Pathways of the Maillard reaction under physiological conditions

Initially investigated as a color formation process in thermally treated foods, nowadays, the relevance of the Maillard reaction in vivo is generally accepted. Many chronic and age-related diseases such as diabetes, uremia, atherosclerosis, cataractogenesis and Alzheimer’s disease are associated with Maillard derived advanced glycation endproducts (AGEs) and α-dicarbonyl compounds as their most important precursors in terms of reactivity and abundance. However, the situation in vivo is very challenging, because Maillard chemistry is paralleled by enzymatic reactions which can lead to both, increases and decreases in certain AGEs. In addition, mechanistic findings established under the harsh conditions of food processing might not be valid under physiological conditions. The present review critically discusses the relevant α-dicarbonyl compounds as central intermediates of AGE formation in vivo with a special focus on fragmentation pathways leading to formation of amide-AGEs.     

The non-oxidative degradation of ascorbic acid at physiological conditions

The degradation of L-ascorbate (AsA) and its primary oxidation products, L-dehydroascorbate (DHA) and 2,3-L-diketogulonate (2, 3-DKG) were studied under physiological conditions. Analysis determined that L-erythrulose (ERU) and oxalate were the primary degradation products of ASA regardless of which compound was used as the starting material. The identification of ERU was determined by proton decoupled (13)C-nuclear magnetic resonance spectroscopy, and was quantified by high performance liquid chromatography, and enzymatic analysis. The molar yield of ERU from 2,3-DKG at pH 7.0 37 degrees C and limiting O(2)97%. This novel ketose product of AsA degradation, was additionally qualitatively identified by gas-liquid chromatography, and by thin layer chromatography. ERU is an extremely reactive ketose, which rapidly glycates and crosslinks proteins, and therefore may mediate the AsA-dependent modification of protein (ascorbylation) seen in vitro, and also proposed to occur in vivo in human lens during diabetic and age-onset cataract formation.     

The importance of the Maillard reaction in ophtalmology

Non enzymatic glycosylation( glycation) of proteins, described by L. C. Maillard in 1912, results in the formation of advanced glycation end products (AGE-s). These exhibit a number of harmful reactions, increasing with age and involved in several age-associated pathologies. In ocular pathology, their role was demonstrated at several levels of age-associated eye-diseases, such as the rigidification of cornea, in the separation of vitreous fibers from the hyaluronan jelly, which might result in retinal detachment. AGE-s are involved also in retinal microvascular alterations in diabetics as well as in age-related macular degeneration. We compared the cytotoxic effect of several AGE-s on human skin fibroblasts and corneal keratocytes. Keratocytes were shown to be much more resistant to the cytotoxic effect of several AGE-products than fibroblasts. This higher resistance of keratocytes to the free radical mediated cytotoxic effect of AGE-s might be the result of the constant exposure of cornea to UV-light possibly mediating the appearance of more efficient protective mechanisms during evolution.     

Inhibitory mechanisms of Maillard reaction products

In order to determine the inhibitory mechanism of antibacterial Maillard reaction products (MRP), heated mixtures containing arginine and xylose (AX) or histidine and glucose (HG) were studied for their mutagenic effect, using the Salmonella mutagenic test system ('Ames test'), with regard to their effect on the uptake of serine, glucose and oxygen and for their effect on iron solubility. It was found that the MRPs tested had little or no mutagenic effect, while an inhibitory effect on the uptake of serine, glucose and oxygen was observed. The MRPs also had an effect on the solubility of iron in nutrient broth and in phosphate buffer. While the AX mixture reduced the solubility of iron it was increased by the HG mixture. The reduction of iron solubility in the presence of the AX mixture was much greater at pH 8 than at pH 5. The present data suggest that the antibacterial effect of the MRPs tested is primarily due to the interaction between MRPs and iron, resulting in reduced oxygen uptake.     

The pathogenic role of Maillard reaction in the aging eye

The proteins of the human eye are highly susceptible to the formation of advanced glycation end products (AGEs) from the reaction of sugars and carbonyl compounds. AGEs progressively accumulate in the aging lens and retina and accumulate at a higher rate in diseases that adversely affect vision such as, cataract, diabetic retinopathy and age-related macular degeneration. In the lens AGEs induce irreversible changes in structural proteins, which lead to lens protein aggregation and formation of high-molecular-weight aggregates that scatter light and impede vision. In the retina AGEs modify intra- and extracellular proteins that lead to an increase in oxidative stress and formation of pro-inflammatory cytokines, which promote vascular dysfunction. This review outlines recent advances in AGE research focusing on the mechanisms of their formation and their role in cataract and pathologies of the retina. The therapeutic action and pharmacological strategies of anti-AGE agents that can inhibit or prevent AGE formation in the eye are also discussed.     

