Abstract: Enzymatic saccharification of biomass for reducing
sugar production is one of the crucial processes in biofuel production
through biochemical conversion. In this study, enzymatic
saccharification of dilute potassium hydroxide (KOH) pre-treated
Tetraselmis suecica biomass was carried out by using cellulase
enzyme obtained from Trichoderma longibrachiatum. Initially, the
pre-treatment conditions were optimised by changing alkali reagent
concentration, retention time for reaction, and temperature. The T.
suecica biomass after pre-treatment was also characterized using
Fourier Transform Infrared Spectra and Scanning Electron
Microscope. These analyses revealed that the functional group such
as acetyl and hydroxyl groups, structure and surface of T. suecica
biomass were changed through pre-treatment, which is favourable for
enzymatic saccharification process. Comparison of enzymatic
saccharification of untreated and pre-treated microalgal biomass
indicated that higher level of reducing sugar can be obtained from
pre-treated T. suecica. Enzymatic saccharification of pre-treated T.
suecica biomass was optimised by changing temperature, pH, and
enzyme concentration to solid ratio ([E]/[S]). Highest conversion of
carbohydrate into reducing sugar of 95% amounted to reducing sugar
yield of 20 (wt%) from pre-treated T. suecica was obtained from
saccharification, at temperature: 40°C, pH: 4.5 and [E]/[S] of 0.1
after 72 h of incubation. Hydrolysate obtained from enzymatic
saccharification of pretreated T. suecica biomass was further
fermented into biobutanol using Clostridium saccharoperbutyliticum
as biocatalyst. The results from this study demonstrate a positive
prospect of application of dilute alkaline pre-treatment to enhance
enzymatic saccharification and biobutanol production from
microalgal biomass.
Abstract: The present study was designed to investigate the
cardio protective role of chronic oral administration of alcoholic
extract of Terminalia arjuna in in-vivo ischemic reperfusion injury
and the induction of HSP72. Rabbits, divided into three groups, and
were administered with the alcoholic extract of the bark powder of
Terminalia arjuna (TAAE) by oral gavage [6.75mg/kg: (T1) and
9.75mg/kg: (T2), 6 days /week for 12 weeks]. In open-chest
Ketamine pentobarbitone anaesthetized rabbits, the left anterior
descending coronary artery was occluded for 15 min of ischemia
followed by 60 min of reperfusion. In the vehicle-treated group,
ischemic-reperfusion injury (IRI) was evidenced by depression of
global hemodynamic function (MAP, HR, LVEDP, peak LV (+) & (-
) (dP/dt) along with depletion of HEP compounds. Oxidative stress
in IRI was evidenced by, raised levels of myocardial TBARS and
depletion of endogenous myocardial antioxidants GSH, SOD and
catalase. Western blot analysis showed a single band corresponding
to 72 kDa in homogenates of hearts from rabbits treated with both the
doses. In the alcoholic extract of the bark powder of Terminalia
arjuna treatment groups, both the doses had better recovery of
myocardial hemodynamic function, with significant reduction in
TBARS, and rise in SOD, GSH, catalase were observed. The results
of the present study suggest that the alcoholic extract of the bark
powder of Terminalia arjuna in rabbit induces myocardial HSP 72
and augments myocardial endogenous antioxidants, without causing
any cellular injury and offered better cardioprotection against
oxidative stress associated with myocardial IR injury.
Abstract: We investigated oxidative DNA damage caused by
radio frequency radiation using 8-oxo-7, 8-dihydro-2'-
deoxyguanosine (8-oxodG) generated in mice tissues after exposure
to 900 MHz mobile phone radio frequency in three independent
experiments. The RF was generated by a Global System for Mobile
Communication (GSM) signal generator. The radio frequency field
was adjusted to 25 V/m. The whole body specific absorption rate
(SAR) was 1.0 W/kg. Animals were exposed to this field for 30 min
daily for 30 days. 24 h post-exposure, blood serum, brain and spleen
were removed and DNA was isolated. Enzyme-linked
immunosorbent assay (ELISA) was used to measure 8-oxodG
concentration. All animals survived the whole experimental period.
The body weight of animals did not change significantly at the end of
the experiment. No statistically significant differences observed in
the levels of oxidative stress. Our results are not in favor of the
hypothesis that 900 MHz RF induces oxidative damage.
