Characterization and acid-induced gelation of butter oil emulsions produced from heated whey protein dispersions

Whey protein isolate was dispersed at 4% or 8% (w/v) and heated at neutral pH to produce protein polymers. Butter oil, up to 20%, was homogenized in heated whey protein dispersions at pressure ranging from 10 to 120 MPa. Emulsion gelation was induced by acidification with glucono-δ-lactone. Whey protein polymers produced finely dispersed emulsions with fat droplet diameter ranging from 340 to 900 nm. Homogenization pressure was the main factor influencing droplet size. At low fat volume fraction, the emulsions exhibited Newtonian behaviour. As fat content increased, shear thinning behaviour developed as a result of depletion flocculation. Emulsion consistency index increased with protein and fat concentrations. Increasing homogenization pressure had no effect on Newtonian emulsions but promoted flocculation and significantly increased the consistency of high fat emulsions. Protein concentration was the main factor explaining emulsion gel hardness and syneresis. Syneresis decreased with increasing fat content in the gel.     

The effect of protein profile and preheating on denaturation of whey proteins and development of viscosity in milk protein beverages during heat treatment

The effect of preheat temperature (63 or 77 °C for 30 s; final heat 120 °C for 30 s) and casein to whey protein ratio on the physical characteristics of 3.3%, w/w, dairy protein beverages was investigated. Dispersions preheated at 77 °C had lower viscosity than dispersions preheated at 63 °C. Casein‐containing dispersions had significantly lower levels of α‐lactalbumin denaturation than whey protein‐only dispersions. A higher proportion of casein improved the thermal stability of protein dispersions. Overall, alteration of preheat temperature and casein to whey protein ratio can influence dairy beverage quality, with increasing levels of casein reducing physical changes due to heat treatment.     

Antihypertensive activity of milk fermented by Enterococcus faecalis strains isolated from raw milk

A total of 231 microorganisms were isolated from raw cow milk samples and the angiotensin-converting enzyme-inhibitory (ACEI) activity of the resultant fermented milk produced with the isolated microorganisms was assayed. Forty-six of these microorganisms were selected on the basis of high ACEI activity. Four Enterococcus faecalis strains stood out as producers of fermented milk with potent ACEI activity (IC₅₀ (the protein concentration that inhibits 50% of ACE activity): 34–59 μg mL⁻¹). Single doses (5 mL kg⁻¹) of the whey fraction obtained from these fermented milk samples were administered to spontaneously hypertensive rats (SHR) and to normotensive Wistar-Kyoto (WKY) rats in order to investigate their possible antihypertensive activity. Highly significant decreases in the systolic blood pressure (SBP) and in the diastolic blood pressure (DBP) were observed when the fermented milk was administered to SHR. Nevertheless, the fermented milk did not modify the SBP and the DBP of the WKY rats. Raw cow milk is an excellent source of wild lactic acid bacteria able to produce fermented milk with antihypertensive activity and antihypertensive activity of milk fermented by Enterococcus faecalis strains was associated with peptides different from Ile-Pro-Pro and Val-Pro-Pro.     

美国乳清蛋白在即饮饮料中的应用

乳清在乳制品加工中的应用前景非常广阔,本刊特邀美国乳品出口协会合作开辟专栏,就乳清在食品中应用的最新资讯进行专题系列报道。     

Antihypertensive effect of bioactive peptides produced by protease-facilitated lactic acid fermentation of milk

