Preparation of lactose‐free milk by using salt‐fractionated almond (Amygdalus communis) β‐galactosidase

β‐galactosidase was isolated from almond (Amygdalus communis) extract by ammonium sulfate precipitation. Almond proteins precipitated by using ammonium sulfate and then dialysed exhibited 5.3‐fold purification of β‐galactosidase, and the yield of enzyme preparation was 96.5%. The partially purified β‐galactosidase exhibited pH and temperature optima at pH 5.5 and 50 °C, respectively. The enzyme was significantly stable against heat, pH, calcium and magnesium ions and D ‐galactose. The almond β‐galactosidase preparation exhibited over 89% activity even after 2 months storage at 4 °C. Hydrolysis of lactose in milk and whey was performed in a stirred batch process by using this enzyme preparation. These observations indicated that the hydrolysis of lactose increased continuously with time. The enzyme could hydrolyse 94% of lactose in buffer solution and whey whereas 90% of lactose hydrolysis was achieved in milk. The main aim of the present study was to prepare lactose‐free milk, which must be free from contamination, and the process should be inexpensive. Copyright © 2007 Society of Chemical Industry     

A Simple and Fast Detection Method for Bovine Milk Residues in Foods: A 2‐Site Monoclonal Antibody Immunochromatography Assay

The ingredient declaration on food labels assumes paramount importance in the protection of food‐allergic consumers. China has not implemented Food allergen labeling. A gold immunochromatography assay (GICA) was developed using 2 monoclonal antibodies (mAb) against the milk allergen β‐lactoglobulin in this study. The GICA was specific for pure milk samples with a sensitivity of 0.2 ng/mL. Milk protein traces extracted from 110 food products were detected by this method. The labels of 106 were confirmed by our GICA method: 57 food samples originally labeled as containing milk were positive for β‐lactoglobulin and 49 food samples labeled as not containing milk were negative for β‐lactoglobulin. However, 3 food samples falsely labeled as containing milk were found to contain no β‐lactoglobulin whereas 1 food sample labeled as not containing milk actually contained β‐lactoglobulin. First, these negatives could be because of the addition of a casein fraction. Second, some countries demand that food manufacturers label all ingredients derived from milk as “containing milk” even though the ingredients contain no detectable milk protein by any method. Our GICA method could thus provide a fast and simple method for semiquantitatation of β‐lactoglobulin in foods. Practical Application: The present method provides a fast, simple, semiquantitative method for the determination of milk allergens in foods.     

Effects of Different β‐D‐Glycosidases on Bound Aroma Compounds in Muscat Grape Determined by HS‐SPME and GC‐MS

Important “floral” aromas naturally occur in grapes predominantly as flavourless glycoconjugate precursors. Since these aroma compounds can be released by hydrolysis, different glycosidase enzymes can potentially contribute different aromas to wines. In this paper, we first established a procedure for profiling the free and bound volatile compounds in grape using GC‐MS combined with headspace solid‐phase microextraction (HS‐SPME). Comparison of the free and bound aroma compounds revealed that non‐volatile glycosides, known as aroma precursors, occur in high concentrations in musts. Among all compounds identified, 11 were fully quantified according to established standard calibration curves, while others were semi‐quantified. Using three different glycosidase enzymes, a total of 38 bound volatile compounds were identified in Muscat grape, including terpenes, higher alcohols, C‐6 compounds, and phenols, among others. The different enzymes had significant effects on the varietal aroma. Principal component analysis indicated that the characteristic aroma hydrolyzed by the commercial enzyme AR2000 was clearly different from that produced by other enzymes.     

Glycation of the Major Milk Allergen β‐Lactoglobulin Changes Its Allergenicity by Alterations in Cellular Uptake and Degradation

1 Scope

During food processing, the Maillard reaction (МR) may occur, resulting in the formation of glycated proteins. Glycated proteins are of particular importance in food allergies because glycation may influence interactions with the immune system. This study compared native and extensively glycated milk allergen β‐lactoglobulin (BLG), in their interactions with cells crucially involved in allergy.

2 Methods and results

BLG was glycated in MR and characterized. Native and glycated BLG were tested in experiments of epithelial transport, uptake and degradation by DCs, T‐cell cytokine responses, and basophil cell degranulation using ELISA and flow cytometry. Glycation of BLG induced partial unfolding and reduced its intestinal epithelial transfer over a Caco‐2 monolayer. Uptake of glycated BLG by bone marrow–derived dendritic cells (BMDC) was increased, although both BLG forms entered BMDC via the same mechanism, receptor‐mediated endocytosis. Once inside the BMDC, glycated BLG was degraded faster, which might have led to observed lower cytokine production in BMDC/CD4⁺ T‐cells coculture. Finally, glycated BLG was less efficient in induction of degranulation of BLG‐specific IgE sensitized basophil cells.

3 Conclusions

This study suggests that glycation of BLG by MR significantly alters its fate in processes involved in immunogenicity and allergenicity, pointing out the importance of food processing in food allergy.     

