Compositional Analysis of Whey Powders Using Near Infrared Diffuse Reflectance Spectroscopy

Compositional analysis data for 47 whey powders (including sweet whey, acid whey, demineralized delactosed whey and whey protein concentrates) were correlated with near infrared reflectance (NIR) measurements. The most useful wavelengths for determining constituent concentrations were chosen from 19 pre-selected wavelengths by using linear regression analysis. Results indicate that samples to be used for NIR calibrations should be similar in composition and should represent a limited range of constituent concentrations. Individual NIR calibrations need to be developed for each type of whey powder to be analyzed.     

Stability of Whey Proteins during Thermal Processing: A Review

Whey proteins have many benefits due to their high nutritional value and their various applications in food products. A drawback of whey proteins is their instability to thermal processing, which leads to their denaturation, aggregation, and, under some conditions, gelation. As thermal processing is a major treatment in the processing of milk and milk products, its influence on whey proteins has been extensively studied. Understanding the mechanisms involved during each stage of denaturation and aggregation of whey proteins is critical to devising ways of improving their stability. These aspects are reviewed in this paper. Also covered are approaches to preventing or reducing heat‐induced aggregation of whey proteins. Inhibition of aggregate formation has considerable potential for alleviating the problems that arise from the instability of whey proteins.     

ASSESSMENT of NEAR INFRARED (NIR) DIFFUSE REFLECTANCE ANALYSIS FOR MEASURING MOISTURE and WATER ACTIVITY IN RAISINS

Near Infrared (NIR) Analysis was applied to assess its potential for measuring water activity (a_w) and moisture in California Seedless Raisins in moisture equilibrium and nonequilibrium conditions. Samples covered a wide range of anticipated moisture contents. an NIR calibration for a_w was made for whole raisins that were fully equilibrated. For ground raisins, NIR calibrations were made both a_w and vacuum oven moisture, covering a large range of moisture contents and equilibrium conditions. Direct measurement of a_w by a dewpoint device gave highly repeatable results, especially for ground raisins. Vacuum oven analysis of moisture was less precise than a_w measurements. Full spectrum NIR calibrations indicated that NIR could predict a_w of raisins in ground form, irrespective of their state of hydration beyond three hours following processing. A much larger sample population will be needed to establish precise calibrations for vacuum oven moisture.     

近红外快速检测奶粉乳清粉成分研究

采用Perten8620近红外奶粉分析仪检测奶粉的近红外光谱特性，建立了奶粉近红外检测的校准模型。结果表明，近红外检测技术无需称重和化学试剂，可在1min以内快速检测奶粉和乳清粉的蛋白、脂肪、水分等指标，其准确性和精度可以达到在国际允许的误差范围内。根据近红外技术可能在短时间内收集大量样品的光谱图，开发了新检测项目的校准曲线。     

High pressure and the enzymatic hydrolysis of soybean whey proteins

The effect of high-pressure (HP) treatment on the hydrolysis of soybean whey proteins by trypsin, chymotrypsin and pepsin was studied. The experiments were carried out at atmospheric pressure (0.1 MPa, for 30 min, at 37 °C) and at HP (100 and 200 MPa for 15 min at 37 °C) before or during the reaction of hydrolysis. The extent of hydrolysis was measured by OPA method and in the extracts from TCA. The results showed that HP treatments increased the hydrolysis in the three enzymes used and 100 MPa was the better pressure to enhance the hydrolysis. Polyacrylamide gel electrophoreses (SDS–PAGE), showed five peptides lower than 14 kDa after hydrolysis by chymotrypsin and trypsin, and 11 peptides by pepsin. Soybean whey proteins, which are industrially discarded, could be used as a source of peptides, with applications as base in some diets.     

Enzymatic Hydrolysis of Milk Proteins Under Alkaline and Acidic Conditions

The pH-stat and osmometric methods were adopted for control of hydrolysis of casein and whey proteins by chymotrypsin, trypsin and pepsin. Experiments in alkaline medium showed that the mean pK values determined for amino groups in milk protein hydrolysates at 52°C were 7.11 for casein hydrolysates and 7.18 for whey protein hydrolysates. During hydrolysis in acidic medium the osmotic coefficients determined for casein and whey proteins enabled calculation of the calibration factor for the osmometer (1.05) which could be assumed as constant. Results will enable monitoring of DH during hydrolysis of milk proteins in such systems.     

