Effects of protein oxidation on the texture and water-holding of meat: a review

Abstract

Protein oxidation readily occurs in postmortem muscle during storage and processing. Over the past decade new analytical methods have been developed and new aspects of protein oxidation in meat have been studied, such as the reaction mechanism, and impacts on eating quality and nutritional value. It is now evident that amino acid side chains in myofibrillar proteins undergoes modifications due to oxidative stress. In turn this will lead to formation of new protein-protein cross-links in structural proteins, however, also the overall level of fixed-charge groups attached to the peptide backbones is modified. Meat texture and water-holding are important quality attributes and they are affected by the oxidation of structural proteins. Different mechanisms have been suggested to explain the oxidation-induced quality changes, focusing mainly on reduced proteolysis and formation of cross-links. This review explores the current understanding of protein oxidation in fresh meat in relation to texture and water-holding. The consequences of protein oxidation at molecular level in relation to oxidation-induced cross-linking and changes in net charges of myofibrillar proteins, and the impacts on texture and water-holding are discussed.     

Characterization of Textural Properties and Changes of Myofibrillar and Sarcoplasmic Proteins in Salame Felino During Ripening

Textural properties and changes of myofibrillar and sarcoplasmic proteins of salame Felino, a typical Italian dry-cured fermented sausage, were studied during ripening. Hardness, chewiness, and gumminess increased over the ripening period. Adhesiveness showed a gradual but significant decrease after 14 days and further at 28 days of ripening. A remarkable increase of the proteolysis index and a significant decrease of myofibrillar and sarcoplasmic protein solubility were observed. Hardness showed a significant negative correlation with myofibrillar protein solubility. Sarcoplasmic proteins appeared to be more susceptible to degradation.     

Properties of refrigerated ground beef treated with potassium lactate and sodium diacetate

BACKGROUND: Ground meat is product that perishes easily and therefore various preservatives are applied to prolong its shelf life. The aim of this study was to investigate the effect of potassium lactate and sodium diacetate (Protec K‐DI preparation) on general proteolytic activity, protein degradation, texture, colour and water‐holding capacity of vacuum‐packaged ground beef. RESULTS: The activity of endogenous proteolytic enzymes, and hence the rate of protein hydrolysis changed significantly (P < 0.05) during refrigerated storage. The proteolysis was more intensive in the meat with the preservative than in a control sample. According to the results of SDS‐PAGE, the level of myosin in ground beef decreased with increasing storage time, whereas the preservative did not have a significant effect (P > 0.05) on these changes. However, its impact was the most significant in the case of tropomyosin and troponin T content, as well as changes of 30 kDa protein content. The preservative as well as the increase in storage time influenced an increase in content of polypeptides, peptides and some amino acids in the samples, and had an inhibiting effect on unfavourable changes in ground beef hardness. The preservative addition allowed the meat to maintain desired meat colour, and the main pigment during refrigerated storage was oxymyoglobin (MbO₂). CONCLUSION: Results showed that the Protec K‐DI preparation can be useful in minced meat production as a good stabiliser of colour and texture despite the increase observed in proteolytic activity after the preservative addition. Copyright © 2007 Society of Chemical Industry     

Changes in the ultrastructure and texture of prawn muscle (Macrobrachuim rosenbergii) during cold storage

Changes in the ultrastructure and proteolytic activity of prawn muscle were determined during storage at 5 °C, in order to better understand changes in physical and sensory texture measurements. Progressive deterioration of myofibril structure was observed during refrigeration of prawn for 14 d. The loss of density and order in Z-line alignment was first detected after 3 d of storage. Progressive disruption of Z-line, I-bands and M-lines was observed after 4-6 d of storage. Muscle degradation included pronounced disruption of the mitochondria as revealed by swollen cristae, loss of cristae material, and membrane breakage. Along with ultrastructural changes, decreased shear force values and mean textures scores were measured. An initial shear force value of 18.21 N/g decreased to 14.50, 12.46, and 10.79 N/g on days 3, 6, and 14, respectively. Mean texture scores indicated that prawn muscle maintained firm texture during 0-3 d of storage, and became soft during 4-6 d of storage. After 6 d of storage, the prawn texture was very soft. Increased deterioration of the muscle ultrastructure coincided with the increase of proteolytic microorganisms and salt soluble muscle proteins found by Sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It is concluded that the weakening of the ultrastructure is related to proteolytic activity and results in a more soft texture in prawns.     

