Abstract: A. niger XP isolated from Vietnam produces very low amount of acidic phytase with optimal pH at 2.5 and 5.5. The phytase production of this strain was successfully improved through gene cloning and expression. A 1.4 - kb DNA fragment containing the coding region of the phyA gene was amplified by PCR and inserted into the expression vector pPICZαA with a signal peptide α- factor, under the control of AOX1 promoter. The recombined plasmid was transformed into the host strain P. pastoris KM71H and X33 by electroporation. Both host strains could efficiently express and secret phytase. The multicopy strains were screened for over expression of phytase. All the selected multicopy strains of P. pastoris X33 were examined for phytase activity, the maximum phytase yield of 1329 IU/ml was obtained after 4 days of incubation in medium BMM. The recombinant protein with MW of 97.4 KW showed to be the only one protein secreted in the culture broth. Multicopy transformant P. pastoris X33 supposed to be potential candidate for producing the commercial preparation of phytase.
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: MicroRNAs (miRNAs) are small, non-coding and
regulatory RNAs about 20 to 24 nucleotides long. Their conserved
nature among the various organisms makes them a good source of
new miRNAs discovery by comparative genomics approach. The
study resulted in 21 miRNAs of 20 pre-miRNAs belonging to 16
families (miR156, 157, 158, 164, 165, 168, 169, 172, 319, 390, 393,
394, 395, 400, 472 and 861) in evergreen spruce tree (Picea). The
miRNA families; miR 157, 158, 164, 165, 168, 169, 319, 390, 393,
394, 400, 472 and 861 are reported for the first time in the Picea. All
20 miRNA precursors form stable minimum free energy stem-loop
structure as their orthologues form in Arabidopsis and the mature
miRNA reside in the stem portion of the stem loop structure. Sixteen
(16) miRNAs are from Picea glauca and five (5) belong to Picea
sitchensis. Their targets consist of transcription factors, growth
related, stressed related and hypothetical proteins.
Abstract: Lectins have a good scope in current clinical
microbiology research. In the present study evaluated the
antimicrobial activities of a D-galactose binding lectin (PnL) was
purified from the annelid, Perinereis nuntia (polychaeta) by affinity
chromatography. The molecular mass of the lectin was determined to
be 32 kDa as a single polypeptide by SDS-PAGE under both reducing
and non-reducing conditions. The hemagglutinating activity of the
PnL showed against trypsinized and glutaraldehyde-fixed human
erythrocytes was specifically inhibited by D-Gal, GalNAc,
Galβ1-4Glc and Galα1-6Glc. PnL was evaluated for in vitro
antibacterial screening studies against 11 gram-positive and
gram-negative microorganisms. From the screening results, it was
revealed that PnL exhibited significant antibacterial activity against
gram-positive bacteria. Bacillus megaterium showed the highest
growth inhibition by the lectin (250 μg/disc). However, PnL did not
inhibit the growth of gram-negative bacteria such as Vibrio cholerae
and Pseudomonas sp. PnL was also examined for in vitro antifungal
activity against six fungal phytopathogens. PnL (100 μg/mL) inhibited
the mycelial growth of Alternaria alternata (24.4%). These results
indicate that future findings of lectin applications obtained from
annelids may be of importance to life sciences.
Abstract: In April 2009, a new variant of Influenza A virus
subtype H1N1 emerged in Mexico and spread all over the world. The
influenza has three subtypes in human (H1N1, H1N2 and H3N2)
Types B and C influenza tend to be associated with local or regional
epidemics. Preliminary genetic characterization of the influenza
viruses has identified them as swine influenza A (H1N1) viruses.
Nucleotide sequence analysis of the Haemagglutinin (HA) and
Neuraminidase (NA) are similar to each other and the majority of
their genes of swine influenza viruses, two genes coding for the
neuraminidase (NA) and matrix (M) proteins are similar to
corresponding genes of swine influenza. Sequence similarity between
the 2009 A (H1N1) virus and its nearest relatives indicates that its
gene segments have been circulating undetected for an extended
period. Nucleic acid sequence Maximum Likelihood (MCL) and
DNA Empirical base frequencies, Phylogenetic relationship amongst
the HA genes of H1N1 virus isolated in Genbank having high
nucleotide sequence homology.
In this paper we used 16 HA nucleotide sequences from NCBI for
computing sequence relationships similarity of swine influenza A
virus using the following method MCL the result is 28%, 36.64% for
Optimal tree with the sum of branch length, 35.62% for Interior
branch phylogeny Neighber – Join Tree, 1.85% for the overall
transition/transversion, and 8.28% for Overall mean distance.
Abstract: Certain tRNA synthetases have developed highly accurate molecular machinery to discriminate their cognate amino acids. Those aaRSs achieve their goal via editing reaction in the Connective Polypeptide 1 (CP1). Recently mutagenesis studies have revealed the critical importance of residues in the CP1 domain for editing activity and X-ray structures have shown binding mode of noncognate amino acids in the editing domain. To pursue molecular mechanism for amino acid discrimination, molecular modeling studies were performed. Our results suggest that aaRS bind the noncognate amino acid more tightly than the cognate one. Finally, by comparing binding conformations of the amino acids in three systems, the amino acid binding mode was elucidated and a discrimination mechanism proposed. The results strongly reveal that the conserved threonines are responsible for amino acid discrimination. This is achieved through side chain interactions between T252 and T247/T248 as well as between those threonines and the incoming amino acids.
