Abstract: β-Glucosidase is an important enzyme for production
of ethanol from lignocellulose. With hydrolytic activity on
cellooligosaccharides, especially cellobiose, β-glucosidase removes
product inhibitory effect on cellulases and forms fermentable sugars.
In this study, β-glucosidase encoding gene (BGL1) from traditional
starter yeast Saccharomycosis fibuligera BMQ908 was cloned and
expressed in Pichia pastoris. BGL1 of S. fibuligera BMQ 908 shared
98% nucleotide homology with the closest GenBank sequence
(M22475) but identity in amino-acid sequences of catalytic domains.
Recombinant plasmid pPICZαA/BGL1 containing the sequence
encoding BGL1 mature protein and α-factor secretion signal was
constructed and transformed into methylotrophic yeast P. pastoris by
electroporation. The recombinant strain produced single extracellular
protein with molecular weight of 120 kDa and cellobiase activity of
60 IU/ml. The optimum pH of the recombinant β-glucosidase was 5.0
and the optimum temperature was 50°C.
Abstract: Candida spp. are common and aggressive pathogens.
Because of the growing resistance of Candida spp. to current
antifungals, novel targets, found in Candida spp. but not in humans
or other flora, have to be identified. The alternative oxidase (AOX)
is one such possibility. This enzyme is insensitive to cyanide, but is
sensitive to compounds such as salicylhydroxamic acid (SHAM),
disulfiram and n-alkyl gallates. The growth Candida albicans was
inhibited by SHAM (Ki = 9-15 mM) and cyanide (Ki = 2-4 mM),
albeit to differing extents. The rate of O2 uptake was inhibited by
less than 10% by 25 mM SHAM and by about 90% by 250 μM
KCN. Although SHAM substantially inhibited the growth of C.
albicans, it is unlikely that the inhibition of AOX was the cause.
Salicylhydroxamic acid is used therapeutically in the treatment of
urinary tract infections and urolithiasis, but it also has some potential
in the treatment of C. albicans infection.
Abstract: Hepatocellular carcinoma, also called hepatoma, most
commonly appears in a patient with chronic viral hepatitis. In
patients with a higher suspicion of HCC, such as small or subtle
rising of serum enzymes levels, the best method of diagnosis
involves a CT scan of the abdomen, but only at high cost. The aim of
this study was to increase the ability of the physician to early detect
HCC, using a probabilistic neural network-based approach, in order
to save time and hospital resources.
Abstract: Enzymatic hydrolysis of starch from natural sources
finds potential application in commercial production of alcoholic
beverage and bioethanol. In this study the effect of starch
concentration, temperature, time and enzyme concentration were
studied and optimized for hydrolysis of cassava (Manihot esculenta)
starch powder (of mesh 80/120) into glucose syrup by immobilized
(using Polyacrylamide gel) a-amylase using central composite
design. The experimental result on enzymatic hydrolysis of cassava
starch was subjected to multiple linear regression analysis using
MINITAB 14 software. Positive linear effect of starch concentration,
enzyme concentration and time was observed on hydrolysis of
cassava starch by a-amylase. The statistical significance of the model
was validated by F-test for analysis of variance (p < 0.01). The
optimum value of starch concentration temperature, time and enzyme
concentration were found to be 4.5% (w/v), 45oC, 150 min, and 1%
(w/v) enzyme. The maximum glucose yield at optimum condition
was 5.17 mg/mL.
Abstract: Lutein is a dietary oxycarotenoid which is found
to reduce the risks of Age-related Macular Degeneration
(AMD). Supercritical fluid extraction of lutein esters from
marigold petals was carried out and was found to be much
effective than conventional solvent extraction. The
saponification of pre-concentrated lutein esters to produce free
lutein was studied which showed a composition of about 88%
total carotenoids (UV-VIS spectrophotometry) and 90.7%
lutein (HPLC). The lipase catalyzed hydrolysis of lutein esters
in conventional medium was investigated. The optimal
temperature, pH, enzyme concentration and water activity
were found to be 50°C, 7, 15% and 0.33 respectively and the
activity loss of lipase was about 25% after 8 times re-use in at
50°C for 12 days. However, the lipase catalyzed hydrolysis of
lutein esters in conventional media resulted in poor
conversions (16.4%).
