Anti-Oxidative and Anti-Aging Activities of Porcine By-Product Collagen Hydrolysates Produced by Commercial Proteases: Effect of Hydrolysis and Ultrafiltration.

To investigate methods for improving the processing of porcine waste, porcine skin was hydrolyzed using different commercially available proteases (Alcalase, Flavorzyme, Neutrase, Bromeline, Protamex, and Papain) under several optimal conditions. Following enzymatic hydrolysis, the collagen hydrolysates (CHs) were fractionated by molecular weight (3 kDa) via membrane ultrafiltration. The CHs were analyzed for physical properties (pH, protein recovery, free amino group content, molecular weight distribution, and amino composition) as well as for functional properties (antioxidant activities and anti-aging activities). Among the CHs, CHs hydrolyzed by Alcalase (CH-Alcalase) exhibited the highest degree of hydrolysis compared to other CHs. Both "CH-Alcalase" and "CH-Alcalase < 3 kDa" fractions showed a considerably high antioxidant activity and collagenase inhibition activity. Therefore, resulting bioactives have potential for development as antioxidants and anti-aging ingredients in the food, cosmetics, and pharmaceuticals, from animal by-products.

Production of low molecular collagen peptides-loaded liposomes using different charged lipids.

To encapsulate water soluble collagen peptides, liposomes loaded with peptides were assembled using a combination of thin film hydration and ultrasonication emulsification techniques. The influence of lipid charge, duration and power of ultrasonication, and collagen peptide concentration were evaluated. Layered liposomes loaded with collagen peptides, charged lipids, chitosan (+) or low-methoxyl pectin (-) were produced using the layer-by-layer electrostatic deposition method. For the liposomes loaded with collagen peptides, the most efficient and dependable manufacturing method was variation of the ultrasonication duration, which was capable of producing smaller sizes (through increasing ultrasonication duration) and liposomes loaded with peptides with >60% encapsulation efficiency. For layered liposomes loaded with collagen peptides, charged lipids were determined to be more effective in the production of smaller liposomes than charged biopolymers. In addition, layered and non-layered liposomes loaded with peptides with a particle size <100nm were physically stable during storage, regardless of storage temperature and time.

Retraction Note to: Effect of High Pressure Homogenization on the Physicochemical Properties of Natural Plant-based Model Emulsion Applicable for Dairy Products.

[This retracts the article DOI: 10.5851/kosfa.2015.35.5.630.].

Effect of Sub- and Super-critical Water Treatment on Physicochemical Properties of Porcine Skin.

Super- and sub-critical water treatments have been of interest as novel methods for protein hydrolysis. In the present study, we studied the effect of sub-critical water (Sub-H2O, 300℃, 80 bar) treatment as well as super-critical water (Super-H2O, 400℃, 280 bar) treatment on the physicochemical properties of porcine skin (PS), which has abundant collagen. Porcine skin was subjected to pre-thermal treatment by immersion in water at 70℃, and then treated with sub- or super-critical water. Physicochemical properties of the hydrolysates, such as molecular weight distribution, free amino acid content, amino acid profile, pH, color, and water content were determined. For the molecular weight distribution analysis, 1 kDa hydrolyzed porcine skin (H-PS) was produced by Super-H2O or Sub-H2O treatment. The free amino acid content was 57.18 mM and 30.13 mM after Sub-H2O and Super-H2O treatment, respectively. Determination of amino acid profile revealed that the content of Glu (22.5%) and Pro (30%) was higher after Super-H2O treatment than after Sub-H2O treatment, whereas the content of Gly (28%) and Ala (13.1%) was higher after Sub-H2O treatment. Super-H2O or Sub-H2O treatment affected the pH of PS, which changed from 7.29 (Raw) to 9.22 (after Sub-H2O treatment) and 9.49 (after Super-H2O treatment). Taken together, these results showed that Sub-H2O treatment was slightly more effective for hydrolysis than Super-H2O was. However, both Sub-H2O and Super-H2O treatments were effective processing methods for hydrolysis of PS collagen in a short time and can be regarded as a green chemistry technology.

Changes in Quality Characteristics of Pork Patties Containing Multilayered Fish Oil Emulsion during Refrigerated Storage.

