updated information related to the latest advances in meat animals breeding and technologies for meat preservation, processing, and packaging. In addition, new relevant aspects of nutritional value, quality and safety of meat as well as methodologies for authenticity and traceability are provided with a compilation of chapters written by a select group of the most experienced and knowledgeable people in the meat field. This book covers essential information and latest advances and developments, from the initial meat animal’s growth and development to the time of slaughter and to the processing technologies, packaging and distribution till consumption of its meat. Relevant aspects of its composition, nutritional value, eating quality, consumer acceptance, safety and sustainability issues are also covered. Key Features Includes new information on improved added value of meat by-products for increased sustainability Presents best practices sustainable animal production and meat processing Provides the latest developments in organic meat and meat products and on cell-cultured meat and future market opportunities

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 <3 kDa, and the highest BC values were obtained for peptide fractions >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.

Food-derived bioactive peptides are receiving an increased interest for their contribution to human health. Current trends are addressed towards the mass production of bioactive peptides. Modern in silico strategies based on simulation using bioinformatics tools are providing large amounts of data in comparison to the empirical approaches traditionally used. Novel green technologies such as ultrasound, microwave, hydrostatic pressure, pulsed electric field and subcritical water are being applied for improving the extraction yield and increasing the enzymatic hydrolysis in the production of bioactive peptides. Continuous reactors based on the use of membranes or immobilised enzymes are being developed for increased hydrolysis rates and larger production. Although much research has been performed at laboratory scale, further research is needed to overcome the challenges related to large-scale production of bioactive peptides.

Blood is a meat by-product rich in proteins with properties that can be improved after hydrolysis, making it a sustainable alternative for use in the generation of bioactive peptides. The objective of this study was to identify dipeptidyl peptidase IV (DPP-IV) inhibitory peptides obtained from different chicken blood hydrolysates prepared using combinations of four different enzymes. Best results were observed for AP (2% Alcalase + 5% Protana Prime) and APP (2% Alcalase + 5% Protana Prime + 3% Protana UBoost) hydrolysates obtaining inhibition values of 60.55 and 53.61%, respectively, assayed at a concentration of 10 mg/mL. Free amino acids were determined to establish the impact of exopeptidase activity in the samples. A total of 79 and 12 sequences of peptides were identified by liquid chromatography and mass spectrometry in tandem (LC-MS/MS) in AP and APP samples, respectively. Nine of the identified peptides were established as potential DPP-IV inhibitory using in silico approaches and later synthesized for confirmation. Thus, peptides GPF, IGL, and GGGW showed good DPP-IV inhibitory activity with IC50 values of 0.94, 2.22, and 2.73 mM, respectively. This study confirmed the potential of peptides obtained from chicken blood hydrolysates to be used as DPP-IV inhibitors and, therefore, in the control or modulation of type 2 diabetes.

[EN] 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 petroleumbased 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.

The processing of dry-cured ham results in the generation of small peptides by the action of endogenous enzymes on muscle proteins. Common proteomic workflows involve previous separation techniques based on liquid chromatography which are expensive and time-consuming. In this study, a convenient proteomic approach based on MALDI-ToF is proposed for the first time for the detection of dipeptides in Spanish dry-cured ham. Dipeptides AH, AL, DD, EV, and VF were identified in hams of 18 and 24 months of dry-curing. This work provides insights on the efficiency of a new peptidomic workflow for the short peptide identification from a complex food matrix and permits to evaluate the sample in terms of the presence of taste-related and bioactive dipeptides.

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.

