Effect of lactose on cross‐linking of milk proteins during heat treatments

The major types of nondisulphide cross‐linking which cause milk protein aggregation were investigated in milk, with and without lactose, heated at 95 °C for up to 8 h. Compared with the milk containing no lactose, the milk containing lactose showed a smaller increase in pH, a larger increase in pH 4.6 soluble nitrogen, much smaller increase in lysinoalanine (LAL) and a much higher percentage of cross‐linked proteins. It was concluded that cross‐linking in milk products containing lactose occurs mainly via Maillard reaction products, and in milk products with no lactose, it occurs mainly via isopeptide linkages such as in LAL.     

The dual effects of Maillard reaction and enzymatic hydrolysis on the antioxidant activity of milk proteins

The objective of this study was to determine the enhanced effects on the biological characteristics and antioxidant activity of milk proteins by the combination of the Maillard reaction and enzymatic hydrolysis. Maillard reaction products were obtained from milk protein preparations, such as whey protein concentrates and sodium caseinate with lactose, by heating at 55°C for 7 d in sodium phosphate buffer (pH 7.4). The Maillard reaction products, along with untreated milk proteins as controls, were hydrolyzed for 0 to 3 h with commercial proteases Alcalase, Neutrase, Protamex, and Flavorzyme (Novozymes, Bagsværd, Denmark). The antioxidant activity of hydrolyzed Maillard reaction products was determined by reaction with 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, their 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, and the ability to reduce ferric ions. Further characteristics were evaluated by the o-phthaldialdehyde method and sodium dodecyl sulfate-PAGE. The degree of hydrolysis gradually increased in a time-dependent manner, with the Alcalase-treated Maillard reaction products being the most highly hydrolyzed. Radical scavenging activities and reducing ability of hydrolyzed Maillard reaction products increased with increasing hydrolysis time. The combined products of enzymatic hydrolysis and Maillard reaction showed significantly greater antioxidant activity than did hydrolysates or Maillard reaction products alone. The hydrolyzed Maillard reaction products generated by Alcalase showed significantly higher antioxidant activity when compared with the other protease products and the antioxidant activity was higher for the whey protein concentrate groups than for the sodium caseinate groups. These findings indicate that Maillard reaction products, coupled with enzymatic hydrolysis, could act as potential antioxidants in the pharmaceutical, food, and dairy industries.     

Demonstration of the presence of aminoreductone formed during the Maillard reaction in milk

The Maillard reaction is a common chemical reaction that occurs in food and it generates multiple reaction products. Aminoreductone (AR) is one of the early-stage Maillard reaction products. At present the formation of AR has only been demonstrated in a model system consisting of a monosaccharide or disaccharide and an amino group-containing compound. There is no direct evidence to show the presence of AR in food. In this study, we demonstrated the formation and presence of AR in milk using a combination of 2,4-dinitrophenylhydrazine (DNP) and Cu²⁺. A DNP derivative of AR oxidised by Cu²⁺ was isolated and its detailed structure was identified by NMR analysis. We thus directly demonstrated the formation and presence of AR in milk.     

The effect of transglutaminase on the properties of milk gels and processed cheese

The aim of this study was to investigate the influence of transglutaminase (TG) on rennet coagulation properties, and the impact on the properties of processed cheese. The TG was added before, at the same time as, and after the addition of rennet. The parameters moisture, total solids and proteins and physical parameters (syneresis index, wheying‐off and rheological behaviour) were measured. The manufacturing of the milk gels treated with transglutaminase after rennet addition was shown to be an effective means of improving the physical properties (reduced synereseis index and increased consistency index) of processed cheese, possible by the occurrence of enzymatic cross‐linking.     

The preparation of an oligochitosan‐glycosylated and cross‐linked caseinate obtained by a microbial transglutaminase and its functional properties

A glycosylated and cross‐linked caseinate (GCC) with glucosamine amount of 4.74 g/kg protein was generated from caseinate and oligochitosan by a microbial transglutaminase. The applied temperature, pH and molar ratio of acyl donor/acceptor were 37 °C, 7.5 and 1:3, respectively; while caseinate concentration, transglutaminase addition and reaction time selected from single‐factor trials were 50 g/L, 10 kU/kg protein and 3 h, respectively. Electrophoretic analysis revealed the cross‐linking and glycosylation of caseinate. Compared with caseinate, GCC showed improved solubility in pH 4–11, higher digestibility in vitro and water binding capacity, about 3‐fold, but lower surface hydrophobicity and oil binding capacity (34%).     