This study was performed to determine the effect of multilayered fish oil (FO) emulsion without or with trans-cinnamaldehyde on pork patties. Multilayered FO (-primary, -secondary, -tertiary) emulsions were prepared using a layer-by-layer deposition technique with Tween 20, chitosan, and low methoxyl pectin, and were added to pork patties at the same concentration. Pork patties were then stored for 20 d in a refrigerator (5℃) to study changes in quality. The results showed that the pH value of all samples significantly decreased but cooking loss increased during storage (p<0.05). However, water-holding capacity and moisture content showed no remarkable difference between treatments and storage periods (p>0.05). All pork patties containing multilayered FO (treated samples) showed higher values for lightness and significantly lower values for yellowness compared to control pork patties (untreated sample). Lipid oxidation was higher in treated pork patties than in control pork patties during storage. In addition, lipid oxidation and total viable bacterial count in pork patties decreased as the number of coating layers increased. However, hardness, cohesiveness, and springiness of all samples showed no significant change during storage (p>0.05) as compared to fresh pork patties. Furthermore, these did not remarkable change with addition of trans-cinnamaldehyde in all pork patties. From our results, we suggest that FO emulsion did not affect the texture characteristics of fresh pork patties, indicating that it could be used to improve the quality of pork patties by contributing high-quality fat such as unsaturated fatty acids.

Effect of Porcine Collagen Peptides on the Rheological and Sensory Properties of Ice Cream.

The effects of low molecular-weight collagen peptides derived from porcine skin were investigated on the physicochemical and sensorial properties of chocolate ice cream. Collagen peptides less than 1 kDa in weight were obtained by sub-critical water hydrolysis at a temperature of 300℃ and a pressure of 80 bar. Ice cream was then prepared with gelatin powder and porcine skin hydrolysate (PSH) stabilizers mixed at seven different ratios (for a total of 0.5 wt%). There was no significant difference in color between the resulting ice cream mixtures. The increase in apparent viscosity and shear thinning of the ice cream was more moderate with PSH added than with gelatin. Moreover, the samples containing more than 0.2 wt% PSH had enhanced melting resistance, while the mixture with 0.2 wt% PSH had the lowest storage modulus at -20℃ and the second highest loss modulus at 10℃, indicating that this combination of hydrocolloids leads to relatively softer and creamier chocolate ice cream. Among the seven types of ice creams tested, the mixture with 0.2 wt% PSH and 0.3 wt% gelatin had the best physicochemical properties. However, in sensory evaluations, the samples containing PSH had lower chocolate flavor scores and higher off-flavor scores than the sample prepared with just 0.5 wt% gelatin due to the strong off-flavor of PSH.

Effects of Ethanol Addition on the Efficiency of Subcritical Water Extraction of Proteins and Amino Acids from Porcine Placenta.

In a previous study, hydrolysates of porcine placenta were obtained and the extraction efficiency for proteins and amino acids was compared between sub- and super-critical water extraction systems; optimum efficiency was found to be achieved using subcritical water (170℃, 10 bar). In this study, the effects of adding ethanol to the subcritical water system were investigated. The lowest-molecular-weight extraction product detected weighed 434 Da, and the efficiency of extraction for low-molecular-weight products was increased when either the concentration of ethanol was decreased, or the extraction time was lengthened from 10 min to 30 min. The highest concentration of free amino acids (approximately 8 mM) was observed following 30 min extraction using pure distilled water. The concentration of free amino acids was significantly lower when ethanol was added or a shorter extraction time was used (p<0.05). Color change of the solution following extraction was measured. There were no significant differences in color between lysates produced with different extraction times when using distilled water (p>0.05); however, using different extraction times produced significant differences in color when using 20% or 50% ethanol solution for subcritical extraction (p<0.05). The range of pH for the hydrolysate solutions was 6.4-7.5. In conclusion, the investigated extraction system was successful in the extraction of ≤ 500 Da hydrolysates from porcine placenta, but addition of ethanol did not yield higher production of low-molecular-weight hydrolysates than that achieved by DW alone.