Whey—pollution problem and potential utilization

The high biochemical oxygen demand (BOD₅) of whey poses a major world-wide disposal and pollution problem for the dairy industry, for which an effective and permanent solution is urgently needed. Biological waste water treatment technologies can assist in the safe disposal of whey or whey permeate within the federal environment specifications, but only at substantial cost. One alternative is use of whey, or whey permeate rich in lactose and protein, in processes in which saleable products contribute wholly or partially to the costs. Production of whey proteins by ultrafiltration, lactose hydrolysis products, and the use of whole whey or whey permeate as a fermentation feedstock are possible options. The present situation is discussed, together with projections for commercial potential, limitations of the biotechnology of whey processing, the end products and nutritional aspects.     

Identification and characterization of novel α-amylase and α-glucosidase inhibitory peptides from camel whey proteins

This study explores the inhibitory properties of camel whey protein hydrolysates (CWPH) toward α-amylase (AAM) and α-glucosidase (AG). A general full factorial design (3 × 3) was applied to study the effect of temperature (30, 37, and 45°C), time (120, 240, and 360 min), and enzyme (pepsin) concentration (E%; 0.5, 1, and 2%). The results showed that maximum degree of hydrolysis was obtained when hydrolysis was carried out at higher temperature (45°C; P < 0.05), compared with lower temperatures of 30 and 37°C. Electrophoretic pattern displays degradation of all protein bands upon hydrolysis by pepsin at various hydrolysis conditions applied. All the 27 CWPH generated showed significant AAM and AG inhibitory potential as indicated by their lower IC₅₀ values (mg/mL) compared with intact whey proteins. In total 196 peptides were identified from selected hydrolysates and 15 potential peptides (PepSite score > 0.8; http://pepsite2.russelllab.org/) were explored via in silico approach. Novel peptides PAGNFLMNGLMHR, PAVACCLPPLPCHM, MLPLMLPFTMGY, and PAGNFLPPVAAAPVM were identified as potential inhibitors for both AAM and AG due to their high number of binding sites and highest binding probability toward the target enzymes. CCGM and MFE, as well as FCCLGPVPP were identified as AG and AAM inhibitory peptides, respectively. This is the first study that reports novel AG and AAM inhibitory peptides from camel whey proteins. The future direction for this research involves synthesis of these potential AG and AAM inhibitory peptides in a pure form and investigate their antidiabetic properties in the in vitro, as well as in vivo models. Thus, CWPH can be considered for potential applications in glycaemic regulation.     

Shrimp processing assessed by low field nuclear magnetic resonance, near infrared spectroscopy, and physicochemical measurements--the effect of polyphosphate content and length of prebrining on shrimp muscle

The effect of using polyphosphates during prebrining and the effect of prebrining time of cold water shrimp (Pandalus borealis) was studied with low field nuclear magnetic resonance (LF-NMR) transverse relaxation time measurements (benchtop and unilateral) and near infrared (NIR) spectroscopy with the aim of improving shrimp processing. Strong calibrations were obtained for moisture content and water-holding capacity (WHC) using the NIR technique. Multivariate analysis indicated significant correlations between benchtop NMR parameters and moisture content and WHC, as measured with physicochemical methods and NIR spectroscopy. Significant correlations were also observed between NMR parameters and muscle pH, protein content, and phosphate content. The study showed that LF-NMR contribute to improved understanding of the shrimp brining process and to improved process control on-line or at-line, especially in combination with NIR measurements. However, optimization of the unilateral device is necessary.     

Peptides released from acid goat whey by a yeast-lactobacillus association isolated from cheese microflora

