Interactions between whey proteins and salivary proteins as related to astringency of whey protein beverages at low pH

Whey protein beverages have been shown to be astringent at low pH. In the present study, the interactions between model whey proteins (β-lactoglobulin and lactoferrin) and human saliva in the pH range from 7 to 2 were investigated using particle size, turbidity, and ζ-potential measurements and sodium dodecyl sulfate-PAGE. The correlation between the sensory results of astringency and the physicochemical data was discussed. Strong interactions between β-lactoglobulin and salivary proteins led to an increase in the particle size and turbidity of mixtures of both unheated and heated β-lactoglobulin and human saliva at pH ∼3.4. However, the large particle size and high turbidity that occurred at pH 2.0 were the result of aggregation of human salivary proteins. The intense astringency in whey protein beverages may result from these increases in particle size and turbidity at these pH values and from the aggregation and precipitation of human salivary proteins alone at pH <3.0. The involvement of salivary proteins in the interaction is a key factor in the perception of astringency in whey protein beverages. At any pH, the increases in particle size and turbidity were much smaller in mixtures of lactoferrin and saliva, which suggests that aggregation and precipitation may not be the only mechanism linked to the perception of astringency in whey protein.     

Variation in bull beef quality due to ultimate muscle pH is correlated to endopeptidase and small heat shock protein levels

This study set out to determine if ultimate pH (pH_u) affected the performance of intracellular small heat shock protein and endopeptidase dynamics in muscle during beef ageing. Longissimus dorsi muscles from 39 Angus or Limousin × Angus bulls were examined to see if pH_u achieved at 22 h post mortem (rigor) affected tenderness and water holding capacity of beef. Samples were segregated into three pH_u groups termed high (pH > 6.3), intermediate (5.7 < pH < 6.3) and low (pH < 5.7) pH_u beef. More than 30% of bull beef did not achieve acceptable tenderness at 8 days post mortem with this ageing regime. No significant differences in calpain or cathepsin enzyme levels due to meat pH were observed until after 22 h post mortem, but low pH_u beef had elevated caspase 3/7 activity soon after slaughter. At 22 h post mortem, greater levels of μ-calpain enzyme were found in the high and intermediate pH_u beef and cathepsin B levels were superior in the low pH_u beef after 2 days post mortem. Different rates of desmin and troponin T protein degradation were also observed in aged bull beef. Both proteins were degraded within 6 h post mortem for high pH_u beef, but took >3 days post mortem for intermediate pH_u beef. High levels of alpha β-crystallin (aβC) at 22 h post mortem coincided with delayed muscle protein degradation for low pH_u beef. Our results support the hypothesis that aβC shields myofibrils and buffers against endopeptidase degradation of beef structure during ageing.     

Physical and chemical stability of β-carotene-enriched nanoemulsions: Influence of pH, ionic strength, temperature, and emulsifier type

The enrichment of foods and beverages with carotenoids may reduce the incidences of certain chronic diseases. However, the use of carotenoids in foods is currently limited because of their poor water-solubility, high melting point, low bioavailability, and chemical instability. The potential of utilising oil-in-water (O/W) nanoemulsions stabilised by a globular protein (β-lactoglobulin) for encapsulating and protecting β-carotene was examined. The influence of temperature, pH, ionic strength, and emulsifier type on the physical and chemical stability of β-carotene enriched nanoemulsions was investigated. The rate of colour fading due to β-carotene degradation increased with increasing storage temperature (5-55 °C), was faster at pH 3 than pH 4-8, and was largely independent of ionic strength (0-500 mM of NaCl). β-Lactoglobulin-coated lipid droplets were unstable to aggregation at pH values close to the isoelectric point of the protein (pH 4 and 5), at high ionic strengths (NaCl >200 mM, pH 7), and at elevated storage temperatures (55 °C). β-Carotene degradation was considerably slower in β-lactoglobulin-stabilised nanoemulsions than in Tween 20-stabilised ones. These results provide useful information for facilitating the design of delivery systems to encapsulate and stabilise β-carotene for application within food, beverage, and pharmaceutical products.     

