Free Lysine Content Research Articles

Maternal dietary protein and amino acid intake is not associated with the amino acid composition of human milk in an affluent environment.

Amino acids (AA) are essential nutrients in human milk (HM) and critical for infant growth and development. Several maternal lifestyle factors have been suggested to influence HM AA composition, with possible consequences for the breastfed infant. Whether maternal dietary protein and AA intake is associated with AA concentrations in HM is still largely unknown. Therefore, the aim of this study was to investigate the association between maternal dietary AA intake and AA concentrations in HM over the first month postpartum. Data from the observational longitudinal Amsterdam Mother's Milk study were used, consisting of 123 lactating women in their first postpartum month. HM samples were collected three times, on day 10, 17 and 24 postpartum. Maternal dietary protein and AA intake on these collection days was assessed using three 24-h recalls. HM protein-bound and free AA (BAA and FAA, respectively) were analysed by liquid chromatography. Associations between maternal AA intake and AA concentrations in HM were assessed using linear mixed models. Maternal intake was negatively associated with milk concentrations of free arginine (-0·0003; P = 0·01) and free lysine (-0·0004; P = 0·03) and was positively associated with free glutamine (0·002; P = 0·03) and free threonine (0·0008; P = 0·03). However, these associations were attenuated after correction for multiple testing. Both the quality and quantity of dietary protein intake in lactating women do not seem to influence the amino composition of their breast milk when living in an affluent environment.

Sulfonamides stimulate ROS formation upon glycation of human carbonic anhydrase II

Advanced glycation end products are the most important species of glycation pathway, and cause disorders such as oxidative stress and diabetes. Sulfonamide compounds, which are generally known as widespread inhibitors, are potential agents used in different drug products, which can readily enter biological matrices. In the present work, the structure and activity of human carbonic anhydrase II studied in the presence of glucose as well as four sulfonamide agents from different views. These included enzyme kinetics, free lysine content, fluorescence spectroscopy, circular dichroism, and ROS measurement. Our results indicated that upon glycation, the structure of HCA II collapses and 8 to 13 lysine residues will be more available based on ligand incubation. Secondary and tertiary structural changes were also observed in the presence and absence of sulfonamide agents using fluorescence and circular dichroism methods, respectively. These spectroscopic data also showed a remarkable increase in hydrophobicity and decrease in α-helix contents during glycation, especially after 35 days of incubation. ROS assay was studied in the presence of glucose and sulfonamide compounds, that demonstrated the role of sulfonamide compounds in ROS formation in the presence of glucose in a synergistic manner. Overall, our data indicated that sulfonamides act as a stimulant factor upon prolonged interaction with HCA II and may intensify the complications of some disorders, such as diabetes and other conformational diseases.

Lysine-rich rice partially enhanced the growth and development of skeletal system with better skeletal microarchitecture in young rats

Rice is the primary staple food for half of the world's population but is low in lysine content. Previously, we developed transgenic rice with enhanced free lysine content in rice seeds (lysine-rich rice), which was shown safe for consumption and improved the growth in rats. However, the effects of lysine-rich rice on skeletal growth and development remained unknown. In this study, we hypothesized that lysine-rich rice improved skeletal growth and development in weaning rats. Male weaning Sprague–Dawley rats received lysine-rich rice (HFL) diet, wild-type rice (WT) diet, or wild-type rice with various contents of lysine supplementation diet for 70 days. Bone microarchitectures were examined by microcomputed tomography, bone strength was investigated by mechanical test, and dynamics of bone growth were examined by histomorphometric analysis. In addition, we explored the molecular mechanism of lysine and skeletal growth through biochemical testing of growth hormone, bone turnover marker, and amino acid content of rat serum analysis, as well as in a cell culture system. Results indicated that the HFL diet improved rats' bone growth, strength, and microarchitecture compared with the WT diet group. In addition, the HFL diet increased the serum essential amino acids, growth hormone (insulin-like growth factor-1), and bone formation marker concentrations. The cell culture model showed that lysine deficiency reduced insulin-like growth factor-1 and Osterix expression, Akt/mammalian target of rapamycin signaling, and matrix mineralization, and inhibited osteoblast differentiation associated with bone growth. Our findings showed that lysine-rich rice improved skeletal growth and development in weaning rats. A further increase of rice lysine content is highly desirable to fully optimize bone growth and development.

