Cold storage of porcine plasma treated with microbial transglutaminase under high pressure. Effects on its heat-induced gel properties

The objective of this work was to study the heat-induced gelling properties, at acid pH, of porcine plasma previously treated with microbial transglutaminase (MTGase) under high pressure (HP), when kept under refrigeration conditions for different times (setting time). The results indicated that, although the cross-linking activity of MTGase was enhanced under pressure, consequently, improving the thermal gel texture, the most significant effects, particularly on gel hardness, were obtained by keeping the treated plasma solutions under refrigeration for at least 2 h before gelation. On the whole, under such conditions, increases of approximately 60% of this textural parameter, calculated on the basis of the values corresponding to the heat-induced non-treated plasma gels at pH 5.5, were achieved. However, from the SDS–PAGE profiles, it can be suggested that mechanisms other than polymerisation by MTGase explain the beneficial effects of the treated plasma cold storage on gel texture. In contrast, the setting time had no effects on the water-holding capacity of heat-induced plasma gels at acid pH value, although this gel property was slightly enhanced by submitting porcine plasma solutions to the combined treatment (MTGase plus HP), with improvements being in accordance with the better-structured network of these heat-induced plasma gels.     

Effect of transglutaminase on heat-induced gel properties at acid pH of mixtures of plasma and haemoglobin hydrolysate from porcine blood

The purpose of the present study was to determine the effects of treating protein solutions prepared by mixing different percentages of haemoglobin hydrolysate and plasma from porcine blood with microbial transglutaminase (MTGase) on the properties of thermal-induced gels at pH 5.5. Solutions were adjusted to pH 7.0 just before their incubation with different MTGase concentrations. After the enzymatic treatment, solutions were lowered to pH 5.5 and submitted to the thermal gelation process. The results indicate an overall better gelling ability of the lowest HbH- content mixtures relative to those containing the highest HbH-content ones, with a clearly different behaviour regarding the MTGase effects.     

Effect of pH on the gel properties and secondary structure of fish myosin

The relationships between gel properties and the secondary structures of silver carp myosin were investigated at pH 5.5–9.0 using dynamic rheological measurement, circular dichroism and scanning electron microscopy. The gel properties of fish myosin were strongly pH and temperature dependent. During heating at 1 °C/min, myosin formed gels in the pH range 5.5–7.5, but not at pH 8.0–9.0. α-Helix was the predominant structure at pH 7.0. The α-helix fraction declined with increasing temperature and the pH away from 7.0, whilst the other secondary structure fractions increased. The α-helix structure of myosin was more susceptive to acid-treatment than alkali-treatment. As pH increased, the gelation rate and gel strength decreased, and the water-holding capacity (WHC) showed an increasing trend followed by a plateau. High β-sheet and β-turn fractions prior to heating could improve G′ at 90 °C, but they depressed the WHC. A compact and uniform gel of fish myosin was obtained at pH 7.0.     

Effects of combining microbial transglutaminase and high pressure processing treatments on the mechanical properties of heat-induced gels prepared from arrowtooth flounder (Atheresthes stomias)

The objective of this study was to evaluate the effect of setting conditions (25 °C for 2 h or 40 °C for 30 min) and combining of microbial transglutaminase (MTGase) and high pressure processing (HPP) on the mechanical properties of heat induced gels obtained from paste from arrowtooth flounder (Atheresthes stomias). Treatments included fish paste control without added MTGase, fish paste incubated with MTGase but not pressurized (MTGase + cooking), fish paste incubated with MTGase and pressurized at 600 MPa for 5 min (MTGase + HPP + cooking) and fish paste pressurized at 600 MPa for 5 min and incubated with MTGase (HPP + MTGase + cooking). The controls and the treated samples were then subjected to one of two thermal treatments: 90 °C for 15 min or 60 °C for 30 min before cooking at 90 °C for 15 min. Samples of fish paste heated at 60 °C before cooking could not be used to prepare gels for texture profile analysis (TPA). TPA showed that pressurization improved the mechanical properties of gels made from paste treated with MTGase and set at 25 °C. The opposite was observed for samples set at 40 °C. Setting at 40 °C appeared to induce proteolytic degradation of myofibrillar proteins.     