Dry-cured ham-derived dipeptides, generated along a dry-curing process, are of high importance since they play a role in flavor development of dry-cured ham. The objective of this study was to analyze the residues of the less-studied metabotropic glutamate receptor 1 (mGluR1) implicated in the recognition of umami dry-cured ham dipeptides by molecular docking simulation using the AutoDock Suite tool. AH, DA, DG, EE, ES, EV, and VG (and glutamate) were found to attach the enzyme with inhibition constants ranging from 12.32 µM (AH) to 875.75 µM (ES) in the case if Rattus norvegicus mGluR1 and 17.44 µM (VG) to 294.68 µM (DG) in the case of Homo sapiens, in the open–open conformations. Main interactions were done with key receptor residues Tyr74, Ser186, Glu292, and Lys409; and Ser165, Ser186, and Asp318, respectively, for the two receptors in the open–open conformations. However, more residues may be involved in the complex stabilization. Specifically, AH, EE and ES relatively established a higher number of H-bonds, but AH, EV, and VG presented relatively lower Ki values in all cases. The results obtained here could provide information about structure and taste relationships and constitute a theoretical reference for the interactions of novel umami food-derived peptides.

[EN] 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, triand 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.

"This document is the unedited Author's version of a Submitted Work that was subsequently accepted for publication in Journal of Agricultural and Food Chemistry, copyright © American Chemical Society after peer review. To access the final edited and published work see https://pubs.acs.org/doi/10.1021/acs.jafc.9b08297", [EN] There is a wide variety of peptides released from food proteins that are able to exert a relevant benefit for human health, such as angiotensin-converting enzyme inhibition, antioxidant, anti-inflammatory, hypoglucemic, or antithrombotic activity, among others. This manuscript is reviewing the recent advances on enzymatic mechanisms for the hydrolysis of proteins from foods of animal origin, including the types of enzymes and mechanisms of action involved, the strategies followed for the isolation and identification of bioactive peptides through advanced proteomic tools, and the assessment of bioactivity and its beneficial effects. Specific applications in fermented and/or ripened foods where a significant number of bioactive peptides have been reported with relevant in vivo physiological effects on laboratory rats and humans as well as the hydrolysis of animal food proteins for the production of bioactive peptides are also reviewed.

There is a wide variety of peptides released from food proteins that are able to exert a relevant benefit for human health, such as angiotensin-converting enzyme inhibition, antioxidant, anti-inflammatory, hypoglucemic, or antithrombotic activity, among others. This manuscript is reviewing the recent advances on enzymatic mechanisms for the hydrolysis of proteins from foods of animal origin, including the types of enzymes and mechanisms of action involved, the strategies followed for the isolation and identification of bioactive peptides through advanced proteomic tools, and the assessment of bioactivity and its beneficial effects. Specific applications in fermented and/or ripened foods where a significant number of bioactive peptides have been reported with relevant in vivo physiological effects on laboratory rats and humans as well as the hydrolysis of animal food proteins for the production of bioactive peptides are also reviewed.

To investigate the oxidative effect of CPF on chicken breast proteins, peptidomics and proteomics analysis were used. For this, chicken breast samples were exposed to CPF and subjected to simulated gastrointestinal digestion. The identification of oxidized peptides from digested and undigested proteins were performed by LC MS/MS (nanoESI qQTOF). Prior to mass analyses undigested proteins were trypsinated. Data were analysed using MASCOT and ProteinPilot v 4.5 software. In this study, 90 and 107 oxidized peptides from digested proteins of control and exposed samples were identified, respectively. These peptides corresponding to 12 oxidized proteins. Meanwhile, 260 and 324 oxidized peptides from undigested proteins (control and exposed samples) were identified, which corresponding to 19 and 17 proteins, respectively. Collagen was protein more susceptible to oxidation promoted by chlorpyrifos in digested and undigested proteins. Identification of these oxidized proteins from simulated digestion provides an important insight about the impact of substances like certain veterinary drugs at trace concentrations on the nutritional value of chicken breast meat.

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.

The agro-food industry generates large amounts of waste that contribute to environmental contamination. Animal fat waste constitutes some of the most relevant waste and the treatment of such waste is quite costly because environmental regulations are quite strict. Part of such costs might be reduced through the generation of bioenergy. Biodiesel constitutes a valid renewable source of energy because it is biodegradable, non-toxic and has a good combustion emission profile and can be blended up to 20% with fossil diesel for its use in many countries. Furthermore, up to 70% of the total cost of biodiesel majorly depends on the cost of the raw materials used, which can be reduced using animal fat waste because they are cheaper than vegetable oil waste. In fact, 6% of total feedstock corresponded to animal fat in 2019. Transesterification with alkaline catalysis is still preferred at industrial plants producing biodiesel. Recent developments in heterogeneous catalysts that can be easily recovered, regenerated and reused, as well as immobilized lipases with increased stability and resistance to alcohol denaturation, are promising for future industrial use. This manuscript reviews the available processes and recent advances for biodiesel generation from animal fat waste.