乳及乳制品加工中的美拉德反应

针对乳及乳制品在加工和储藏过程中由于发生美拉德反应不仅使乳制品的营养物质受到破坏，还产生对人体有害的物质，在很大程度上影响的乳制品的质量，对乳制品中美拉德反应的历程，产生的标志物及其测定方法等进行了的综述。     

Characterisation and quantification of the reaction(s) catalysed by transglutaminase using the o‐phthaldialdehyde reagent

The potential application of the o‐phthaldialdehyde (OPA) reagent for quantification of the type and extent of the reaction(s) catalysed by transglutaminase (TGase) during incubation with sodium caseinate (NaCN) was investigated. Initial studies were performed to ensure that NH₃, a by‐product of TGase activity, could be determined with the OPA reagent in trichloroacetic acid (TCA) supernatants of NaCN solutions. The detectable concentration of exogenously added NH₃ (at NH₃ concentrations > 10 mM ) was found to decrease during extended incubation at 23, 37 and 50°C and at either pH 7.0 or 8.0 in 4% w/v NaCN solutions, even when taking into account the evaporation of water from the sample. The recovery of NH₃ from 12% w/v TCA supernatants of NaCN solutions spiked with 5 mM NH₃ at 23°C and pH 7.0 was found to be 88%. The release of NH₃ and the decrease in ε‐amino groups on incubating NaCN with TGase was subsequently quantified using the OPA reagent. Incubation of NaCN (4% w/v) with TGase at 23°C resulted in progressive increases and decreases, respectively, in NH₃ and ε‐amino group concentration with increased incubation time. These changes were dependent on TGase : NaCN. It was estimated that approximately 20% of the available Lys residues in NaCN were involved in TGase‐catalysed cross‐links. However, the observed decrease in ε‐amino group concentration was higher than expected. This may be due to concealment of noncross‐linked amino groups in polymerised NaCN, making them unavailable for reaction with the OPA reagent.     

The effects of using transglutaminase,exopolysaccharide‐producing starter culture and milk powder on the physicochemical,sensory and texture properties of low‐fat set yoghurt

This study focused on the effects of transglutaminase (TGase), exopolysaccharide (EPS)‐producing starter culture or skim milk powder addition on some properties of low‐fat set yoghurt. The incorporation of TGase to yoghurt did not affect the acidity of the experimental samples. However, the proteolytic activity of the starter culture significantly decreased in TGase‐treated yoghurt. Addition of TGase to yoghurt with a low dry matter content (11.08%) resulted in low serum separation and gel firmness and viscosity close to those of yoghurt with a higher dry matter content (14.81%). According to the sensory results, it would be possible to produce an acceptable product with a low dry matter and low fat content using TGase. The results shown that the use of EPS‐producing starter cultures for the purpose of improving the texture properties of low dry matter content yoghurt could not be recommended.     

Effects of cross‐linking on the rheological and thermal properties of sweet potato starch

Sweet potato starches were modified with three different concentrations of phosphorus oxychloride (POCl₃) (0.01, 0.02, and 0.03%, based on dry weight of starch) as a cross‐linking agent. The effects of crosslinking on rheological and thermal properties of sweet potato starch (SPS) pastes were evaluated. Cross‐linking considerably reduced the swelling power, consistency index (K), apparent viscosity (η_a), and yield stress (σ_oc) values of SPS, which significantly decreased with increase in POCl₃ concentration. The gelatinization temperature (T_p) and enthalpy (ΔH) values of the cross‐linked SPS, which were determined using differential scanning calorimetry, were higher than those of native SPS. Storage modulus (G′), loss modulus (G″), and complex viscosity (η*) of the cross‐linked SPS pastes determined using small deformation oscillatory rheometry, were higher than native starch, and they also decreased with increase in POCl₃ concentration from 0.01 to 0.03%. The tan δ (ratio of G″/G′) values (0.15–0.19) of the cross‐linked SPS samples were much lower than that (0.37) of the native SPS, indicating that the elastic properties of the SPS pastes were strongly influenced by modifications from cross‐linking. Finally, Cox–Merz plots showed that η* was much higher than η_a for the cross‐linked SPS pastes.     

The quality of set‐style yoghurt responsible to partial lactose hydrolysis followed by protein cross‐linking of the skimmed milk

Skimmed milk used for set‐style yoghurt production was treated with lactase at 0.1 g/kg for 30 min to give partial lactose hydrolysis and then treated with horseradish peroxidase and glucose oxidase at 200 and 6 kU/kg protein to result in protein cross‐linking. Two treatments conferred higher apparent viscosity on the milk, but led to the yoghurt prepared from it with insignificantly different chemical compositions to the counterparts (P > 0.05). The prepared yoghurt also showed decreased syneresis (about 17.7%), higher apparent viscosity and viscoelastic modulus, firmer texture and finer microstructure. This ternary enzyme system is a potential approach to improving the quality of set‐style yoghurt.     