Milk was fermented for up to 5 h at 43 °C with two lactic acid bacteria (Streptococcus thermophilus, Lactobacillus bulgaricus). A protease, flavourzyme, was added at the beginning of fermentation. The whey fraction was separated from the fermented milk and freeze-dried. During the 5 h of fermentation, the soluble protein content increased from 4.9 to 57.4 mg/g and peptide content increased from 2.1 to 32.8 mg/g, while inhibition of angiotensin I-converting enzyme (ACE) increased by a decrease of IC₅₀ from 0.708 to 0.266 mg/ml, respectively. The whey was fractionated into four fractions by size exclusion chromatography on a Sephadex G-15 column. The fourth fraction of the whey showed the highest inhibitory efficiency ratio (IER) being 1329%/mg/ml. The amino acid sequence of the inhibitory peptide was Tyr-Pro-Tyr-Tyr, of which the IC₅₀ was 90.9 μM. The whey showed mixed-type inhibition kinetics, while Captopril, the positive control showed competitive inhibition on ACE. Their K_i values were 0.188 mg/ml and 0.0067 μg/ml, respectively. The systolic blood pressure (SBP) and diastolic blood pressure (DBP) was reduced to 15.9 and 15.6 mm Hg, respectively, in spontaneously hypertensive rat (SHR), after 8 weeks of oral administration of diluted whey (peptide concentration 4.9 mg/ml).     

Sugar beet pectin as a natural carrier for curcumin,a water-insoluble bioactive for food and beverage enrichment: Formation and characterization

Food and beverages enrichment with water-insoluble health-promoting nutraceuticals is important, but technologically challenging. Sugar beet pectin (SBP) is a natural dietary fiber with high emulsifying capacity. However, its potential as a natural encapsulator of hydrophobic nutraceuticals for beverage enrichment, has hardly been explored. Curcumin is a potent antioxidant with numerous attributed health benefits, but very low aqueous-solubility. We herein explored SBP as a carrier for solubilization and protection of curcumin (CUR). Using spectrofluorimetry, the CUR-SBP binding constant determined was (6.74 ± 0.5) ∙ 10⁵M⁻¹. As CUR:SBP molar ratio increased from 14:1 to 140:1, CUR encapsulation capacity increased from 14.5 ± 0.8 to 127.4 ± 0.4 mg _{CUR/g SBP}, and encapsulation efficiency moderately decreased from ~100% to 86 ± 7%. The encapsulation in SBP dramatically decreased CUR particle size, from >17 μm to <0.5 μm, in average, and conferred substantial protection to CUR during simulated shelf-life, decreasing the decay rate constant ~7 fold. Therefore, SBP is a potent natural encapsulator for hydrophobic nutraceuticals for food and even clear beverage enrichment.     

Application of multiple sensory evaluations to produce fermented beverages made from sole whey using Kluyveromyces marxianus

Various fermented whey-based beverages have been developed to reduce waste. However, approaches to increase the preference of whey-based beverages are required because of the low sensory acceptability of whey. Here, we identified the better starting material (whey type), between raw whey (RW) and demineralised whey (DMW), and determined the optimal initial concentration using multiple sensory evaluations to develop acceptable fermented beverages made from sole whey with pure cultured Kluyveromyces marxianus (i.e. without additional ingredients and processing methods). Acceptance tests showed that fermented beverages made from DMW were superior to RW as the starting material. The amounts of ethanol produced were 5.0%, 7.6% and 9.5% v/v from the different initial DMW concentrations of 10%, 15% and 20% w/v, respectively. We observed a significant positive correlation (r_s = 0.32, P < 0.05) between the assessment attributes, strength of taste as alcohol beverage and overall acceptability, indicating that higher concentrations of DMW yielded a more desirable product.     

Factors regulating astringency of whey protein beverages

A rapidly growing area of whey protein use is in beverages. There are 2 types of whey protein-containing beverages: those at neutral pH and those at low pH. Astringency is very pronounced at low pH. Astringency is thought to be caused by compounds in foods that bind with and precipitate salivary proteins; however, the mechanism of astringency of whey proteins is not understood. The effect of viscosity and pH on the astringency of a model beverage containing whey protein isolate was investigated. Trained sensory panelists (n = 8) evaluated the viscosity and pH effects on astringency and basic tastes of whey protein beverages containing 6% wt/vol protein. Unlike what has been shown for alum and polyphenols, increasing viscosity (1.6 to 7.7 mPa·s) did not decrease the perception of astringency. In contrast, the pH of the whey protein solution had a major effect on astringency. A pH 6.8 whey protein beverage had a maximum astringency intensity of 1.2 (15-point scale), whereas that of a pH 3.4 beverage was 8.8 (15-point scale). Astringency decreased between pH 3.4 and 2.6, coinciding with an increase in sourness. Decreases in astringency corresponded to decreases in protein aggregation as observed by turbidity. We propose that astringency is related to interactions between positively charged whey proteins and negatively charged saliva proteins. As the pH decreased between 3.4 and 2.6, the negative charge on the saliva proteins decreased, causing the interactions with whey proteins to decrease.     