The use of edible biopolymers and natural additives obtained from food processing by-products is a sustainable strategy for food packaging applications. Gelatin is a biopolymer with great potential as a coating due to its low cost, high availability, and technological and functional properties. Among them, gelatin can be used as a carrier of bioactive compounds such as antioxidants, which can retard oxidation processes and thus extend the shelf-life of highly-perishable products. This study evaluated the effect of gelatin coating enriched with antioxidant tomato by-products hydrolysate (TBPH) on the quality of pork meat during cold storage. Results showed that TBPH obtained from Alcalase hydrolysis presented antioxidant activity with good stability against cooking. Additionally, chromatographic and mass spectrometry techniques, as well as in silico analysis, were used for the peptidomic characterisation of TBPH. The application of enriched gelatin coating on meat led to some physicochemical changes including increased weight loss and colour differences; however, the pH and water activity, which control meat spoilage, were maintained during storage. Moreover, coating prevented lipid oxidation of meat, and enriched-coated meat presented high antioxidant activity after cooking. These results suggest the positive role of gelatin coating enriched with TBPH in extending the shelf-life of meat during storage

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.

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.

"This document is the unedited Author s version of a Submitted Work that was subsequently accepted for publication in Journal of Agricultural and Food Chemistry, copyright © American Chemical Society after peer review. To access the final edited and published work see https://pubs.acs.org/doi/10.1021/acs.jafc.8b05888", [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.

[EN] 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.

[ES] La alta actividad proteolítica ocurrida en el jamón curado durante el periodo de producción es la responsable de la liberación de péptidos de diversos tamaños y aminoácidos capaces de aportar unas propiedades sensoriales características. Una de las enzimas implicadas en este proceso es la dipeptidil peptidasa 4 o DPP 4 cuya actividad proteolítica genera una serie de dipéptidos determinados. La DPP 4, además, es la encargada de la degradación de las hormonas incretinas responsables de la liberación de insulina, lo que la convierte en una enzima de interés para el diseño de tratamientos contra la diabetes tipo 2. En estudios anteriores, se ha demostrado que muchos de los dipéptidos presentes en el jamón curado inhiben a la DPP 4, confiriéndole propiedades antidiabéticas. El principal objetivo de este trabajo consiste en la caracterización de la DPP 4 mediante un estudio de cinética enzimática. Los ensayos de actividad enzimática se realizaron mediante un método fluorimétrico, es decir, se midió la fluorescencia liberada por la enzima DPP 4 durante la hidrólisis de un sustrato con un grupo fluorescente. La enzima presentó una cinética enzimática que se ajusta a la ecuación de MichaelisMenten con una constante de Michaelis, KM de 30,92 µM. También se estudió el efecto inhibidor de los dipéptidos Ala-Ala, Ala-Leu, Pro-Ala y Ala-Pro, como dipéptidos producto generados por la DPP 4 en el jamón curado. Se observó una inhibición por producto de tipo mixta con una constante de disociación para Ala-Pro y Pro-Ala muy similar, siendo sus valores 1,46 y 1,37 mM, respectivamente., [EN] The proteolytic activity occurred in dry-cured ham during the ripening is responsible for the release of peptides and amino acids that provide sensory properties. Dipeptidyl peptidase IV or DPP IV is one of the enzymes involved in this process whose proteolytic activity gives rise to several dipeptides. Additionally, DPP IV degrades two incretin hormones involved in the release of insulin. Consequently, this enzyme could be used for designing diabetes type 2 treatments. In previous studies, it has been shown that many of these dipeptides inhibit DPP IV acquiring antidiabetic properties. The main purpose of the research was the characterization and enzyme kinetic study of DPP IV. The enzyme activity was determined through a fluorimetric method. A substrate with a fluorescent group is degraded by the enzyme DPP IV increasing the fluorescence measured. The enzyme kinetic followed the equation of Michaelis-Menten with a Michaelis constant, KM of 30.92 µM. This assay was carried out at different concentrations of Ala-Ala, Ala-Leu, Pro-Ala and Ala-Pro (as products generated by DPP IV in Spanish dry-cured ham). Assuming a mixed product inhibition, the dissociation constant Ki for Ala-Pro and Pro-Ala were 1.46 and 1.37 mM, respectively.

