Determination Of Amino Acid Profile Research Articles

Determination of Amino acid Profile and Some Characteristics of Collagen Extracted from Skin and Bone of Luciobarbus esocinus (Heckel, 1843)

Acid soluble collagens (ASC) from bone (ASC-B) and skin (ASC-S) of mangar (Luciobarbus esocinus (Heckel, 1843) were extracted, characterized, and their amino acid profiles were determined. To best of our knowledge, the current study is the first research that used this species as a source of collagen. Both ASC-S and ASC-B from mangar skin and bone contained glycine as the major amino acid and high amounts of proline, hydroxyproline, alanine, and glutamic acid. On the basis of dry weight, yields of ASC-S and ASC-B were 9.38 and 3.71%, respectively. Furthermore, fourier transform infrared spectroscopy proved that both collagens were integrated and native. Additionally, the results of XRD demonstrated that both of the collagens reserved their helical structures. The screened collagens had prominent absorptions at 230 nm by UV-Vis spectra. Additionally, the SEM studies have shown that both ACS-S and ASC-B have porous and fibrous nature. According to the UV–Vis and FTIR results, extracted collagens were characterized as type I collagen based on their amino acid composition. According to the obtained results, the collagen isolated from mangar can potentially be an alternative source of vertebrate collagens for use in the diet and other industries such as medical and pharmaceutical industries.

Degradation of rice protein by mushroom proteases and determination of amino acid profiles

Degradation of rice protein by mushroom proteases and determination of amino acid profiles

Amino Acid Profiling of the Leaves of Senna Obtusifolia and Biochemical Changes during Fermentation for Production of ''Kawal'', a Traditional Food Condiment

Kawal is made by a natural or uncontrolled fermentation of Senna obtusifolia leaves. It is a food condiment, and appreciated because of its particular flavor. It is also used as a meat or fish substitute. Kawal is an important source of protein in the diet of low-income families. The present work was done to determine the total amino acid content in raw and fermented Senna obtusifolia leaves. The high performance liquid chromatography (HPLC) with The Pico-Tag method was used for the analysis and determination of amino acid profiles. The results obtained showed that the kawal protein is rich in essential amino acids and semi-essential for humans, with different concentrations. Leucine gave a maximum value of 208.56 mg/100 g (DM) and 538,29 mg/100 g (DM); valine showed the value of 173,46 mg/100 g (DM) and 476.25 mg/100 g (DM), for lysine this value is 205.58 mg/100 g (DM) and 456.13 mg/100 g (DM). In the background, we have histidine, threonine, tyrosine, isoleucine and phenylalanine, whose best concentrations are between 157.36 mg/100 g and 397.93 mg/100 g (DM) in fermented leaves. For semi-essential amino acids, the fermented leaves have concentrations of between 121.96 mg/100 g and 1260.92 mg/100 g (DM).

Determination of amino acid profile for argininosuccinic aciduria disorder using High-Performance Liquid Chromatography with fluorescence detection.

Argininosuccinic aciduria is an autosomal, recessive amino acid disorder that is caused by a deficiency of the argininosuccinate lyase enzyme. Citrulline is the most significant marker to detect this disorder. We used the High-performance liquid chromatography with fluorescence detection with 450 nm emission and 330 nm excitation wavelengths, 15 mmol/L potassium dihydrogen phosphate and 5 mmol/L dipotassium hydrogen phosphate as Mobile Phase A, and 50 mL water, 250 mL acetonitrile, and 200 mL methanol as Mobile Phase B in gradient mode with flow rate of 1.2 mL/min. The citrulline concentration was 22 µmol/L in healthy infants and 220 µmol/L in infants suffering from the disorder.

Alterations in some physiological processes of Bactrocera oleae Rossi (Diptera: Tephritidae) following pyridalyl treatment

Pyridalyl is one of the recently discovered insecticides which has shown toxicity against several insect pests with slight- or non-lethal effects against natural enemies. Cytotoxicity and prevention of protein synthesis have been reported as pyridalyl mode of action. In the current study, adults of Bacterocera oleae, as a major pest of olive, were fed on protein hydrolysate diet containing LC50 concentration of pyridalyl to determine the possible mode of action through gene expression and amino acid profile. Results showed that treated flies had significantly higher gene expressions of cytochrome P450, insulin receptor-like, antioxidant enzymes (catalase, peroxidase, superoxide dismutase) and the enzymes involved in cell death; caspase1, caspase3, caspase8 and caspase9 compared to control. Moreover, determination of amino acid profile in the whole body of both control and pyridalyl-treated B. oleae revealed the significant higher amounts of glutamic acid, glutamine, histidine, threonine, citrulin, arginine, taurine, alanine, tyrosine, α-amino butric acid, methionine and isoleucine in the treated adults while the control flies had the higher amounts of serine, glycine and tryptophane. No significant differences were found in the amounts of asparagine and phenylalanine between control and treatments. These results are in accordance with earlier findings on mode of action of pyridalyl through activation of cytochrome P450 system and alterations in protein synthesis machinery although here other mechanisms were also reported including activation of insulin receptor and antioxidant enzymes. Moreover, it is the first report on cytotoxicity through apoptosis with engagement of caspases.