Effect of metabolic activity of lactic acid bacteria and propionibacteria on cheese protein digestibility and fatty acid profile

This study aimed at investigating the influence of different variants of bacterial starter cultures on the metabolism of the bacteria used, cheese protein digestibility, and fatty acid profile. The results revealed that lactic acid bacteria had a significant effect on the proportions of fatty acids in cheeses, with saturated fatty acids being predominant in in all cheese variants. Fatty acid proportions are complex and depend on the type of cheese culture and monoculture used. Additionally, the analysis of fatty acid composition showed variations in the proportion of saturated and unsaturated fatty acids, impacting the values of atherogenic and thrombogenic indices. Notably, the atherogenic index was highest in samples of mature cheeses obtained from a typical mesophilic cheese culture, whereas it was lowest in samples of fresh milk and mature cheeses obtained from a mesophilic cheese culture and monocultures of Lacticaseibacillus casei and Propionibacterium. The study also highlighted the influence of lactobacilli on the content of available free lysine, glycine, and methionine in cheese proteins. Mature cheeses obtained with Propionibacterium and L. casei starter cultures exhibited higher free lysine and glycine content compared with fresh cheeses and those obtained solely with the cheese culture. Additionally, mature cheeses obtained with starter cultures of mesophilic cheese culture, Propionibacterium, and L. casei had the highest free methionine content. Based on these findings, it is evident that the choice of cheese making cultures and monocultures can significantly affect the fatty acid composition and amino acid content of cheese and fresh milk, potentially bearing important health implications.

Genetic control of grain amino acid composition in a UK soft wheat mapping population.

Wheat (Triticum aestivum L.) is a major source of nutrients for populations across the globe, but the amino acid composition of wheat grain does not provide optimal nutrition. The nutritional value of wheat grain is limited by low concentrations of lysine (the most limiting essential amino acid) and high concentrations of free asparagine (precursor to the processing contaminant acrylamide). There are currently few available solutions for asparagine reduction and lysine biofortification through breeding. In this study, we investigated the genetic architecture controlling grain free amino acid composition and its relationship to other traits in a Robigus × Claire doubled haploid population. Multivariate analysis of amino acids and other traits showed that the two groups are largely independent of one another, with the largest effect on amino acids being from the environment. Linkage analysis of the population allowed identification of quantitative trait loci (QTL) controlling free amino acids and other traits, and this was compared against genomic prediction methods. Following identification of a QTL controlling free lysine content, wheat pangenome resources facilitated analysis of candidate genes in this region of the genome. These findings can be used to select appropriate strategies for lysine biofortification and free asparagine reduction in wheat breeding programs.

Potassium-Induced Changes in Muscle Free Amino Acid Concentrations in Chicks

Two experiments with purified crystalline amino acid diets were conducted to determine the effect of dietary potassium level on chick breast muscle free amino acid concentrations. In both experiments, concentrations of muscle free glutamine, lysine and arginine increased as dietary potassium increased from 0.12 to 0.18% and decreased as dietary potassium exceeded 0.18%. As dietary potassium increased, muscle potassium concentration increased linearly with an equivalent linear decrease in muscle sodium concentration. Peak concentrations of muscle free glutamine, lysine and arginine occurred in muscle with a K+:Na+ ratio of approximately 1.4. Concentrations of the three amino acids were less in muscle with K+:Na+ ratios either less than or greater than 1.4. Breast muscle concentrations of free histidine decreased slightly, while muscle free carnosine increased substantially when dietary potassium exceeded 0.18%.amino acids glutamine lysine arginine potassium

Effects of low-protein diet and feed restriction on mRNA expression of cationic amino acid transporters in porcine skeletal muscles.

