Fractionation and identification of a novel hypocholesterolemic peptide derived from soy protein Alcalase hydrolysates

A novel hypocholesterolemic peptide was fractionated by gradient ethanol elution from a macroporous adsorption resin (MAR DA201-C), and then separated on Sephadex G-15 and RP-HPLC from a soy protein hydrolysate (SAPH DH 18%). Identification of the hypocholesterolemic peptide structure was accomplished with HPLC–MS. The peptide with the highest hypocholesterolemic activity was found in 75% ethanol fraction among the four fractions from gradient ethanol elution with MAR DA201-C. The calculated average hydrophobicity by amino acid composition of each ethanol eluted fraction suggested that the peptides could be separated in terms of hydrophobicity with MAR DA201-C. Four peaks were obtained with further fractionation on Sephadex G-15, the highest cholesterol micellar solubility inhibition rate, 81.3%, was obtained in Peak 2, corresponding to the molecular weight fraction of 300–800 Da. Fifteen main peaks were obtained with RP-HPLC fractionation, the highest cholesterol micellar solubility inhibition rate (94.3%) was in Peak 7. The amino acid sequence of this peptide was identified as WGAPSL with LC–MS and amino acid composition analysis.     

Comparison of a modified spectrophotometric and the pH-stat methods for determination of the degree of hydrolysis of whey proteins hydrolysed in a tangential-flow filter membrane reactor

A procedure was developed to determine the degree of hydrolysis (DH) of whey protein hydrolysates (WPH) during hydrolysis in either 3 kDa or 10 kDa tangential-flow filter (TFF) enzymatic membrane reactors (EMR). Protease N Amano G (IUB 3.4.24.28, Bacillus subtilis) was used to hydrolyse an initial 5% (w v^?1) aqueous solution of whey protein isolate (86.98% protein) at pH 7.0 and 55 °C with continuous recirculation and simultaneous removal of hydrolysates through the TFF, in single- or two-stage operation. The DH in the permeate and the retentate were determined as the concentration of the free α-NH₂ using 2,4,6-trinitrobenzene 1-sulphonic acid (TNBS) and compared to the pH-stat method. In the new method, the DH of the permeate, the retentate and for the total EMR process could be quantified together or independently. The pH-stat method exaggerated the DH in the EMR because of the leakage of the alkali. The TNBS method was more reliable for DH estimation in the EMR.     

Taste modification of amino acids and protein hydrolysate by α-cyclodextrin

The objective of this work was to characterize the formation of amino acid inclusion complexes with α-cyclodextrin (α-CD) and evaluate the influence of added α-CD on the taste perception of amino acids and hydrolyzed soy protein at pH 4.5. The formation of the inclusion complexes of phenylalanine, tryptophane, tyrosine, isoleucine, proline and histidine with α-CD were detected by nuclear magnetic resonance techniques (ROESY and DOSY) and the sensory characteristics of soy hydrolysates were judged by a panel of trained tasters. It was concluded that these amino acids form inclusion complexes with α-CD and the order of affinity for the α-CD cavity is phenylalanine ≈ tryptophane > proline > isoleucine ≈ tyrosine ≈ histidine. α-CD alters the bitter taste perception of the amino acids and reduces the bitter taste from hydrolyzed soy protein. These results indicate a potential use of α-CD for debittering protein hydrolysates in acidic beverages.     

Effect of limited hydrolysis of soy protein on the interactions with polysaccharides at the air–water interface