Zinc‐loaded whey protein nanoparticles prepared by enzymatic cross‐linking and desolvation

Zinc‐loaded whey protein nanoparticles were prepared by enzymatic cross‐linking whey protein followed by ethanol desolvation. Whey protein isolate (WPI) was denatured by heating (80 °C for 15 min) at pH 7.0 and then cross‐linked by transglutaminase at 50 °C for 4 h while stirring. Transglutaminase was inactivated by heating at 90 °C for 10 min, and then, ZnSO₄·7H₂O (1–10 mm ) was added. Zinc‐loaded whey protein nanoparticles were formed by adding ethanol at one to five times the volume of the protein solution at pH 9.0. The desolvated solutions were diluted by adding distilled water at ratio of 1:100 (w/v) immediately after desolvation. Dynamic light scattering (DLS) data showed that the particle size of zinc‐loaded whey protein nanoparticles increased with the amount of zinc and the volume of ethanol. Scanning electron microscopy micrographs revealed an almost spherical morphology for zinc‐loaded whey protein nanoparticles. The zinc loading efficiency was determined ranging from 76.7% to 99.2%. In vitro test data showed that the zinc release rate was low in simulated gastric fluid but high in simulated intestinal fluid. The results indicated that enzymatic cross‐linked whey protein nanoparticles may be used as a good vehicle to deliver zinc as a supplement.     

Combined effect of autoclaving‐cooling and cross‐linking treatments of normal corn starch on the resistant starch formation and physicochemical properties

Normal corn starch (NCS) was treated by both autoclaving‐cooling and cross‐linking to produce resistant starch (RS). The RS yield, crystalline structure, as well as other physicochemical properties of the modified starch were investigated. The yield of RS was 12.2% by four autoclaving‐cooling cycles, and it could be further increased up to 31.0% by the subsequent cross‐linking using sodium trimetaphosphate/sodium tripolyphosphate. The scanning electron microscopy images clearly illustrated that the granular structure of native starch was disrupted and a continuous network with irregular shape was formed after autoclaving‐cooling cycles. The subsequent chemical cross‐linking appeared to make the network structure more compact and dense. X‐ray diffraction patterns showed that B‐ and V‐types coexisted in all the modified corn starches, and all these modified starches exhibited very low viscosity which remained almost constant regardless of temperature changes.     

Gel‐enhancing effect and protein cross‐ linking ability of tilapia sarcoplasmic proteins

BACKGROUND: Tilapia (Oreochromis niloticus) sarcoplasmic proteins contain substantial transglutaminase (TGase) activity. The enzyme catalyzes the protein cross‐linking reaction, resulting in a more elastic gel. The objective was to investigate the gel‐enhancing effect of sarcoplasmic proteins from tilapia as related to TGase activity. RESULTS: Total TGase activity of sarcoplasmic proteins concentrate (SpC) increased about 3.6‐fold after ultrafiltration using 30 kDa membrane, but specific activity remained unchanged, indicating minimal TGase purification by ultrafiltration. Addition of 1 mg mL^?1 SpC containing 40 units TGase activity induced cross‐linking of tilapia actomyosin, and the extent of cross‐linking increased with added level of SpC. Myosin heavy chain (MHC) and troponin were preferably cross‐linked by tilapia SpC, while actin and tropomyosin were not affected. Higher retention of MHC was observed concomitantly with greater content of cross‐linked protein when SpC was added to lizardfish surimi. Lizardfish surimi with 10 g kg^?1 SpC added and pre‐incubated at 37 °C for 1 h exhibited 91.6% and 26.7% increase in breaking force and deformation, respectively, when compared to the control. CONCLUSIONS: Residual TGase activity in SpC played an important role in catalyzing the protein cross‐linking and enhancing actomyosin gelation. SpC could be a potential ingredient for improving textural properties of fish protein gel. Copyright © 2007 Society of Chemical Industry     

Preventive effect of fermented Maillard reaction products from milk proteins in cardiovascular health