Protein oxidation in emulsified cooked burger patties with added fruit extracts: Influence on colour and texture deterioration during chill storage

The influence of protein oxidation, as measured by the dinitrophenylhydrazine (DNPH) method, on colour and texture changes during chill storage (2 °C, 12 days) of cooked burger patties was studied. Extracts from arbutus-berries (Arbutus unedoL., AU), common hawthorns (Crataegus monogynaL., CM), dog roses (Rosa caninaL., RC) and elm-leaf blackberries (Rubus ulmifoliusSchott., RU) were prepared, added to burger patties (3% of total weight) and evaluated as inhibitors of protein oxidation and colour and texture changes. Negative (no added extract, C) and positive control (added quercetin; 230 mg/kg, Q) groups were also considered. The significant increase of protein carbonyls during chill storage of control burger patties reflect the intense oxidative degradation of the muscle proteins. Concomitantly, an intense loss of redness and increase of hardness was found to take place in burger patties throughout refrigerated storage. Most fruit extracts as well as Q significantly reduced the formation of protein carbonyls and inhibited colour and texture deterioration during chill storage. Likely mechanisms through which protein oxidation could play a major role on colour and texture changes during chill storage of burger patties are discussed. Amongst the extracts, RC was most suitable for use as a functional ingredient in processed meats since it enhanced oxidative stability, colour and texture properties of burger patties with no apparent drawbacks.     

Raman spectroscopic study of the effects of microbial transglutaminase on heat-induced gelation of pork myofibrillar proteins and its relationship with textural characteristics

The effects of microbial transglutaminase (MTG) on heat-induced gelation of pork myofibrillar proteins (PMP) structural changes, textural properties were studied by Raman spectroscopy and texture profile analysis (TPA), respectively. And the relationships between the structural changes and textural characteristics were estimated by principal component analysis (PCA). Changes in the Raman spectra were interpreted as the occurrence of secondary structural changes in myofibrillar proteins with MTG added. Modifications in the amide I (1600-1700 cm^− 1) regions indicated a significant (p < 0.05) decrease in ??-helix content, accompanied by a significant (p < 0.05) increase in ??-sheets, ??-turns and random coil content due to the addition of the enzyme. Obvious texture property changes were also determined by TPA. All these changes showed a strong, irreversible heat-induced gel formed due to the addition of MTG. The application of a dimensionality reducing technique such as PCA proved to be useful to determine the most influential properties of heat-induced gel. Significant (p < 0.05) correlations were found between these structural changes and the textural characteristics (hardness) in PMP system with the addition of MTG by PCA. The hardness was related positively to fraction of ??-sheet, ??-turns and random coil, and negatively to normalized intensity of 760 cm^− 1 and fraction of ??-helix. The samples are closely grouped in a cluster defined by level of MTG.     

Broad‐spectrum inhibition of proteolytic enzymes by allicin and application in mitigating textural deterioration of ice‐stored grass carp (Ctenopharyngodon idella) fillets

The inhibitory effect of allicin on proteolytic enzymes and textural deterioration of ice‐stored grass carp (Ctenopharyngodon idella) fillets was investigated. The results of in vitro study showed that allicin inhibited the activity of cathepsin B, L and D, calpain and collagenase in crude extract of grass carp muscle. Among endogenous enzymes, cathepsin B, L and collagenase were the most susceptible to allicin. Proteolysis of myofibrillar proteins by either crude enzyme or cathepsin B and L was almost prevented by allicin when employed at a concentration higher than 100 mm . After storage of 21 days, shear force of fillets treated with allicin at 10–100 mm was 39–51% higher than that of control. Myofibrillar proteins of fillets during storage were well protected against degradation when allicin concentration increased to 100 mm , as evidenced by SDS‐PAGE. Therefore, allicin could be a potential broad‐spectrum inhibitor to retard softening of fish fillets via mitigating myofibrillar proteolysis by endogenous enzymes especially cathepsin B and L during ice‐storage.     