Effect of NaCl Concentration on the Emulsifying Properties of Myofibrilla Protein in the Soybean Oil and Fish Oil Emulsion.

The aim of the present work was to investigate the effect of NaCl concentration on the emulsifying and rheological properties of porcine myofibrillar protein (MF)-stabilized soybean oil and fish oil emulsion (SO-EMs and FO-EMs). Emulsions (EMs) were prepared from 1% MF with 10% SO or FO at various NaCl concentration (0-0.5 M). The emulsifying ability index (EAI) of the EMs increased with increasing NaCl concentration for both oil types. Conversely, increasing NaCl manifested decrease in the emulsion stability index (ESI). In addition, creaming index (CI) also increased with NaCl concentration. From the microscopic observation, droplets of the EMs were more aggregated at relatively higher NaCl concentrations, especially for FO-EMs. All EMs had a gel-like structure owing to G' > G" from the rheological analysis. Comparing the oil types, the emulsifying capacity of SO-EMs was more stable than that of FO-EMs at all NaCl concentrations as determined from the CI value and microscopic observation. Therefore, it can be concluded that SO-EMs and FO-EMs are more stable at relatively lower concentrations of NaCl. In addition, the dispersed stability of SO-EMs was better than that of FO-EMs at the same concentration of NaCl.

Quality Characteristics of Beef by Different Cooking Methods for Frozen Home Meal Replacements.

Blanching beef for use in home meal replacements (HMR) is an important process that determines the final quality of the beef after the cooking process. Thermal pretreatment also minimizes the change in quality during the main cooking process or storage. In this study, beef samples were washed and sliced, then treated by immersion in boiling water (1-10 min), steaming (1-10 min), or pan-frying in oil (30-240 s). The color after each thermal treatment showed higher L* and b* values and lower a* values compared with the raw beef, except for the pan-frying thermal treatment. The total color difference (∆E) and pH value were significantly increased by panfrying (p<0.05). There was no significant difference in the shear force of the beef samples, except for the sample pan-fried for 210 s. The nutritional content of beef was measured as the moisture, protein, fat, and ash contents, which were 69.96, 16.64, 3.49, and 1.13%, respectively, in raw beef. After thermal treatment, the crude protein and fat contents were increased, whereas the moisture and ash contents decreased. The mineral content, including Na, Mg, Fe, and Ca was highest after pan-frying. The heat treatment decreased microorganisms in all the samples. The total bacteria count in raw beef was 4.5-4.7 Log CFU/g, whereas the bacteria count decreased to 2.2-2.8 Log CFU/g after blanching. Thermophilic bacteria, coliform, mold, and yeast not detected in any thermally treated sample.

Effect of Soy Protein Hydrolysates Prepared by Subcritical Water Processing on the Physicochemical Properties of Pork Patty during Chilled Storage.

The present study was carried out to investigate the effects of soy protein hydrolysates (SPHs) addition on the quality characteristics of pork patties. The SPHs was prepared by subcritical water process (SWP) at 180℃ without holding time and mixed with the pork patty components at varying concentrations (0-3%), and the patties were stored at 4℃ for 14 d. As quality parameters, instrumental color, thiobarbituric acid-reactive substances (TBARS), pH, water holding capacity (WHC) and shear force were measured at the end of storage. Regardless of SPHs concentration, the addition of SPHs significantly manifested low L* and high a* values compared to those of untreated control (p<0.05). For b* value, addition of SPHs in the 0.5-1.5% was unaffected, while >2.0% of SPHs caused significantly lower b* than control (p<0.05). The color changes in pork patties with and without SPHs were also identified in visual appearance where the pork patties containing 0.5-2.0% showed bright red color which was comparable to brownish color of control and patties containing >2.5% SPHs. Lipid oxidation was delayed by the addition of 0.5-1.5% SPHs, while it was accelerated by the addition of 3% SPHs. The pH of patties increased with increasing concentration of SPHs, whereas there were no significant differences in WHC and shear force of patties. Consequently, the results indicated that the addition of 0.5-1.5% SPHs had a potential advantage in suppressing oxidative deterioration of fat-containing meat products during chilled storage.

Effect of High Pressure Homogenization on the Physicochemical Properties of Natural Plant-based Model Emulsion Applicable for Dairy Products.