A comparison of micellar casein and β‐casein as sources of basic peptides through tryptic hydrolysis and their enrichment using two‐stage ultrafiltration

For the application of functional peptides in innovative products, these peptides have to be individually available. This may be accomplished by a well‐considered combination of a pure protein precursor, a selective cleaving enzyme, and an advanced separation technology. In this proof‐of‐principle approach, micellar casein and β‐casein were subjected to tryptic hydrolysis and the generated casein peptides were identified and characterised. The basic target peptides, β‐casein f(177‐183) and β‐casein f(170‐176), were then enriched. A permeate containing 35% of the target peptides was produced after two‐stage ultrafiltration of micellar casein tryptic hydrolysate. In comparison, enrichment up to 58% was achieved starting from β‐casein tryptic hydrolysate.     

The initial stage of high‐pressure induced β‐lactoglobulin aggregation: the long‐run simulation

Research objective of this study was to clarify how the initial stage of high‐pressure induced aggregation of β‐lactoglobulin takes place. For this purpose a special simulation method was developed. Distinctive features of this approach are: the lowest resolution model of protein particles, the local interaction potential and the abandonment of the continuous simulation of particle trajectories. Relatively short molecular dynamics trajectories are used only in order to find the average time step between the collisions. Results of particle collisions that occur with some probability, are separately (‘statically’) modelled using a random variable. This allows the analysis of the process which takes place within 10²–10³ s real‐time, with an existing probability of successful collision of 10^?10–10^?11. Modelling results confirm that at the initial stage of aggregation of 0.5–2% solutions with a neutral pH‐value only dimers as well as trimers arise due to SH/S–S interaction. In addition it was shown that aggregation follows the general principles of the reaction‐limited aggregation of colloids. The proposed approach could further be used in research projects examining the aggregation of β‐lactoglobulin or similar systems.     

Effect of In Vitro Digestion on Free α‐Dicarbonyl Compounds in Balsamic Vinegars

We investigated the influence of an in vitro simulated digestion process on the content of the free α‐dicarbonyl compounds most frequently found in food. A Glyoxal (GO), methylglyoxal (MGO), and diacetyl (DA) aqueous standard mixture and 2 brands of balsamic vinegar were analyzed before and after exposure to digestive enzymes. A strong matrix effect required adoption of validated RP‐HPLC‐DAD standard addition methods. The results showed that the digestive enzymes markedly alter the concentrations of the exogenous free α‐dicarbonyl compounds ingested with food; the extent of such changes varied with the α‐dicarbonyl compound itself and the diet components, which determined important but different food matrix effects also during digestion. The data also indicate that digestion can reduce the bioavailability of the toxic α‐dicarbonyl compounds ingested with food. However, no firm conclusions can be drawn about a putative positive influence of digestion on the toxic potential of dietary α‐dicarbonyl compounds, because their reaction in the presence of digestive enzymes likely gives rise to advanced glycation end products, which are involved in the development of chronic diseases.     

In vitro β‐Carotene Bioaccessibility and Lipid Digestion in Emulsions: Influence of Pectin Type and Degree of Methyl‐Esterification

Citrus pectin (CP) and sugar beet pectin (SBP) were demethoxylated and fully characterized in terms of pectin properties in order to investigate the influence of the pectin degree of methyl‐esterification (DM) and the pectin type on the in vitro β‐carotene bioaccessibility and lipid digestion in emulsions. For the CP based emulsions containing β‐carotene enriched oil, water and pectin, the β‐carotene bioaccessibility, and lipid digestion were higher in the emulsions with pectin with a higher DM (57%; “CP57 emulsion”) compared to the emulsions with pectin with a lower DM (30%; “CP30 emulsion”) showing that the DM plays an important role. In contrast, in SBP‐based emulsions, nor β‐carotene bioaccessibility nor lipid digestion were dependent on pectin DM. Probably here, other pectin properties are more important factors. It was observed that β‐carotene bioaccessibility and lipid digestion were lower in the CP30 emulsion in comparison with the CP57, SBP32, and SBP58 emulsions. However, the β‐carotene bioaccessibility of CP57 emulsion was similar to that of the SBP emulsions, whereas the lipid digestion was not. It seems that pectin type and pectin DM (in case of CP) are determining which components can be incorporated into micelles. Because carotenoids and lipids have different structures and polarities, their incorporation may be different. This knowledge can be used to engineer targeted (digestive) functionalities in food products. If both high β‐carotene bioaccessibility and high lipid digestion are targeted, SBP emulsions are the best options. The CP57 emulsion can be chosen if high β‐carotene bioaccessibility but lower lipid digestion is desired.     

Interaction of β‐casein with (−)‐epigallocatechin‐3‐gallate assayed by fluorescence quenching: effect of thermal processing temperature

The interactions between the flavan‐3‐ol (?)‐epigallocatechin‐3‐gallate (EGCG) and bovine β‐casein in phosphate‐buffered saline (PBS) of pH 6.5 subjected to thermal processing at various temperatures (25–100 °C) were investigated using fluorescence quenching. The results indicated that different temperatures had different effects on the structural changes and EGCG‐binding ability of β‐casein. At temperatures below 60 °C, the β‐casein–EGCG interaction changed little (P > 0.05) with increasing temperature. At temperatures above 80 °C, native assemblies of β‐casein in solution dissociated into individual β‐casein molecules and unfolded, as demonstrated by a red shift of the maximum fluorescence emission wavelength (λ_max) of up to 8.8 nm. The highest quenching constant (K_q) and the number of binding sites (n) were 0.92 (±0.01) × 10¹³ m ^?1 s^?1 and 0.73 (±0.02) (100 °C), respectively. These results provide insight into the potential of interactions between β‐casein–EGCG that may modulate bioactivity or bioavailability to be altered during thermal process.     