Endogenous proteases in giant freshwater prawn (Macrobrachium rosenbergii): changes and its impacts on texture deterioration during frozen storage

Effects of endogenous enzymes on texture deterioration of Macrobrachium rosenbergii during frozen storage (−18 °C) were investigated. The prawn was pretreated with iodoacetic acid (IAA) solution to inhibit cysteine proteases and/or phenylmethylsulfonyl fluoride (PMSF) solution to inhibit trypsin before frozen storage, and sterile distilled water was used as the control. After 16 weeks of cryopreservation, the shear force of the groups treated with enzyme inhibitors was significantly higher than that of sterile distilled water-treated group (control; P < 0.05), while significantly fewer TCA-dissolved peptides, α-amino nitrogen and Myofibril fragmentation index (MFI) value were observed in all inhibitor-treated groups (P < 0.05). Compared with PMSF treatment, IAA or IAA + PMSF treatment effectively alleviated softening and remarkably reduced the accumulation of TCA-soluble peptides and AAN (α-amino nitrogen) during frozen storage, with a lower MFI value and a higher sodium dodecyl sulphate-polyacrylamide gel electrophoresis band intensity. Therefore, endogenous cysteine proteases in muscle play a crucial part in texture deterioration of giant freshwater prawn during cryopreservation.     

Addition of sardine to hake minces and subsequent effect on dimethylamine and formaldehyde formation

Formaldehyde formation and reaction with muscle proteins in lean fish species during frozen storage is considered to be a major factor affecting texture and functionality deterioration. Formaldehyde formation and reaction with muscle compounds was reduced in lean fish minces and model systems when lipids with different degrees of oxidation were added. In order to increase the lipid content and slow down functional and textural changes, hake (Merluccius merluccius) and sardine (Sardina pilchardus) minces mixed in the ratios 3:1; 1:1 and 1:3 (w/w) were stored at ?20 °C and studied for 1 year. Dimethylamine formation and, by deduction, formaldehyde formation increased. However, less free formaldehyde was detected, probably owing to reaction with muscle compounds in the mixed minces. Nevertheless, addition of sardine minces improved the texture, protein solubility and viscosity of the mixed minces compared with the hake minces. In the mixed lots, formation of large protein aggregates was delayed or prevented. This suggests that in the mixed minces formaldehyde reacted with proteins in a different way from that in lean fish or reacted with other muscle components not directly involved in textural changes. © 2002 Society of Chemical Industry     

Sponge cake microstructure,starch retrogradation and quality changes during frozen storage

The effect of frozen storage (6 months) on the microstructure (microscopy, flatbed scanning and image analysis), quality (texture, moisture and specific volume) and starch retrogradation of the cake crumb were evaluated. After 2 months of storage, texture (firmness, cohesiveness and resilience) was significantly (P < 0.05) affected and starch retrogradation was observed, while by the fourth month, the crystallinity increased and crumb fractures were noticeable. Additionally, the shrinkage of starch granules was observed as the starch circularity (Sc) values significantly decreased (P < 0.05) by the sixth month of storage. Although structural changes were not detected by image analysis, it was demonstrated that cake microstructure damage is related to physical changes because the Sc was significantly correlated (P < 0.05) with moisture and specific volume and therefore with the cake quality and texture. Moreover, sugar re‐crystallisation occurred during frozen storage, and it was significantly correlated (P < 0.05) with the deterioration in cake quality.     

Influence of size distribution of proteins,thiol and disulfide content in whole wheat flour on rheological and chapati texture of Indian wheat varieties