The objective of the work was to study the effect of limited hydrolysis of soy protein on the interactions with polysaccharides with and without surface activity at the air–water interface at neutral pH where a limited incompatibility between macromolecules can occur. The surface pressure and phase angle as a function of time were evaluated with a drop tensiometer at 20 °C, pH 7 and ionic strength 0.05 M.Hydrolysates of 2% (H1) and 5.4% (H2) degree of hydrolysis (DH) with neutral protease from Aspergillus oryzae were obtained from a commercial soy protein isolate. The polysaccharides used were: hydroxypropylmethylcellulose (HPMC) as surface active polysaccharide; lambda carrageenan (λC) and locust bean gum (LB) as non-surface active polysaccharides.It was found that increasing DH decreased the surface pressure and increased film viscoelasticity (determined as the phase angle, θ) of soy protein hydrolysates and the nature of protein–polysaccharide interactions was strongly affected by DH. The presence of polysaccharides led to an increase of surface pressure of H1 but when added to H2, HPMC and λC decreased the surface pressure.The less hydrolyzed protein H1 gave rise to a higher surface pressure and film viscoelasticity in combination with the polysaccharides. This result points out that a limited protein hydrolysis was sufficient to improve the surface properties of soy proteins if used in combination with polysaccharides.Polysaccharides used in admixture with hydrolyzed soy proteins could control and improve the stability of foams and emulsions not only by increasing bulk viscosity but also by improving film viscoelasticity.     

Screening of whey protein isolate hydrolysates for their dual functionality: Influence of heat pre-treatment and enzyme specificity

Heat pre-treated and non heat pre-treated whey protein isolate (WPI) were hydrolysed using α-chymotrypsin (chymotrypsin), pepsin and trypsin. The in vitro antioxidant activity, ACE-inhibition activity and surface hydrophobicities of the hydrolysates were measured in order to determine if peptides with dual functionalities were present. Dual functional peptides have both biological (e.g. antioxidant, ACE-inhibition, opioid activities) and technological (e.g. nanoemulsification abilities) functions in food systems. Heat pre-treatment marginally enhanced the hydrolysis of WPI by pepsin and trypsin but had no effect on WPI hydrolysis with chymotrypsin. With the exception of the hydrolysis by trypsin, heat pre-treatment did not affect the peptide profile of the hydrolysates as analysed using size exclusion chromatography, or the antioxidant activity (P > 0.05). Heat pre-treatment significantly affected the ACE-inhibition activities and the surface hydrophobicities of the hydrolysates (P < 0.05), which was a function of the specificity of the hydrolysing enzyme. Extended hydrolysis (up to 24 h) had no significant effect on the DH and the molecular weight profiles (P > 0.05) but in some instances caused a reduction in the antioxidant activity of WPI hydrolysates. The chymotrypsin hydrolysate showed a broad MW size range, and was followed by pepsin and then trypsin. The bioactivities of the hydrolysates generally decreased in the order; chymotrypsin > trypsin > pepsin. This study showed that by manipulating protein conformation with pre-hydrolysis heat treatment, combined with careful enzyme selection, peptides with dual functionalities can be produced from WPI for use as functional ingredients in the manufacture of functional foods.     

Antioxidant activities of enzymatic rapeseed protein hydrolysates and the membrane ultrafiltration fractions

In this study, rapeseed protein isolate was hydrolyzed with various proteases to obtain hydrolysates that were separated by membrane ultrafiltration into four molecular size fractions (<1, 1–3, 3–5, and 5–10 kDa). Alcalase hydrolysis significantly (p < 0.05) produced the highest yield of protein hydrolysate while Flavourzyme produced the least. The <1 kDa fraction was the most abundant after the membrane ultrafiltration of the protein hydrolysates, which indicates that the proteases were efficient at reducing the native rapeseed proteins into low molecular weight peptides. Antioxidant properties of the resulting hydrolysates and membrane fractions were characterized and results showed the Pepsin + Pancreatin (P + P) protein hydrolysate had significantly highest (p < 0.05) scavenging activity against DPPH radical among the unfractionated enzymatic hydrolysates. But the P + P hydrolysate was not as effective as other hydrolysates during long-term inhibition of linoleic acid oxidation. For most of the samples, fractionation into the <1 kDa peptides significantly (p < 0.05) improved DPPH and superoxide scavenging properties when compared to the unfractionated protein hydrolysates. Only the <1 kDa fraction showed ferric reducing antioxidant power and the effect was dose-dependent. Overall, Alcalase and Proteinase K seem to be more efficient proteases to release antioxidant peptides from rapeseed proteins when compared to P + P, Flavourzyme and Thermolysin.     