Abstract: In this study, a synthetic pathway was created by
assembling genes from Clostridium butyricum and Escherichia coli
in different combinations. Among the genes were dhaB1 and dhaB2
from C. butyricum VPI1718 coding for glycerol dehydratase (GDHt)
and its activator (GDHtAc), respectively, involved in the conversion
of glycerol to 3-hydroxypropionaldehyde (3-HPA). The yqhD gene
from E.coli BL21 was also included which codes for an NADPHdependent
1,3-propanediol oxidoreductase isoenzyme (PDORI)
reducing 3-HPA to 1,3-propanediol (1,3-PD). Molecular modeling
analysis indicated that the conformation of fusion protein of YQHD
and DHAB1 was favorable for direct molecular channeling of the
intermediate 3-HPA. According to the simulation results, the yqhD
and dhaB1 gene were assembled in the upstream of dhaB2 to express
a fusion protein, yielding the recombinant strain E. coliBL21
(DE3)//pET22b+::yqhD-dhaB1_dhaB2 (strain BP41Y3). Strain
BP41Y3 gave 10-fold higher 1,3-PD concentration than E. coliBL21
(DE3)//pET22b+::yqhD-dhaB1_dhaB2 (strain BP31Y2) expressing
the recombinant enzymes simultaneously but in a non-fusion mode.
This is the first report using a gene fusion approach to enhance the
biological conversion of glycerol to the value added compound 1,3-
PD.
Abstract: “Garbage enzyme", a fermentation product of kitchen waste, water and brown sugar, is claimed in the media as a multipurpose solution for household and agricultural uses. This study assesses the effects of dilutions (5% to 75%) of garbage enzyme in reducing pollutants in domestic wastewater. The pH of the garbage enzyme was found to be 3.5, BOD concentration about 150 mg/L. Test results showed that the garbage enzyme raised the wastewater-s BOD in proportion to its dilution due to its high organic content. For mixtures with more than 10% garbage enzyme, its pH remained acidic after the 5-day digestion period. However, it seems that ammonia nitrogen and phosphorus could be removed by the addition of the garbage enzyme. The most economic solution for removal of ammonia nitrogen and phosphorus was found to be 9%. Further tests are required to understand the removal mechanisms of the ammonia nitrogen and phosphorus.
Abstract: Empty Fruit Bunches (EFB) and Palm Oil Mill
Effluent (POME) are two main wastes from oil palm industries which
contain rich lignocellulose. Degradation of EFB and POME by
microorganisms will produce hydrolytic enzyme which will degrade
cellulose and hemicellulose during composting process. However,
normal composting takes about four to six months to reach maturity.
Hence, application of fungi into compost can shorten the period of
composting. This study identifies the effect of xylanase and cellulase
produced by Aspergillus niger and Trichoderma virens on
composting process using EFB and POME. The degradation of EFB
and POME indicates the lignocellulolytic capacity of Aspergillus
niger and Trichoderma virens with more than 7% decrease in
hemicellulose and more than 25% decrease in cellulose for both
inoculated compost. Inoculation of Aspergillus niger and
Trichoderma virens also increased the enzyme activities during the
composting period compared to the control compost by 21% for both
xylanase and cellulase. Rapid rise in the activities of cellulase and
xylanase was observed by Aspergillus niger with the highest
activities of 14.41 FPU/mg and 3.89 IU/mg, respectively. Increased
activities of cellulase and xylanase also occurred in inoculation of
Trichoderma virens with the highest activities obtained at 13.21
FPU/mg and 4.43 IU/mg, respectively. Therefore, it is evident that
the inoculation of fungi can increase the enzyme activities hence
effectively degrading the EFB and POME.
Abstract: This study has investigated the antidiabetic and
antioxidant potential of Pseudovaria macrophylla bark extract on
streptozotocin–nicotinamide induced type 2 diabetic rats. LCMSQTOF
and NMR experiments were done to determine the chemical
composition in the methanolic bark extract. For in vivo experiments,
the STZ (60 mg/kg/b.w, 15 min after 120 mg/kg/1 nicotinamide, i.p.)
induced diabetic rats were treated with methanolic extract of
Pseuduvaria macrophylla (200 and 400 mg/kg·bw) and
glibenclamide (2.5 mg/kg) as positive control respectively.
Biochemical parameters were assayed in the blood samples of all
groups of rats. The pro-inflammatory cytokines, antioxidant status
and plasma transforming growth factor βeta-1 (TGF-β1) were
evaluated. The histological study of the pancreas was examined and
its expression level of insulin was observed by
immunohistochemistry. In addition, the expression of glucose
transporters (GLUT 1, 2 and 4) were assessed in pancreas tissue by
western blot analysis. The outcomes of the study displayed that the
bark methanol extract of Pseuduvaria macrophylla has potentially
normalized the elevated blood glucose levels and improved serum
insulin and C-peptide levels with significant increase in the
antioxidant enzyme, reduced glutathione (GSH) and decrease in the
level of lipid peroxidation (LPO). Additionally, the extract has
markedly decreased the levels of serum pro-inflammatory cytokines
and transforming growth factor beta-1 (TGF-β1). Histopathology
analysis demonstrated that Pseuduvaria macrophylla has the
potential to protect the pancreas of diabetic rats against peroxidation
damage by downregulating oxidative stress and elevated
hyperglycaemia. Furthermore, the expression of insulin protein,
GLUT-1, GLUT-2 and GLUT-4 in pancreatic cells was enhanced.