Seven lactobacilli and a variety of microflora extracted from twenty five commercial cheeses were grown on unsupplemented acid goat whey and screened for their capacity to hydrolyse whey proteins [alpha-lactalbumin (alpha-la) and beta-lactoglobulin (beta-lg)] and to generate peptides. Fermentations were performed aerobically or anaerobically at 37 degrees C using crude or pre-heated whey (10 min at 65, 75 or 85 degrees C). Under aerobic conditions, growth of lactobacilli was poor and protein hydrolysis did not occur. Anaerobic conditions slightly increased lactobacilli growth but neither beta-lg hydrolysis nor peptide generation were observed. More than 50% of alpha-la was digested into a truncated form of alpha-la (+/- 12 kDa) in crude whey and whey pre-heated at 65 degrees C. Twenty-five microflora extracted from raw milk cheeses were screened for their proteolytic activities on acid goat whey under the conditions previously described. Eight of them were able to hydrolyse up to 50% of alpha-la mainly during aerobic growth on crude or pre-heated whey. The corresponding hydrolysates were enriched in peptides. The hydrolysate involving microflora extracted from Comté cheese after or at 18 months ripening was the only one to exhibit hydrolysis of both alpha-la and beta-lg. Microbiological analysis showed that microorganisms originating from Comté cheese and capable of growth on unsupplemented whey consisted of Candida parapsilosis and Lactobacillus paracasei. Fermentation kinetic profiles suggested that peptides were released from alpha-la hydrolysis. The co-culture of both microorganisms was required for alpha-la hydrolysis that occurred concomitantly with the pH decrease. During whey fermentation, Cand. parapsilosis excrete at least one protease responsible for alpha-la hydrolysis, and Lb. paracasei is responsible for medium acidification that is required for protease activation.     

Inhibitory properties of camel whey protein hydrolysates toward liver cancer cells,dipeptidyl peptidase-IV,and inflammation

This report describes an investigation of camel whey protein hydrolysates (CWPH) produced by gastric and pancreatic enzymes for their in vitro antidiabetic, anticancer, and anti-inflammatory properties. Degree of hydrolysis (DH) ranged from 8.54 to 47.53%, with hydrolysates generated using chymotrypsin for 6 h displaying the highest DH. Reverse phase-HPLC analysis showed that α-lactalbumin underwent complete degradation, with no intact α-lactalbumin detected in CWPH. The CWPH displayed enhanced antidiabetic activity compared with intact whey proteins; with pepsin- and chymotrypsin-generated CWPH displaying greater inhibition of dipeptidyl peptidase IV (DPP-IV), α-glucosidase, and α-amylase compared with trypsin-generated CWPH. The highest antiproliferative effect was observed for CWPH generated by chymotrypsin for 3 h, with only 4.5 to 6.5% viable liver cancer cells (HepG2) remaining when tested at concentrations from 400 to 1,000 µg/mL. The highest anti-inflammatory activity was manifested by CWPH generated by pepsin at 6-h hydrolysis. We report enhanced antiproliferative, antidiabetic, and anti-inflammatory activities upon hydrolysis of camel whey proteins, indicating their potential utilization as bioactive and functional ingredients.     

HPLC-MS/MS-Detection of Caseins and Whey Proteins in Meat Products

Screening methods for the mass spectrometric detection of caseins and whey proteins in meat products have been developed. After tryptic digestion, two α-S1-casein and two β-lactoglobulin marker peptides were measured by HPLC-MS/MS. For matrix calibrations, emulsion-type sausages with different concentrations of milk and whey protein (ppm level) were produced. The limits of detection (LODs) were below 1 ppm for milk protein and about 3 ppm for whey protein. The determination coefficients for the correlation between peak area of the marker peptides and the concentrations of milk and whey proteins were R²≥0.9899.     

分步酶解法制备黄浆水活性肽

黄浆水是传统豆制品点脑成型过程压榨出的废弃物,富含低聚糖、蛋白质等营养成分。该研究通过比较酶种类及用量、酶解温度、酶解时间对黄浆水蛋白质水解度的影响,采用正交试验优化获得黄浆水短肽最佳分步酶解工艺：（1）酸性蛋白酶加酶量2 000 U/g,pH 4.0,温度55 ℃,水解2 h;（2）中性蛋白酶8 000 U/g,pH 6.0,温度50 ℃,水解6 h。在此条件下进行验证,水解度可达25.95%,血管紧张素转化酶体外抑制活性达92.0%。采用酸性蛋白酶和中性蛋白酶分步酶解黄浆水制备短肽,制备条件温和,水解度高,可为豆制品加工废弃物的高值化利用奠定基础。     

大豆乳清蛋白特性及回收利用研究进展

大豆分离蛋白加工过程中产生大量乳清废水,直接排放会造成环境污染和资源浪费。大豆乳清废水中含有大豆乳清蛋白(Soybean Whey Proteins, SWP)、大豆异黄酮、大豆低聚糖等多种营养成分,其中大豆乳清蛋白应用价值极高,富含胰蛋白酶抑制剂、β-淀粉酶、大豆血球凝集素、脂肪氧合酶等多种功能因子。基于此,本文针对大豆乳清蛋白的回收利用,归纳总结了大豆乳清蛋白中的主要组成成分,并对各组分的研究利用以及其功能特性进行总结与分析,同时对大豆乳清蛋白的回收方法及利用进行了梳理,以期为工业生产实践中高值化利用提供理论和技术上的参考。     