High pressure processing assisted enzymatic hydrolysis – An innovative approach for the reduction of soy immunoreactivity

Soybean (Glycine max (L.) MERR.) is recognized as a potent food allergen causing one of the most frequent food allergies worldwide. The effect of high pressure processing (HPP) prior to and during enzymatic hydrolysis using the enzyme preparation Flavourzyme® on the degree of hydrolysis (DH), molecular weight distribution (SDS-PAGE) and β-conglycinin (Gly m5) immunoreactivity of soy protein isolate (SPI) was studied. Enzymatic hydrolysis was carried out at atmospheric pressure (0.1 MPa) and HPP (100–600 MPa) at 50 °C for 15 min. Pressures higher than 300 MPa enhanced the degradation of Gly m5, which was confirmed by SDS-PAGE and LC-MS/MS analyses. The immunoreactivity of the samples was assessed by in vitro sandwich ELISA using mouse monoclonal anti-Gly m5 antibodies. Depending on the antibody tested, the residual immunoreactivity was completely inhibited or significantly impaired up to 99.5% applying HPP during hydrolysis at 400 and 500 MPa. By means of principal component analysis, the beany and green off-flavors characteristic for unprocessed SPI could be reduced by pressure enhanced hydrolysis at 400–500 MPa. The resulting hydrolysates possessed improved protein solubility, foaming activities and oil-binding capacities, which were improved by 45%, 66%, and 210%, respectively. HPP prior to and during enzymatic hydrolysis at 400–500 MPa constitutes an innovative approach for the production of low-allergen food ingredients that combine good taste and enhanced functional properties.Industrial relevanceFood allergy has emerged in the last years as the incidence and prevalence are rising dramatically. Up to now, enzymatic hydrolysis is the only feasible method to mitigate soy allergy. However, the major drawback associated with enzymatic hydrolysis is the incomplete destruction of allergenic epitopes and the formation of a strong bitter taste. This research activity demonstrates that high pressure assisted enzymatic hydrolysis using the enzyme preparation Flavourzyme effectively reduces the immunoreactivity of soy proteins. Degree of hydrolysis analysis, SDS-PAGE, mass spectrometry as well as sandwich ELISA with mouse monoclonal anti-Gly m5 antibodies have been applied to analyze the destruction of allergenic proteins as well as to determine the residual immunoreactivity. This study provides preliminary evidence that this innovative combination process of high pressure and enzymatic hydrolysis has great potential to produce tasty low-allergen soy-based food ingredients with good physicochemical properties, i.e. protein solubility and foamability.     

Peptide–peptide and protein–peptide interactions in mixtures of whey protein isolate and whey protein isolate hydrolysates

The extent of aggregation in whey protein isolate (WPI) hydrolysates induced by Bacillus licheniformis protease was quantified as a function of degree of hydrolysis (DH), temperature and ionic strength. The capacity of the hydrolysates to aggregate added intact protein was also studied. The amount of aggregated material and the size of the aggregated peptides were measured by nitrogen content and size exclusion chromatography, respectively. Aggregation increased with DH up to the practical end point of hydrolysis (DH 6.8%). The aggregates formed under the various conditions studied consisted of peptides with masses ranging from 1.4 to 7.5 kDa. The hydrolysates were also able to aggregate added WPI. The additional amount of aggregated material increased with increasing DH. Peptides involved in peptide–peptide interactions were also involved in protein–peptide interactions. It is hypothesized that hydrophobic interactions dominated peptide–peptide interactions, while protein–peptide interactions depended on the balance between hydrophobic attraction and electrostatic repulsion.     