The findings of this study support the anti-diabetic claims of
Pseudovaria macrophylla bark.
Abstract: The complexity of lignocellulosic biomass requires
a pretreatment step to improve the yield of fermentable sugars. The
efficient pretreatment of corn cobs using microwave and potassium
hydroxide and enzymatic hydrolysis was investigated. The
objective of this work was to characterize the optimal condition of
pretreatment of corn cobs using microwave and potassium
hydroxide enhance enzymatic hydrolysis. Corn cobs were
submerged in different potassium hydroxide concentration at varies
temperature and resident time. The pretreated corn cobs were
hydrolyzed to produce the reducing sugar for analysis. The
morphology and microstructure of samples were investigated by
Thermal gravimetric analysis (TGA, scanning electron microscope
(SEM), X-ray diffraction (XRD). The results showed that lignin
and hemicellulose were removed by microwave/potassium
hydroxide pretreatment. The crystallinity of the pretreated corn
cobs was higher than the untreated. This method was compared
with autoclave and conventional heating method. The results
indicated that microwave-alkali treatment was an efficient way to
improve the enzymatic hydrolysis rate by increasing its
accessibility hydrolysis enzymes.
Abstract: The objective of the present study was to examine the
dose-response relationships between antioxidant parameters and liver
contaminant levels of Kazakhstan light crude oil (KLCO) in albino
rats. The animals were repeatedly exposed, by intraperitoneal
injection, to low dosages (0.5–1.5 ml/kg) of KLCO. Rats exposed to
these doses levels did not show any apparent symptoms of
intoxication. Serum aminotransferases increased significantly
(p
Abstract: Our results showed that treatment with both
cyclooxygenase (COX1 or COX2) inhibitors impair reproduction
parameters of the medaka. Resveratrol (COX1 inhibitor) caused an
decrease in the number of spawning females at the first week of
feeding fish with experimental diets. In the group treated with NS-
398 (COX2 inhibitor) we found the lowest sperm velocity parameters
and decreased linearity of movement. The ovaries of the medaka fed
feed supplemented with Resveratrol or NS-398 were confirmed to
have a lower share of matured oocytes however during the
experiment (four weeks) the number of eggs spawned by females was
similar. Both inhibitors in fish diet (20 mg/kg body weight/day)
caused a decrease in the embryo survival. Our results revealed that
for the medaka female reproduction, activity of both COX enzymes
might be necessary whereas males reproduction competence, as
expressed by sperm motility parameters, might be related to COX2
activity.
Abstract: Oxyleotris marmorata is considered as
undomesticated fish, and its culture occasionally faces a problem of
food deprivation. The present study aims to evaluate alteration of
hematological indices, blood chemical associated with liver function
during 4 weeks of fasting. A non-linear relationships between fasting
days and hematological parameters (red blood cell number; y = -
0.002x2 + 0.041x + 1.249; R2=0.915, P0.05), mean corpuscular
volume; y = -0.180x2 + 2.183x + 149.61; R2=0.732, P>0.05, mean
corpuscular hemoglobin; y = -0.041x2 + 0.862x + 29.864; R2=0.818,
P>0.05 and mean corpuscular hemoglobin concentration; y = -
0.044x2 + 0.711x + 21.580; R2=0.730, P>0.05) were demonstrated.
Significant change in hematocrit (Ht) during fasting period was
observed. Ht elevated sharply increase at the first weeks of fasting
period. Higher Ht also was detected during week 2-4 of fasting time.
The significant reduction of hepatosomatic index was observed (y = -
0.007x2 - 0.096x + 1.414; R2=0.968, P0.05, serum glutamic oxaloacetic transaminase;
y = 0.005x2 – 0.201x2 + 1.297x + 33.256; R2=1, P0.05). Taken together, prolonged fasting has
deleterious effects on hematological indices, liver mass and enzyme
associated in liver function. The marked adverse effects occurred
after the first week of fasting state.
Abstract: 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.
Abstract: This paper aims at overviewing the topics of a research project (CARDIOSENSOR) on the field of health sciences (biomaterials and biomedical engineering). The project has focused on the development of a nanosensor for the assessment of the risk of cardiovascular diseases by the monitoring of C-reactive protein (CRP), which has been currently considered as the best validated inflammatory biomarker associated to cardiovascular diseases. The project involves tasks such as: 1) the development of sensor devices based on field effect transistors (FET): assembly, optimization and validation; 2) application of sensors to the detection of CRP in standard solutions and comparison with enzyme-linked immunosorbent assay (ELISA); and 3) application of sensors to real samples such as blood and saliva and evaluation of their ability to predict the risk of cardiovascular disease.