The effect of enhancement with salt, phosphate and milk proteins on the physical and sensory properties of pork loin

Pork loins were pumped to 110% of original weight with solutions containing 5.5% salt and 3.3% sodium tripolyphosphate, 5.5% salt and 3.3% β-lactoglobulin (β-lg) enriched fraction or 5.5% salt and 3.3% whey protein concentrate 80% (WPC80) for comparison with non-enhanced control loins. The enhancement of pork loins significantly increased (p<0.001) the tenderness and juiciness. Warner-Bratzler shear force values were lower (p<0.001) in enhanced then in non-enhanced control chops. The enhanced pork chops had a slightly higher overall flavour and overall acceptability to the control pork chops. Sensory analysis showed that β-lg enriched fraction and WPC80 can be used as replacement ingredients to reduce the amount of phosphate used in enhancement solutions, as they were comparable to salt/sodium tripolyphosphate (salt/STPP) enhancement solution. Consumers rated the tenderness, juiciness and taste of the enhanced chops significantly (p<0.001) higher then the control chops.     

Effects of dietary alpha-amylase on metabolism and performance of transition dairy cows

Twenty-four multiparous Holstein cows [body weight, 759 kg (SD = 30 kg); body condition score, 3.2 (SD = 0.13)] were used in a randomized complete block design to determine the effect of feeding α-amylase during the transition period on rumen fermentation, key metabolic indicators, and lactation performance. Cows were assigned to either a control diet or the control diet supplemented with α-amylase (662 fungal amylase units per gram, AMA) at 0.1% of diet dry matter (DM). Experimental diets were fed from 21 d before expected calving through 21 d in milk. From 22 to 70 d in milk, all cows were fed a similar lactation cow diet. Average pre- and postpartum DM intakes were 12.4 and 17.8 kg/d, respectively, and did not differ between treatments; however, DM intakes during the last week prepartum decreased to a greater degree in AMA than control cows compared with wk −2. Supplementing diets with α-amylase tended to increase proportions of ruminal butyrate prepartum but not postpartum. Treatment differences were not detected for concentrations of insulin in plasma and lipid and glycogen in liver tissue. Prepartum, concentrations of β-hydroxybutyrate and nonesterified fatty acids were increased in cows fed AMA compared with the control diet. Postpartum, concentrations of glucose in plasma tended to be increased by feeding AMA. Increased plasma β-hydroxybutyrate and nonesterified fatty acids pre- but not postpartum and a tendency for increased plasma glucose postpartum demonstrate shifting reliance from lipid- to carbohydrate-based metabolism postpartum in cows fed α-amylase.     

Interactions between β-lactoglobulin and dextran sulfate at near neutral pH and their effect on thermal stability

The effect of interactions between β-lactoglobulin (β-LG) and dextran sulfate (DS) on thermal stability at near neutral pH was investigated. Samples containing 6% w/w β-LG and DS (M_w = 5–500 kDa) at different biopolymer weight ratios, pH (5.6–6.2), and NaCl concentrations (0–30 mM) were heated at 85 °C for 15 min. Turbidity results showed that the presence of DS at appropriate biopolymer weight ratio and pH significantly lowered the turbidity of heated β-LG. Solutions containing DS:β-LG weight ratios of 0.02 or less showed improved heat stability as indicated by decreased turbidity. Analysis of the unheated mixture by size exclusion chromatography coupled with multi-angle laser light scattering (SEC–MALLS) showed an interaction between β-LG and DS. The size of the aggregates increased as pH decreased. The β-LG–DS aggregates had a greater negative charge as seen from electrophoretic mobility measurement. Addition of 30 mM NaCl inhibited complex formation and the effect of DS on reducing the turbidity of heated β-LG, suggesting that the interaction was electrostatic in nature. Other than charge property, the amount and size of native aggregates appeared to be the major factor in determining how DS altered heat-induced aggregation. The presence of DS decreased denaturation temperature of β-LG, indicating that DS did not improve thermal stability of β-LG by stabilizing its native state but rather by altering its aggregation. The results provide information that will facilitate the application of whey proteins and polysaccharides as functional ingredients in foods and beverages.     