The aim of this study was to determine the dual effect of Maillard reaction and fermentation on the preventive cardiovascular effects of milk proteins. Maillard reaction products (MRP) were prepared from the reaction between milk proteins, such as whey protein concentrates (WPC) and sodium caseinate (SC), and lactose. The hydrolysates of MRP were obtained from fermentation by lactic acid bacteria (LAB; i.e., Lactobacillus gasseri H10, L. gasseri H11, Lactobacillus fermentum H4, and L. fermentum H9, where human-isolated strains were designated H1 to H15), which had excellent proteolytic and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities (>20%). The antioxidant activity of MRP was greater than that of intact proteins in assays of the reaction with 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt and trivalent ferric ions; moreover, the effect of MRP was synergistically improved by fermentation. The Maillard reaction dramatically increased the level of antithrombotic activity and 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) inhibitory effect of milk proteins, but did not change the level of activity for micellar cholesterol solubility. Furthermore, specific biological properties were enhanced by fermentation. Lactobacillus gasseri H11 demonstrated the greatest activity for thrombin and HMGR inhibition in Maillard-reacted WPC, by 42 and 33%, respectively, whereas hydrolysates of Maillard-reacted SC fermented by L. fermentum H9 demonstrated the highest reduction rate for micellar cholesterol solubility, at 52%. In addition, the small compounds that were likely released by fermentation of MRP were identified by size-exclusion chromatography. Therefore, MRP and hydrolysates of fermented MRP could be used to reduce cardiovascular risks.     

乳制品中的Maillard反应标示物及其应用

乳制品中的Maillard反应标示物既可作为乳制品热处理程度的指标，还可用于不同热处理乳制品的鉴别。介绍了在评价乳制品热处理程度以及区分不同乳制品中常用的几种Maillard反应标示物。这些标示物包括糠氨酸、乳果糖、糠醛类化合物、荧光化合物等。此外，还介绍了可溶性乳清蛋白、碱性磷酸酶和乳过氧化物酶等热处理标示物，这些标示物与Maillard反应之间具有较好的相关性，也是评价乳制品热处理程度以及区分不同乳制品时常用的指标，可与Maillard反应标示物联合应用。     

美拉德反应产物的抗突变作用

美拉德反应是食品制备、加工和贮存过程中发生的非酶褐变反应，褐变的发生对产品质量有很大影响，从食品卫生学的角度来看，有些产物对人体健康有极大的危害。本文讨论的是美拉德反应产物有益的一面，即其抗突变的效果。     

Effect of xylose on the molecular and particle size distribution of peanut hydrolysate in Maillard reaction system

BACKGROUND: The Maillard reaction is a complex series of reactions between reducing sugars and amino groups. Changing any of reaction parameters would alter the reaction pathway. This study investigated the effect of xylose concentration on the molecular and particle size distribution of Maillard reaction products (MRPs) derived from peanut hydrolysate and xylose to discuss their formation mechanism. RESULTS: Molecular weight and particle size distribution analyses indicated that both peptide degradation and peptide cross‐linking occurred during the Maillard reaction. Heat treatment would make the high‐molecular‐weight peptides degrade into low‐molecular‐weight peptides and free amino acids. Maillard reaction increased the molecular weight and particle sizes of products as the xylose concentration increased from 1% to 4%. CONCLUSION: The study shows that both peptide degradation and peptide cross‐linking occurred during the Maillard reaction. The thermal degradation product (TDP) and MRPs had significantly different molecular size distribution, and the particle size distribution of TDPs and MRPs had similar change tendency to that of the molecular size distribution. These would provide an insight into the formation mechanism of MRPs. Copyright © 2011 Society of Chemical Industry     

Influence of the Cross‐linking Agent on the Gel Structure of Starch Derivatives

Hydrogels were synthesized through cross‐linking of carboxymethyl starch (CMS; Degree of Substitution DS = 0.45) using polyfunctional carboxylic acids (malic, tartaric, citric, malonic, succinic, glutaric and adipic acid). The syntheses used a cross‐linking agent ratio (ratio of the number of cross‐linking agent molecules to the number of monomer units constituting the polymer) of F_Z = 0.05. After cross‐linking the gels were dried, ground and then hydrogels of a polymer concentration of 4 mass‐% were produced. These CMS‐hydrogels were then rheologically characterized using dynamic oscillatory measurements. From measurements of the plateau region storage modulus G'_P, the network parameters molar mass between two entanglement points M_e (M_e ranging from 9.318 (citric acid) to 281.397 g/mol (tartaric acid)), the cross‐link density ν_e and the distance between two entanglement points ξ were calculated. Using carboxylic acids without other functional groups, a maximum in gel sturdiness is found at a spacer length of two CH₂‐groups. The evaluation of the loss factor tan δ for the CMS‐hydrogels showed that values of tan δ = 0.2 varied only slightly with the frequency ω. Flow curves showed a pseudopIastic flow behavior for all CMS‐hydrogels (the shear viscosity η declining over five decades in the range of the shear rate γ of 10⁻³ to 10³ s⁻¹) The different polyfunctional carboxylic acids have a strong influence on the sturdiness of the synthesized CMS‐hydrogels.