Abstract: Deaminated lesions were produced via nitrosative oxidation of natural nucleobases; uracul (Ura, U) from cytosine (Cyt, C), hypoxanthine (Hyp, H) from adenine (Ade, A), and xanthine (Xan, X) and oxanine (Oxa, O) from guanine (Gua, G). Such damaged nucleobases may induce mutagenic problems, so that much attentions and efforts have been poured on the revealing of their mechanisms in vivo or in vitro. In this study, we employed these deaminated lesions as useful probes for analysis of DNA-binding/recognizing proteins or enzymes. Since the pyrimidine lesions such as Hyp, Oxa and Xan are employed as analogues of guanine, their comparative uses are informative for analyzing the role of Gua in DNA sequence in DNA-protein interaction. Several DNA oligomers containing such Hyp, Oxa or Xan substituted for Gua were designed to reveal the molecular interaction between DNA and protein. From this approach, we have got useful information to understand the molecular mechanisms of the DNA-recognizing enzymes, which have not ever been observed using conventional DNA oligomer composed of just natural nucleobases.
Abstract: Forty-five dairy cows were used to compare the
enzyme activity of alkaline phosphatase (ALP), lactate
dehydrogenase (LDH), α -amylase in the cervical mucus of cows
during spontaneous and induced estrus using progestagen or PGF2 α
and to determine whether these enzymes affect the fertility in cows
with induced estrus, at the time of Al. The animals were assigned to 3
groups (no treatment, a Crestar® for 12 days, a double im injection of
PGF2 α). The cows were artificially inseminated (AI). Cervical
mucus samples were collected from all cows 3 to 5 min before the
AI. The results are summarized as follows: ALP and α -amylase
activity for spontaneous estrus were similar to those for induced
estrus (P>0.05) . LDH activity levels during spontaneous and PGF2 α
induced estrus was significantly lower (P < 0.001) than that in
progestagene induced estrus groups. While no difference was found
between the first and the third groups. Our result showed a significant
difference in LDH activity levels between cows conceived with 2 or
more AI and those conceived with 1 AI. The result of this study
showed that the enzyme activity in cervical mucus is helpful for
detection of ovulation and time of AI.
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: A procedure for the preparation of clarified Pawpaw
Juice was developed. About 750ml Pawpaw pulp was measured into
2 measuring cylinders A & B of capacity 1 litre heated to 400C,
cooled to 200C. 30mls pectinase was added into cylinder A, while
30mls distilled water was added into cylinder B. Enzyme treated
sample (A) was allowed to digest for 5hours after which it was heated
to 900C for 15 minutes to inactivate the enzyme. The heated sample
was cooled and with the aid of a mucillin cloth the pulp was filtered
to obtain the clarified pawpaw juice. The juice was filled into 100ml
plastic bottles, pasteurized at 950C for 45 minutes, cooled and stored
at room temperature. The sample treated with 30mls distilled water
also underwent the same process. Freshly pasteurized sample was
analyzed for specific gravity, titratable acidity, pH, sugars and
ascorbic acid. The remaining sample was then stored for 2 weeks and
the above analyses repeated. There were differences in the results of
the freshly pasteurized samples and stored sample in pH and ascorbic
acid levels, also sample treated with pectinase yielded higher
volumes of juice than that treated with distilled water.
Abstract: Leonotisleonurus a shrub indigenous to Southern
Africa is widely used in traditional medicine to treat a variety of
conditions ranging from skin diseases and cough to epileptic fits and
‘heart problems’. Studies on the aqueous extract of the leaves have
indicated cycloxegenase enzyme inhibitory activity and an
antihypertensive effect.