Abstract: The objective of current issue was to develop a model
of testicular herpes simplex virus (HSV) type I infection for
assessment of viral effect on fertility. 56 male mice were inoculated
intraperitoneally with different concentrations of HSV on 8 day post
partum. It was revealed that the optimal dose was 100 plaque
forming units per mice as it provided testicular infection in 100% of
survivors. HSV proteins were detected both in somatic and germ
cells (spermatogonia, spermatocytes, spermatides). Although DNA
load in testis was descending from 3 to 28 days post infection only
12.5% of infected males had offspring after mating with uninfected
females comparing to 87.5% in control (p=0.012). These results are
the first direct evidence for HSV impact in male sterility. Prepuberal
mice appeared to be a suitable model for investigation of
pathogenesis of virus-associated fertility disorders.
Abstract: Bacillus subtilis strain LB5 produced lipopeptide
antibiotic iturin A-2 in liquid medium. Crude extract
from cell-free supernatant of B. subtilis cultivated broth extracted
with n-butanol showed antifungal activity to conidial germination of
Colletotrichum gloeosporioides. The germination of conidia was
completely inhibited by crude extract. The ultrastructure of conidia
after treated with crude extract was found an accumulation of vesiclelike
material between cell wall and plasma membrane while this
accumulation was not observed in untreated and germinated conidia.
Besides, the cell wall was not affected by crude extract.
Abstract: The present study has been taken to explore the
screening of in vitro antimicrobial activities of D-galactose-binding
sponge lectin (HOL-30). HOL-30 was purified from the marine
demosponge Halichondria okadai by affinity chromatography. The
molecular mass of the lectin was determined to be 30 kDa with a
single polypeptide by SDS-PAGE under non-reducing and reducing
conditions. HOL-30 agglutinated trypsinized and glutaraldehydefixed
rabbit and human erythrocytes with preference for type O
erythrocytes. The lectin was subjected to evaluation for inhibition of
microbial growth by the disc diffusion method against eleven human
pathogenic gram-positive and gram-negative bacteria. The lectin
exhibited strong antibacterial activity against gram-positive bacteria,
such as Bacillus megaterium and Bacillus subtilis. However, it did
not affect against gram-negative bacteria such as Salmonella typhi
and Escherichia coli. The largest zone of inhibition was recorded of
Bacillus megaterium (12 in diameter) and Bacillus subtilis (10 mm in
diameter) at a concentration of the lectin (250 μg/disc). On the other
hand, the antifungal activity of the lectin was investigated against six
phytopathogenic fungi based on food poisoning technique. The lectin
has shown maximum inhibition (22.83%) of mycelial growth of
Botrydiplodia theobromae at a concentration of 100 μg/mL media.
These findings indicate that the lectin may be of importance to
clinical microbiology and have therapeutic applications.
Abstract: Approximate tandem repeats in a genomic sequence are
two or more contiguous, similar copies of a pattern of nucleotides.
They are used in DNA mapping, studying molecular evolution
mechanisms, forensic analysis and research in diagnosis of inherited
diseases. All their functions are still investigated and not well
defined, but increasing biological databases together with tools for
identification of these repeats may lead to discovery of their specific
role or correlation with particular features. This paper presents a new
approach for finding approximate tandem repeats in a given sequence,
where the similarity between consecutive repeats is measured using
the Hamming distance. It is an enhancement of a method for finding
exact tandem repeats in DNA sequences based on the Burrows-
Wheeler transform.
Abstract: Recent research result has shown that two multidelay
feedback systems can synchronize each other under different
schemes, i.e. lag, projective-lag, anticipating, or projectiveanticipating
synchronization. There, the driving signal is significantly
complex due that it is constituted by multiple nonlinear transformations
of delayed state variable. In this paper, a secure communication
model is proposed based on synchronization of coupled multidelay
feedback systems, in which the plain signal is mixed with a complex
signal at the transmitter side and it is precisely retrieved at the receiver
side. The effectiveness of the proposed model is demonstrated and
verified in the specific example, where the message signal is masked
directly by the complex signal and security is examined under the
breaking method of power spectrum analysis.
Abstract: The ability to distinguish missense nucleotide
substitutions that contribute to harmful effect from those that do not
is a difficult problem usually accomplished through functional in
vivo analyses. In this study, instead current biochemical methods, the
effects of missense mutations upon protein structure and function
were assayed by means of computational methods and information
from the databases. For this order, the effects of new missense
mutations in exon 5 of PTEN gene upon protein structure and
function were examined. The gene coding for PTEN was identified
and localized on chromosome region 10q23.3 as the tumor
suppressor gene. The utilization of these methods were shown that
c.319G>A and c.341T>G missense mutations that were recognized in
patients with breast cancer and Cowden disease, could be pathogenic.
This method could be use for analysis of missense mutation in others
genes.
Abstract: Biologically active peptides are of particular interest
in food science and human nutrition because they have been shown to play several physiological roles. In vitro gastrointestinal digestion of lentil and whey proteins in this study produced high angiotensin-I converting enzyme inhibitory activity with 75.5±1.9 and 91.4±2.3%
inhibition, respectively. High ACE inhibitory activity was observed in lentil after 5 days of germination (84.3±1.2%). Fractionation by
reverse phase chromatography gave inhibitory activities as high as
86.3±2.0 for lentil, 94.8±1.8% for whey and 93.7±1.7% at 5th day of germination. Further purification by HPLC resulted in several
inhibitory peptides with IC50 values ranging from 0.064 to 0.164
mg/ml. These results demonstrate that lentil proteins are a good
source of peptides with ACE inhibitory activity that can be released by germination or gastrointestinal digestion. Despite the lower bioactivity in comparison with whey proteins, incorporation of lentil proteins in functional food formulations and natural drugs look promising.