Preservation of Millet Flour by Refrigeration: Changes in Total Protein and Amino Acids Composition During Storage

This work describes refrigeration effects during storage on total protein and amino acids composition of raw and processed flour of two pearl millet cultivars (Ashana and Dembi). The protein content of the whole raw flour was found to be 14.46 and 13.38% for Ashana and Dembi cultivars, respectively. Dehulling of the grains reduced the protein content to 13.38 and 12.67% for the cultivars, respectively. For both cultivars, the protein content of the whole and dehulled raw flour before and after cooking was slightly decreased when the flour was stored for 60 days even after refrigeration. The effect of refrigeration process in combination with the storage period, cooking or dehulling was found to be vary between amino acids and even between cultivars. Regardless of the storage period and processing method, the amino acids content was remained unchanged after refrigeration for both cultivars.

Sensory Evaluation of Meatballs with Jerusalem Artichoke (Helianthus tuberosus L.)

Meat and meat products for human consumption are one of main sources of protein, amino acids, fatty acids, vitamins, and minerals. Popular variety of meat product is meatballs, which can be enriched with valuable product – Jerusalem artichoke powder, made from dried and grinded Jerusalem artichoke tubers, it is raw material with low-calorie, low fat, rich in dietary fibres, minerals, and vitamins. The results of this study indicate that that people could accept the new product - meatballs with Jerusalem artichoke powder and Jerusalem artichoke powder is suitable for meatballs preparation, in result them is possible to improve meatballs sensory and physical properties.

Production of the Protein-Vitamin Complex from Wheat Germ

Wheat germ has a balanced amino acid composition of the protein, which is well digested by enzymes in the gastrointestinal tract of humans, a high content of vitamins, minerals and unsaturated acids. Introduction components grain food products will enrich their biologically important substances, giving these products a number of valuable properties and reducing their caloric. A complex natural system of substances in foods will help replenish the body's need of essential nutrients, increasing its resistance to the harmful effects of the environment, prolong life. In this regard, there was a need for the development of production technology of protein complexes from wheat germ and then applying them in food, particularly in the dairy industry. Experimental studies were conducted to determine the number of herbal supplements on the sensory characteristics of the product. Studies have been conducted to determine the optimal process parameters of water activity and moisture content of the investigational product. 

The Chemical Composition of Yoghurt Enriched with Flakes from Biologically Activated Hullless Barley Grain and Malt Extract

The influence of flakes from biologically activated hull-less barley grain and malt extract on chemical composition of yoghurt was studied. Pasteurized milk, freeze-dried yoghurt culture YF-L811 (Chr. Hansen, Denmark), flakes from biologically activated hull-less barley grain (Latvia) and malt extract (Ilgezeem, Latvia) were used for experiments. Yoghurt samples with and without flakes from biologically activated hull-less barley grain and malt extract were analyzed for content of total solids, total proteins, fats, amino acids and riboflavin. The addition of flakes from biologically activated hull-less barley grain and malt extract allowed increase of nutritional value of yoghurt samples. There was obtained the increase of total proteins (p>0.05) and the decrease of fat (p>0.05). The presence of flakes from biologically activated hull-less barley grain and malt extract in yoghurt samples provided significant increase of amino acids amount (p

A Novel Cytokine Derived Fusion Tag for Over- Expression of Heterologous Proteins in E. coli

We report a novel fusion tag for expressing recombinant proteins in E. coli. The fusion tag is the C-terminus part of the human GMCSF gene comprising 45 amino acids, which aid in over expression of otherwise non expressible genes. Expression of hIFN a2b with this fusion tag also escapes the requirement of rare codons for expression. This is also a first report of a small fusion tag of human origin having affinity to heparin sepharose column facilitating the purification of fusion protein.