In the dairy industry, natural plant-based powders are widely used to develop flavor and functionality. However, most of these ingredients are water-insoluble; therefore, emulsification is essential. In this study, the efficacy of high pressure homogenization (HPH) on natural plant (chocolate or vanilla)-based model emulsions was investigated. The particle size, electrical conductivity, Brix, pH, and color were analyzed after HPH. HPH significantly decreased the particle size of chocolate-based emulsions as a function of elevated pressures (20-100 MPa). HPH decreased the mean particle size of chocolate-based emulsions from 29.01 µm to 5.12 µm, and that of vanilla-based emulsions from 4.18 µm to 2.44 µm. Electrical conductivity increased as a function of the elevated pressures after HPH, for both chocolate- and vanilla-based model emulsions. HPH at 100 MPa increased the electrical conductivity of chocolate-based model emulsions from 0.570 S/m to 0.680 S/m, and that of vanilla-based model emulsions from 0.573 S/m to 0.601 S/m. Increased electrical conductivity would be attributed to colloidal phase modification and dispersion of oil globules. Brix of both chocolate- and vanilla-based model emulsions gradually increased as a function of the HPH pressure. Thus, HPH increased the solubility of plant-based powders by decreasing the particle size. This study demonstrated the potential use of HPH for enhancing the emulsification process and stability of the natural plant powders for applications with dairy products.

Effects of binders combined with glucono-δ-lactone on the quality characteristics of pressure-induced cold-set restructured pork.

This study investigated the effects of binders and glucono-δ-lactone (GdL) on characteristics of pressure-induced (450MPA for 3min) cold-set restructured pork. Isolated soy protein (SP), wheat flour (WF), and κ-carrageenan (CG) were adopted as binders. The addition of binders improved water-binding properties of restructured pork, and the binders diminished the decrease in water binding properties caused by GdL-induced acidification. Pressure-induced restructured pork prepared with binders showed less harder and more cohesive texture than those of the thermal-treated control (TC). The results indicate that pressure-induced cold-set meat restructuring could be achieved when binders and GdL were used in the formulation.

Effect of High Pressure on the Porcine Placenral Hydrolyzing Activity of Pepsin, Trypsin and Chymotrypsin.

This study investigated the effects of protease treatments (trypsin, chymotrypsin, and pepsin) under various pressure levels (0.1-300 MPa) for the characteristics of porcine placenta hydrolysates. According to gel electrophoretic patterns, the trypsin showed the best placental hydrolyzing activity followed by chymotrypsin, regardless of the pressure levels. In particular, the peptide bands of tryptic-digested hydrolysate were not shown regardless of applied pressure levels. The peptide bands of hydrolysate treated chymotrypsin showed gradual decreases in molecular weights (M w) with increasing pressure levels. However, the pepsin did not show any evidences of placental hydrolysis even though the pressure levels were increased to 300 MPa. The gel permeation chromatography (GPC) profiles showed that the trypsin and pepsin had better placental hydrolyzing activities under high pressure (particularly at 200 MPa), with lower M w distributions of the hydrolysates. Pepsin also tend to lower the M w of peptides, while the major bands of hydrolysates being treated at 300 MPa were observed at more than 7,000 Da. There were some differences in amino acid compositions of the hydrolysates, nevertheless, the peptides were mainly composed of glycine (Gly), alanine (Ala), hydroxyproline (Hyp) and proline (Pro). Consequently, the results indicate that high pressure could enhance the placental hydrolyzing activities of the selected proteases and the optimum pressure levels at which the maximum protease activity is around 200 MPa.

Effects of Pressure-shift Freezing on the Structural and Physical Properties of Gelatin Hydrogel Matrices.