We investigated the effects of a low-protein diet and feed restriction on the mRNA expression of cationic amino acid transporters (CATs) in the longissimus dorsi (LD), rhomboideus (RH), and biceps femoris (BF) muscles of pigs. Eighteen piglets were divided into three groups: a control (CP21%), low-protein diet (LP, CP16%), and feed-restricted diet (FR, CP21%, 76% feed intake of control pigs) groups. The expression levels of CAT-1 in the LD and BF muscles of LP pigs were higher than that of control pigs, whereas that of FR pigs showed no difference. The CAT-2A expression levels in the RH muscle of FR pigs were higher than that of control pigs. The free lysine concentrations in all muscles of LP and FR pigs were lower than that of control pigs. To examine the factors that affect CATs mRNA expression, we evaluated the effects of lysine, arginine, insulin-like growth factor-I, and dexamethasone on the expression of CATs in C2C12 myotubes. CAT-1 expression levels increased in lysine and/or arginine deprivation. We show that CAT-1 and CAT-2A expression levels in skeletal muscles differ in response to dietary treatments and CAT-1 expression in skeletal muscles appears to increase in response to low free lysine concentrations.

Lysine Dipeptide Enhances Gut Structure and Whole-Body Protein Synthesis in Neonatal Piglets with Intestinal Atrophy

BackgroundParenteral nutrition (PN) is often a necessity for preterm infants; however, prolonged PN leads to gut atrophy, weakened gut barrier function, and a higher risk of intestinal infections. Peptide transporter-1 (PepT1) is a di- or tripeptide transporter in the gut and, unlike other nutrient transporters, its activity is preserved with the onset of intestinal atrophy from PN. As such, enteral amino acids in the form of dipeptides may be more bioavailable than free amino acids when atrophy is present. ObjectivesIn Yucatan miniature piglets with PN-induced intestinal atrophy, we sought to determine the structural and functional effects of enteral refeeding with lysine as a dipeptide, compared to free L-lysine. MethodsPiglets aged 7-8 days were PN-fed for 4 days to induce intestinal atrophy, then were refed with enteral diets with equimolar lysine supplied as lysyl-lysine (Lys-Lys; n = 7), free lysine (n = 7), or Lys-Lys with glycyl-sarcosine (n = 6; to determine whether competitive inhibition of Lys-Lys uptake would abolish PepT1-mediated effects). The diets provided lysine at 75% of the requirement and were gastrically delivered for a total of 18 hours. Whole-body and tissue-specific protein synthesis, as well as indices for gut structure and barrier function, were measured. ResultsThe villus height, mucosal weight, and free lysine concentration were higher in the Lys-Lys group compared to the other 2 groups (P < 0.05). Lysyl-lysine led to greater whole-body protein synthesis compared to free lysine (P < 0.05). Mucosal myeloperoxidase activity was lower in the Lys-Lys group (P < 0.05), suggesting less inflammation. The inclusion of glycyl-sarcosine with Lys-Lys abolished the dipeptide effects on whole-body and tissue-specific protein synthesis (P < 0.05), suggesting that improved lysine availability was mediated by PepT1. ConclusionsImproved intestinal structure and whole-body protein synthesis suggests that feeding strategies designed to exploit PepT1 may help to avoid adverse effects when enteral nutrition is reintroduced into the compromised guts of neonatal piglets.

Dietary guanidinoacetic acid supplementation improves water holding capacity and lowers free amino acid concentration of fresh meat in finishing pigs fed with various dietary protein levels.

The current study was carried out to detect the effect of dietary guanidinoacetic acid (GAA) supplementation on carcass characteristics and meat quality in finishing pigs fed different dietary crude protein (CP) levels. Sixty-four barrows with an initial body weight of 73.05 ± 2.34 kg were randomly allocated into 1 of 4 dietary treatments in a 2 (100% vs. 125% NRC CP level) × 2 (0 vs. 300 mg/kg GAA) factorial arrangement (n = 7). The feeding trial lasted for 49 d. GAA supplementation significantly reduced drip loss (P = 0.01), free water distribution (T23 peak area ratio) (P = 0.05) and the concentrations of free alanine, threonine, methionine and isoleucine (P < 0.05); but increased total glycine content (P = 0.03) in the longissimus dorsi muscle of finishing pigs regardless of the dietary CP levels. Furthermore, primary myogenic cell differentiation system was employed to investigate the influence of inclusion of GAA on free amino acid concentrations in myotubes (n = 4) and validate the finding in the animal feeding trial. We found that GAA inclusion in culture medium also decreased intracellular concentrations of free alanine, threonine, methionine, isoleucine, valine and proline in differentiated primary myogenic cells in vitro (P < 0.05). Meanwhile, relative to diets with 100% NRC CP level, the intake of diets with 125% NRC CP level improved sarcoplasmic protein solubility, increased the contents of carnosine and total free amino acids as well as flavor amino acids in the longissimus dorsi muscle and decreased backfat thickness at the 6–7th ribs in pigs (P < 0.05). In addition, we observed that the impact of dietary GAA supplementation on the last rib fat thickness, shear force, and free lysine content in the longissimus dorsi muscle was dependent on dietary CP levels (P < 0.05). Collectively, dietary GAA supplementation can reduce drip loss, decrease the concentrations of free amino acids and flavor amino acids of fresh meat independent of dietary CP levels.

