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 Potato starch powder (of
mesh 80/120) into glucose syrup by immobilized (using Sodium
arginate) α-amylase using central composite design. The
experimental result on enzymatic hydrolysis of Potato 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 Potato starch
by α-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, enzyme concentration, temperature, time and
were found to be 6% (w/v), 2% (w/v), 40°C and 80min respectively.
The maximum glucose yield at optimum condition was 2.34 mg/mL.
Abstract: Phytases (myo-inositol hexakisphosphate
phosphohydrolases; EC 3.1.3.8) catalyze the hydrolysis of phytic acid
(myoinositol hexakisphosphate) to the mono-, di-, tri-, tetra-, and
pentaphosphates of myo-inositol and inorganic phosphate.
Therrmophilic bacteria isolated from water sampled from hot spring.
About 120 isolates of bacteria were successfully isolated form hot
spring water sample and tested for extracellular phytase producing.
After 5 passages of the screening on the PSM media, 4 isolates were
found stable in producing phytase enzyme. The 16s RDNA
sequencing for identification of bacteria using molecular technique
revealed that all isolates those positive in phytase producing are
belong to Geobacillus spp. And Anoxybacillus spp. Anoxybacillus
rupiensis UniSZA-7 were identified for their carbon source utilization
using Phenotype Microarray Plate of Biolog and found they utilize
several kind of carbon source provided.
Abstract: Phytases are enzymes used as an important component
in monogastric animals feeds in order to improve phosphorous
availability, since it is not readily assimilated by these animals in the
form of the phytate presented in plants and grains. As these enzymes
are used in industrial activities, they must retain its catalytic activities
during a certain storage period. This study presents information about
the stability of 4 different phytases, produced by four macromycetes
fungi through solid-state fermentation (SSF). There is a lack of data
in literature concerning phytase from macromycetes shelf-life in
storage conditions at room, cooling and freezing temperatures. The 4
phytases from macromycetes still had enzymatic activities around
100 days of storage at room temperature. At cooling temperature in
146 days of studies, the phytase from G. stipitatum was the most
stable with 44% of the initial activity, in U.gds (units per gram of
dried fermented substrate). The freezing temperature was the best
condition storage for phytases from G. stipitatum and T. versicolor.
Each condition provided a study for each mushroom phytase,
totalizing 12 studies. The phytases showed to be stable for a long
period without the addition of additives.
Abstract: To assess the effects of functional protein on osteoblast, Large quantity of high-purity osteoblasts had been cultivated successfully by adopting sequential enzyme digestion. The growth curve of osteoblasts was protracted by cell counting. Proliferation of osteoblasts was assessed by MTT colorimetry. The experimental results show the functional protein can enhance proliferation, the properties of adhesion and discuss the effect of osteopontin on osteoblast.
Abstract: This paper studied the synthesis of monoacylglycerol (monolaurin) by glycerolysis of coconut oil and crude glycerol, catalyzed by Carica papaya lipase. Coconut oil obtained from cold pressed extraction method and crude glycerol obtained from the biodiesel plant in Department of Chemistry, Uttaradit Rajabhat University, Thailand which used oils were used as raw materials for biodiesel production through transesterification process catalyzed by sodium hydroxide. The influences of the following variables were studied: (i) type of organic solvent, (ii) molar ratio of substrate, (iii) reaction temperature, (iv) reaction time, (v) lipase dosage, and (vi) initial water activity of enzyme. High yields in monoacylglycerol (58.35%) were obtained with molar ratio of glycerol to oil at 8:1 in ethanol, temperature was controlled at 45oC for 36 hours, the amount of enzyme used was 20 wt% of oil and initial water activity of enzyme at 0.53.