Five methanol leaf extract fractions (MLEa - MLEe) of L.
leonurus were tested on anaesthetized normotensive male Wistar rats
(AWR) and isolated perfused working rat hearts (IWH). Fraction
MLEc (0.01mg/kg – 0.05mg/kg) induced significant increases in BP
and HR in AWR and positive chronotropic and inotropic effects in
IWH (1.0mg/ml – 5.0mg/ml). Pre-administration of atenolol
(2.0mg/kg) and prazosin (60μg/kg) significantly inhibited MLEc
effect on HR and MAP respectively in vivo, while atenolol
(7.0mg/ml) pre-perfusion significantly inhibited MLEc effect in vitro.
The hypertensive effect of MLEc is probably via β1agonism.
Results also indicate the presence of multiple cardioactive
compounds in L. leonurus.
Abstract: Urinary Tract Infections (UTI) account for an estimated 25-40% nosocomial infection, out of which 90% are associated with urinary catheter, called Catheter associated urinary tract infection (CAUTI). The microbial populations within CAUTI frequently develop as biofilms. In the present study, microbial contamination of indwelling urinary catheters was investigated. Biofilm forming ability of the isolates was determined by tissue culture plate method. Prevention of biofilm formation in the urinary catheter by Pseudomonas aeruginosa was also determined by coating the catheter with some enzymes, gentamycin and EDTA. It was found that 64% of the urinary catheters get contaminated during the course of catheterization. Of the total 6 isolates, biofilm formation was seen in 100% Pseudomonas aeruginosa and E. coli, 90% in Enterococci, 80% in Klebsiella and 66% in S. aureus. It was noted that the biofilm production by Pseudomonas was prolonged by 7 days in amylase, 8 days in protease, 6 days in lysozyme, 7days in gentamycin and 5 days in EDTA treated catheter.
Abstract: Type 2 diabetes mellitus (T2DM) is a complex
metabolic disorder that characterized by the presence of high glucose
in blood that cause from insulin resistance and insufficiency due to
deterioration β-cell Langerhans functions. T2DM is commonly
caused by the combination of inherited genetic variations as well as
our own lifestyle. Metallothionein (MT) is a known cysteine-rich
protein responsible in helping zinc homeostasis which is important in
insulin signaling and secretion as well as protection our body from
reactive oxygen species (ROS). MT scavenged ROS and free
radicals in our body happen to be one of the reasons of T2DM and its
complications. The objective of this study was to investigate the
association of MT1A and MT2A polymorphisms between T2DM and
control subjects among Malay populations. This study involved 150
T2DM and 120 Healthy individuals of Malay ethnic with mixed
genders. The genomic DNA was extracted from buccal cells and
amplified for MT1A and MT2A loci; the 347bp and 238bp banding
patterns were respectively produced by mean of the Polymerase
Chain Reaction (PCR). The PCR products were digested with Mlucl
and Tsp451 restriction enzymes respectively and producing
fragments lengths of (158/189/347bp) and (103/135/238bp)
respectively. The ANOVA test was conducted and it shown that there
was a significant difference between diabetic and control subjects for
age, BMI, WHR, SBP, FPG, HBA1C, LDL, TG, TC and family
history with (P0.05). The genotype
frequency for AA, AG and GG of MT1A polymorphisms was 72.7%,
22.7% and 4.7% in cases and 15%, 55% and 30% in control
respectively. As for MT2A, genotype frequency of GG, GC and CC
was 42.7%, 27.3% and 30% in case and 5%, 40% and 55% for
control respectively. Both polymorphisms show significant difference
between two investigated groups with (P=0.000). The Post hoc test
was conducted and shows a significant difference between the
genotypes within each polymorphism (P=0. 000). The MT1A and
MT2A polymorphisms were believed to be the reliable molecular
markers to distinguish the T2DM subjects from healthy individuals in
Malay populations.