Antioxidant activity of protein hydrolysates obtained from discarded Mediterranean fish species

In this study, five discarded species in the Mediterranean Sea, namely sardine, horse mackerel, axillary seabream, bogue and small-spotted catshark, were evaluated as raw material for obtaining fish protein hydrolysates exhibiting antioxidant activity. The DH of the hydrolysates ranged from 13.2 to 21.0%, with a protein content varying from 60.7 to 89.5%. The peptide profile of all hydrolysates was very similar, except for the hydrolysate of small-spotted catshark. Their lipid content was found to be between 4.6 and 25.3%. The highest DPPH scavenging activity was found for the hydrolysates of sardine and horse mackerel with EC₅₀ values varying from 0.91 to 1.78 mg protein/mL. Sardine and small-spotted catshark hydrolysates exhibited the highest ferrous chelating activity with an EC₅₀ value of 0.32 mg protein/mL. Moreover, sardine and bogue hydrolysates presented the highest reducing power. Finally, a total of six antioxidant peptides were theoretically identified within the structure of myosin and actin proteins from sardine and small-spotted catshark. The potential antioxidant activity exhibited by the hydrolysates suggests that it is feasible to obtain added-value products such as natural antioxidants from these discarded species.     

Effect of degree of hydrolysis on the antioxidant activity of loach (Misgurnus anguillicaudatus) protein hydrolysates

Loach (Misgurnus anguillicaudatus) proteins were hydrolysed by papain and Protamex, the antioxidant activity of loach protein hydrolysates (LPH) was investigated. The results demonstrated that extensive hydrolysis by papain and Protamex led to the browning of the hydrolysates. When the degree of hydrolysis (DH) was 23%, hydrolysates prepared by papain (HA) exhibited the strongest antioxidant activity. The maximum values of the hydroxyl, 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging activities and the reducing power were 56.1%, 95.5%, 2.80 mM and 1.46, respectively. The hydrolysates prepared by Protamex (HB) showed the strongest hydroxyl radical scavenging activity (55.0%) at DH 28%, DPPH radical scavenging activity (92.2%) and ABTS radical scavenging activity (2.81 mM) at DH 23%, and the reducing power (1.17) at DH 33%. At the same DH value, there were significant (p < 0.05) differences between HA and HB. Several antioxidant amino acid residues, especially Trp and His, contributed significantly to the antioxidant activity of the hydrolysates. An increase of peptides with molecular weight below 500 Da was observed as the DH increased for all LPH. The above results indicated that DH and protease greatly influenced the molecular weight and amino acid residue composition of LPH, and further influenced the antioxidant activity.Industrial relevanceLoach has long been employed as a traditional Chinese medicine for the treatment of many kinds of diseases. From our previous work, loach was determined to be a good source of protein (accounts for approximately 17% (w/w) of the body weight). In this work, loach proteins were hydrolyzed by papain and Protamex to specific extent. The effect of DH on the antioxidant activities of hydrolysates was investigated. The results indicated that loach protein hydrolysates were potent antioxidants which were significantly affected by DH. This research is helpful for extensive development of loach product.     

Peptides derived from high oleic acid soybean meals inhibit colon,liver and lung cancer cell growth

Soybean meal, a co-product after oil extraction from seeds, is rich in protein. Our objective was to utilize this co-product, obtain gastrointestinal (GI) resistant peptides from the isolated protein, and test for bioactivity against colon, liver and lung cancer cell lines. N98-4445A, S03-543CR high oleic acid soybean lines, and R95-1705 high protein soybean line were used for this study. Protein isolates were prepared at alkaline pH and hydrolyzed using alcalase enzyme to generate peptide hydrolysates. After determining gastrointestinal resistance of the peptide hydrolysates they were fractionated into definite molecular sizes of < 5 kDa, 5–10 kDa, and 10–50 kDa and tested against human colon (HCT-116, Caco-2), liver (HepG-2) and lung (NCL-H1299) cancer cell lines. MTS, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2 H-tetrazolium, cytotoxicity assay was performed to test in vitro cancer cell viability upon treatment with peptide fractions. The peptide fractions from N98-4445A and S03-543CR lines showed cell growth inhibition of 73% of colon cancer (HCT-116), 70% of liver cancer cells and 68% of lung cancer cells. Dose response showed that the peptides had significant inhibitory effect at higher concentrations (1000 μg/mL to 600 μg/mL) and gradually decreased with decreased dosage (500 μg/mL to 100 μg/mL). Reverse phase HPLC identified three single peptides from the 10–50 kDa fractions of N98-4445A soy line that have potential for enhanced activity. Soybean peptide fractions can thus be a source of bioactivity against colon, liver and lung cancer cell proliferation.     