Functional and Biological Properties of Peptides Obtained by Enzymatic Hydrolysis of Whey Proteins

The study of peptides released by enzymatic hydrolysis of whey proteins has been initially focusing on improving their functional properties in food model systems. Our first study showed that peptides 41 to 60 and 21 to 40 from β-lactoglobulin (β-LG) were responsible for improved emulsifying properties of a tryptic hydrolysate of whey protein concentrate (WPC). Further work showed that adding negatively charged peptides from tryptic hydrolysates of WPC could prevent phase separation of dairy-based concentrated liquid infant formula, as a replacement for carrageenan. Hydrolysis of whey proteins using a bacterial enzyme was also successful in improving heat stability of whey proteins in an acidic beverage. Some tryptic peptides demonstrated improvement in the heat stability and in modifying thermal aggregation of whey proteins. Recent research has shown that whey peptides could trigger some physiological functions. Within the scope of this research our work has led to the development of a whey protein enzymatic hydrolysate that has demonstrated antihypertensive properties when orally administered to spontaneously hypertensive rats and human subjects. Our work then focused on the fractionation of hydrolysates by nanofiltration to prepare specific peptidic fractions; however, peptide/peptide and peptide/protein interactions impaired membrane selectivity. The study of those interactions has lead to the demonstration of the occurrence of interactions between β-LG and its hydrophobic fragment 102–105 (opioid peptide), which probably binds in the central cavity of the protein. This latest result suggests that β-LG could be used as a carrier for the protection of bioactive peptides from gastric digestion. Our work therefore has shown that the enzymatic hydrolysis of whey proteins is not only improving their functional properties, but it is also providing powerful technology in the exploitation of their biological properties for functional foods and nutraceutical applications.     

Combined microwave and enzymatic treatments for ��-lactoglobulin and bovine whey proteins and their effect on the IgE immunoreactivity

Effect of combined microwave (MW) and enzymatic hydrolysis on the human immunoglobulin E (IgE)-binding properties of ??-lactoglobulin (??-lg) and other whey proteins (WP) was investigated. Separated ??-lg and full whey protein isolate (WPI) were hydrolyzed with trypsin, chymotrypsin, mixture of trypsin/chymotrypsin, and pepsin at three microwave power levels: 50?W during 1 and 5?min, 100 and 200?W during 1 and 3?min. The immunoreactivity of the obtained hydrolysates resulting from combined microwave protease treatment was assessed using sera of young patients allergic to bovine whey proteins. The application of microwave treatment at 200?W enhances the hydrolysis of ??-lg by pepsin in 3?min and decreases significantly its immunoreactivity. The extensive hydrolysis of the microwave-treated ??-lg and WPI with trypsin, chymotrypsin, and the mixture of trypsin with chymotrypsin did not have an impact on the IgE binding of the products obtained in all the studied conditions.     

Design of whey protein nanostructures for incorporation and release of nutraceutical compounds in food

Whey proteins are widely used as nutritional and functional ingredients in formulated foods because they are relatively inexpensive, generally recognized as safe (GRAS) ingredient, and possess important biological, physical, and chemical functionalities. Denaturation and aggregation behavior of these proteins is of particular relevance toward manufacture of novel nanostructures with a number of potential uses. When these processes are properly engineered and controlled, whey proteins may be formed into nanohydrogels, nanofibrils, or nanotubes and be used as carrier of bioactive compounds. This review intends to discuss the latest understandings of nanoscale phenomena of whey protein denaturation and aggregation that may contribute for the design of protein nanostructures. Whey protein aggregation and gelation pathways under different processing and environmental conditions such as microwave heating, high voltage, and moderate electrical fields, high pressure, temperature, pH, and ionic strength were critically assessed. Moreover, several potential applications of nanohydrogels, nanofibrils, and nanotubes for controlled release of nutraceutical compounds (e.g. probiotics, vitamins, antioxidants, and peptides) were also included. Controlling the size of protein networks at nanoscale through application of different processing and environmental conditions can open perspectives for development of nanostructures with new or improved functionalities for incorporation and release of nutraceuticals in food matrices.     