Growth effects of lysine in compounded diets for American alligator, Alligator mississippiensis

AbstractNutritional requirements of the American alligator, Alligator mississippiensis, are largely unknown despite the value of farm‐raised alligator as an aquaculture product. We conducted a 10‐month growth trial to determine effects of graded levels of dietary lysine on alligator growth as measured by increase in body length, body weight gain, and free lysine concentration in blood plasma. One‐hundred‐ninety‐two hatchling alligators were fed 50% crude protein, pelletized diets, composed primarily of practical feed ingredients, with 1.5, 2.25, 3.0, or 3.75% dietary lysine. Diets were fed for 326 days under temperature‐controlled, laboratory conditions. Body lengths and body weights were measured at 60‐day intervals, and blood samples were taken at the final weighing. Increases in body length, body weight gain, and free lysine level in blood plasma indicated that a concentration of at least 2.25% dietary lysine (as fed)—about 4.5% of protein—was adequate for growth of alligator under the conditions of this study. Results suggest that the lysine requirement of American alligator is similar to requirements reported for other aquaculture species, including some fishes, crustaceans, and amphibians.

Characterization of Glyoxal Modified LDL: Role in the Generation of Circulating Autoantibodies in Type 2 Diabetes Mellitus and Coronary Artery Disease.

This study aims to investigate the role of glyoxal modified LDL in the immunopathology of diabetes and cardiovascular disease. Glycoxidation of proteins is widely studied in relation to diabetes and cardiovascular disease. This study probed the glyoxal mediated modifications in LDL, analyzed the immunogenicity of the glycated LDL and ascertained the presence of circulating antibodies against modified LDL in patients with type 2 diabetes mellitus (T2DM), coronary artery disease (CAD) and patients with both (T2DM+CAD). Glyoxal mediated modifications in LDL were studied by multiple spectroscopic techniques, high-performance liquid chromatography and electron microscopy. Immunization studies were carried in New Zealand rabbits. The presence of antibodies against glyoxal modified LDL in immunized rabbits and human subjects was analyzed by ELISA. Glyoxal altered the structural integrity of LDL and led to the formation of AGEs. It decreased the alpha-helix content of LDL; increased β sheet formation, increased carbonyl content and decreased free lysine and arginine content. Modified LDL showed aggregation, generation of of Nε- (Carboxymethyl) lysine and the formation of amorphous type aggregates. It exhibited high antigenicity and generated a specific immune response that shared common antigenic determinants with other glycated proteins. Direct binding data showed the presence of anti-glyoxal modified LDL antibodies in patients with T2DM, CAD and patients with both T2DM and CAD. Further analysis in competitive binding assay revealed specific binding characteristics of auto-antibodies. Sera from patients with T2DM+CAD exhibited the highest binding with glyoxal modified LDL. Glyoxal-modified LDL has neo-antigenic determinants that cause the generation of circulating antibodies in diabetes and coronary artery disease. The study might have potential relevance in biomarker development.