This study investigates the effects of the gelatin concentration (10-40%, w/v), freezing temperatures (from -20℃ to -50℃) and freezing methods on the structural and physical properties of gelatin matrices. To freeze gelatin, the pressure-shift freezing (PSF) is being applied at 0.1 (under atmospheric control), 50 and 100 MPa, respectively. The freezing point of gelatin solutions decrease with increasing gelatin concentrations, from -0.2℃ (10% gelatin) to -6.7℃ (40% gelatin), while the extent of supercooling did not show any specific trends. The rheological properties of the gelatin indicate that both the storage (G') and loss (G") moduli were steady in the strain amplitude range of 0.1-10%. To characterize gelatin matrices formed by the various freezing methods, the ice crystal sizes which were being determined by the scanning electron microscopy (SEM) are affected by the gelatin concentrations. The ice crystal sizes are affected by gelatin concentrations and freezing temperature, while the size distributions of ice crystals depend on the freezing methods. Smaller ice crystals are being formed with PSF rather than under the atmospheric control where the freezing temperature is above -40℃. Thus, the results of this study indicate that the PSF processing at a very low freezing temperature (-50℃) offers a potential advantage over commercial atmospheric freezing points for the formation of small ice crystals.

Effects of Concentration and Reaction Time of Trypsin, Pepsin, and Chymotrypsin on the Hydrolysis Efficiency of Porcine Placenta.

This study investigated the effects of three proteases (trypsin, pepsin and chymotrypsin) on the hydrolysis efficiency of porcine placenta and the molecular weight (Mw) distributions of the placental hydrolysates. Because placenta was made up of insoluble collagen, the placenta was gelatinized by applying thermal treatment at 90 ℃ for 1 h and used as the sample. The placental hydrolyzing activities of the enzymes at varying concentrations and incubation times were determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and gel permeation chromatography (GPC). Based on the SDS-PAGE, the best placental hydrolysis efficiency was observed in trypsin treatments where all peptide bands disappeared after 1 h of incubation as compared to 6 h of chymotrypsin. Pepsin hardly hydrolyzed the placenta as compared to the other two enzymes. The Mw distribution revealed that the trypsin produced placental peptides with Mw of 106 and 500 Da. Peptides produced by chymotrypsin exhibited broad ranges of Mw distribution (1-20 kDa), while the pepsin treatment showed Mw greater than 7 kDa. For comparisons of pre-treatments, the subcritical water processing (37.5 MPa and 200 ℃ of raw placenta improved the efficiency of tryptic digestions to a greater level than that of a preheating treatment (90 ℃ for 1 h). Consequently, subcritical water processing followed by enzymatic digestions has the potential of an advanced collagen hydrolysis technique.

Effects of Partial Substitutions of NaCl with KCl, CaSO4 and MgSO4 on the Quality and Sensorial Properties of Pork Patties.

This study investigated the effects of NaCl replacers (KCl, CaSO4, and MgSO4) on the quality and sensorial properties of pork patty. In the characteristics of spray-dried salt particles, KCl showed the largest particle size with low viscosity in solution. Meanwhile CaSO4 treatment resulted in the smallest particle size and the highest viscosity (p<0.05). In comparison of the qualities of pork patties manufactured by varying level of Na replacers, MgSO4 treatment exhibited low cooking loss comparing to control (p<0.05). Textural properties of KCl and MgSO4 treatments showed similar pattern, i.e., low level of the replacers caused harder and less adhesive texture than those of control (p<0.05), whereas the hardness of these products was not different with control when the replacers were added more than 1.0%. The addition of CaSO4 also manifested harder and less adhesive than control (p<0.05), but the textural properties of CaSO4 treatment was not affected by level of Ca-salt. Eventually, sensorial properties indicated that KCl and CaSO4 influenced negative effects on pork patties. In contrast, MgSO4 showed better sensorial properties in juiciness intensity, tenderness intensity as well as overall acceptability than control, reflecting that MgSO4 was an effective Na-replacer in meat product formulation.

Effects of NaCl Replacement with Gamma-Aminobutyric acid (GABA) on the Quality Characteristics and Sensorial Properties of Model Meat Products.