Aerobic degradation of adamantanes at highly acidic conditions

Biodegradation of alkyl-substituted adamantane derivatives (1-methyl, 1,3-dimethyl-, and 1,3,5-trimethyladamantane) by slow-growing bacteria Mycobacterium AG_S10 was studied. The process was carried out under extremely acidic conditions (pH 2.5). Bacterial strain AG_S10 was able to utilize these alicyclic hydrocarbons with a high degree of condensation and diamond-like structure, which are usually resistant to microbial transformation. Efficiency of alkyaldamantane biodegradation by the cells growing with these substrates as the sole carbon and energy sources was affected significantly by their aggregate state, which depended on molecular structure. Compared to the solid 1-methyladamantane, 1,3-dimethyladamantane, which is liquid under normal conditions, was a preferable substrate. Adamantanes in the gas condensate were generally more resistant to bacterial degradation than such markers as normal and isoprenoid alkanes. Moreover, biodegradation had no significant effect on relative distribution of the tested С₁₁–С₁₃ alkyladamantanes.     

Prion inactivation by the Maillard reaction

Since variant Creutzfeldt-Jakob disease (vCJD) has been suspected to be attributable to the infectious agents associated with bovine spongiform encephalopathy (BSE), it is important to prevent the transmission of pathogenic forms of prion protein (PrP(Sc)) through contaminated feeding materials such as meat and bone meal (MBM). Here, we demonstrate that the Maillard reaction employing a formulation of glucose in combination with sodium hydrogen carbonates effectively reduced the infectivity (approximately 5.9-log reduction) of a scrapie-infected hamster brain homogenate. In addition to a bioassay, a protein misfolding cyclic amplification (PMCA) technique, in which PrP(Sc) can be amplified in vitro, was used as a rapid test for assessing PrP(Sc) inactivation. The PMCA analysis also indicated that the PrP(Sc) level in the infected material significantly decreased following the Maillard reaction. Therefore, the Maillard reaction can be employed for the decontamination of large amounts of byproducts such as MBM.     

The Maillard reaction and oxidative stress during aging of soybean seeds

The chemical reactions that may lead to the loss of seed viability were investigated both during the accelerated aging and natural aging of soybeans ( Glycine max Merrill cv. Chippewa 64). Under conditions of accelerated aging (36°C and 75% RH), fluorescence of soluble proteins accumulated, which was closely correlated with the loss of seed germinability and vigor. We were able to show this correlation by using partially purified proteins for the assay. Fluorescence also increased in seeds under good storage conditions (5°C for up to 21 years), although there was a less significant correlation between seed viability and the accumulation of fluorescent products during the time of natural aging. The rise in protein fluorescence is interpreted as an increase of Maillard products. The carbonyl content of soluble proteins (a measure of the oxidative damage) did not change significantly during either accelerated aging or natural aging: however the elimination of carbonyls during germination seemed to be hindered in seeds that had poor germination. The Maillard reaction may be a consequence of the formation of reducing sugars through a gradual hydrolysis of oligosaccharides during aging. Preliminary evidence from the natural aging study showed that, when seeds were in the glassy state, the sugar hydrolysis was inhibited. These results suggest that the Maillard reaction and oxidative reaction may play an important role in seed deterioration.     

Fractionating recalcitrant lignocellulose at modest reaction conditions

Effectively releasing the locked polysaccharides from recalcitrant lignocellulose to fermentable sugars is among the greatest technical and economic barriers to the realization of lignocellulose biorefineries because leading lignocellulose pre-treatment technologies suffer from low sugar yields, and/or severe reaction conditions, and/or high cellulase use, narrow substrate applicability, and high capital investment, etc. A new lignocellulose pre-treatment featuring modest reaction conditions (50 degrees C and atmospheric pressure) was demonstrated to fractionate lignocellulose to amorphous cellulose, hemicellulose, lignin, and acetic acid by using a non-volatile cellulose solvent (concentrated phosphoric acid), a highly volatile organic solvent (acetone), and water. The highest sugar yields after enzymatic hydrolysis were attributed to no sugar degradation during the fractionation and the highest enzymatic cellulose digestibility ( approximately 97% in 24 h) during the hydrolysis step at the enzyme loading of 15 filter paper units of cellulase and 60 IU of beta-glucosidase per gram of glucan. Isolation of high-value lignocellulose components (lignin, acetic acid, and hemicellulose) would greatly increase potential revenues of a lignocellulose biorefinery.     

Maillard reaction rate in various glassy matrices

The Maillard Reaction (MR) rate below the glass transition temperature (T(g)) for various model glassy food systems was studied at temperatures between 40 degrees C and 70 degrees C. As a sample, freeze-dried glucose and lysine systems embedded in various glassy matrices (e.g., polyvinylpyrrolodone and trehalose) were used, and the MR rate below the T(g) was compared among the various glassy matrices. The extent of MR was estimated spectrophotometrically from the optical density at 280 nm (OD(280)), and the MR rate (k(280)) was determined as a pseudo zero order reaction rate from the time course of OD(280). Although k(280) was described by the Arrhenius plot, the temperature dependence of k(280) was almost the same and the intercept was different among the matrices. From the comparison of k(280), it was suggested that the MR rate in glassy matrix was affected not only by the T(g), but also by the hydrogen bonding between MR reactants and glassy matrix.     