Determination of amino acid profile of mare milk produced in the highlands of the Kyrgyz Republic during the milking season

This study was carried out to determine the influence of milking season on amino acid (AA) profile and chemical composition of milk samples from Kyrgyz native breed mares under traditional pasture conditions. Milk samples were collected monthly from May to August 2014 from mares grazing at 2,200m above sea level. The AA composition was determined by precolumn derivatization with diethyl ethoxymethylenemalonate in HPLC and 18 AA were determined in mare milk. The analytical parameters of linearity (0.01–4μg/mL), precision of the method (0.26–4.88% relative standard deviation), derivatization procedure (0.82–2.80% relative standard deviation), and instrument precision (0.08–1.00% relative standard deviation) were determined. The most abundant AA were glutamic acid (0.393–480g/100g of milk), leucine (0.192–0.230g/100g of milk), and lysine (0.192–0.230g/100g of milk). The amount of ornithine was small (0.002–0.015g/100g of milk). The percentages of essential AA in the protein of mare milk were 46, 46, 51, and 48% for May, June, July, and August, respectively, which demonstrate the high biological value of milk during the whole milking season.

Effect of Sub- and Super-critical Water Treatment on Physicochemical Properties of Porcine Skin.

Super- and sub-critical water treatments have been of interest as novel methods for protein hydrolysis. In the present study, we studied the effect of sub-critical water (Sub-H2O, 300℃, 80 bar) treatment as well as super-critical water (Super-H2O, 400℃, 280 bar) treatment on the physicochemical properties of porcine skin (PS), which has abundant collagen. Porcine skin was subjected to pre-thermal treatment by immersion in water at 70℃, and then treated with sub- or super-critical water. Physicochemical properties of the hydrolysates, such as molecular weight distribution, free amino acid content, amino acid profile, pH, color, and water content were determined. For the molecular weight distribution analysis, 1 kDa hydrolyzed porcine skin (H-PS) was produced by Super-H2O or Sub-H2O treatment. The free amino acid content was 57.18 mM and 30.13 mM after Sub-H2O and Super-H2O treatment, respectively. Determination of amino acid profile revealed that the content of Glu (22.5%) and Pro (30%) was higher after Super-H2O treatment than after Sub-H2O treatment, whereas the content of Gly (28%) and Ala (13.1%) was higher after Sub-H2O treatment. Super-H2O or Sub-H2O treatment affected the pH of PS, which changed from 7.29 (Raw) to 9.22 (after Sub-H2O treatment) and 9.49 (after Super-H2O treatment). Taken together, these results showed that Sub-H2O treatment was slightly more effective for hydrolysis than Super-H2O was. However, both Sub-H2O and Super-H2O treatments were effective processing methods for hydrolysis of PS collagen in a short time and can be regarded as a green chemistry technology.

Fifteen years experience: Egyptian metabolic lab

BackgroundInborn errors of metabolism (IEM) are single gene disorders responsible for abnormalities in the synthesis or catabolism of proteins, carbohydrates and fats by means of defective enzymes or transport proteins which results in a block of the metabolic pathway and accumulation of metabolites in different tissues. This study shows the most common diagnosed inherited inborn errors of metabolism among the Egyptian population. Prior to 1995, the diagnosis of inherited metabolic disorders in Egypt was very limited and diagnosed mainly on clinical suspicion. In 1995, The Biochemical Genetics Unit at The National Research Centre has been established as a part of The Human Genetics Department and later on in 2003 it was developed into The Biochemical Genetics Department by applying advanced techniques and equipments and providing early diagnosis for the metabolic disorders which led to better outcome in our patients. Material and methodsWe have retrospectively reviewed a total of 12,148 cases suspected to have inborn errors of metabolism (IEM) with different age groups. They had been referred from several diagnostic centers and hospitals in Egypt to The Department of Biochemical Genetics at The National Research Centre. The diagnosis of these disorders was confirmed by qualitative determination of amino acid profile, quantitative determination of phenylalanine and galactose levels using dried blood spots (DBSs), quantitative determination of urinary glycosaminoglycans (GAGs), two-dimensional electrophoretic separation of GAGs in urine and the assay for lysosomal enzymes activities in plasma and leukocytes. ResultsOut of the total number of cases; 1041 (8.6%) patients were proved to have metabolic disorders. Those patients were classified as: 722 patients (69.4%) with lysosomal storage disorders, 302 patients (29%) with amino acid disorders and 17 patients (1.6%) with galactosemia. ConclusionThis study illustrates the experience of the reference metabolic lab in Egypt over 15years. The lab began metabolic disorder screening by using simple diagnostic techniques like thin layer chromatography and colored tests in urine which by time updated and upgraded the methods to diagnose a wide range of disorders. This study shows the most common diagnosed inherited inborn errors of metabolism among the Egyptian population.