Angiotensin-converting enzyme inhibitory activity of milk protein hydrolysates: Effect of substrate,enzyme and time of hydrolysis

Nine milk protein substrates were hydrolysed in vitro with five proteases for various times (0, 3, 6, and 24 h), and the angiotensin-converting enzyme (ACE)-inhibitory activity of hydrolysates was assessed. Overall, the casein substrates gave rise to hydrolysates with significantly higher ACE-inhibitory activity than the whey protein (WP) substrates (85% vs. 79%). No significant difference between 3 and 24 h of hydrolysis was found. A reasonable correlation was found between the ACE inhibition of the 6 h hydrolysates determined in vitro and estimated by in silico modelling. The highest ACE-inhibitory activity was found in hydrolysates made with thermolysin followed by proteinase K, trypsin, pepsin and Bacillus licheniformis protease. The IC₅₀ values for thermolysin hydrolysates of caseins and WPs were 45–83 and 90–400 μg mL⁻¹, respectively, with α-lactalbumin giving the highest inhibitory activity. Thermolysin, proteinase K and trypsin were useful for the release of highly potent ACE-inhibitory peptides from both WPs and caseins.     

Assessment of the effects of soy protein isolates with different protein compositions on gluten thermosetting gelation

Although soy proteins are known to have a deleterious effect on gluten thermosetting gelation, the causes are still poorly understood. Different sources of soy protein isolates (SPI) were used to investigate the interactions between gluten and soy proteins during hydro-thermal treatments. Commercial SPI and isolates prepared from soybean lines with different subunit composition were used to study the influence of protein denaturation and subunit composition on thermoset gel formation. Rapid Visco Analyser analysis showed that replacement of gluten with more than 1% SPI decreased the peak viscosity and interfered with formation of thermoset gels. However, peak viscosity was higher for 11% gluten + 2% SPI than for 11% gluten alone, suggesting a cooperative effect. After heating and cooling, 11% gluten + 2% SPI rich in A1 and A2 subunits formed a coherent thermoset gel suggesting that the cysteine residue content of soy proteins can affect gel formation.     

Identification and characterization of antioxidant peptides from sweet potato protein hydrolysates by Alcalase under high hydrostatic pressure

Sweet potato protein hydrolysates (SPPH) were generated by Alcalase under high hydrostatic pressure (HHP, 100, 200 and 300 MPa). HHP significantly improved the degree of hydrolysis (DH) and antioxidant activity, and increased the < 3 kDa fraction content of SPPH (P < 0.05). SPPH by Alcalase at 300 MPa for 60 min exhibited the highest DH and antioxidant activity and was separated into three fractions by ultrafiltration. The most active fraction FIII (< 3 kDa) was further separated into fifty four fractions by semi-preparative RP-HPLC and measured using the ORAC assay. In addition, more active fractions were examined by LC–MS/MS, and diverse peptides were identified, matching sequences of Sporamins A and B. To evaluate the structure–activity dependences, twenty sequences were synthesized, of which the antioxidant activity was assessed. Five peptides showed good activity: HDSASGQY ≥ YYMVSA ≥ HDSESGQY ~ YYIVS ~ RYYDPL, with the contribution of His and Tyr.Industrial relevanceThis study will give a novel technique for using industrial waste slurry, a byproduct in the process of sweet potato starch manufacturing, which contains various bioactive components (such as protein, minerals, etc.) since most of them are normally discarded. The present study is focused on assessing the effects of enzymatic hydrolysis by Alcalase under high hydrostatic pressure (HHP) on the release of antioxidant peptides from sweet potato protein (SPP). The results of this work provide a potential application of enzymatic hydrolysis assisted by HHP on the development of ingredients from SPP in functional foods.     