Intervention against the Maillard reaction in vivo

The field of Maillard/glycation reactions in vivo has grown enormously during the past 20 years, going from 25 to 500 publications per year. It is now well recognized that many of the “advanced” products form oxidatively or anaerobically and can have deleterious effects on macromolecular and biological function. The feasibility of developing pharmacological agents with beneficial in vivo properties, based on in vitro inhibition of glycation, has been surprisingly successful. This Editorial sets the stage for a series of articles by experts in the field, who have made key contributions to our understanding of the Maillard reaction in vivo.     

Micellization of casein-graft-dextran copolymer prepared through Maillard reaction

Casein is almost insoluble at around pH 4.6, which is its isoelectric point (pI). Grafting copolymer, casein-g-dextran, was prepared through the Amadori rearrangement of the Maillard reaction. The copolymer has a reversible pH sensitive property: micellization at the pI of casein forming a casein core and dextran shell structure and dissociation when pH differs from the pI. The micelles produced at pH 4.6 have a spherical shape and their size is dependent on the Maillard reaction: reaction time, molar ratio of casein to dextran, and molecular weight of dextran used. Typically, the hydrodynamic diameter of the micelles is about 100 nm and the critical micelle concentration is about 10 mg/L. The micelles are very stable in aqueous solution and can be stored as lyophiled powder. The micelles are able to encapsulate hydrophobic compounds such as pyrene.     

Formation of flavour compounds in the Maillard reaction

This paper discusses the importance of the Maillard reaction for food quality and focuses on flavour compound formation. The most important classes of Maillard flavour compounds are indicated and it is shown where they are formed in the Maillard reaction. Some emphasis is given on the kinetics of formation of flavour compounds. It is concluded that the essential elements for predicting the formation of flavour compounds in the Maillard reaction are now established but much more work needs to be done on specific effects such as the amino acid type, the pH, water content and interactions in the food matrix. It is also concluded that most work is done on free amino acids but hardly anything on peptides and proteins, which could generate peptide- or protein-specific flavour compounds.     

Lack of effect of copper on advanced Maillard reaction and glucose autoxidation at physiological concentrations of albumin

Summary

This study examines the possible action of copper on advanced glycation. Copper has been shown to induce fluorescence due to advanced-glycated-end-products (AGEs) on albumin incubated with glucose, and this was interpreted as activation of the glucose or Amadori product (AP) autoxidation. We glycated albumin (60 g/L) to several levels with increasing concentrations of glucose. The dialysed glucose-free glycated albumin was then incubated with 1.5 μmol/L copper or 1 mmol/L diethylenetriaminepentaacetic acid (DTPA), plus or minus glucose. The production of AP, measured as furosine, was similar whether DTPA or copper was present in the incubation medium. It linearly increased as a function of time and glucose concentration in both cases up to a maximum (furosine around 20 mmol/g protein), indicating saturation of the free NH₂ residues on the protein. The fluorescence due to AGEs increased linearly over time for glycated albumin incubated without glucose, and exponentially when glucose was added to the incubation medium. This fluorescence was also unaffected by DTPA or copper for a glucose concentration below 125 mmol/L and initial furosine below 10 mmol/g. However copper caused a slight activation in samples with very high glucose (1.25 mol/L) and furosine (30–40 mmol/g) concentrations. We therefore find no effect of copper in this experiment, because the copper concentration is lower and the albumin higher than that used in previous studies. In these conditions, albumin chelates copper and inhibits its oxidative activity. The protein concentrations used in most in vitro studies showing a copper effect were below 10 g/L with copper often above 10 μmol/L, so that copper may act oxidatively. As the lens and arterial wall have high protein concentrations, copper should have no action on protein glycation in vivo, unless altered protein structure impedes the inactivation of copper by chelation.     

Pyridinium-carbaldehyde: active Maillard reaction product from the reaction of hexoses with lysine residues

Besides the formation of the aminotriazine N6-[4-(3-amino-1,2,4-triazin-5-yl)-2,3-dihydroxybutyl]-L-lysine, the reaction of [1-13C]D-glucose with lysine and aminoguanidine leads to the generation of 6-[2-([[amino(imino)methyl]hydrazono]methyl)pyridinium-1-yl]-L-norleucine (14-13C1). The dideoxyosone N6-(2,3-dihydroxy-5,6-dioxohexyl)-L-lysine was shown to be a precursor in the formation of 14-13C1, which proceeds via the reactive carbonyl intermediate 6-(2-formylpyridinium-1-yl)-L-norleucine (13-13C1). In order to study the reactivity of 13-13C1, the model compound 1-butyl-2-formylpyridinium (18) was prepared in a two-step procedure starting from 2-pyridinemethanol. The reaction of the pyridinium-carbaldehyde 18 with L-lysine yielded the Strecker analogous degradation product 2-(aminomethyl)-1-butylpyridinium and another compound, which was shown to be as 1-butyl-2-[(2-oxopiperidin-3-ylidene)methyl]pyridinium. Reaction of 18 with the C-H acidic 4-hydroxy-5-methylfuran-3(2H)-one leads to the formation of the condensation product 1-butyl-2-[hydroxy-(4-hydroxy-5-methyl-3-oxofuran-2(3H)-ylidene)methyl]-pyridinium.