Molecular Characterization of Free Radicals Decomposing Genes on Plant Developmental Stages

Biochemical and molecular analysis of some antioxidant enzyme genes revealed different level of gene expression on oilseed (Brassica napus). For molecular and biochemical analysis, leaf tissues were harvested from plants at eight different developmental stages, from young to senescence. The levels of total protein and chlorophyll were increased during maturity stages of plant, while these were decreased during the last stages of plant growth. Structural analysis (nucleotide and deduced amino acid sequence, and phylogenic tree) of a complementary DNA revealed a high level of similarity for a family of Catalase genes. The expression of the gene encoded by different Catalase isoforms was assessed during different plant growth phase. No significant difference between samples was observed, when Catalase activity was statistically analyzed at different developmental stages. EST analysis exhibited different transcripts levels for a number of other relevant antioxidant genes (different isoforms of SOD and glutathione). The high level of transcription of these genes at senescence stages was indicated that these genes are senescenceinduced genes.

Effect of Drought Stress on Nitrogen Components in Corn

An attempt was made to study of nitrogen components response of corn (Zea mays L.) to drought stress. A farm research was done in RCBD as split-plot with four replications in Khorramabad, west Iran. Drought stress levels as irrigation regimes after 75 (control), 100, and 120 (stress) mm cumulative evaporation were in main plots, and four seed corn varieties include 500 (medium maturity), 647, 700, and 704 (long maturity) were as subplots. Soluble protein, nitrate and proline amino acid were measured in shoot and root at flowering stage, and grain yield was measured in harvesting stage. As the drought progressed, the amount of nitrate and proline followed an increasing trend, but soluble protein decreased in shoot and root. The highest amount of nitrate and proline was observed in longer maturity varieties than shorter ones, but decrease yield of long maturity varieties was higher than medium maturity varieties in drought condition, because of long duration of stress.

Genome-Wide Analysis of BES1/BZR1 Gene Family in Five Plant Species

Brassinosteroids (BRs) regulate cell elongation, vascular differentiation, senescence, and stress responses. BRs signal through the BES1/BZR1 family of transcription factors, which regulate hundreds of target genes involved in this pathway. In this research a comprehensive genome-wide analysis was carried out in BES1/BZR1 gene family in Arabidopsis thaliana, Cucumis sativus, Vitis vinifera, Glycin max and Brachypodium distachyon. Specifications of the desired sequences, dot plot and hydropathy plot were analyzed in the protein and genome sequences of five plant species. The maximum amino acid length was attributed to protein sequence Brdic3g with 374aa and the minimum amino acid length was attributed to protein sequence Gm7g with 163aa. The maximum Instability index was attributed to protein sequence AT1G19350 equal with 79.99 and the minimum Instability index was attributed to protein sequence Gm5g equal with 33.22. Aliphatic index of these protein sequences ranged from 47.82 to 78.79 in Arabidopsis thaliana, 49.91 to 57.50 in Vitis vinifera, 55.09 to 82.43 in Glycin max, 54.09 to 54.28 in Brachypodium distachyon 55.36 to 56.83 in Cucumis sativus. Overall, data obtained from our investigation contributes a better understanding of the complexity of the BES1/BZR1 gene family and provides the first step towards directing future experimental designs to perform systematic analysis of the functions of the BES1/BZR1 gene family.

Mutational Effect to Particular Interaction Energy of Cycloguanil Drug to Plasmodium Plasmodium Falciparum Dihydrofolate Reductase Enzymes

In order to find the particular interaction energy between cylcloguanil and the amino acids surrounding the pocket of wild type and quadruple mutant type PfDHFR enzymes, the MP2 method with basis set 6-31G(d,p) level of calculations was performed. The obtained interaction energies found that Asp54 has the strongest interaction energy to both wild type and mutant type of - 12.439 and -11.250 kcal/mol, respectively and three amino acids; Asp54, Ile164 and Ile14 formed the H-bonding with cycloguanil drug. Importantly, the mutation at Ser108Asn was the key important of cycloguanil resistant with showing repulsive interaction energy.