Antioxidant activity and characterization of whey protein-based beverages: Effect of shelf life and gastrointestinal transit on bioactivity

Whey proteins can exhibit antioxidant activity. The objectives of this study were to formulate model whey based beverages with well-established antioxidants (plant polyphenols, vitamins and astaxanthin) to investigate (1) the antioxidant shelf life over a 24-week period and (2) the antioxidant activity after upper gastrointestinal transit. Pilot scale processing (pasteurization, ultra-high temperature or spray drying) was used to prepare beverages which were representative of current product formats. In vitro gastrointestinal digestion of test samples was performed using the standardised INFOGEST method and antioxidant activity of samples was determined using ABTS, FRAP and ORAC. Results from the antioxidant shelf life study provided evidence that powder products functionality was preserved. Whey beverages (pasteurised or spray dried) increased or maintained antioxidant activity during gastrointestinal transit. Combination of whey with additional antioxidant ingredients increased the bioactivity of formulated products; however, this greater bioactivity was altered after gastrointestinal transit, depending on processing type and antioxidant methodology.Industrial relevanceWhey protein-based antioxidant beverages could benefit the elderly consumer to meet their increased protein requirements and boost their antioxidant status. Consumer's acceptance for whey protein-based beverages often improves with clear formulations. This work generated whey protein-based UHT beverages with greater stability and clarity than pasteurised formulations. A novel combination of plant and marine antioxidants increased antioxidant activity of whey protein-based formulations. Furthermore, to suit export markets this work generated spray dried whey protein formulations that did not alter antioxidant potential.     

Identifying Key Attributes for Protein Beverages

This study identified key attributes of protein beverages and evaluated effects of priming on liking of protein beverages. An adaptive choice‐based conjoint study was conducted along with Kano analysis to gain insight on protein beverage consumers (n = 432). Attributes evaluated included label claim, protein type, amount of protein, carbohydrates, sweeteners, and metabolic benefits. Utility scores for levels and importance scores for attributes were determined. Subsequently, two pairs of clear acidic whey protein beverages were manufactured that differed by age of protein source or the amount of whey protein per serving. Beverages were evaluated by 151 consumers on two occasions with or without priming statements. One priming statement declared “great flavor,” the other priming statement declared 20 g protein per serving. A two way analysis of variance was applied to discern the role of each priming statement. The most important attribute for protein beverages was sweetener type, followed by amount of protein, followed by type of protein followed by label claim. Beverages with whey protein, naturally sweetened, reduced sugar and ≥15 g protein per serving were most desired. Three consumer clusters were identified, differentiated by their preferences for protein type, sweetener and amount of protein. Priming statements positively impacted concept liking (P < 0.05) but had no effect on overall liking (P > 0.05). Consistent with trained panel profiles of increased cardboard flavor with higher protein content, consumers liked beverages with 10 g protein more than beverages with 20 g protein (6.8 compared with 5.7, P < 0.05). Protein beverages must have desirable flavor for wide consumer appeal.     