Effects of combined high-pressure homogenization and enzymatic treatment on extraction yield,hydrolysis and function properties of peanut proteins

Peanut protein isolate (PPI) was extracted by high-pressure homogenization (HPH) under 0.1 MPa (atmospheric pressure) and 40 or 80 MPa (high pressure). Effects of Alcalase (a proteolytic enzyme) on the enzymatic hydrolysis of PPI and the antioxidant activity of the PPI hydrolysates were investigated. The molecular weight distributions of the PPI hydrolysates were analyzed using Sephadex G-25 gel filtration chromatography while the antioxidant activities, including reducing power, 1,1-dipheny-2-picrylhydrazyl (DPPH) radical-scavenging activity and hydroxyl free radical-scavenging activity of the PPI hydrolysates were evaluated. The extraction yields of PPI by HPH under 0.1, 40 and 80 MPa were 16.84, 30.65 and 39.86%, respectively, which showed that HPH treatment improved the PPI extraction. The HPH treatment increased the degree of hydrolysis of PPI and significantly increased the reducing power and hydroxyl radical­scavenging activity. Furthermore, the molecular weight distributions of the PPI hydrolysates appeared principally over the range of 1000–5000 Da, while the HPH treatment enhanced the production of small peptides, which was in agreement with the high PPI hydrolysis degree. These results suggest that HPH treatment in combination with enzymatic hydrolysis could modify PPI properties and increase the antioxidant activities of the PPI hydrolysates.Industrial relevanceThis study was focused to evaluate the effects of high-pressure homogenization (HPH) in combination with enzymatic hydrolysis on extraction yield and enzymatic hydrolysis of PPI and antioxidant activity of the PPI hydrolysates. This study indicated the possibility of improving the availability of PPI by HPH treatment via increasing extraction yield and enzymatic hydrolysis of the PPI, which can provide a better utilization of the peanut by-product.     

Functional properties and Angiotensin-I converting enzyme inhibitory activity of soy–whey proteins and fractions

Soy proteins when prepared to high purity can confer good functional properties and the whey by-product is a potential source for bioactivity. In this study, we determined the protein, moisture, fiber, solubility, foaming, emulsion properties, as well as Angiotensin-I converting enzyme (ACE-I) inhibitory activity of prepared soy–whey proteins and its fractions. The soy–whey proteins were fractionated into < 5, > 5, > 10, and > 50 kDa using ultrafiltration. The expanded AACC methods were used to determine protein, moisture, and fiber analyses of the whey and its fractions. Solubility method was conducted to determine the protein solubility profile of the soy–whey and its fractions at varying pHs. Turbidimetric method was used to evaluate emulsifying activity (EA) and emulsion stability (ES). There were significant differences observed in moisture, protein and salt contents between unfractionated, > 50 kDa and smaller sized fractions. No significant differences were observed with phytic acid and total dietary fiber contents among all samples. The unfractionated whey protein and > 50 kDa fraction showed better solubility than other fractions. Unfractionated whey protein had the highest foam capacity (42.7 mL) while the fraction > 5 kDa showed the greatest foaming stability (46 min). The highest emulsion activity (0.33 ± 0.1) and stability (825.1 ± 45.1) was obtained with the > 50 kDa fraction while the unfractionated whey protein had the highest ACE-I inhibition activity. The findings indicate that soy–whey protein fraction (> 50 kDa) had good solubility, emulsion activity and stability, while the unfractionated whey protein exhibited the strongest ACE-I inhibition percentage (19%).     