A Novel Strategy for Oriented Protein Immobilization

A new strategy for oriented immobilization of proteins was proposed. The strategy contains two steps. The first step is to search for a docking site away from the active site on the protein surface. The second step is trying to find a ligand that is able to grasp the targeted site of the protein. To avoid ligand binding to the active site of protein, the targeted docking site is selected to own opposite charges to those near the active site. To enhance the ligand-protein binding, both hydrophobic and electrostatic interactions need to be included. The targeted docking site should therefore contain hydrophobic amino acids. The ligand is then selected through the help of molecular docking simulations. The enzyme α-amylase derived from Aspergillus oryzae (TAKA) was taken as an example for oriented immobilization. The active site of TAKA is surrounded by negatively charged amino acids. All the possible hydrophobic sites on the surface of TAKA were evaluated by the free energy estimation through benzene docking. A hydrophobic site on the opposite side of TAKA-s active site was found to be positive in net charges. A possible ligand, 3,3-,4,4- – Biphenyltetra- carboxylic acid (BPTA), was found to catch TAKA by the designated docking site. Then, the BPTA molecules were grafted onto silica gels and measured the affinity of TAKA adsorption and the specific activity of thereby immobilized enzymes. It was found that TAKA had a dissociation constant as low as 7.0×10-6 M toward the ligand BPTA on silica gel. The increase in ionic strength has little effect on the adsorption of TAKA, which indicated the existence of hydrophobic interaction between ligands and proteins. The specific activity of the immobilized TAKA was compared with the randomly adsorbed TAKA on primary amine containing silica gel. It was found that the orderly immobilized TAKA owns a specific activity twice as high as the one randomly adsorbed by ionic interaction.

Regulatory Effects of Carbon Sources on Tabtoxin Production (A β-lactam Phytotoxin of Pseudomonas syringae pv. tabaci)

The effects of divers carbon substrates were investigated for the tabtoxin production of an isolated pathogenic Pseudomonas syringae pv. tabaci, the causal agent of wildfire of tobacco and are discussed in relation to the bacterium growth. The isolated organism was grown in batch culture on Woolley's medium (28°C, 200 rpm, during 5 days). The growth has been measured by the optical density (OD) at 620 nm and the tabtoxin production quantified by Escherichia coli (K-12) bioassay technique. The growth and the tabtoxin production were both influenced by the substrates (sugars, amino acids, organic acids) used, each, as a sole carbon source and as a supplement for the same amino acids. The most significant quantities of tabtoxin were obtained in presence of some amino acids used as sole carbon source and/or as supplement.

An Information Theoretic Approach to Rescoring Peptides Produced by De Novo Peptide Sequencing

Tandem mass spectrometry (MS/MS) is the engine driving high-throughput protein identification. Protein mixtures possibly representing thousands of proteins from multiple species are treated with proteolytic enzymes, cutting the proteins into smaller peptides that are then analyzed generating MS/MS spectra. The task of determining the identity of the peptide from its spectrum is currently the weak point in the process. Current approaches to de novo sequencing are able to compute candidate peptides efficiently. The problem lies in the limitations of current scoring functions. In this paper we introduce the concept of proteome signature. By examining proteins and compiling proteome signatures (amino acid usage) it is possible to characterize likely combinations of amino acids and better distinguish between candidate peptides. Our results strongly support the hypothesis that a scoring function that considers amino acid usage patterns is better able to distinguish between candidate peptides. This in turn leads to higher accuracy in peptide prediction.

Molecular Dynamics and Circular Dichroism Studies on Aurein 1.2 and Retro Analog

Aurein 1.2 is a 13-residue amphipathic peptide with antibacterial and anticancer activity. Aurein1.2 and its retro analog were synthesized to study the activity of the peptides in relation to their structure. The antibacterial test result showed the retro-analog is inactive. The secondary structural analysis by CD spectra indicated that both of the peptides at TFE/Water adopt alpha-helical conformation. MD simulation was performed on aurein 1.2 and retro-analog in water and TFE in order to analyse the factors that are involved in the activity difference between retro and the native peptide. The simulation results are discussed and validated in the light of experimental data from the CD experiment. Both of the peptides showed a relatively similar pattern for their hydrophobicity, hydrophilicity, solvent accessible surfaces, and solvent accessible hydrophobic surfaces. However, they showed different in directions of dipole moment of peptides. Also, Our results further indicate that the reversion of the amino acid sequence affects flexibility .The data also showed that factors causing structural rigidity may decrease the activity. Consequently, our finding suggests that in the case of sequence-reversed peptide strategy, one has to pay attention to the role of amino acid sequence order in making flexibility and role of dipole moment direction in peptide activity. KeywordsAntimicrobial peptides, retro, molecular dynamic, circular dichroism.

