UHT milk contains multiple forms of αS1-casein that undergo degradative changes during storage

Milk proteins are susceptible to chemical changes during processing and storage. We used proteomic tools to analyse bovine α_S1-casein in UHT milk. 2-D gels of freshly processed milk α_S1-casein was presented as five or more spots due to genetic polymorphism and variable phosphorylation. MS analysis after phosphopeptide enrichment allowed discrimination between phosphorylation states and genetic variants. We identified a new alternatively-spliced isoform with a deletion of exon 17, producing a new C-terminal sequence, K¹⁶⁴SQVNSEGLHSYGL¹⁷⁷, with a novel phosphorylation site at S¹⁷⁴. Storage of UHT milk at elevated temperatures produced additional, more acidic α_S1-casein spots on the gels and decreased the resolution of minor forms. MS analysis indicated that non-enzymatic deamidation and loss of the N-terminal dipeptide were the major contributors to the changing spot pattern. These results highlight the important role of storage temperature in the stability of milk proteins and the utility of proteomic techniques for analysis of proteins in food.     

Effects of deamidation on aggregation and emulsifying properties of barley glutelin

The structure, solubility and emulsifying properties of the deamidated barley glutelin were investigated. The SDS–PAGE, size exclusion chromatography combined with a laser photometer (SEC-LP) and Fourier transform infrared spectroscopy (FTIR) results revealed that glutelin underwent a rapid hydrolysis and unfolding even at the initial stage of deamidation, leading to remarkably improved glutelin solubility at both acidic and neutral pHs. The deamidated glutelin demonstrated a strong tendency to form soluble aggregates of very large molecular weight (10⁶–10⁷ g/mol) probably due to disulphide-crosslinking and hydrophobic interactions. These aggregates played a major role in stabilising the emulsions at a broad range of deamidation levels (2.2–5.6% to 43%). In this way, undesirable property changes which result from excessive deamidation can be avoided, leading to wider applications of deamidated barley glutelin as an emulsifying ingredient in food and non-food applications.     

Effects of enzymatic modification of wheat protein on the formation of pyrazines and other volatile components in the Maillard reaction

Enzymatically hydrolysed wheat gluten hydrolysate (WGH) was deamidated using glutaminase to produce deamidated wheat gluten hydrolysate (DWGH). Volatile components were analysed in WGH and DWGH thermally reacted with glucose or fructose. In the reaction system containing glucose, 19 pyrazines, 2 furans, and 5 sulphur-containing components were detected in WGH, while 34 pyrazines, 4 furans, and 7 sulphur-containing components were found in DWGH. In the system containing fructose, 24 pyrazines, 3 furans, and 6 sulphur-containing components were identified in the thermal reaction of WGH, whereas 36 pyrazines, 4 furans, and 8 sulphur-containing components were found in DWGH. The volatile components increased in DWGH, both qualitatively and quantitatively, mainly due to free ammonia released by deamidation. More volatiles were also developed in WGH and DWGH with fructose than with glucose. It was found that ammonia released from wheat protein via deamidation participated in the generation of diverse volatile components including pyrazines in the Maillard reaction.     

Continuous production of wheat gluten peptide with foaming properties using immobilized enzymes

Gluten peptides are prepared using limited hydrolysis of wheat gluten, resulting in improved solubility and physico-chemical properties. In general, the hydrolysis is performed using a batch-wise process. In this study, we developed a bioreactor system that can continuously produce gluten peptides with foaming properties. Two kinds of acid protease, pepsin and rapidase, were immobilized on porous chitosan beads. The partial deamidation of gluten in advance increased initial velocity of hydrolysis by immobilizing protease and also worked to enhance foaming properties. A packed-bed reactor filled with immobilized protease was designed, and operating parameters were optimized. The optimized conditions were as follows: pH, 3.0; temperature, 40 °C; substrate concentration, 40 mg/ml; and space velocity, 2.0 h^–1. Based on these results, a bench-scale reactor was manufactured to determine the stability of continuous operation. The half-life of the reactor was approximately 45 days, and both productivity and quality were stable and excellent.     