Functional Properties of Heat Induced Gels from Liquid and Spray-Dried Porcine Blood Plasma as Influenced by pH

The properties of gels obtained from porcine blood plasma were studied under different pH conditions. Gels from liquid and spray-dried plasma were prepared and analyzed for water holding capacity (WHC), texture, and microstructure at pH 7.4, 6, 5.5 and 4.5. The denaturation extent of proteins was also determined by differential scanning calorimetry (DSC). All properties studied were dependent on pH. The WHC and consistency of gels decreased when pH decreased. These results correlated with microstructural changes observed by SEM. Spray drying affected the consistency of gels. The penetration force of the gel from dehydrated plasma was always lower than that prepared from liquid plasma where the pH was the same, but neither the WHC nor the micro-structure of gels were affected.     

Gelling properties of white shrimp (Penaeus vannamei) meat as influenced by setting condition and microbial transglutaminase

The properties of white shrimp (Penaeus vannamei) gel added with different levels of microbial transglutaminase (MTGase) and subjected to setting at 25 °C for 2 h or 40 °C for 30 min, prior to heating at 90 °C for 20 min were studied. Breaking force of gels with and without setting increased with increasing MTGase amount added (P<0.05). However, no changes in deformation in all samples were noticeable (P>0.05). Directly heated gels showed the lower breaking force than those with prior setting at all MTGase levels added (P<0.05). Generally, gels prepared by setting at 25 °C exhibited the greater breaking force than those set at 40 °C, possibly associated with the appropriate protein structure for cross-linking at 25 °C and greater degradation at 40 °C as evidenced by a greater trichloroacetic acid soluble peptide content (P<0.05). Sodium dodecyl sulfate polyacrylamide gel electrophoretic study revealed that myosin heavy chain (MHC) underwent polymerization to a higher extent in the presence of MTGase, but the strengthening effect on gel was dependent on setting temperature. Regardless of setting condition, microstructure of gel added with MTGase was finer with a smaller void, compared with those of gel without MTGase. Therefore, setting temperature played an essential role in gel property of white shrimp meat added with MTGase.     

Freshness assessment of ray fish stored in ice by biochemical,chemical and physical methods

Ray fish were caught, filleted, and stored in ice. Fillets were analysed for 18 days to determine the chemical, biochemical and physical changes and their relation to the muscle eating quality. Trimethylamine (TMA-N), total volatile bases (TVB-N), ATP content and breakdown products, K value, pH, texture, water-holding capacity (WHC) and colour changes were monitored. At the beginning of the study, the ray fish muscle showed a low concentration of ATP and a high value of inosine 5′-monophosphate (IMP). Regarding to the signs of freshness and deterioration, K value presented an exponential increase (r2 = 0.95) with an initial value of 4.7% and a final value of 47.5%. Furthermore, the TBV-N and TMA-N significantly increased (P < 0.05) during the storage in ice. As for the physical analysis whereas the texture changed (P < 0.05); pH and the WHC were not affected (P < 0.05). The overall results of this study indicated that the edible quality of ray fish muscle was maintained during at least 15 days of ice storage.     

Postmortem changes in cazon fish muscle stored on ice

The biochemical, chemical and physical postmortem changes and their relation to the cazon muscle eating quality were studied during an 18 day storage period at 0 °C (ice). Content of ATP and breakdown products, K value, pH, trimethylamine (TMA-N), total volatile bases (TVB-N), water-holding capacity (WHC), colour and texture changes were examined. At the beginning of the study, the cazon muscle showed a low concentration of ATP and a high value of IMP. Regarding to the signs of freshness and deterioration, K value presented a linear increase (r² = 0.97) with an initial value of 1.05% and a final value of 58.9%. The TBV-N and TMA-N significantly increased (P < 0.05). As for the physical analysis whereas the pH and the WHC changed (P < 0.05); texture was not affected (P < 0.05). The overall results of this study indicated that the edible quality of cazon fish muscle was maintained during at least 18 days of ice storage.     