The influence of protein composition, as measured by size-exclusion high performance liquid chromatography (SE-HPLC), on rheological properties and chapati texture was investigated in the whole wheat flours of eight Indian wheat cultivars grown at a single location. Proteins were extracted using two-step procedure: extraction with buffer containing 0.5% SDS (SDS buffer), followed by sonication. The results showed that SDS buffer extracted 72–90% of the total flour protein in different varieties and 7–11% protein was extracted from the remaining residues by sonication. The proteins extracted were fractionated by SE-HPLC into large polymeric proteins (>130 kDa), small polymeric proteins (80–130 kDa) and monomeric proteins (10–80 kDa). Total polymeric protein content in the flour protein showed a significant positive correlation with dough hardness (r = 0.71, p < 0.05) and positive correlation with chapati texture (r = 0.58, p < 0.05). Of the SDS extractable polymeric proteins, large polymeric protein in flour protein had significant positive correlation to dough hardness (r = 0.89, p < 0.05) and chapati cutting force, which reflects the chapati texture (r = 0.70, p < 0.05). Protein disulfide content showed positive correlation to dough hardness (r = 0.66, p < 0.05) and texture of chapati (r = 0.58, p < 0.05) while protein thiol content showed significant negative correlation to chapati texture (r = −0.77, p < 0.05). Thus, the results indicate that high proportion of SDS extractable large polymeric protein in flour protein increases the toughness of chapati texture while flours having high thiol content decrease the toughness of chapati.     

EFFECTS OF MUSCLE PROTEASES, ENDOGENOUS PROTEASE INHIBITORS AND MYOFIBRIL FRAGMENTATION ON POSTMORTEM AGING OF GOAT MEAT

The present study was conducted to evaluate the extent of postmortem proteolysis in longissimus dorsi, biceps femoris, semimembranosus and semitendinosus goat muscles on postmortem aging at an ambient (27C) temperature. The activities of calpains and calpastatin were determined after separation on a (diethylamino)ethyl–Sephacel column (Sigma, St. Louis, MO) and cathepsin (B, B + L and H) by carboxymethyl–Sepharose column (Sigma). The results showed that the decrease in calpain I and calpastatin activities was significantly higher than that of calpain II. Cathepsin B, B + L, H and cystatin were found to fall by 30–80% after 12 h, whereas cathepsin D decreased significantly in all the muscles. The disappearance of titin 1 and nebulin, and the appearance of a 30‐kDa component were confirmed by Western blot analysis. The appearance of the 30‐kDa component reported here explains the time‐induced structural changes of myofibrils. The Z‐line degradation had occurred by 6 h postmortem. Cathepsins are not stable compared to calpains during postmortem aging, and both enzymes may play a significant role in the proteolysis of myofibrillar proteins at ambient temperature.     

Proteins and proteolytic activity changes during refrigerated storage in sea bass (Dicentrarchus labrax L.) muscle after high-pressure treatment

Contrary to other preservation methods like thermal treatments, high pressure can destroy microorganisms without affecting the nutritional quality, color, or food texture. The firm texture of fish flesh is an important quality parameter. During the refrigerated storage, the tissue becomes softer and the muscle is deteriorated by different proteases. The aim of this study was to study the modification of the fish muscle proteins after high-pressure treatment during the refrigerated storage and to evaluate the effect of high-pressure treatment level on the post-mortem protein changes and enzyme activities. The calpain activity decreased with the high-pressure treatment and evolved differently during the refrigerated storage, depending on the level of pressurisation. Its inhibitor, the calpastatin was not affected by high pressure, but its inhibiting potential decreased during the post-mortem storage. The activities of cathepsins were modified by the high-pressure treatment and the time of storage, but depended according to their class. The electrophoresis profiles showed that sarcoplasmic proteins were modified according both the high-pressure treatment and the period of storage. For the myofibrillar proteins, the only changes were due to the high-pressure treatment.     

Influence of inulin/oligofructose on the acid‐induced cold aggregation and gelation of preheated soy proteins

BACKGROUND: In recent years inulin‐type prebiotics have attracted much attention due to consumers' awareness of the health benefits of functional foods. Currently no information is available about the possible texture‐modifying effect of these non‐ionizable polar carbohydrates in different soy‐based food systems. In this study, the effect of inulin/oligofructose on the cold aggregation and gelation of preheated soy protein isolate (SPI) and its fractions (7S, 11S, and their mixture), induced by glucono‐δ‐lactone (GDL), were evaluated by turbidity (A₆₀₀) and dynamic rheological measurements. RESULTS: Oligofructose significantly delayed the aggregation of all protein samples and decreased the end‐point optical density of 11S fraction and SPI. Inulin, a long‐chain fructan, only delayed the aggregation of 7S globulin and reduced the capacity of aggregation (A₆₀₀) of SPI. While oligofructose showed no significant effect, the addition of 5% (w/v) inulin enhanced the gelation of SPI and the 7S/11S mixture, which was demonstrated by the increase in gel storage modulus up to 13.6% and 10.1% (P < 0.05), respectively. CONCLUSION: Inulin was found to enhance the viscoelastic properties of GDL‐induced cold‐set soy protein gels. It is expected that ‘functional’ cold‐set gel products with improved texture can be prepared from preheated soy proteins and inulin. Copyright © 2009 Society of Chemical Industry     