Abstract: Fertilization plays an important role in crop growth and soil improvement. This study was conducted to determine the best fertilization system for wheat production. Experiments were arranged in a complete block design with three replications in two years. Main plots consisted of six methods of fertilization including (N1): farmyard manure; (N2): compost; (N3): chemical fertilizers; (N4): farmyard manure + compost; (N5): farmyard manure + compost + chemical fertilizers and (N6): control were arranged in sub plots. The addition of compost or farm yard manure significantly increased the soil microbial biomass carbon in comparison to the chemical fertilizer. The dehydrogenase, phosphatase and urease activities in the N3 treatment were significantly lower than in the farm yard manure and compost treatments.
Abstract: In this study, we investigated the effects of ginger and
L-carnitine on the reproductive performance of male rats with respect
to semen parameters, male sex hormones and the testicular
antioxidant system. A total of sixty mature male albino rats were
divided into four groups of fifteen rats. The control group received
saline, whereas the other three groups received ginger (100 mg kg-1 d-
1.), L-carnitine (150 mg kg-1 d-1.) or a combination of both ginger
(100 mg kg-1 d-1.) and L-carnitine (150 mg kg-1 d-1.) via a stomach
tube daily for one month. At the end of the treatment period, the rats
were sacrificed, and their sperm characteristics (count, motility and
viability), antioxidant enzyme factors levels (reduced glutathione,
catalase, superoxide dismutase and total antioxidant capacity) and sex
hormone levels (testosterone, Follicle stimulating hormone(FSH) and
luteinizing hormone (LH) were analysed. Our results showed that the
three experimental treatments improved sperm parameters,
antioxidant enzyme activity and testosterone hormone levels; the
most pronounced positive effects were observed in the group that
received a combination of both ginger and L-carnitine. Therefore, the
administration of a combination of ginger and L-carnitine may be
beneficial for improving male sexual performance.
Abstract: The methanolic extracts from seeds of tamarind
(Tamarindus indica) was prepared by Soxhlet apparatus extraction
and evaluated for total phenolic content by Folin-Ciocalteu method.
Then, methanolic extract was screened biological activities (In vitro)
for anti-melanogenic activity by tyrosinase inhibition test, antiinflammation
activity by cyclooxygenase 1 (COX-1) and
cyclooxygenase 2 (COX-2) inhibition test, and cytotoxic screening
test with Vero cells. The results showed that total phenolic content,
which contained in extract, was contained 27.72 mg of gallic acid
equivalent per g of dry weight. The ability to inhibit tyrosinase
enzyme, which exerted by Tamarind seed extracts (1 mg/ml) was
52.13 ± 0.42 %. The extract was not possessed inhibitory effect to
COX-1 and COX-2 enzymes and cytotoxic effect to Vero cells. The
finding is concludes that tested seed extract was possessed
antimelanogenic activity with non-toxic effects. However, there was
not exhibited anti-inflammatory activity. Further studies include the
use of advance biological models to confirm this biological activity,
as well as, the isolation and characterization of the purified
compounds that it was contained.
Abstract: The p53 tumor suppressor gene plays two important
roles in genomic stability: blocking cell proliferation after DNA
damage until it has been repaired, and starting apoptosis if the
damage is too critical. Codon 72 exon4 polymorphism (Arg72Pro) of
the P53 gene has been implicated in cancer risk. Various studies have
been done to investigate the status of p53 at codon 72 for arginine
(Arg) and proline (Pro) alleles in different populations and also the
association of this codon 72 polymorphism with various tumors. Our
objective was to investigate the possible association between P53
Arg72Pro polymorphism and susceptibility to colorectal cancer
among Isfahan and Chaharmahal Va Bakhtiari (a part of south west
of Iran) population. We investigated the status of p53 at codon 72 for
Arg/Arg, Arg/Pro and Pro/Pro allele polymorphisms in blood
samples from 145 colorectal cancer patients and 140 controls by
Nested-PCR of p53 exon 4 and digestion with BstUI restriction
enzyme and the DNA fragments were then resolved by
electrophoresis in 2% agarose gel. The Pro allele was 279 bp, while
the Arg allele was restricted into two fragments of 160 and 119 bp.