Characterisation of a thermostable family 42 β-galactosidase (BgalC) family from Thermotoga maritima showing efficient lactose hydrolysis

A β-galactosidase gene (TM_1195) of Thermotoga maritima was cloned and expressed in Escherichia coli. The recombinant β-galactosidase (BgalC), belonging to glycosyl hydrolase (GH) family 42, was purified to homogeneity with 23.4-fold purification and a recovery of 36.6%. Its molecular mass was estimated to be 78 kDa by SDS–PAGE. BgalC exhibited maximum activity at an optimal pH of 5.5 and an optimum temperature of 80 °C. The enzyme displayed important properties, such as stability over a broad pH range of 5.0–9.0 and thermostability up to 75 °C. K_m values of BgalC for p-nitrophenyl-β-galactopyranoside (pNPGal), o-nitrophenyl-β-galactopyranoside (oNPGal) and lactose were 1.21, 7.31 and 6.5 mM, respectively. BgalC was efficient in complete removal of lactose from milk. BgalC is significantly one of the few β-galactosidases from family 42 displaying significant hydrolysis of lactose. These properties make BgalC an ideal candidate for commercial use, in the production of lactose-free milk.     

Proteolysis development in enzyme-modified Cheddar cheese using natural and recombinant enzymes of Lactobacillus rhamnosus S93

Proteolysis in enzyme-modified cheese was investigated with natural crude enzyme or recombinant aminopeptidase, both derived from Lactobacillus rhamnosus S93 in the presence of a commercial proteinase, Neutrase. For production of enzyme-modified cheeses, a cheese slurry was produced and pre-incubated with Neutrase. Natural enzyme or recombinant aminopeptidase (50 units 200 g⁻¹ slurry) was added alone or in combination to the cheese slurries, which were then incubated anaerobically under vacuum at 37 °C for 1, 3 and 6 d. The greatest levels of phosphotungstic acid soluble nitrogen and free amino acids were observed in the enzyme-modified cheese containing natural enzyme followed by the one treated with a combination of the natural enzyme and recombinant aminopeptidase. The enzyme-modified cheese containing the recombinant aminopeptidase alone resulted in the complete disappearance of proline after 1 d of maturation time.     

Characterisation of lipoxygenase from pea seeds (Pisum sativum var. Telephone L.)

Lipoxygenase (LOX) from pea seeds (Pisum sativum var. Telephone L.) was extracted and studied of biochemical properties. The molecular mass of purified lipoxygenase was 93 kDa. The effects of substrate specificity, pH, and sensibility to various inhibitors: caffeic acid, ferulic acid, benzoic acid, catechin, quercetin and kaempferol of LOX were investigated. Lipoxygenase showed the highest activity toward linoleic acid and the lowest toward oleic acid as substrates. Kinetic studies indicated that V_max of the LOX activity was 151.5 U/min and corresponding K_m value of 0.44 × 10⁻³ M. Optimum pH of lipoxygenase was reported at 5.5. Caffeic acid was the most effective inhibitor and kaempferol was the least effective.     

Transglutaminase treatment of pea proteins: Effect on physicochemical and rheological properties of heat-induced protein gels

Rheological properties of heat-induced pea protein isolate (PPI) gels with added microbial transglutaminase (MTGase) were studied under various reaction conditions. A positive linear relationship was observed between level of MTGase used (0 to 0.7% w/w) and shear stress and shear strain of heat-set commercial pea protein isolate (PPIc) gels at 92 °C following incubation at 50 °C. Use of MTGase allowed for preparation of PPIc gels of similar strength and elasticity as commercial soy protein isolate gels and commercial meat bologna. MTGase treatment did not alter thermal properties of PPI gels. The shear stress and strain of PPIc gels were also improved following low temperature (4 °C) incubation of PPI with MTGase. Enhancement of shear strain or gel elasticity of heat-induced PPI gels with MTGase has not been reported before and provides opportunities for extending the properties of pea proteins when developing new food products.     