Abstract: In the present study, we aimed to design the
intrauterine and extrauterine exposure to 1800 MHz GSM-like RF
radiation and investigate its possible bio-effects on infant female
rabbits. Totally thirty-six New Zealand White female rabbits, onemonth
old, were randomly divided into four groups which are
composed of 9 rabbits; i. Group I [Intrauterine (IU) exposure(-);
Extrauterine (EU) exposure (-)], Group II [IU exposure (-); EU
exposure (+)], Group III [IU exposure(+);EU exposure(-)], Group IV
[IU exposure (+);EU exposure(+)]. The master regulatory enzymes
activities of pentose phosphate pathway (glucose-6-phosphate
dehydrogenase, G-6PD; 6-phosphogluconate dehydrogenase, 6-
PGDH) and glutathione-dependent metabolism (glutathione
peroxidase, GSH-Px; glutathione reductase, GR; glutathione Stransferase,
GST, thioredoxin reductase, TRx) were analyzed in liver
tissues of young female rabbits. Decreased G-6PD, 6-PGD, GSH-Px,
GR activities were found in Group III compared to Group I (p
Abstract: Protective effect of ethanolic extract of polyherbal formulation (PHF) was studied on carbon tetrachloride induced liver damage on carbon tetrachloride induced liver damage. Treatment of rats with 250mg /kg body weight of ethanolic extract of PHF protected rats against carbon tetrachloride liver injury by significant lowerering 5’ nucleotidase (5’NT), Gamma Glutamyl transferase (GGT), Glutamate dehdyrogenasse (GDH) and Succinate Dehydrogenase (SDH) levels compared to control. Normalization in these enzyme levels indicates strong hepatoprotective property of PHF extract.
Abstract: In vitro gastro-duodenal digestion model was used to investigate the changes of emulsions under digestion conditions. Oil in water emulsions stabilized by whey proteins (2%) and stabilized by whey proteins (2%) with addition of carboxymethyl cellulose (0.75%) as gelling agent of continuous phase were prepared at pH7. Both emulsions were destabilized under gastric conditions; however the protective role of carboxymethyl cellulose was indicated by recording delay of fat digestibility of this emulsion. In the presence of carboxymethyl cellulose whey proteins on the interfacial surface of droplets were more resistant to gastric degradation causing limited hydrolysis of fat due to the poor acceptability of lipids for the enzymes. Studies of emulsions using in vivo model supported results from in vitro studies. Lower content of triglycerides in blood serum and higher amount of fecal fat of rats were determined when rats were fed by diet containing emulsion made with whey proteins and carboxymethyl cellulose.
Abstract: Two commercial proteases from Bacillus
licheniformis (Alcalase 2.4 L FG and Alcalase 2.5 L, Type DX) were
screened for the production of Z-Ala-Phe-NH2 in batch reaction.
Alcalase 2.4 L FG was the most efficient enzyme for the C-terminal
amidation of Z-Ala-Phe-OMe using ammonium carbamate as
ammonium source. Immobilization of protease has been achieved by
the sol-gel method, using dimethyldimethoxysilane (DMDMOS) and
tetramethoxysilane (TMOS) as precursors (unpublished results). In
batch production, about 95% of Z-Ala-Phe-NH2 was obtained at
30°C after 24 hours of incubation. Reproducibility of different
batches of commercial Alcalase 2.4 L FG preparations was also
investigated by evaluating the amidation activity and the entrapment
yields in the case of immobilization. A packed-bed reactor (0.68 cm
ID, 15.0 cm long) was operated successfully for the continuous
synthesis of peptide amides. The immobilized enzyme retained the
initial activity over 10 cycles of repeated use in continuous reactor at
ambient temperature. At 0.75 mL/min flow rate of the substrate
mixture, the total conversion of Z-Ala-Phe-OMe was achieved after 5
hours of substrate recycling. The product contained about 90%
peptide amide and 10% hydrolysis byproduct.
Abstract: Extraction of laccase produced by L. polychrous in an
aqueous two-phase system, composed of polyethylene glycol and
phosphate salt at pH 7.0 and 250C was investigated. The effect of
PEG molecular weight, PEG concentration and phosphate
concentration was determined. Laccase preferentially partitioned to
the top phase. Good extraction of laccase to the top phase was
observed with PEG 4000. The optimum system was found in the
system containing 12% w/w PEG 4000 and 16% w/w phosphate salt
with KE of 88.3, purification factor of 3.0-fold and 99.1% yield.