5-Aminolevulinic Acid-Loaded Gel, Sponge Collagen to Enhance the Delivery Ability to Skin

Topical photodynamic therapy (PDT) with 5-aminolevulinic acid (ALA) is an alternative therapy for treating superficial cancer, especially for skin or oral cancer. ALA, a precursor of the photosensitizer protoporphyrin IX (PpIX), is present as zwitterions and hydrophilic property which make the low permeability through the cell membrane. Collagen is a traditional carrier; its molecular composed various amino acids which bear positive charge and negative charge. In order to utilize the ion-pairs with ALA and collagen, the study employed various pH values adjusting the net charge. The aim of this study was to compare a series collagen form, including solution, gel and sponge to investigate the topical delivery behavior of ALA. The in vivo confocal laser scanning microscopy (CLSM) study demonstrated that PpIX generation ability was different pattern after apply for 6 h. Gel type could generate high PpIX, and archived more deep of skin depth.

New Malate Dehydrogenase-Glutamate Oxaolacetate Aminotransferase Glutamate Oxaloacetate Aminotransferase Enzyme System from Cereals and its Bioengineering Application

Malate dehydrogenase-glutamate oxaloacetate aminotransferase (MDh-GOAT) enzyme complex (the EC) was isolated and purified from wheat and rise, their some main physicchemical properties were studied. Michael-s constants of the EC MDh-GOAT to malate, glutamate and NAD were investigated. This kinetic results show a high relationship to glutamate. Taking into account important role of the the EC in catabolism of glutamate – the central amino acid of a nitric exchange, there is a sharp necessity of deeper studying of this enzyme complex. Therefore the basic purpose of the work is studying the basic physical and chemical properties of this enzyme complex discovered by us, which would be very important for understanding the mechanisms of reaction catalyzed by the EC.

Expression of Leucaena Leucocephala de Wit Chitinase in Transgenic Koshihikari Rice

The cDNA encoding the 326 amino acids of a Class I basic chitinase gene from Leucaena leucocephala de Wit (KB3, Genbank accession: AAM49597) was cloned under the control of CaMV35S promoter in pCAMBIA 1300 and transferred to Koshihikari. Calli of Koshihikari rice was transformed with agrobacterium with this construct expressing the chitinase and β- glucouronidase (GUS). The frequencies of calli 90 % has been obtained from rice seedlings cultured on NB medium. The high regeneration frequencies, 74% was obtained from calli cultured on regeneration medium containing 4 mg/l BAP, and 7 g/l phytagel at 25°C. Various factors were studied in order to establish a procedure for the transformation of Koshihikari Agrobacterium tumefaciens. Supplementation of 50 mM acetosyringone to the medium during coculivation was important to enhance the frequency to transient transformation. The 4 week-old scutellum-derived calli were excellent starting materials. Selection medium based on NB medium supplement with 40 mg/l hygromycin and 400 mg/l cefotaxime were an optimized medium for selection of transformed rice calli. The percentage of transformation 70 was obtained. Recombinant calli and regenerated rice plants were checked the expression of chitinase and gus by PCR, northern blot gel, southern blot gel, and gus assay. Chitinase and gus were expressed in all parts of recombinant rice. The rice line expressing the KB3 chiitnase was more resistant to the blast fungus Fusarium monoliforme than control line.