Authentication of Whey Protein Powders by Portable Mid‐Infrared Spectrometers Combined with Pattern Recognition Analysis

The objective of this study was to develop a simple and rapid method to differentiate whey protein types (WPC, WPI, and WPH) used for beverage manufacturing by combining the spectral signature collected from portable mid‐infrared spectrometers and pattern recognition analysis. Whey protein powders from different suppliers are produced using a large number of processing and compositional variables, resulting in variation in composition, concentration, protein structure, and thus functionality. Whey protein powders including whey protein isolates, whey protein concentrates and whey protein hydrolysates were obtained from different suppliers and their spectra collected using portable mid‐infrared spectrometers (single and triple reflection) by pressing the powder onto an Attenuated Total Reflectance (ATR) diamond crystal with a pressure clamp. Spectra were analyzed by soft independent modeling of class analogy (SIMCA) generating a classification model showing the ability to differentiate whey protein types by forming tight clusters with interclass distance values of >3, considered to be significantly different from each other. The major bands centered at 1640 and 1580 cm^?1 were responsible for separation and were associated with differences in amide I and amide II vibrations of proteins, respectively. Another important band in whey protein clustering was associated with carboxylate vibrations of acidic amino acids (～1570 cm^?1). The use of a portable mid‐IR spectrometer combined with pattern recognition analysis showed potential for discriminating whey protein ingredients that can help to streamline the analytical procedure so that it is more applicable for field‐based screening of ingredients.     

Effect of high pressure - low temperature processing on composition and colloidal stability of casein micelles and whey proteins

The aim of this study was to identify the impact of high pressure treatments at sub-zero temperatures (high pressure - low temperature; HPLT) on milk proteins. Whey protein solutions, micellar casein dispersions and two mixtures (micellar caseins:whey proteins, 80:20 and 20:80, w/w) were pressure treated (100–600 MPa) at pH 7.0 or 5.8 at −15 °C, −35 °C and ambient temperature. Solubility data showed that whey proteins could only be affected by HPLT treatments at pH 7.0 if caseins were present, while effects could be induced at pH 5.8 without the presence of caseins. The caseins formed on the one hand large aggregates (flocs) and on the other hand the solubility was increased by the creation of smaller micelles. The formation of flocs could only be observed for HPLT treated samples, which indicates the formation of different protein interactions in milk protein based samples compared with common HP treatments.     

Sequential preheating and transglutaminase pretreatments improve stability of whey protein isolate at pH 7.0 during thermal sterilization

Whey protein isolate (WPI) is a potential ingredient to manufacture shelf-stable transparent beverages if proteins are heat stable, i.e., without causing turbidity, precipitation and gelation after the required thermal processing to obtain commercial sterility (138 °C for 8 s or longer). However, information is lacking about stability of WPI during heating at 138 °C. Furthermore, novel technology and mechanistic understanding on how to produce clear products after heating systems with >5% WPI, particularly with salt, is needed. In this work, 5% w/v WPI was pretreated by microbial transglutaminase (mTGase) at three levels for 1–15 h at 50 °C, with and without prior preheating at 80 °C for 15 min. Heat stability of the pretreated samples at pH 7.0 and 0, 50, and 100 mM NaCl was evaluated at 138 °C. Samples pretreated by mTGase for a greater extent demonstrated improved heat stability. Samples subjected to sequential preheating and mTGase pretreatments produced clear dispersions even after heating at 138 °C for 30 min in the presence of 0 and 50 mM NaCl at pH 7. All pretreatments increased the magnitude of zeta-potential and resistance against thermal denaturation. The sequentially-pretreated WPI was the most heat-resistant, having decreased hydrodynamic diameter (<36 nm) during extended heating at 138 °C and 50 mM NaCl. The present study demonstrates the feasibility of using sequential preheating and mTGase pretreatments to develop sterilized beverage products with a high content (5% w/v) of whey protein and yet of transparent appearance at ambient temperatures.     