Shelf Life Extension of Toasted Groundnuts through the Application of Cassava Starch and Soy Protein-Based Edible Coating

The use of cassava starch and soy protein concentrate edible coatings containing 20% glycerol in extending the shelf life of toasted groundnut during ambient (27 ± 1°C) storage for 14 days was studied. Chemical indices of oxidative rancidity and sensory parameters were evaluated using standard procedures. Moisture uptake, peroxide and thiobarbituric acid values of uncoated groundnuts were higher than 100% cassava starch coated groundnuts while toasted groundnuts coated with 50:50 (cassava starch:soy protein concentrate) had the lowest values. Toasted groundnuts coated with 50:50 (Cassava starch:soy protein concentrate) film had higher colour, taste, texture and overall acceptability scores than toasted groundnuts coated with 100% cassava starch film and control. The use of 50:50 (cassava starch:soy protein concentrate) edible coatings on toasted groundnut extended the shelf life of toasted groundnuts for 14 days compared to uncoated toasted groundnuts which developed objectionable taste after second day of storage at ambient (27 ± 1°C) condition.     

Anti-obesity effects of epigallocatechin-3-gallate,orange peel extract,black tea extract,caffeine and their combinations in a mouse model

The anti-obesity effects of epigallocatechin-3-gallate (EGCG), orange peel extract (OPE), black tea extract (BTE), and caffeine (CF) in female CF-1 mice were studied. Female CF-1 mice were fed high-fat diets containing 0.1% EGCG, 0.2% OPE, 0.2% BTE and 0.05% caffeine alone and in combination for 10 weeks. The body weight gain and weights of abdominal fat and brown adipose tissue were significantly reduced in mice whose diets contained OPE, BTE, caffeine, OPE + BTE and OPE + CF. Notably, mice fed a high-fat diet supplemented daily with 0.2% OPE + 0.2% BTE + 0.05% CF prevented body weight gain by 48.8%, parametrial fat pad weight by 88.2%, retroperitoneal fat pad weight by 82.8% and brown adipose tissue by 63.7% compared with mice fed a high-fat diet. On the basis of these findings, it was concluded that oral feeding of orange peel extract, black tea extract and caffeine had anti-obesity effects by suppressing body weight gain and adipose tissue formation.     

Physical,Chemical and Sensory Qualities of Roselle Water Extract-coagulated Tofu Compared with Tofu from Two Natural Coagulants

Roselle calyces (Hibiscus sabdariffa) aqueous extracts were used in coagulating soymilk at four different concentrations (2.5%, 5%, 10% and 20%). The physical, chemical and sensory qualities of the tofu preparations were compared with fermented maize liquor and Calotropis procera extract coagulated tofus. pH of roselle extracts was acidic in nature (2.01 – 3.74) which was attributed to the ability to coagulate soy proteins. pH, titratable acidity of the roselle coagulated tofus ranged from 5.32 – 6.26 and 0.16 – 0.43% respectively and the yield and protein content ranged from 87.3 – 95.9 g and 42.6 – 46.3 g/100 g respectively. The yield of roselle coagulated tofu increased with increase in concentration of the roselle extracts. Roselle extract when utilized at 2.5% concentration will yield tofu that is acceptable and comparable to tofu coagulated with fermented maize steep water in terms of appearance, flavour and overall acceptability at p > 0.05. Roselle extract at 5% and 10% also yielded tofus that were acceptable in terms of all of the attributes tested.     

Effect of soy protein substitution for sodium caseinate on the transglutaminate-induced cold and thermal gelation of myofibrillar protein

The influence of soy protein isolate (SPI) substitution for sodium caseinate (SC) on the properties of cold-set (4 °C) and heat-induced gels of pork myofibrillar protein (MP) incubated with microbial transglutaminase (TG) was investigated. The strength of cold-set MP–SC gels (formed in 0.45 M, NaCl, 50 mM phosphate buffer, pH 6.25) increased with time of TG incubation, but those gels with more than 66% SPI substituted for SC had a >26% reduced strength (P < 0.05). Upon cooking, both incubated and non-incubated protein sols were quickly transformed into highly elastic gels, showing up to 6000 Pa in storage modulus (G′) at the final temperature (72 °C). However, no differences (P < 0.05) in G′ were observed between heated samples with SPI and SC. Myosin heavy chain, casein and soy proteins gradually disappeared with TG incubation, contributing to MP gel network formation. Both cold-set and heat-induced gels had a compact protein matrix, attributable to protein cross-linking by TG.