Some properties of the enzyme such as thermal stability, effect of
heavy metal ions and kinetic constants were also presented in this
work. The thermal stability decreased sharply with high temperature
above 60 0C. The enzyme was inhibited by Cd2+, Pb2+, Zn2+ and
Cu2+. The Vmax and Km values of the enzyme were 74.70
μmol/min/ml and 9.066 mM respectively.
Abstract: 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.
Abstract: Glutathione S-transferase was purified from human
erythrocytes and effects of some polyphenols were investigated on
the enzyme activity. The purification procedure was performed on
Glutathione-Agarose affinity chromatography after preparation of
erythrocytes hemolysate with a yield of 81%. The purified enzyme
showed a single band on the SDS-PAGE. The effects of some
poliphenolic compounds such as catechin, dopa, dopamine, progallol
and catechol were examined on the in vitro GST activity. Catechin
was determined to be inhibitor for the enzyme, but others were not
effective on the enzyme as inhibitors or activators. IC50 value -the
concentration of inhibitor which reduces enzyme activity by 50%-
was estimated to be 10 mM. Ki constants were also calculated as 6.38
± 0,70 mM with GSH substrate, and 3.86 ± 0,78 mM with CDNB
substrate using the equations of graphs for the inhibitor, and its
inhibition type was determined as non-competitive.
Abstract: Sol-gel immobilization of enzymes, which can improve considerably their properties, is now one of the most used techniques. By deposition of the entrapped lipase on a solid support, a new and improved biocatalyst was obtained, which can be used with excellent results in acylation reactions. In this paper, lipase B from Candida antarctica was double immobilized on different adsorbents. These biocatalysts were employed in the kinetic resolution of several aliphatic secondary alcohols in organic medium. High total recovery yields of enzymatic activity, up to 560%, were obtained. For all the studied alcohols the enantiomeric ratios E were over 200. The influence of the reaction medium was studied for the kinetic resolution of 2-pentanol.
Abstract: Palm oil could be converted to cocoa butter equivalent by lipase-catalyzed interesterification. The objective of this research was to investigate the structure modification of palm oil to cocoa butter equivalent using Carica papaya lipase –catalyzed interesterification. The study showed that the compositions of cocoa butter equivalent were affected by acyl donor sources, substrate ratio, initial water of enzyme, reaction time, reaction temperature and the amount of enzyme. Among three acyl donors tested (methyl stearate, ethyl stearate and stearic acid), methyl stearate appeared to be the best acyl donor for incorporation to palm oil structure. The best reaction conditions for cocoa butter equivalent production were : substrate ratio (palm oil : methyl stearate, mol/mol) at 1 : 4, water activity of enzyme at 0.11, reaction time at 4 h, reaction temperature at 45 ° C and 18% by weight of the enzyme. The chemical and physical properties of cocoa butter equivalent were 9.75 ± 0.41% free fatty acid, 44.89 ± 0.84 iodine number, 193.19 ± 0.78 sponification value and melting point at 37-39 °C.
Abstract: The antioxidant compounds are needed for the food, beverages, and pharmaceuticals industry. For this purpose, an appropriate method is required to measure the antioxidant properties in various types of samples. Spectrophotometric method usually used has some weaknesses, including the high price, long sample preparation time, and less sensitivity. Among the alternative methods developed to overcome these weaknesses is antioxidant biosensor based on superoxide dismutase (SOD) enzyme. Therefore, this study was carried out to measure the SOD activity originating from Deinococcus radiodurans and to determine its kinetics properties. Carbon paste electrode modified with ferrocene and immobilized SOD exhibited anode and cathode current peak at potential of +400 and +300mv respectively, in both pure SOD and SOD of D. radiodurans. This indicated that the current generated was from superoxide catalytic dismutation reaction by SOD. Optimum conditions for SOD activity was at pH 9 and temperature of 27.50C for D. radiodurans SOD, and pH 11 and temperature of 200C for pure SOD. Dismutation reaction kinetics of superoxide catalyzed by SOD followed the Lineweaver-Burk kinetics with D. radiodurans SOD KMapp value was smaller than pure SOD. The result showed that D. radiodurans SOD had higher enzyme-substrate affinity and specificity than pure SOD. It concluded that D. radiodurans SOD had a great potential as biological recognition component for antioxidant biosensor.