High hydrostatic pressure assisted enzymatic hydrolysis of whey proteins

Whey proteins, due to their high nutritional value, are generally hydrolyzed to reduce the allergenicity and used as ingredients in many special products, such as infant formulae, geriatric products, highly energetic supplements or dietetic foods or in foods produced to prevent nutrition related diseases, like food intolerances and allergies.The aim of this work was to assess the applicability of innovative technologies, such as high hydrostatic pressure (HHP) processes, to assist the enzymatic hydrolysis of target proteins, namely whey protein concentrate (WPC-80), in order to modify their antigenicity. Experiments were carried out to verify the effectiveness of HHP technology to accelerate whey protein hydrolysis reaction with selected enzymes (α-chymotrypsin, bromelain), and to affect the protein allergenic power. To this purpose, different HHP treatments were carried out at several pressure levels (100, 200, 300 and 400 MPa) and the untreated whey protein samples were used as control. A defined enzyme/substrate ratio of 1/10 w/w was used in the experiments, while the treatment time was changed from 0 to 30 min (0, 5, 15, or 30 min).The experimental data demonstrated that High Hydrostatic Pressure (HHP) induced WPC-80 unfolding at the highest value of the pressure applied (400 MPa) as indicated by the higher exposure of free sulfhydryl groups. When HHP was used in combination with enzymatic hydrolysis, the degree of hydrolysis increased not only with the pressure level applied but also with the processing time. These results suggested that, even if the exposure of hidden epitopes upon protein unfolding increased the antigenicity of whey proteins, further peptide bonds cleavage also took place after hydrolysis. This effect could modify whey proteins antigenic sequences, and thus their antigenic power.     

Stability of protein‐stabilised β‐carotene nanodispersions against heating,salts and pH

BACKGROUND: Milk proteins are used in a wide range of formulated food emulsions. The stability of food emulsions depends on their ingredients and processing conditions. In this work, β‐carotene nanodispersions were prepared with selected milk‐protein products using solvent‐displacement method. The objective of this work was to evaluate the stability of these nanodispersions against heating, salts and pH. RESULTS: Sodium caseinate (SC)‐stabilised nanodispersions possessed the smallest mean particle size of 17 nm, while those prepared with whey‐protein products resulted in larger mean particle sizes (45–127 nm). Formation of large particles (mean particle size of 300 nm) started after 1 h of heating at 60 °C in nanodispersions prepared with SC. More drastic particle size changes were observed in nanodispersions prepared with whey protein concentrate and whey protein isolate. The SC‐stabilised nanodispersions were fairly stable against Na⁺ ions at concentrations below 100 mmol L^?1, but drastic aggregation occurred in ≥ 50 mmol L^?1 CaCl₂ solutions. Aggregation was also observed in whey protein‐stabilised nanodispersions after the addition of NaCl and CaCl₂ solutions. All sample exhibited the smallest mean particle size at neutral pH, but large aggregates were formed at both ends of extreme pH and at pH around the isoelectric point of the proteins. CONCLUSION: The nanodispersions prepared with SC were generally more stable against thermal processing, ionic strength and pH, compared to those prepared with whey proteins. The stable β‐carotene nanodispersions showed a good potential for industrial applications. Copyright © 2008 Society of Chemical Industry     

Enzymatic proteolysis,under high pressure of soybean whey: Analysis of peptides and the allergen Gly m 1 in the hydrolysates

Soybean (Glycine max) whey was hydrolyzed with Alcalase, Neutrase, Corolase 7089 and Corolase PNL during high pressure (HP) treatment at 100, 200 and 300 MPa and at atmospheric pressure for 15 min. The protein content and the degree of hydrolysis were determined. Furthermore, the allergen Gly m 1 in the treated soybean whey and the hydrolysates was detected. The results showed that HP treatments increased the hydrolysis by the four proteases used. Pressure at 200 and 300 MPa proved to be better pressures to enhance the proteolysis. The immunochemical response of soybean whey to anti-Gly m 1 monoclonal antibodies decreased after the HP treatments and this decrease was greater after the combined treatment of high pressure and enzymatic hydrolysis. Soybean whey proteins hydrolysed at high pressure could be used as sources of peptides with low antigenicity when incorporated as food ingredients.