High-pressure processing enhances saltiness perception and sensory acceptability of raw but not of cooked cured pork loins-leveraging salty and umami taste.

The salt (NaCl) content in processed meats must be reduced because of its adverse effects on cardiovascular health. However, reducing salt in meat products typically leads to a lower taste intensity and, thus, consumer acceptability. Industry interventions must reduce salt content while maintaining taste, quality, and consumer acceptability. In this context, high-pressure processing (HPP) has been proposed to enhance saltiness perception, though there are contradictory reports to date. The present work aimed to conduct a targeted experiment to ascertain the influence of HPP (300/600 MPa) and cooking (71°C) on saltiness perception and sensory acceptability of meat products. HPP treatment (300/600 MPa) did enhance those two sensory attributes (approx. +1 on a 9-point hedonic scale) in raw (uncooked) cured pork loins but did not in their cooked counterparts. Further, the partition coefficient of sodium (PNa+), as an estimate of Na+ binding strength to the meat matrix, and the content of umami-taste nucleotides were investigated as potential causes. No effect of cooking (71°C) and HPP (300/600 MPa) could be observed on the PNa+ at equilibrium. However, HPP treatment at 300 MPa increased the inosine-5'-monophosphate (IMP) content in raw cured pork loins. Finally, hypothetical HPP effects on taste-mediating molecular mechanisms are outlined and discussed in light of boosting the sensory perception of raw meat products as a strategy to achieve effective salt reductions while keeping consumer acceptability.

Investigation of Pulsed Electric Field Conditions at Low Field Strength for the Tenderisation of Beef Topside.

Tenderness is the most critical eating quality trait of meat, and consequently, processing interventions for meat tenderisation have significant economic relevance. The objective of this study was to investigate pulsed electric field (PEF) conditions for the tenderisation of beef topside. The PEF settings included combinations of three field strengths (0.25, 0.50 and 1.00 kV/cm), two frequencies (20 and 100 Hz) and three treatment times (10, 30 and 50 ms). The effect of PEF on meat quality parameters (pH, drip loss, shear force, cook loss and colour) immediately after treatment and after storage (1 and 14 days at 4 °C) was evaluated. PEF did not affect meat tenderness after 1 day of chilled storage but resulted in a 5-10% reduction in the shear force in some cases (0.25-0.5 kV/cm) compared to the untreated control after 14 days of storage. Other quality traits (cook loss and colour) were not impaired. Thus, we concluded that PEF technology is a possible intervention to improve meat tenderness of beef topside after 2 weeks of storage.

High-pressure processing of meat: Molecular impacts and industrial applications.

High-pressure processing (HPP) has been the most adopted nonthermal processing technology in the food industry with a current ever-growing implementation, and meat products represent about a quarter of the HPP foods. The intensive research conducted in the last decades has described the molecular impacts of HPP on microorganisms and endogenous meat components such as structural proteins, enzyme activities, myoglobin and meat color chemistry, and lipids, resulting in the characterization of the mechanisms responsible for most of the texture, color, and oxidative changes observed when meat is submitted to HPP. These molecular mechanisms with major effect on the safety and quality of muscle foods are comprehensively reviewed. The understanding of the high pressure-induced molecular impacts has permitted a directed use of the HPP technology, and nowadays, HPP is applied as a cold pasteurization method to inactive vegetative spoilage and pathogenic microorganisms in ready-to-eat cold cuts and to extend shelf life, allowing the reduction of food waste and the gain of market boundaries in a globalized economy. Yet, other applications of HPP have been explored in detail, namely, its use for meat tenderization and for structure formation in the manufacturing of processed meats, though these two practices have scarcely been taken up by industry. This review condenses the most pertinent-related knowledge that can unlock the utilization of these two mainstream transformation processes of meat and facilitate the development of healthier clean label processed meats and a rapid method for achieving sous vide tenderness. Finally, scientific and technological challenges still to be overcome are discussed in order to leverage the development of innovative applications using HPP technology for the future meat industry.

Biochemical and Nutritional Changes during Food Processing and Storage.

Domestic food processing goes a long way back in time, for example, heat for cooking was used 1 [...].

Application of pulsed electric fields in meat and fish processing industries: An overview.