Abstract: Hypertension is characterized with stress on the heart and blood vessels thus increasing the risk of heart attack and renal diseases. The Renin angiotensin system (RAS) plays a major role in blood pressure control. Renin is the enzyme that controls the RAS at the rate-limiting step. Our aim is to develop new drug-like leads which can inhibit renin and thereby emerge as therapeutics for hypertension. To achieve this, molecular dynamics (MD) simulation and receptor-based pharmacophore modeling were implemented, and three rennin-inhibitor complex structures were selected based on IC50 value and scaffolds of inhibitors. Three pharmacophore models were generated considering conformations induced by inhibitor. The compounds mapped to these models were selected and subjected to drug-like screening. The identified hits were docked into the active site of renin. Finally, hit1 satisfying the binding mode and interaction energy was selected as possible lead candidate to be used in novel renin inhibitors.
Abstract: Matrix metalloproteinases (MMP) are a class of
structural and functional related enzymes involved in altering the
natural elements of the extracellular matrix. Most of the MMP
structures are cristalographycally determined and published in
WorldWide ProteinDataBank, isolated, in full structure or bound to
natural or synthetic inhibitors. This study proposes an algorithm to
replace missing crystallographic structures in PDB database. We
have compared the results of a chosen docking algorithm with a
known crystallographic structure in order to validate enzyme sites
reconstruction there where crystallographic data are missing.
Abstract: Visfatin and apelin are two new adipokines that recently gained a special interest in diabetes research. This study was conducted to study the interplay between these two adipokines and their correlation with other inflammatory and biochemical parameters in type 2 diabetic (T2D) postmenopausal women with CAD. Visfatin and apelin were measured by enzyme-linked immunoassay (ELISA). Visfatin was found to be significantly higher in the following groups: T2D patients without CAD, non-obese and obese T2D patients with CAD when compared to control group. Apelin was found to be significantly lower in non-obese and obese T2D patients with CAD when compared to control group. Visfatin and apelin were found to be significantly associated with each other and with other biochemical parameters. The current study provides evidence for the interplay between visfatin and apelin through the inflammatory milieu characteristic of T2D and their possible role in the pathogenesis of CAD complication of T2D.
Abstract: The kinetic properties of enzymes are often reported
using the apparent KM and Vmax appropriate to the standard
Michaelis-Menten enzyme. However, this model is inappropriate to
enzymes that have more than one substrate or where the rate
expression does not apply for other reasons. Consequently, it is
desirable to have a means of estimating the appropriate kinetic
parameters from the apparent values of KM and Vmax reported for each
substrate. We provide a means of estimating the range within which
the parameters should lie and apply the method to data for glutamate
dehydrogenase from the nematode parasite of sheep Teladorsagia
circumcincta.
Abstract: An experiment was conducted to study the effects of different types of probiotic on Sucrase enzyme activity of the small intestine mucosa in male broilers. The experimental design was arranged as randomized completely blocks in 4 × 2 factorial arrangement of treatment. 180 male broilers of Ross 308 commercial hybrid were designated into 4 groups. Three replicates of 15 birds were assigned to each treatment. Control treatments (diet contained no probiotic) were fed according to the NRC as base diet and three treatment groups were fed from the same diet plus three different types of probiotics. Birds were slaughtered after 21 and 42 days and different segments of small intestine (at 1,10,30,50,70 and 90% of total length the small intestine) were taken from each replicates (N=2) Sucrase enzyme activities were measured and recorded. Obtained data were analyzed by Spss (P