Biochemical Characteristics of Sorghum Flour Fermented and/or Supplemented with Chickpea Flour

Sorghum flour was supplemented with 15 and 30% chickpea flour. Sorghum flour and the supplement were fermented at 35 oC for 0, 8, 16, and 24 h. Changes in pH, titrable acidity, total soluble solids, protein content, in vitro protein digestibility and amino acid composition were investigated during fermentation and/or after supplementation of sorghum flour with chickpea. The pH of the fermenting material decreased sharply with a concomitant increase in the titrable acidity. The total soluble solids remained unchanged with progressive fermentation time. The protein content of sorghum cultivar was found to be 9.27 and that of chickpea was 22.47%. The protein content of sorghum cultivar after supplementation with15 and 30% chickpea was significantly (P ≤ 0.05) increased to 11.78 and 14.55%, respectively. The protein digestibility also increased after fermentation from 13.35 to 30.59 and 40.56% for the supplements, respectively. Further increment in protein content and digestibility was observed when supplemented and unsupplemented samples were fermented for different periods of time. Cooking of fermented samples was found to increase the protein content slightly and decreased digestibility for both supplements. Amino acid content of fermented and fermented and cooked supplements was determined. Supplementation was found to increase the lysine and therionine content. Cooking following fermentation decreased lysine, isoleucine, valine and sulfur containg amino acids.

Quality Properties of Fermented Mugworts and Rapid Pattern Analysis of Their Volatile Flavor Components by Electric Nose Based On SAW (Surface Acoustic Wave) Sensor in GC System

The changes in quality properties and nutritional components in two fermented mugworts (Artemisia capillaries Thumberg, Artemisiaeasiaticae Nakai) were characterized followed by the rapid pattern analysis of volatile flavor compounds by Electric Nose based on SAW(Surface Acoustic Wave) sensor in GC system. There were remarkable decreases in the pH and small changes in the total soluble solids after fermentation. The L (lightness) and b (yellowness) values in Hunter's color system were shown to be decreased, whilst the a (redness) value was increased by fermentation. The HPLC analysis demonstrated that total amino acids were increased in quantity and the essential amino acids were contained higher in A. asiaticaeNakai than in A. capillaries Thumberg. While the total polyphenol contents were not affected by fermentation, the total sugar contents were dramatically decreased. Scopoletinwere highly abundant in A. capillarisThumberg, however, it was not detected in A. asiaticaeNakai. Volatile flavor compounds by Electric Nose showed that the intensity of several peaks were increased much and seven additional flavor peaks were newly produced after fermentation. The flavor differences of two mugworts were clearly distinguished from the image patterns of VaporPrintTM which indicate that the fermentation enables the two mugworts to have subtle flavor differences.

Cloning and Expression of D-Threonine Aldolase from Ensifer arboris NBRC100383

D-erythro-cyclohexylserine (D chiral unnatural β-hydroxy amino acid expected for the synthesis of drug for AIDS treatment. To develop a continuous bioconversion system with whole cell biocatalyst of D-threonine aldolase (D genes for the D-erythro-CHS production, D-threonine aldolase gene was amplified from Ensifer arboris 100383 by direct PCR amplication using two degenerated oligonucleotide primers designed based on genomic sequence of Shinorhizobium meliloti Sequence analysis of the cloned DNA fragment revealed one open-reading frame of 1059 bp and 386 amino acids. This putative D-TA gene was cloned into NdeI and EcoRI (pEnsi His-tag sequence or BamHI (pEnsi-DTA[2]) sequence of the pET21(a) vector. The expression level of the cloned gene was extremely overexpressed by E. coli BL21(DE3) transformed with pEnsi-DTA[1] compared to E. coli BL21(DE3) transformed with pEnsi-DTA[2]. When the cells expressing the wild used for D-TA enzyme activity, 12 mM glycine was successfully detected in HPLC analysis. Moreover, the whole cells harbouring the recombinant D-TA was able to synthesize D-erythro of 0.6 mg/ml in a batch reaction.

A General Model for Amino Acid Interaction Networks

In this paper we introduce the notion of protein interaction network. This is a graph whose vertices are the protein-s amino acids and whose edges are the interactions between them. Using a graph theory approach, we identify a number of properties of these networks. We compare them to the general small-world network model and we analyze their hierarchical structure.