Structural and ultrastructural changes of full-cycle cultured Pacific bluefin tuna (Thunnus orientalis) muscle slices during chilled storage

Background: This study examined the structural and ultrastructural changes of dorsal and ventral muscle tissues of full‐cycle cultured Pacific bluefin tuna (PBT), Thunnus orientalis Temminck & Schlegel 1844, cut into slices simulating sashimi and placed in chilled storage for varying periods. Structural and ultrastructural changes were determined in order to understand the physical texture by breaking strength measurement. Results: Progressive deterioration of myofibril structure was observed during chilled storage (4 °C) of PBT muscle slices over 5 days post mortem . Muscle degradation included detachment between myofibres, detachment of the plasmalemma, disruption of mitochondria, loss of Z‐line density and alignment, cementation of myofibrils, loss of the hexagonal arrangement of thick versus thin myofilaments and migration of subsarcolemmal nuclei to intermyofibrillar spaces. Conclusion: Loss of myofibre‐myofibre adhesion, detachment of the plasmalemma and disruption of other components did not lower the breaking strength of PBT muscle. This provides evidence that the muscle breaking strength of PBT is not only associated with the detachment of myofibres or detachment of the plasmalemma. Other factors that produce cement‐like substances, such as cementation of the myofibrillar components and degradation of the sarcoplasmic reticulum, may also increase breaking strength. Copyright © 2011 Society of Chemical Industry     

Proteome changes in the insoluble protein fraction of bovine Longissimus dorsi muscle as a result of low-voltage electrical stimulation

Changes induced by low-voltage electrical stimulation (ES; 0-95 V for 8 s; 95 V for 32 s) in the insoluble protein fraction of bovine longissimus dorsi (LD) muscle at 1 and 24h post-ES were investigated by proteomics. Protein abundance patterns from ten Norwegian Red (NRF) young bulls were compared, and significant changes due to ES were found by rotation test and partial least square (PLS) regression analyses. Five protein spots showed lower abundance in ES samples at both sampling times, and in addition, 10 proteins at 1 h post-ES and 13 proteins at 24 h post-ES changed significantly in abundance due to ES. Reduced abundance of full-length structural proteins in ES samples indicates an accelerated proteolysis due to ES. Moreover, increased abundance of small heat shock proteins indicates earlier initiation of stress responses due to ES. These findings provide a better understanding of the biochemical processes taking place as a result of ES during post mortem storage of meat.     

Ultrasound Pretreatment as an Useful Tool to Enhance Egg White Protein Hydrolysis: Kinetics,Reaction Model,and Thermodinamics

The impact of ultrasound waves generated by probe‐type sonicator and ultrasound cleaning bath on egg white protein susceptibility to hydrolysis by alcalase compared to both thermal pretreatment and conventional enzymatic hydrolysis was quantitatively investigated. A series of hydrolytic reactions was carried out in a stirred tank reactor at different substrate concentrations, enzyme concentrations, and temperatures using untreated, and pretreated egg white proteins (EWPs). The kinetic model based on substrate inhibition and second‐order enzyme deactivation successfully predicts the experimental behavior providing an effective tool for comparison and optimization. The ultrasound pretreatments appear to greatly improve the enzymatic hydrolysis of EWPs under different conditions when compare to other methods. The apparent reaction rate constants for proteolysis (k₂) are 0.009, 0.011, 0.053, and 0.045 min^?1 for untreated EWPs, and those pretreated with heat, probe‐type sonicator, and ultrasound cleaning bath technologies, respectively. The ultrasound pretreatment also decreases hydrolysis activation (E_a) and enzyme deactivation (E_d) energy, enthalpy (ΔH), and entropy (ΔS) of activation and for the probe‐type sonication this decrease is 61.7%, 61.6%, 63.6%, and 32.2%, respectively, but ultrasound has little change in Gibbs free energy value in the temperature range of 318 to 338 K. The content of sulfhydryl groups and ζ potential show a significant increase (P < 0.05) for both applied ultrasound pretreatments and the reduction of particle size distribution are achieved, providing some evidence that the ultrasound causes EWP structural changes affecting the proteolysis rate.     