The market demand for new meat and fish products with enhanced physicochemical and nutritional properties attracted the interest of the food industry and academia to investigate innovative processing approaches such as pulsed electric fields (PEF). PEF is an emerging technology based on the application of electrical currents between two electrodes thus inducing electroporation phenomena and enabling a non-invasive modification of the tissues' structure. This review provides an overview of the current knowledge on the use of PEF processing in meat and fish to enhance the physicochemical and nutritional changes, as a preservation method, as well as for improving the extraction of high added-value compounds. PEF treatment had the ability to improve several processes such as preservation, tenderization, and aging. Besides, PEF treatment could be used as a useful strategy to increase water holding properties of fish products as well as for fish drying. Finally, PEF could be also used in both meat and fish foods for by-products valorization, due to its potential to enhance the extraction of high added-value compounds. However, more studies are warranted to completely define specific treatments that can be consistently applied in the industry. This review provides the directions for this purpose in the near future.

Effect of cooking and simulated gastrointestinal digestion on the activity of generated bioactive peptides in aged beef meat.

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 effect of high pressure on the functional properties of pork myofibrillar proteins.

Complementary methodologies were used to analyse the pressure-induced modification and functionality of myofibrillar proteins from pork meat pressurised at 200, 400, 600, or 800 MPa (10 min, 5 or 20 °C). Pressure at 400 MPa was found to be the threshold for loss of solubility, and the structural proteins, myosin and actin, lost their native solubility due to aggregation. The results from the extraction of proteins with different reagents targeting the disruption of specific molecular interactions suggested that pressure-induced aggregation was caused mainly by hydrogen bonding during pressurisation and not hydrophobic interactions nor disulphide cross-links. Furthermore, the soluble proteins were exposed to remarkable structural changes already at 200 MPa and lost their native functionality. The modification of the proteins in pressurised meat affected the water binding sites of the myofibrillar proteins and, thereby, the interactions between proteins and water molecules, and distribution between myofibrillar and extra-myofibrillar compartments.

Effect of electrohydraulic shockwave treatment on tenderness, muscle cathepsin and peptidase activities and microstructure of beef loin steaks from Holstein young bulls.

Hydrodynamic pressure processing (HDP) or shockwave treatment improved tenderness (18% reduction in Warner-Bratzler shear force (WBSF) of beef loin steaks. Endogenous muscle proteolyic activities (cathepsins and peptidases) and protein fragmentation of sarcoplasmic and myofibrillar proteins detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) were not influenced by HDP. However, microstructure changes were clearly detected using confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). Specifically a disruption of the structure at the muscle fiber bundles and an increased endomysium space were observed. The present paper supports the evidence of physical disruption of the muscle fibers as a cause behind the tenderness improvement. The paper discusses the possible mechanisms responsible for the meat tenderisation induced by HDP treatment.

Mechanisms of radical formation in beef and chicken meat during high pressure processing evaluated by electron spin resonance detection and the addition of antioxidants.

The generation of radicals during high pressure (HP) processing of beef loin and chicken breast was studied by spin trapping and electron spin resonance detection. The pressurization resulted in a higher level of spin adducts in the beef loin than in the chicken breast. It was shown that radicals were formed in the sarcoplasmic and myofibrillar fractions as well as in the non-soluble protein fraction due to the HP treatment, indicating that other radicals than iron-derived radicals were formed, and most likely protein-derived radicals. The addition of iron as well as the natural antioxidants caffeic acid, rosemary extract, and ascorbic acid resulted in an increased formation of radicals during the HP treatment, whereas addition of ethylendiamintetraacetic acid (EDTA) reduced the radical formation. This suggests that iron-species (protein-bound or free) catalyses the formation of radicals when meat systems are submitted to HP.

New developments in shockwave technology intended for meat tenderization: Opportunities and challenges. A review.

Meat tenderness is an important quality parameter determining consumer acceptance and price. Meat tenderness is difficult to ensure in the global meat chain because the production systems are not always aiming at this purpose (ex.: cattle derived from milk production) and by the existence within the carcass of "tough" primals. Different methods can be used by the meat industry to improve meat tenderness each with its advantages and drawbacks. The application of hydrodynamic pressure or shockwaves has showed outstanding improvements by reducing the Warner Bratzler Shear Force by 25% or more. However, the technology has not penetrated into the market as first systems were based on the use of explosives and further developments seemed to lack the robustness to fulfill industrial requirements. The present paper describes the main challenges to construct a prototype for the continuous treatment of meat by shockwaves based on electrical discharges under water. Finally, improvements on the tenderness of meat by using the novel prototype are presented.