Comparative study of the phenolics,antioxidant and metagenomic composition of novel soy whey-based beverages produced using three different water kefir microbiota

Water kefir microbiota was used to develop novel soy whey-based beverages that have antioxidant activity. In the present study, comparative phenolics, antioxidant and metagenomic composition of the soy whey beverages fermented using three different water kefir microbiota, named WKFS-A, WKFS-B and WKFS-C were investigated. WKFS-B beverage had the highest concentrations of isoflavone aglycones (208.73 ± 2.78 mg L⁻¹) and phenolic acids (132.33 ± 3.41 mg L⁻¹) compared with WKFS-A (193.88 ± 1.15 mg L⁻¹) and (91.73 ± 2.34 mg L⁻¹) and WKFS-C (160.63 ± 1.76 mg L⁻¹) and (97.13 ± 2.63 mg L⁻¹), respectively. The WKFS-B also showed higher DPPH and ABTS radical scavenging activity and ferric reducing antioxidant power compared with WKFS-A and WKFS-C beverages. Microbial species diversity index analysis showed that a higher concentration of isoflavone aglycones, phenolic acids and increased antioxidant activity in the WKFS-B beverage correlates with the higher relative abundance of Lactobacillus genus. This study thus revealed that Lactobacillus dominated water kefir microbiota produces soy whey beverages with high phenolic acids, isoflavone aglycones and antioxidant activity.     

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.     

Use of ultrafiltration and supercritical fluid extraction to obtain a whey buttermilk powder enriched in milk fat globule membrane phospholipids

Whey buttermilk, a by-product from whey cream processing to butter, is rich in milk fat globule membrane (MFGM) constituents, which have technological and potential health properties. The objective of this work was to produce a dairy ingredient enriched in MFGM material, especially phospholipids, from whey buttermilk. Whey buttermilk was concentrated by ultrafiltration (10×) and subsequently diafiltered (5×) (10 kDa molecular mass cutoff membrane) at 25 °C and the final retentate was spray-dried. The whey buttermilk powder was submitted to supercritical extraction (350 bar, 50 °C) using carbon dioxide. The membrane filtration removed most of the lactose and ash from the whey buttermilk, and the supercritical extraction extracted exclusively non-polar lipids. The final powder contained 73% protein and 21% lipids, of which 61% were phospholipids. This ingredient, a phospholipids-rich dairy powder, could be used as an emulsifier in different food systems.     

Rheological properties and structure of inulin–whey protein gels

The rheological properties, structure and synergistic interactions of whey proteins (1–7%) and inulin (20% and 35%) were studied. Gelation of whey proteins was induced with Na⁺. Inulin was dissolved in preheated whey protein solutions (80 °C, 30 min). Inulin gel formation was strongly affected by whey proteins. The presence of whey proteins at a level allowing for protein gel network formation (7%) significantly increased the G′ and G″ values of the gels. Scanning electron micrographs showed a thick structure for the mixed gel. Whey proteins at low concentrations (1–4%) were not able to form a gel; further, these low concentrations partly or wholly impaired formation of a firm inulin gel. Although interactions between inulin and whey proteins may be concluded from hydrophobicity measurements, the use of an electrophoretic technique did not show any inulin–whey protein complexes.     

Effect of fermentation on biochemical and sensory characteristics of sorghum flour supplemented with whey protein

Changes in pH, titrable acidity, protein, non-protein nitrogen, total soluble solids, protein fractions and in vitro protein digestibility were investigated during fermentation and/or after supplementation of sorghum flour with whey protein. The pH of the fermenting material decreased sharply with a concomitant increase in the titrable acidity. The total soluble solids increased with progressive fermentation time. The crude protein and non-protein nitrogen both increased with fermentation time. The protein content and fractions were significantly (p ⩽ 0.05) increased after supplementation with whey protein. The albumin plus globulin fraction increased significantly (p ⩽ 0.05) during the first 8 h of fermentation after supplementation with 5% whey protein. Other fraction contents were observed to fluctuate during the fermentation time. Supplementation of the cultivar flour with 10% whey protein greatly increased the protein content as well as the albumin plus globulin fraction while other fractions were significantly decreased. The in vitro protein digestibility was significantly (p ⩽ 0.05) improved during fermentation and even after supplementation. Sensory evaluation of locally processed sorghum products (Kisra, Asida and Nasha) before and after supplementation showed no difference between the supplemented samples and the control ones as judged by trained panellists.