The objective of this study was to investigate the lipid content and fatty acid profile in six edible organs, namely heart, brain, pancreas, kidney, liver, and tongue from commercial crossbred pigs. Results indicate that total lipid and fatty acid composition were significantly affected by organ type. Large differences in lipid contents were found, ranging from 1.4 to 6.8 g/100 g of tissue. Brain had the highest lipid content followed by tongue, pancreas, liver, kidney, and heart. The fatty acid profile revealed low relative proportions of saturated fatty acids (SFA) in heart. In contrast, higher amounts of un­saturated fatty acids were obtained for heart (PUFA), and tongue (MUFA). Heart and kidney had the most favourable PUFA/SFA ratio. However, the ratio of n-6 and n-3 PUFA in by-products from pigs was above to the nutritional recommendation for human diet, except in brain. Moreover, variations in fatty acid profile clearly highlight the differences between the organs as showed by principal component analysis. Our findings indicate that some edible offal from pig were relatively rich in SFA and, therefore, should be consumed moderately if integrated into a varied well-balanced diet.

Food-derived bioactive peptides are promising compounds for the prevention and treatment of cardiovascular diseases, the main cause of mortality in developed countries. The aim of this work was to determine the in vitro anti-inflammatory, antioxidant, and angiotensin I-converting enzyme (ACE-I) inhibitory activities of twenty-four peptides that were identified in Spanish dry-cured hams. For the first time, some peptides such as PSNPP, HCNKKYRSEM and FNMPLTIRITPGSKA showed anti-inflammatory activity expressed as platelet-activating factor-acetylhydrolase, autotaxin, and lipoxygenase inhibition. Peptides MDPKYR and TKYRVP were the strongest antioxidants, whereas GGVPGG, TKYRVP, and HCNKKYRSEM showed the highest ACE-I inhibitory activity. Additionally, several peptides such as KPVAAP, MDPKYR, TKYRVP, HCNKKYRSEM exerted more than one of the assayed activities, increasing their health-enhancing potential. More studies are needed to evaluate the bioavailability of such peptides and their in vivo effect. This would contribute to consider dry-cured ham as a source of peptides beneficial for cardiovascular health.

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 minin

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.

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.

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.

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.

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.

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.

Dry-cured ham has been described as a good source of bioactive peptides and taste-active compounds. Some of them are dipeptides and tripeptides that are released in a large amount from different muscle proteins due to the action of exopeptidases during the dry-cured ham processing. The potential of dipeptides and tripeptides to exert bioactivities and impart taste characteristics to dry-cured ham has been evaluated using the BIOPEP database, since in silico approaches are a time- and cost-effective alternative to empirical approaches. Most of the studied dipeptides and tripeptides showed ACE and DPP inhibitory activities as well as imparted bitter taste. In fact, more than one bioactivity and/or taste could be assigned to a given peptide sequence, and there could be a correlation between both, like ACE inhibitory and bitter EA, EI and LG peptides. Furthermore, several dipeptides such as EK, KP, LA, PL, PP, RG, and VE, among others, were found to be multifunctional (ACE and DPP IV inhibitory) which would be determined by their structure, sequence and amino acid composition. In silico analysis evidences the relevance of dipeptides and tripeptides in the bioactivity and taste of dry-cured hams, but further empirical assays including in vitro and in vivo studies are necessary to confirm such theoretical results. Possible degradation of the small peptides during gastrointestinal digestion and intestinal absorption as well as interactions with the food matrix could reduce their bioavailability and bioaccessibility, and modify their biological activities.