Sugars,acids, ethyl β-d-glucopyranose and a methyl inositol in sea buckthorn (Hippophaë rhamnoides) berries

Sea buckthorn berry is a rich source of nutrients and bioactive components beneficial for human health. Sugars and acids play an important role in determining the sensory properties of the berry. Sugars, acids, ethyl β-d-glucopyranose and a methyl inositol were analysed in berries of three subspecies (Hippophaë rhamnoides ssp. sinensis, rhamnoides and mongolica) collected from China, Finland and Russia over four consecutive years. Fructose and glucose were the major sugars, and the dominating acids were malic and quinic acids. Origin and harvesting date have significant impacts on sugars, acids and sugar/acid ratio in the berry. During the harvesting period, the sugar content followed different changing patterns in berries of different subspecies. Ethyl glucose dominated in the sugar fraction of ssp. rhamnoides but existed only in trace amounts in the other two subspecies. In ssp. rhamnoides, the level of ethyl glucose increased during the harvesting period; the increase was accompanied by a decrease in glucose content, indicating the presence of a biochemical pathway converting glucose into its derivatives. A methyl inositol was identified for the first time in sea buckthorn with higher levels found in ssp. sinensis than in the other two subspecies. The levels of ethyl glucose and methyl inositol may be important sensory and nutritional quality factors of sea buckthorn berry. The data presented by this study provide important chemotaxonomic information characterising different subspecies of sea buckthorn and useful guidance for breeding, harvesting, and industrial utilisation of sea buckthorn.     

Galacto-oligosaccharide synthesis by immobilized Aspergillus oryzae β-galactosidase

Aspergillus oryzae β-galactosidase was immobilized by three different techniques, namely adsorption on celite, covalent coupling to chitosan and aggregation by cross-linking. These techniques were compared in terms of the yield of immobilized preparation, enzymatic characteristics, stability and efficiency in oligosaccharide synthesis. Immobilization led to increase in K_m in each case. Immobilization on chitosan gave maximum enzyme yield and oligosaccharide synthesis. At 60 °C, the chitosan-immobilized enzyme was stabilized (by 1.6-fold) due to protection effect of the matrix. However, at 65 °C, the t_1/2 of cross-linked enzyme aggregates (CLEA) of β-galactosidase was 1.07 h as compared to 0.79 h in the case of free enzyme. Both chitosan-immobilized enzyme and CLEA were used for oligosaccharide synthesis. Using 20% (w/v) lactose, the chitosan-immobilized enzyme gave maximum oligosaccharide yield (17.3% of the total sugar) as compared to free enzyme (10.0%) in 2 h at 40 °C. CLEA were instead found effective in lactose hydrolysis yielding 78% monosaccharide in 12 h.     

Aβ/Cu-catalyzed oxidation of cholesterol in 1,2-dipalmitoyl phosphatidylcholine liposome membrane

The amyloid β protein with 42 amino acid residues (Aβ), which is a causative protein of Alzheimer's disease (AD), forms the complex with copper (II) to induce the cholesterol oxidase-like activity by the proton transfer from the cholesterol. In this study, the oxidation of cholesterol by Aβ/Cu complex was investigated on the surface of the zwitterionic phospholipid liposome including the bound water advantageous for the enhancement of the proton transfer. The bound water was pooled by the formation of cholesterol-rich domain within liposomes. The resulting reactivity was enhanced by the proton transfer mediated by the bound water.     

Preparation,characterisation and activity of the inclusion complex of paeonol with β-cyclodextrin

The inclusion complex of β-cyclodextrin (β-CD) and paeonol (2′-hydroxy-4′-methoxyacetophenone, PAE) was synthesised and characterised by thermal gravimetric analysis (TGA) and two-dimensional rotating frame spectroscopy (2D ROESY). The antioxidant activity and tyrosinase inhibition activity were also studied. The TGA results indicated that the thermal stability of PAE was improved when it was included with β-CD. Based on the 2D ROESY analysis, an inclusion structure of the PAE–β-CD complex was proposed, in which PAE penetrated β-CD in a tilted manner due to the interaction of intermolecular hydrogen bonds between PAE and β-CD. The complex of PAE with β-CD increased the antioxidant activity and tyrosinase-inhibiting activity of PAE.