Microscale analysis of amino acids using gas chromatography–mass spectrometry after methyl chloroformate derivatization

To conduct studies of stable isotope incorporation and dilution in growing plants, a rapid microscale method for determination of amino acid profiles from minute amounts of plant samples was developed. The method involves solid-phase ion exchange followed by derivatization and analysis by gas chromatography–mass spectrometry (GC–MS). The procedure allowed the eluent to be derivatized directly with methyl chloroformate without sample lyophilization or other evaporation procedures. Sample extraction and derivatization required only ca. 30 min and quantification of the 19 amino acids eluted from the cation exchange solid-phase extraction step from a single cotyledon (0.4 mg fresh weight) or three etiolated 7-day-old Arabidopsis seedlings (0.1 mg fresh weight) was easily accomplished in the selected ion monitoring mode. This method was especially useful for monitoring mass isotopic distribution of amino acids as illustrated by Arabidopsis seedlings that had been labeled with deuterium oxide and 15N salts. Sample preparation was facile, rapid, economical, and the method is easily modified for integration into robotic systems for analysis with large numbers of samples.

Role of Glutamine Depletion in Directing Tissue-specific Nutrient Stress Responses to L-Asparaginase

Role of Glutamine Depletion in Directing Tissue-specific Nutrient Stress Responses to L-Asparaginase

Separation of twenty underivatized essential amino acids by capillary zone electrophoresis with contactless conductivity detection.

Twenty underivatized essential amino acids were separated using capillary zone electrophoresis and consequently detected with contactless conductivity detection (CCD). A simple acidic background electrolyte (BGE) containing 2.3 M acetic acid and 0.1% w/w hydroxyethylcellulose (HEC) allowed the electrophoretic separation and sensitive detection of all 20 essential amino acids in their underivatized cationic form. The addition of HEC to the BGE suppressed both, electroosmotic flow and analyte adsorption on the capillary surface resulting in an excellent migration time reproducibility and a very good analyte peak symmetry. Additionally, the HEC addition significantly reduced the noise and long-term fluctuations of the CCD baseline. The optimized electrophoretic separation method together with the CCD was proved to be a powerful technique for determination of amino acid profiles in various natural samples, like beer, yeast, urine, saliva, and herb extracts.

Characterization of Saccharomyces Sp. through determination of amino acid profiles by capillary gas chromatography

AbstractThe Saccharomyces strains uvarum, montuliensis, bayanus, capensis and florentinus were grown in liquid media prepared from a 1% aqueous glucose solution to which 0.05% of ammonium sulfate or urea were added as the only nitrogen supply. The amino acids exuded during incubation were isolated by cation exchange and derivatized as the corresponding N,O‐heptafluorobutyryl isopropyl esters. Separation was achieved on a highly deactivated glass capillary column coated with OV‐101. A characteristic amino acid profile was obtained for each of the species studied, allowing the construction of a chemotaxonomic tree, on the basis of present or absent acids.

Determination of amino acid profiles in biological samples by gas chromatography

Based on solvent-free gas chromatography of the n-propyl. N-acetyl derivatives, procedures are described for the routine determination of amino acid profiles in biological samples including protein hydrolysates, plant tissue extracts, urines and sera. Derivatization is carried out in a 25-mifn two-step procedure. Use of a basic acylation environment allows derivatization of arginine and histidine. Chromatography using capsule injection on to a polar column with temperature programming gives resolution of 21 common amino acids within 15 min. Norleucine used as the internal standard provides quantitative capability. Sensitivity of the method permits quantitation at the nanomote level. Recoveries of amino acids added to sera ranged over 90–99%; an exception was arginine which gave 78%. Typical reproducibility data indicate that a coefficient of variation of 2–5% is attainable.