Flash dietary methionine supply over growth requirements in pigs: Multi-facetted effects on skeletal muscle metabolism

Dietary methionine affects protein metabolism, lean gain and growth performance and acts in the control of oxidative stress. When supplied in large excess relative to growth requirements in diets for pigs, positive effects on pork quality traits have been recently reported. This study aimed to decipher the molecular and biochemical mechanisms affected by a dietary methionine supply above growth requirements in the loin muscle of finishing pigs. During the last 14 days before slaughter, crossbred female pigs (n = 15 pigs/diet) were fed a diet supplemented with hydroxy-methionine (Met5; 1.1% of methionine) or not (CONT, 0.22% of methionine). Blood was sampled at slaughter to assess key metabolites. At the same time, free amino acid concentrations and expression or activity levels of genes involved in protein or energy metabolism were measured in the longissimus lumborum muscle (LM). The Met5 pigs exhibited a greater activity of creatine kinase in plasma when compared with CONT pigs. The concentrations of free methionine, alpha-aminobutyric acid, anserine, 3-methyl-histidine, lysine, and proline were greater in the LM of Met5 pigs than in CONT pigs. Expression levels of genes involved in protein synthesis, protein breakdown or autophagy were only scarcely affected by the diet. Among ubiquitin ligases, MURF1, a gene known to target creatine kinase and muscle contractile proteins, and OTUD1 coding for a deubiquitinase protease, were up-regulated in the LM of Met5 pigs. A lower activity of citrate synthase, a reduced expression level of ME1 acting in lipogenesis but a higher expression of PPARD regulating energy metabolism, were also observed in the LM of Met5 pigs compared with CONT pigs. Principal component analysis revealed that expression levels of many studied genes involved in protein and energy metabolism were correlated with meat quality traits across dietary treatments, suggesting that subtle modifications in expression of those genes had cumulative effects on the regulation of processes leading to the muscle transformation into meat. In conclusion, dietary methionine supplementation beyond nutritional requirements in pigs during the last days before slaughter modified the free amino acid profile in muscle and its redox capacities, and slightly affected molecular pathways related to protein breakdown and energy metabolism. These modifications were associated with benefits on pork quality traits.

Investigations on the formation of Maillard reaction products in sweet cookies made of different cereals

In this study, the content of Maillard reaction products from its initial, intermediate and final stage (5-hydroxymethylfurfural, α-dicarbonyl compounds, furosine, N-ε-carboxymethyllysine and N-ε-carboxyethyllysine) was measured in sweet cookies made of wholegrain flour of eight genotypes of small-grain cereals (bread wheat, durum wheat, soft wheat, hard wheat, triticale, rye, hulless barley and hulless oat) and four corn genotypes (white-, yellow- and red-colored standard seeded corn and blue-colored popping corn). Furthermore, the effect of the initial content of sugars, total proteins, free and total lysine in flour on the formation of Maillard reaction products was investigated using the principle component analysis. 3-deoxyglucosone was the predominant α-dicarbonyl compound in all cereal cookies and the highest content was measured in those made from flour of different colored corn genotypes (on average, 98.35, 151.28 and 172.85 mg/kg after baking for 7, 10 and 13 min, respectively). Heating dough at 180 °C for 7, 10 and 13 min differently affected the content of 5-hydroxymethylfurfural and α-dicarbonyl compounds in the cereal cookies. The 5-hydroxymethylfurfural content gradually increased, while a reduction in 3-deoxyglucosone content was observed in the cookies baked for 13 min except for those made from soft wheat, hulless oat, red- and blue-colored corn. After 7 min of heating, the content of furosine measured in the cereal cookies reached its maximum (from 320.9 mg/kg in yellow-colored corn-based cookies to 585.7 mg/kg in hulless oat-based cookies), while N-ε-carboxymethyllysine and N-ε-carboxyethyllysine showed the opposite trend. The highest content of advanced glycation end products was detected in cookies also made from hulless oat flour rich in proteins (16.80%) and total lysine (10670.3 mg/kg). The interrelationship analysis showed that the initial content of sugars in flour of cereals affected 5-hydroxymethylfurfural and 3-deoxyglucosone formation in the cookies. In addition, a high correlation between protein-bound Maillard reaction products in the cookies and the total proteins and the total lysine content in the flours was found.