Emulsion preparation with modified oat bran: Optimization of the emulsification process for microencapsulation purposes

The objective of this study was to develop an oil in water (o/w) emulsion using modified oat bran (MOB). Test emulsions containing 5% rapeseed oil were prepared by varying the content of MOB and maintaining the solid content constant (20%) by using corn syrup solids (DE34) as filler. A central composite (CC) design was applied as a tool to obtain a stable and low viscosity emulsion. The effect of concentration of MOB, homogenization pressure and homogenization time on the emulsion stability (at 25 °C for 26 h), viscosity and particle size were determined. The concentration of MOB in the recipe significantly influenced the emulsion properties whereas the homogenization pressure and the homogenization time had no statistically significant influence on the emulsion properties. Further experiments, however, showed that increasing the homogenization pressure decreased the emulsion viscosity. Emulsions prepared using a MOB concentration of 1.55% and homogenized at 60 MPa for 10 min, exhibited excellent stability, low viscosity, small enough mean particle size and narrow particle size distribution. Modified oat bran containing deamidated oat protein possesses an excellent capability to form stable emulsions which might be suitable for microencapsulation.     

Effects of succinylation and deamidation on functional properties of oat protein isolate

The effects of two different modification methods (deamidation and succinylation) on the functional properties (solubility, water- and oil-binding capacity, foaming capacity and stability, emulsion activity and stability) of oat protein isolates were evaluated. Protein isolates extracted from defatted oat flour at alkaline pH were acylated by 0.20 g/g of succinic anhydride. The protein isolate was also modified using a mild acidic treatment (HCl, 0.5 N). Succinylation and deamidation improved solubility and emulsifying activity of the native protein isolate. Foaming capacity of oat protein isolate increased after deamidation, whereas succinylation decreased it. The deamidated and succinylated proteins had lower foam and emulsion stabilities than had their native counterpart. Water- and oil-binding capacity, in both modified oat proteins, was higher than those of the native oat protein isolate.     

Microbial transglutaminase-induced structural and rheological changes of cationic and anionic myofibrillar proteins

This study investigated the effects of microbial transglutaminase (TG) on structural changes in porcine myofibrillar protein (MP) under varying pH (2.0–6.0) and two ionic strength conditions (0.1 M versus 0.6 M NaCl). Lowering the pH below the isoelectric point (pI) of myosin induced protein unfolding as revealed by surface hydrophobicity and differential scanning calorimetry. Although the MP solubility at the low ionic strength (0.1 M NaCl) was maximal at pH 3.0, both SDS-PAGE profiles and dynamic rheology indicated TG could not cross-link MP under this condition. Based on the carboxyl group content, the TG-catalyzed deamidation was dominant at a pH lower than the pI of myosin (pH 5.0) while cross-linking occurred at higher pH. Moreover, deamidation had no effect on rheological properties of MP. The results indicate that the TG reaction was governed by the pH of substrate protein, and the reaction intensity was related to the solubility of protein.     

Effects of glutaminase deamidation on the structure and solubility of rice glutelin

The effects of glutaminase on deamidation of rice glutelin were investigated. Water-insoluble rice glutelin was deamidated to the extent of deamidation degree 52.29% in 200 mmol/L sodium phosphate buffer (pH 7.0) at 37 °C for 48 h. Zeta potential analysis indicated that the glutamines of rice glutelin were deamidated into glutamic residues. Size exclusion chromatography results demonstrated that glutaminase deamidation broke the hydrophobic, hydrogen and some intermolecular disulfide bonds in rice glutelin and thereby rearranged the molecular weight distributions without serious cleavage of the peptide bonds. Fourier transform infrared analysis revealed the transformation of α-helix to random coil and β-turn by deamidation and suggested that deamidated rice glutelin maintained more flexible or extended forms. Solubility properties of rice glutelin in mildly acid (pH 5) and neutral buffers (pH 7) were remarkably improved by glutaminase deamidation. These new features of deamidated rice glutelin suggested that glutaminase could be a potential tool for enhancing the usability of rice protein in the food industry.