Quality considerations with high pressure processing of fresh and value added meat products.

Pressure can be applied by high hydrostatic pressure, better known as high pressure processing (HPP), or by hydrodynamic pressure (HDP) in the form of shockwaves to alter quality parameters, such as shelf-life and texture of meat and meat products. The aim of this review is to give an overview of the use of pressure in the meat industry and to highlight its usage as a method to inactivate microorganisms but also a novel strategy to alter the structure and the quality parameters of meat and meat products. Benefits and possibilities of the technologies are presented, as well as how to overcome undesired product changes caused by HPP. The use of hydrodynamic shockwaves is briefly described and a promising newly developed industrial prototype for the generation of shockwaves by underwater explosion is presented.

Kinetics of the formation of radicals in meat during high pressure processing.

The kinetics of the formation of radicals in meat by high pressure processing (HPP) has been described for the first time. A threshold for the radicals to form at 400 MPa at 25 °C and at 500 MPa at 5 °C has been found. Above this threshold, an increased formation of radicals was observed with increasing pressure (400-800 MPa), temperature (5-40 °C) and time (0-60 min). The volume of activation (ΔV(#)) was found to have the value -17 ml mol(-1). The energy of activation (E(a)) was calculated to be 25-29 kJ mol(-1) within the pressure range (500-800 MPa) indicating high independence on the temperature at high pressures whereas the reaction was strongly dependent at atmospheric pressure (E(a)=181 kJ mol(-1)). According to the effect of the processing conditions on the reaction rate, three groups of increasing order of radical formation were established: (1) 55 °C at 0.1 MPa, (2) 500 and 600 MPa at 25 °C and 65 °C at 0.1 MPa, and (3) 700 MPa at 25 °C and 75 °C at 0.1 MPa. The implication of the formation of radicals as initiators of lipid oxidation under HPP is discussed.

Purification and characterisation of proteases A and D from Debaryomyces hansenii.

The proteases A (PrA; EC. 3.4.23.25) and D (PrD; EC. 3.4.24.37) of Debaryomyces hansenii CECT 12487 were characterised after their isolation by fractionation with protamine sulfate followed by three chromatographic separations, which included two anion exchange and one gel filtration chromatographic steps. The whole procedures for PrA and PrD resulted in 1349 and 2560 purification-fold with a recovery yield of 1.4 and 1.3%, respectively. PrA was active at acidic-neutral pH with an optimum pH between 5.0 and 6.0. PrD was active at neutral-basic pH with an optimum pH between 7.0 and 8.0. The molecular mass of the native PrA was 55 kDa and (being) 42 kDa in denaturing conditions. Polyclonal-antibodies raised against PrA from Saccharomyces cerevisiae cross-reacted with the corresponding PrA from D. hansenii. PrD showed a native molecular mass of 68 kDa and 65 kDa in denaturing conditions. PrA was an aspartic protease effectively inhibited by pesptatin A while PrD was classified as a metallo protease inhibited by 1,10-phenantroline and affected by some divalent cations such as zinc, cadmium and magnesium. The homology of the PrA to the lisosomal cathepsin D suggests its possible participation in the ripening of fermented meat products.

Protease (PrA and PrB) and prolyl and arginyl aminopeptidase activities from Debaryomyces hansenii as a function of growth phase and nutrient sources.

The effects of nutrient sources and growth phase of Debaryomyces hansenii on the protease (PrA and PrB) and aminopeptidase (prolyl-[PAP] and arginyl-[AAP] aminopeptidases) activities were investigated. These activities were also monitored during growth on a whole sarcoplasmic muscle protein extract (WSPE) and on an equivalent medium but free of compounds under 10 kDa (SPE>10 kDa). The levels of specific protease and aminopeptidase activities were higher when cells were grown in urea and dipeptides than when grown in either ammonium or free amino acids as nitrogen sources. The level of each aminopeptidase (PAP or AAP) activity was preferentially induced by its own substrate (ProLeu or LysAla), suggesting a role in the utilization of exogenous peptides. Higher specific activities for all proteolytic enzymes were detected when using acetate as carbon source. The time course experiments carried out on urea or sarcoplasmic protein-containing media revealed an increase in all activities during transition and advanced stages of stationary phase of growth. In muscle protein extracts, the absence of low molecular mass nutrients (SPE>10 kDa) initially induced the production of PrA, PrB, and AAP activities, possibly involved in the breakdown of muscle oligopeptides.