Mechanistic insight into glycation inhibition of human serum albumin by vitamin B9: Multispectroscopic and molecular docking approach

Advanced glycation end products (AGEs) formation produces free radicals that play a role in diabetes mellitus; hence inhibition of glycation plays a part in minimizing diabetes-related complications. This study was intended to examine the AGEs formation of HSA upon prolonged incubation of 28 days at 37 °C and further investigate the antiglycation potential of folic acid (FA). FA shows a significant binding affinity to the HSA with a binding constant (K) of 104 M−1. The evaluation of enthalpy change (∆H0) and entropy change (∆So) implied that the HSA-FA complex is stabilized primarily by hydrophobic interaction and hydrogen bonding. Molecular docking analysis depicted that FA binds with HSA in subdomain IIA (Sudlow's site I) with a binding energy of −7.0 kcal mol−1. AGEs were characterized by free lysine and thiol groups, carbonyl content, and AGEs specific fluorescence. The presence of FA significantly decreased glycation from free lysine and carbonyl content estimation and AGEs specific fluorescence. Multispectroscopic observations and molecular docking and examination of various biomarkers demonstrate the antiglycation activity of FA and its capacity to prevent disease progression in diabetes.

Withdrawal Notice: Characterization of Glyoxal Modified LDL: Role in the Generation of Circulating Autoantibodies in Type 2 Diabetes Mellitus and Coronary Artery Disease

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Roles of DNA Methylation in Cold Priming in Tartary Buckwheat.

Plants experience a wide array of environmental stimuli, some of which are frequent occurrences of cold weather, which have priming effects on agricultural production and agronomic traits. DNA methylation may act as an epigenetic regulator for the cold response of Tartary buckwheat (Fagopyrum tataricum). Combined with long-term field observation and laboratory experiments, comparative phenome, methylome, and transcriptome analyses were performed to investigate the potential epigenetic contributions for the cold priming of Tartary buckwheat variety Dingku1. Tartary buckwheat cv. Dingku1 exhibited low-temperature resistance. Single-base resolution maps of the DNA methylome were generated, and a global loss of DNA methylation was observed during cold responding in Dingku1. These sites with differential methylation levels were predominant in the intergenic regions. Several hundred genes had different DNA methylation patterns and expressions in different cold treatments (cold memory and cold shock), such as CuAO, RPB1, and DHE1. The application of a DNA methylation inhibitor caused a change of the free lysine content, suggesting that DNA methylation can affect metabolite accumulation for Tartary buckwheat cold responses. The results of the present study suggest important roles of DNA methylation in regulating cold response and forming agronomic traits in Tartary buckwheat.

A biochemical & biophysical study on in-vitro anti-glycating potential of iridin against d-Ribose modified BSA

Non-enzymatic protein glycation results in the formation of advanced glycation end products (AGEs) leads to the pathogenesis of long-term diabetic complications. Iridin (ID), an antioxidant, plays an important role in protecting against oxidative stress and could therefore be an efficacious anti-glycating regimen. Herein, we assessed the anti-glycating potential of ID against d-ribose induced glycation of bovine serum albumin (BSA) by various biophysical and biochemical techniques. Our results from several physicochemical assays advocated that ID was able to evidently prevent the AGEs generation via reducing hyperchromicity, early glycation products (EGPs), carbonyl content (CC), hydroxymethyl furfural (HMF) content, production of fluorescent AGEs, protection against loss of secondary structure (i.e. α-helix and β-sheets) of proteins, increasing the free lysine and free arginine content, reduced binding of congo red (CR), and reduced thioflavin T (ThT) and 8-aninilo-1-napthalene sulphonate (ANS)-specific fuorescence in glycated-BSA (Gly-BSA). On the basis of these findings, we concluded that ID possesses the significant anti-glycation potential and may be established as a remarkable anti-AGEs therapeutic agent. Further in-vivo and clinical studies are still warranted to uncover the therapeutic effects of ID against age-related as well as metabolic diseases.

2'-Deoxyribose Mediated Glycation Leads to Alterations in BSA Structure Via Generation of Carbonyl Species.