Provitamin‐A and xanthophyll carotenoids in vegetables and food grains of nutritional and medicinal importance

This study reports carotenoid composition of vegetables (n = 56), cereals (n = 12), pulses and legumes (n = 12), analysed by HPLC. It was hypothesised that food grains, like vegetables may be good sources of carotenoids. Amongst vegetables, higher level (mg/100 g dry weight) of lutein (210–419) was detected in green/red/capsicum and yellow zucchini, whilst zeaxanthin was dominant in kenaf (4.59). β‐Carotene (mg/100 g dry weight) was higher in green capsicum and kenaf (48,159) whilst carrot, ivy gourd and green capsicum contain α‐carotene (22–110). Amongst food grains, chickpea, split red gram and flaxseed contain higher levels (μg/100 g dry weight) of lutein (185–200) whilst zeaxanthin level was highest in puffed chickpea (1.8). Red unpolished parboiled rice was richest (μg/100 g dry weight) in β‐carotene (67.6) whilst whole black gram contained higher levels of α‐carotene (52.7). Thus, results indicate that chickpea and red unpolished parboiled rice are good sources of carotenoids. These carotenoid‐rich vegetables and grains may be exploited to meet the lutein and β‐carotene requirement.     

Oxidatively induced chemical changes and interactions of mixed myosin, β‐lactoglobulin and soy 7S globulin

The effects of oxidation on the chemical characteristics of myosin, β‐lactoglobulin and soy 7S globulin were investigated in a free radical‐generating system (FeCl₃/H₂O₂/ascorbate). Oxidised myosin exhibited a marked increase (>10‐fold) in carbonyls, a small increase in amines and conversion of some free sulphhydryls to disulphide bonds. Oxidised β‐lactoglobulin and 7S globulin also showed a major increase in the carbonyl content (five‐ and threefold respectively), but other chemical changes were relatively minor. In the oxidised myosin/β‐lactoglobulin composites, some cross‐linked aggregates were formed. Aggregation was also evidenced in the myosin/7S globulin composites exposed to the oxidising agents. The results indicated that oxidation enhanced interactions of the non‐muscle proteins with myosin apparently through the modification of amino acid side chains. These physicochemical changes may influence the functionality of processed muscle foods. © 2000 Society of Chemical Industry     

Effects of low‐temperature blanching on tissue firmness and cell wall strengthening during sweet potato flour processing

Free starch rate has been one of the most important criterions to evaluate the quality of sweet potato flour. Low‐temperature blanching (LTB) of sweet potatoes before steam cooking has shown significant increase in tissue firmness and cell wall strengthening. This research indicated that pectin methylesterase (PME) activity decreased by 87.8% after 30 min of blanching in water at 60 °C, while polygalacturonase (PG) and β‐amylase activity decreased 69.4% and 7.44%, respectively, under the same condition. Both PME and β‐amylase played important roles in tissue firmness. Further studies of tissue firmness and methyl esterification showed that the combination of LTB and Ca²⁺ could increase the activity of PME and significantly enhance the pectin gel hardness to strengthen the cell walls and decrease free starch rate from 12.83% to 7.28%.     

Emulsifying Properties of Legume Proteins Compared to β‐Lactoglobulin and Tween 20 and the Volatile Release from Oil‐in‐Water Emulsions

The emulsifying properties of plant legume protein isolates (soy, pea, and lupin) were compared to a milk whey protein, β‐lactoglobulin (β‐lg), and a nonionic surfactant (Tween 20). The protein fractional composition was characterized using sodium dodecyl sulfate–polyacrylamide gel electrophoresis analysis. The following emulsion properties were measured: particle diameter, shear surface ζ‐potential, interfacial tension (IT), and creaming velocity. The effect of protein preheat treatment (90 °C for 10 min) on the emulsifying behavior and the release of selected volatile organic compounds (VOCs) from emulsions under oral conditions was also investigated in real time using proton transfer reaction‐mass spectrometry. The legume proteins showed comparable results to β‐lg and Tween 20, forming stable, negatively charged emulsions with particle diameter d_3,2 < 0.4 μm, and maintained stability over 50 d. The relatively lower stability of lupin emulsions was significantly correlated with the low protein surface hydrophobicity and IT of the emulsion. After heating the proteins, the droplet size of pea and lupin emulsions decreased. The VOC release profile was similar between the protein‐stabilized emulsions, and greater retention was observed for Tween 20‐stabilized emulsions. This study demonstrates the potential application of legume proteins as alternative emulsifiers to milk proteins in emulsion products.