Protease B from Debaryomyces hansenii: purification and biochemical properties.

The protease B (PrB; EC. 3.4.21.48) of Debaryomyces hansenii CECT 12487 was purified by selective fractionation with protamine sulfate followed by three chromatographic separations. The whole procedure resulted in 324-fold purification with a recovery yield of 1.0%. PrB was active at neutral-basic pH ranging from 6.0 to 12.0 with an optimum at pH 8.0. The molecular mass of the denatured enzyme was 30 kDa. Polyclonal-antibodies raised against PrB from Saccharomyces cerevisiae cross-reacted with the corresponding 30-kDa protein from D. hansenii. The serine protease inhibitor 3,4-DCI and sulphydryl group reagents markedly reduced the enzyme activity. The Km against N-succinyl-Leu-Tyr-7-amido-4-methylcoumarin was 1.79 mM. The presence of endogenous inhibitor for PrB was detected in cell-free extracts of D. hansenii although their inhibitory effect was lost after incubation at 25 degrees C for 20 h. PrB was able to hydrolyze muscle sarcoplasmic proteins by in vitro assays. This is the first endopeptidase purified and characterized from the yeast D. hansenii, whose possible contributions to meat fermentation processes are discussed.

Purification and properties of an arginyl aminopeptidase from Debaryomyces hansenii.

A metallo arginyl aminopeptidase (EC 3.4.11.6) activated by Co(2+) was isolated from Debaryomyces hansenii CECT 12487. The enzyme was purified after precipitation with protamine sulphate, followed by a weak anion exchange chromatography, gel filtration chromatography and a strong anion exchange chromatography. The arginyl aminopeptidase (AAP) was purified 337 folds, with a 18% recovery. The AAP appeared to be a dimer with a molecular mass of 101 kDa. The enzyme was active in the pH range from 6 to 9. The optimal activity was detected at pH 7.0 and at 37 degrees C. AAP activity was inhibited by typical aminopeptidase inhibitors (puromycin and bestatin), reducing agents (DTT), chelating agents (EDTA, EGTA and phenantroline) and sulphydryl groups reagents (iodoacetate). Ca(2+), Mn(2+) and Co(2+) activated the enzyme, while Cu(2+), Cd(2+), Hg(2+) and Mg(2+) inhibited it. The K(m) values calculated for Arg-AMC (7-amido-4-methylcoumarin) and Leu-AMC were 0.071 and 0.094 mM, respectively. The enzyme showed maximum specificity for basic amino acids (Arg and Lys), but was also able to hydrolyze non-charged amino acids (Leu, Met and Ala) and, at a minor rate, aromatic amino acids (Phe and Tyr). AAP showed higher activity when an acid residue was located at the C-terminal position of dipeptides. The described purification of an arginyl aminopeptidase from the yeast D. hansenii can contribute to the lack of knowledge about the exopeptidase activity in one of the yeasts more frequently isolated in sausage and to understand its role during the ripening of a fermented sausage.

Purification and characterization of a prolyl aminopeptidase from Debaryomyces hansenii.

A prolyl aminopeptidase (PAP) (EC 3.4.11.5) was isolated from the cell extract of Debaryomyces hansenii CECT12487. The enzyme was purified by selective fractionation with protamine and ammonium sulfate, followed by two chromatography steps, which included gel filtration and anion-exchange chromatography. The PAP was purified 248-fold, with a recovery yield of 1.4%. The enzyme was active in a broad pH range (from 5 to 9.5), with pH and temperature optima at 7.5 and 45 degrees C. The molecular mass was estimated to be around 370 kDa. The presence of inhibitors of serine and aspartic proteases, bestatin, puromycin, reducing agents, chelating agents, and different cations did not have any effect on the enzyme activity. Only iodoacetate, p-chloromercuribenzoic acid, and Hg(2+), which are inhibitors of cysteine proteases, markedly reduced the enzyme activity. The K(m) for proline-7-amido-4-methylcoumarin was 40 micro M. The enzyme exclusively hydrolyzed N-terminal-proline-containing substrates. This is the first report on the identification and purification of this type of aminopeptidase in yeast, which may contribute to the scarce knowledge about D. hansenii proteases and their possible roles in meat fermentation.