Raman spectroscopy study of the structural effect of microbial transglutaminase on meat systems and its relationship with textural characteristics

Raman spectroscopy and texture analysis (TPA) studies were carried out to determine the effect of adding different levels of microbial transglutaminase (MTGase) to meat systems. This addition produced a significant (p < 0.05) increase in hardness, springiness and cohesiveness in the meat systems. Raman spectroscopy analysis revealed the occurrence of secondary structural changes in meat proteins due to MTGase. Modifications in the amide I (1650–1680 cm⁻¹) and amide III (1200–1300 cm⁻¹) regions indicated a significant (p < 0.05) decrease in α-helix content, accompanied by a significant (p < 0.05) increase in β-sheets and turns due to the addition of the enzyme to meat systems. Significant (p < 0.05) correlations were found between these secondary structural changes in meat proteins and the textural properties (hardness, adhesiveness, springiness and cohesiveness) of meat systems.     

Proteolytic regulation of the extent of dietary proteins with skin grape proanthocyanidin and anthocyanidin's interactions

During technological processing, proanthocyanidins and anthocyanidins could be partly lost due to the complexation phenomena, affecting food and beverage nutritional properties, organoleptic properties and health-promoting potentials. A common issue is encountered when processing food and beverage which is binding of phenolics to dietary proteins. The present investigation aims at evaluating the proteolysis contribution, using pure protease (Pepsin, 3000 units g⁻¹), to protein–anthocyanidin and protein–proanthocyanidin interactions. Bovine serum albumin (BSA), ovalbumin (OVA) and casein (CAS) dietary protein models were used. High-performance liquid chromatography coupled to diode array detector (HPLC-DAD) and size exclusion chromatography analyses proved that pepsin treatment significantly (P > 0.05) decreased the ratio of flavonoids’ interaction with tested proteins . The proteolysis reduced anthocyanidin interactions with CAS, OVA and BSA by 64.88%, 57.37% and 42.87% respectively. Similarly, proanthocyanidins interaction with CAS, OVA and BSA were reduced by 34.23%, 13.74% and 2.39% respectively. This study provides the basis to develop innovative technologies to limit protein–flavonoid complexation during food and beverage processing.     

Effects of different <Emphasis Type="Italic">Lactobacillus</Emphasis> and <Emphasis Type="Italic">Enterococcus</Emphasis> strains and chemical acidification regarding degradation of gluten proteins during sourdough fermentation

Dough quality and baking performance of wheat dough are significantly affected by the qualitative and quantitative composition of the gluten. Therefore, the degradation was studied of specific fractions of gluten proteins in sourdough as affected by starter cultures. Doughs were fermented for 0, 5, and 24 h at 30 °C after addition of Lactobacillus sakei, L. plantarum, L. sanfranciscensis or Enterococcus faecalis. Chemically acidified doughs were used as controls. All doughs were analyzed quantitatively for their content of albumins, globulins, gliadins, glutenins, and glutenin macropolymer by means of a combined extraction/HPLC procedure. Protein degradation during sourdough fermentation was primarily due to acidic proteases present in flour. While L. sakei, L. plantarum and L. sanfranciscensis were mostly non-proteolytic, E. faecalis clearly contributed to gluten proteolysis. Single gluten protein types were clearly different in their resistance to proteolytic activities of the dough system and E. faecalis, and, in contrast to total glutenins, the amounts of gluten macropolymer were significantly reduced already after 5 h of incubation. When longer fermentation times were applied, gluten was substantially degraded. The strongest decrease was found for the glutenin fraction leading to an increase of alcohol soluble oligomeric proteins in the gliadin fraction. The extent of the decrease of monomeric gliadins was strongest for the γ-type followed by the α- and the ω-types. This indicates that dough properties residing in specific types of gluten fractions can be influenced by the duration of fermentation and the application of proteolytic strains.