Emulsifying and foaming properties of Phaseolus vulgaris and coccineus proteins

Protein isolates from two Phaseolus cultivars, common bean (Phaseolus vulgaris L.) and scarlet runner bean (Phaseolus coccineus L.), were prepared by wet extraction methods (isoelectric precipitation – 4000 rpm, ultrafiltration, extraction with NaCl 2%, and isoelectric precipitation – 9900 rpm). The protein isolates were characterized by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and then evaluated for their solubility. The emulsion stability of emulsions produced at pH 7.0 and 5.5 with 1% or 2% or 3% w/v protein isolate was evaluated by average droplet size diameter, viscosity and creaming measurements. Emulsions with 1% protein content were unstable through storage. Emulsions with 3% w/v protein isolate concentration, extracted by ultrafiltration at pH 5.5 from both cultivars, were flocculated; this was more pronounced for coccineus isolates. The foaming properties, for the respective foams, were investigated. Foams with 1% w/v protein showed little foaming ability Ultrafiltration isolates produced more foam, which was especially stable at pH 5.5.     

The impact of concentration,temperature and pH on dynamic rheology of psyllium gels

Structural aspects of the psyllium gum prepared from the seed husk of the plant of Plantago ovata Forsk was characterized by dynamic rheology and microscopy. Dynamic rheological properties of psyllium gel in the linear viscoelastic region, as a function of concentration (2, 2.5 and 3% w/w), temperature (5–95 °C) and pH (2.5–10) were investigated. Mechanical spectra of the psyllium gels were obtained by frequency sweep measurement classified into that of weak gels because G′ was larger than G″ throughout the tested frequency range and the separation of the two moduli (tan δ) was greater than 0.1. The phase angle increased with temperature and a peak associated with gel melting appeared at about 40 °C. All gels at different pH presented a typical weak gel spectrum. Scanning electron microscopy showed porous structures with different pore-size distribution for psyllium gels under different conditions in terms of concentration, pH and temperature.     

Role of general stress-response alternative sigma factors σ(S) (RpoS) and σ(B) (SigB) in bacterial heat resistance as a function of treatment medium pH

This investigation aimed to determine the role of general stress-response alternative sigma factors σ^S (RpoS) and σ^B (SigB) in heat resistance and the occurrence of sublethal injuries in cell envelopes of stationary-phase Escherichia coli BJ4 and Listeria monocytogenes EGD-e cells, respectively, as a function of treatment medium pH. Given that microbial death followed first-order inactivation kinetics (R² > 0.95) the traditional D_T and z values were used to describe the heat inactivation kinetics.Influence of rpoS deletion was constant at every treatment temperature and pH, making a ΔrpoS deletion mutant strain approximately 5.5 times more heat sensitive than its parental strain for every studied condition. Furthermore, the influence of the pH of the treatment medium on the reduction of the heat resistance of E. coli was also constant and independent of the treatment temperature (average z value = 4.9 °C) in both parental and mutant strains.L. monocytogenes EGD-e z values obtained at pH 7.0 and 5.5 were not significantly different (p > 0.05) in either parental or the ?sigB deletion mutant strains (average z value = 4.8 °C). Nevertheless, at pH 4.0 the z value was higher (z = 8.4 °C), indicating that heat resistance of both L. monocytogenes strains was less dependent on temperature at pH 4.0. At both pH 5.5 and 7.0 the influence of sigB deletion was constant and independent of the treatment temperature, decreasing L. monocytogenes heat resistance approximately 2.5 times. In contrast, the absence of sigB did not decrease the heat resistance of L. monocytogenes at pH 4.0.The role of RpoS in protecting cell envelopes was more important in E. coli (4 times) than SigB in L. monocytogenes (1.5 times). Moreover, the role of σ^S in increasing heat resistance seems more relevant in enhancing the intrinsic resilience of the cytoplasmic membrane, and to a lesser extent, outer membrane resilience.Knowledge of environmental conditions related to the activation of alternative sigma factors σ^S and σ^B and their effects on heat resistance would help us to avoid and/or identify situations that increase bacterial stress resistance. Therefore, more efficient food preservation processes might be designed.     