This study investigated the effects of γ-aminobutylic acid (GABA) on the quality and sensorial properties of both the GABA/NaCl complex and model meat products. GABA/NaCl complex was prepared by spray-drying, and the surface dimensions, morphology, rheology, and saltiness were characterized. For model meat products, pork patties were prepared by replacing NaCl with GABA. For characteristics of the complex, increasing GABA concentration increased the surface dimensions of the complex. However, GABA did not affect the rheological properties of solutions containing the complex. The addition of 2% GABA exhibited significantly higher saltiness than the control (no GABA treatment). In the case of pork patties, sensory testing indicated that the addition of GABA decreased the saltiness intensity. Both the intensity of juiciness and tenderness of patties containing GABA also scored lower than the control, based on the NaCl reduction. These results were consistent with the quality characteristics (cooking loss and texture profile analysis). Nevertheless, overall acceptability of the pork patties showed that up to 1.5%, patties containing GABA did not significantly differ from the control. Consequently, the results indicated that GABA has a potential application in meat products, but also manifested a deterioration of quality by the NaCl reduction, which warrants further exploration.

Effect of NaCl/Monosodium Glutamate (MSG) Mixture on the Sensorial Properties and Quality Characteristics of Model Meat Products.

Sodium chloride is an important ingredient added to most of foods which contributes to flavor enhancement and food preservation but excess intake of sodium chloride may also cause various diseases such as heart diseases, osteoporosis and so on. Therefore, this study was carried out to investigate the effect of monosodium glutamate (MSG) as a salty flavor enhancer on the quality and sensorial properties of the NaCl/MSG complex and actual food system. For characterizing the spray-dried NaCl/MSG complex, surface dimension, morphology, rheology, and saltiness intensity were estimated by increasing MSG (0-2.0%) levels at a fixed NaCl concentration (2.0%). MSG levels had no effect of the characteristics of the NaCl/MSG complex, although the addition of MSG increased the surface dimension of the NaCl/MSG complex significantly (p<0.05). Furthermore, the effect of MSG on enhancing the salty flavor was not observed in the solution of the NaCl/MSG complex. In the case of an actual food system, model meat products (pork patties) were prepared by replacing NaCl with MSG. MSG enhanced the salty flavor, thereby increasing overall acceptability of pork patties. Replacement of NaCl with MSG (<1.0%) did not result in negative sensorial properties of pork patties, although quality deterioration such as high cooking loss was found. Nevertheless, MSG had a potential application in meat product formulation as a salty flavor enhancer or a partial NaCl replacer when meat products were supplemented with binding agents.

Effect of Novel Quick Freezing Techniques Combined with Different Thawing Processes on Beef Quality.

This study investigated the effect of various freezing and thawing techniques on the quality of beef. Meat samples were frozen using natural convection freezing (NF), individual quick freezing (IQF), or cryogenic freezing (CF) techniques, followed by natural convection thawing (NCT) or running water thawing (RT). The meat was frozen until the core temperature reached -12℃ and then stored at -24℃, followed by thawing until the temperature reached 5℃. Quality parameters, such as the pH, water binding properties, CIE color, shear force, and microstructure of the beef were elucidated. Although the freezing and thawing combinations did not cause remarkable changes in the quality parameters, rapid freezing, in the order of CF, IQF, and NF, was found to minimize the quality deterioration. In the case of thawing methods, NCT was better than RT and the meat quality was influence on the thawing temperature rather than the thawing rate. Although the microstructure of the frozen beef exhibited an excessive loss of integrity after the freezing and thawing, it did not cause any remarkable change in the beef quality. Taken together, these results demonstrate that CF and NCT form the best combination for beef processing; however, IQF and NCT may have practical applications in the frozen food industry.

Emulsion properties of pork myofibrillar protein in combination with microbial transglutaminase and calcium alginate under various pH conditions.

In this study, the effects of microbial transglutaminase (MTG) and calcium alginate (CA) systems in combination with soybean oil on the emulsion properties of porcine myofibrillar protein (MP) were evaluated under various pH conditions. MTG was shown to improve emulsifying capacity and creaming stability, which increased with increasing pH values up to 6.5. The CA did not influence emulsifying capacity, but it improved the creaming stability of the MP-stabilized emulsions. Both MTG and CA enhanced the rheological properties, but their effects on the physical characteristics of the protein evidenced an opposite trend in relation to pH, i.e., the MTG system improved both the emulsion and gelling properties with increasing pH, whereas the CA system was effective when the pH was lowered. By combining the two MP gelling systems, a stable and pH-insensible emulsion could be produced.