The non-enzymatic glycosylation is a very common phenomenon in the physiological conditions which is mediated by distinct chemical entities containing reactive carbonyl species (RCS) and participates in the modification of various macromolecules particularly proteins. To date, various carbonyl species, i.e., glucose, fructose, D-ribose and methylglyoxal have been used frequently to assess the in-vitro non-enzymatic glycosylation. Similarly, 2'-Deoxyribose is one of the most abundant reducing sugar of the living organisms which forms the part of deoxyribonucleic acid and may react with proteins leading to the production of glycation intermediates, advanced glycation end products (AGEs) and highly reactive RCS. Thymidine phosphorylase derived degradation of thymidine contributes to the formation of 2'-Deoxyribose, therefore, acting as a major source of cellular 2'- Deoxyribose. Since albumin is a major serum protein which plays various roles including binding and transporting endogenous and exogenous ligands, it is more prone to be modified through different physiological modifiers; therefore, it may serve as a model protein for in-vitro experiments to study the effect of 2'Deoxyribose mediated modifications in the protein. In this study, Bovine Serum Albumin (BSA) was glycated with 50 and 100 mM 2'-Deoxyribose followed by examining secondary and tertiary structural modifications in BSA as compared to its native (unmodified) form by using various physicochemical techniques. We evident a significant modification in 2'-Deoxyribose-glycated BSA which was confirmed through increased hyperchromicity, keto amine moieties, carbonyl and hydroxymethylfurfural content, fluorescent AGEs, altered secondary structure conformers (α helix and β sheets), band shift in the amide-I region and diminished free lysine and free arginine content. These modifications were reported to be higher in 100 mM 2'-Deoxyribose-glycated BSA than 50 mM 2'- Deoxyribose-glycated BSA. Our findings also demonstrated that the rate of glycation is positively affected by the increased concentration of 2'-Deoxyribose. The results of the performed study can be implied to uncover the phenomenon of serum protein damage caused by 2'-Deoxyribose leading towards diabetic complications and the number of AGE-related diseases.

Exploitation of sourdough lactic acid bacteria to reduce raffinose family oligosaccharides (RFOs) content in breads enriched with chickpea flour

Wheat baked goods fortification with chickpea flour can improve their overall nutritional quality. However, some antinutritional factors, such as raffinose family oligosaccharides (RFOs), of this legume limit its supplementation in bread. The reduction of some of these drawbacks can be achieved using sourdough technology. Therefore, the aim of this study was to obtain a sourdough bread possessing the beneficial effect of chickpea flour addition together with a reduced content of RFOs, thanks to the activity of selected lactic acid bacteria. Sixty sourdough lactobacilli were screened for the capability of growing in a medium with raffinose as unique carbohydrates source. The most promising strains were inoculated in liquid chickpea sourdoughs. Lactobacillus plantarum M8 decreased up to 63.22% the initial amount of RFOs hence it was selected for the manufacture of wheat breads with different percentage of pre-fermented chickpea flour (0, 5, 10 and 20% on total flour). Chickpea flour addition significantly (p > 0.05) increased free amino acids, lysine and total phenolic content of breads compared to a control bread made only with wheat flour. Furthermore, RFOs total amount decreased in breads with increased percentage of pre-fermented chickpea flour in the recipe, confirming the ability of the selected L. plantarum M8 strain to reduce RFOs.

Evaluating the rumen-protected lysine stability in forage-based total mixed rations in vitro and determining the lysine Brix value.

Three rumen-protected lysine (RPL) products (AjiPro® -L, LysiPEARLTM , and Feedtech Bypass LysineTM : A, B, and C, respectively) were tested for stability in two forage-based total mixed rations (TMR1, 41.3% dry matter (DM), and TMR2, 49.5% DM) (experiment 1) and for Brix value (experiment 2). In experiment 1, each RPL product (2g each) and TMR diet (200g) were mixed and stored in plastic bags at 20°C for 0, 1, 3, 6, 12, 24, and 48hr. In experiment 2, each RPL product (2g) was dispensed into ion-exchanged water (20ml) and kept at 20°C for 0, 0.5, 1, 3, 6, 12, 24, and 48hr. At each time point, free lysine (Lys) content and Brix values of extracts were measured, and Lys release (LR, %) was calculated. All RPL products LR% varied with varying diets DM and increased with increasing of time exposed to diets; it was highest in C, followed by B, and then A. Water LR% positively correlated with that from diets and with Brix values of Lys dissociated in water. Our results indicated that Lys dissociation from RPL products is affected by diet DM content. Brix value may be used as a potential marker for RPL protection efficacy.