Abstract: Through anther and microspore culture methods, complete homozygous plants can be produced within a year as compared to the long inbreeding method. Isolated microspore culture is one of the most important techniques for rapid development of haploid plants. The efficiency of this method is influenced by several factors such as cultural conditions, growth regulators, plant media, pretreatments, physical and growth conditions of the donor plants, pollen isolation procedure, etc. The main purpose of this study was to improve the isolated microspore culture protocol in order to increase the efficiency of embryoids, its regeneration and reducing albinisms. Under this study we have tested mainly three different microspore isolation procedures by glass rod, homozeniger and by blending and found the efficiency on gametic embryogenesis. There are three types of media viz. washing, pre-culture and induction was used. The induction medium as AMC (modified MS) supplemented by 2, 4-D (2.5 mg/l), kinetin (0.5 mg/l) and higher amount of D-Manitol (90 g/l) instead of sucrose and two types of amino acids (L-glutamine and L-serine) were used. Out of three main microspore isolation procedure by homogenizer isolation (P4) showed best performance on ELS induction (177%) and green plantlets (104%) compared with other techniques. For all cases albinisims occurred but microspore isolation from excised anthers by glass rod and homogenizer showed lesser numbers of albino plants that was also one of the important findings in this study.
Abstract: Venison is well known as a traditional meat type in Europe and it is lower in calories, cholesterol and fat content than common cuts of beef, pork or lamb. The aim of the current research was to determine content of amino acids (LVS ISO 13903:2005) in different types of marinades marinated venison during storage. Beef as a control was analyzed for comparison of obtained results. The meat (2x3x2cm) pieces were marinated in two different types of marinades: red wine and tomato sauce marinade. The prepared meat samples were stored (marinated) at 4±2ºC temperature for 48±1h. Marinated meat was placed in polypropylene trays, hermetically sealed with high barrier polymer film under modified atmosphere (C02 40%+N2 60%) without and with iron based oxygen scavenger sachets (Mitsubishi Gas Chemical Europe Ageless®), all samples were compared with packed marinated products in air ambiance. Results of current research show that changes of amino acids content in marinated venison mainly depend on packaging conditions.
Abstract: Some Chromium (III) complexes were synthesized
with three amino acids: L Glutamic Acid, Glycine, and L-cysteine as
the ligands, in order to provide a new supplement containing Cr(III)
for patients with type 2 diabetes mellitus. The complexes have been
prepared by refluxing a mixture of Chromium(III) chloride in
aqueous solution with L-glutamic acid, Glycine, and L-cysteine after
pH adjustment by sodium hydroxide. These complexes were
characterized by Infrared and Uv-Vis spectrophotometer and
Elemental analyzer. The product yields of four products were 87.50
and 56.76% for Cr-Glu complexes, 46.70% for Cr-Gly complex and
40.08% for Cr-Cys complex respectively. The predicted structure of
the complexes are [Cr(glu)2(H2O)2].xH2O, Cr(gly)3..xH2O and
Cr(cys)3.xH2O., respectively.
Abstract: The present study was carried out to evaluate the
nutritional value of sorghum flour during processing of injera
(unleavened thick bread). The proximate composition of sorghum
flour before and after fermentation and that of injera was determined.
Compared to the raw flour and fermented one, injera had low protein
(11.55%), ash (1.57%) and fat (2.40%) contents but high in fiber
content. Moreover, injera was found to have significantly (P ≤ 0.05)
higher energy (389.08 Kcal/100g) compared to raw and fermented
sorghum flour. Injera contained lower levels of anti-nutritional
factors (polyphenols, phytate and tannins) compared to raw and
fermented sorghum. Also it was found to be rich in Ca
(4.75mg/100g), Fe (3.95 mg/100g), and Cu (0.7 mg/100g) compared
to that of raw and fermented flour. Moreover, both the extractable
minerals and protein digestibility were high for injera due to low
amount of anti-nutrients. Injera was found to contain an appreciable
amount of amino acids except arginine and tyrosine.
Abstract: The main objective of this study was to determine the effects
of vermicompost and amino acids on the qualitative and quantitative yield of
chamomile. The experiment was conducted during the growing season of
2010 at the Alborz Medical Research Center. The Treatment groups consisted
of vermicompost (0, 5, 10, 15 and 20 tons/ha) and the sprays of amino acids
(budding stag, flowering stage, and budding + flowering stage). The
experimental design was a factorial experiment based on Randomized
Complete Block Design (RCBD) with three replications. The present results
have shown that the highest plant height, flower head diameter, fresh and dry
flower yield and significant essential oil content were obtained by using 20-
ton vermicompost per hectare. Effects of amino acids were similar to those
seen in vermicompost treatment and all measured traits were seen to be
significant after the spray of amino acids at the budding + flowering stage).
Abstract: The aim of the study was to determine how different
ripening processes (traditional vs. industrial) influenced the
proteolysis in traditional Serbian dry-fermented sausage Petrovská
klobása. The obtained results indicated more intensive pH decline
(0.7 units after 9 days) in industrially ripened products (I), what had a
positive impact on drying process and proteolytic changes in these
samples. Thus, moisture content in I sausages was lower at each
sampling time, amounting 24.7% at the end of production period
(90 days). Likewise, the process of proteolysis was more pronounced
in I samples, resulting in higher contents of non-protein nitrogen
(NPN) and free amino acids nitrogen (FAAN), as well as in faster
and more intensive degradation of myosin (≈220 kDa), actin (≈45
kDa) and other polypeptides during processing. Consequently, the
appearance and accumulation of several protein fragments were
registered.
Abstract: 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.
Abstract: 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.
Abstract: 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
Abstract: 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.
Abstract: 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.
Abstract: 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.
Abstract: 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.
Abstract: 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.
Abstract: 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.
Abstract: 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.
Abstract: 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.
Abstract: 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.
Abstract: 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.
Abstract: 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.