Physicochemical and functional changes in jumbo squid (Dosidicus gigas) mantle muscle during ice storage

Physicochemical and functional changes of jumbo squid (Dosidicus gigas) mantle muscle stored in ice for 15 days were evaluated. Proximate analysis, pH, protein solubility (PS) and SDS-PAGE profile changes were monitored for muscle proteins, while texture profile analysis (TPA) (gel strength, elasticity and cohesiveness), water holding capacity (WHC) and colour changes were monitored for gels produced from this muscle. Fresh muscle chemical composition varied according to fishing season and/or physiological status presented by specimens. pH and PS were stable (p ? 0.05) throughout the study with a slight tendency to decrease towards the end of study. SDS-PAGE revealed no drastic changes in proteins influencing gelation (i.e., myosin). TPA and WHC of gels decreased towards the end of experiment; however, due to sampling variability, no significant differences (p ? 0.05) were found over storage time. Changes in gel colour were observed with hue angle decreasing towards yellow hues. Improved post-capture management of mantle muscle preserved its integrity and functionality for up to two weeks.     

Simultaneous application of microbial transglutaminase and high hydrostatic pressure to improve heat induced gelation of pork plasma

The effects of treating porcine plasma with microbial tranglutaminase (MTGase) under high hydrostatic pressure (HHP) were studied as a means of improving its gel-forming properties when subsequently heated at pH 5.5, near the pH of meats. Plasma containing varying levels of commercial MTGase was pressurized (400MPa, room temperature, pH 7) for different times, and adjusted to pH 5.5 prior to heating to induce gelation. MTGase-treatment under HHP led to greater enhancement of heat-induced plasma gel properties as compared to control samples. The greatest improvements were achieved by pressurising plasma with 43.3U MTGase/g protein for 30min, thereby achieving recoveries of 49% and 63% in fracture force (gel strength) and fracture distance (gel deformability) of the subsequently heat-induced gels, respectively, relative to gel properties obtained by heating untreated plasma at physiological conditions (pH 7.5).     

Effect of different gas stunning methods on Manchega suckling lamb meat packed under different modified atmospheres

Forty-nine Manchega breed male suckling lambs were used in this experiment. The effect of CO₂ concentration and exposure time at stunning [80% CO₂ for 90 s (G1); 90% CO₂ for 90 s (G2); 90% CO₂ for 60 s (G3); 80% CO₂ for 60 s (G4)] plus an electrically stunned control group (G5) was assessed for pH, colour (L^∗, a^∗, b^∗, C^∗ and h^∗), water holding capacity (WHC), drip loss (DL), cooking loss (CL) and shear force (SF) in samples packed under two different types of modified atmospheres (MA: MA A: 70%O₂ + 30%CO₂; MA B: 69.3%N₂ + 30%CO₂ + 0.7%CO) at 7, 14 and 21 d post-packaging. The lowest pH was found in G4 and in G5. The highest WHC and the lowest CL were found in G2 and G3 groups (P < 0.05). Modified atmospheres did not affect on pH, WHC, CL and DL, although a significant effect (P < 0.001) on colour was found at all the analysis times. Both the type of stunning and the modified atmosphere affected SF values.     

Mechanical and gelling properties of comminuted sausages containing chicken MDM

The present investigation focuses on the physicochemical properties, rheological behaviors and texture of raw and cooked emulsions containing different mechanically deboned meat (MDM) from chicken. MDMs were produced from the neck, backs and thighs. The texture and mechanical profile analyses were performed using a small deformation dynamic oscillation in a shear and instrumental texture analyzer. The mechanical spectra of the raw and cooked MDM emulsion gels were classified as weak gels based on their frequency sweep and tan δ results. Both the G′ and G″ values increased with increasing frequency in the temperature of 4 and 10 °C. The MDM from thighs had the lowest water holding capacity (WHC) and emulsifying (EC) values, while the MDM from backs had significant values of 2.41 and 128.87, respectively. The cooked emulsion containing backs showed the highest hardness and cohesiveness values, while the emulsion containing necks and thighs had the lowest texture parameters.