Among the 145 colorectal cancer cases 49 cases (33.79%) were
homozygous for the Arg72 allele (Arg/Arg), 18 cases (12.41%) were
homozygous for the Pro72 allele (Pro/Pro) and 78 cases (53.8%)
found in heterozygous (Arg/Pro).
In conclusion, it can be said that p53Arg/Arg genotype may be
correlated with possible increased risk of this kind of cancers in south
west of Iran.
Abstract: 17α-ethynylestradiol (EE2) is a synthetic estrogen
used as a key ingredient in an oral contraceptives pill. EE2 is an
endocrine disrupting compound, high in estrogenic potency.
Although EE2 exhibits low degree of biodegradability with common
microorganisms in wastewater treatment plants (WWTPs), this
compound can be biotransformed by ammonia-oxidizing bacteria
(AOB) via a co-metabolism mechanism in WWTPs. This study
aimed to investigate the effect of real wastewater on
biotransformation of EE2 by AOB. A preliminary experiment on the
effect of nitrite and pH levels on abiotic transformation of EE2
suggested that the abiotic transformation occurred at only pH
Abstract: The present work represents an investigation of the
hydrolysis of hull-less pumpkin (Cucurbita Pepo L.) oil cake protein
isolate (PuOC PI) by pepsin. To examine the effectiveness and
suitability of pepsin towards PuOC PI the kinetic parameters for
pepsin on PuOC PI were determined and then, the hydrolysis process
was studied using Response Surface Methodology (RSM). The
hydrolysis was carried out at temperature of 30°C and pH 3.00. Time
and initial enzyme/substrate ratio (E/S) at three levels were selected
as the independent parameters. The degree of hydrolysis, DH, was
mesuared after 20, 30 and 40 minutes, at initial E/S of 0.7, 1 and 1.3
mA/mg proteins. Since the proposed second-order polynomial model
showed good fit with the experimental data (R2 = 0.9822), the
obtained mathematical model could be used for monitoring the
hydrolysis of PuOC PI by pepsin, under studied experimental
conditions, varying the time and initial E/S. To achieve the highest
value of DH (39.13 %), the obtained optimum conditions for time
and initial E/S were 30 min and 1.024 mA/mg proteins.
Abstract: Effect of viscosity of media on kinetic parameters of the coupled enzyme system NADH:FMN-oxidoreductase–luciferase was investigated with addition of organic solvents (glycerol and sucrose), because bioluminescent enzyme systems based on bacterial luciferases offer a unique and general tool for analysis of the many analytes and enzymes in the environment, research and clinical laboratories and other fields. The possibility of stabilization and increase of activity of the coupled enzyme system NADH:FMN-oxidoreductase–luciferase activity in vicious aqueous-organic mixtures have been shown.
Abstract: In this paper we have proposed a methodology to
develop an amperometric biosensor for the analysis of glucose
concentration using a simple microcontroller based data acquisition
system. The work involves the development of Detachable
Membrane Unit (enzyme based biomembrane) with immobilized
glucose oxidase on the membrane and interfacing the same to the
signal conditioning system. The current generated by the biosensor
for different glucose concentrations was signal conditioned, then
acquired and computed by a simple AT89C51-microcontroller. The
optimum operating parameters for the better performance were found
and reported. The detailed performance evaluation of the biosensor
has been carried out. The proposed microcontroller based biosensor
system has the sensitivity of 0.04V/g/dl, with a resolution of
50mg/dl. It has exhibited very good inter day stability observed up to
30 days. Comparing to the reference method such as HPLC, the
accuracy of the proposed biosensor system is well within ± 1.5%.
The system can be used for real time analysis of glucose
concentration in the field such as, food and fermentation and clinical
(In-Vitro) applications.