Abstract: Characterization and evaluation of the activity of Vespa basalis DPP-IV, which expressed in Spodoptera frugiperda 21 cells. The expression of rDPP-IV was confirmed by SDS–PAGE, Western blot analyses, LC-MS/MS and measurement of its peptidase specificity. One-step purification by Ni-NTA affinity chromatography and the total amount of rDPP-IV recovered was approximately 6.4mg per liter from infected culture medium; an equivalent amount would be produced by 1x109 infected Sf21 insect cells. Through the affinity purification led to highly stable rDPP-IV enzyme was recovered and with significant peptidase activity. The rDPP-IV exhibited classical Michaelis–Menten kinetics, with kcat/Km in the range of 10-500 mM-1×S-1 for the five synthetic substrates and optimum substrate is Ala-Pro-pNA. As expected in inhibition assay, the enzymatic activity of rDPP-IV was significantly reduced by 80 or 60% in the presence of sitagliptin (a DPP-IV inhibitor) or PMSF (a serine protease inhibitor), but was not apparently affected by iodoacetamide (a cysteine protease inhibitor).
Abstract: This study was designed to determine effect of
supplemented tomato pomace and fobrolytic enzyme on egg
production and egg quality. A total of 40 CP brown laying hens (95
week old) were used in completely randomized design in 2x2
factorial arrangement with or without enzyme supplementation. Four
dietary treatments included: Control (C), Fibrolytic enzyme (FE),
10% Tomato pomace (TP), and Fibrolytic enzyme + 10 % Tomato
pomace (FE+TP). Each of the four dietary treatments was fed up to
30 days (10 birds/treatment). Live performance, egg production, egg
weight and quality were determined for whole period. Dietary
treatments had no effect (P>0.05) on live performance, egg weight,
yolk color, and egg production. Therefore, laying hens fed diets with
fibrolytic enzyme were significantly (P
Abstract: Stochastic models of biological networks are well established in systems biology, where the computational treatment of such models is often focused on the solution of the so-called chemical master equation via stochastic simulation algorithms. In contrast to this, the development of storage-efficient model representations that are directly suitable for computer implementation has received significantly less attention. Instead, a model is usually described in terms of a stochastic process or a "higher-level paradigm" with graphical representation such as e.g. a stochastic Petri net. A serious problem then arises due to the exponential growth of the model-s state space which is in fact a main reason for the popularity of stochastic simulation since simulation suffers less from the state space explosion than non-simulative numerical solution techniques. In this paper we present transition class models for the representation of biological network models, a compact mathematical formalism that circumvents state space explosion. Transition class models can also serve as an interface between different higher level modeling paradigms, stochastic processes and the implementation coded in a programming language. Besides, the compact model representation provides the opportunity to apply non-simulative solution techniques thereby preserving the possible use of stochastic simulation. Illustrative examples of transition class representations are given for an enzyme-catalyzed substrate conversion and a part of the bacteriophage λ lysis/lysogeny pathway.
Abstract: The traditional method for essential oil extraction from agarwood (Aquilaria Crassna) is to soak it in water and follow with hydrodistillation. The effect of various agarwood pretreatments: ethanol, acid, alkaline, enzymes, and ultrasound, and the effect of subcritical water extraction(SWE) was studied to compare with the traditional method. The major compositions of agarwood oil from hydrodistillation were aroma compounds as follow: aristol-9-en-8- one (21.53%), selina-3, 7(11)-diene (12.96%), τ-himachalene (9.28%), β-guaiene (5.79%), hexadecanoic acid (4.90%) and guaia- 3,9-diene (4.21%). Whereas agarwood oil from pretreatments with ethanol and ultrasound, and SWE got fatty acid compounds. Extraction of agarwood oil using these pretreatments could improve the agarwood oil yields up to 2 times that of the traditional method. The components of the pretreated sample with diluted acid (H2SO4) at pH 4 gave quite similar results as the traditional method. Therefore, the enhancement of essential oil from agarwood depends on requirement of type of extracted oil that involved extraction methods.
Abstract: Wheat gluten hydrolyzates (WGHs) and anchovy fine
powder hydrolyzates (AFPHs) were produced at 300 MPa using
combinations of Flavourzyme 500MG (F), Alcalase 2.4L (A),
Marugoto E (M) and Protamex (P), and then were compared to those
produced at ambient pressure concerning the contents of soluble solid
(SS), soluble nitrogen and electrophoretic profiles. The contents of SS
in the WGHs and AFPHs increased up to 87.2% according to the
increase in enzyme number both at high and ambient pressure. Based
on SS content, the optimum enzyme combinations for one-, two-,
three- and four-enzyme hydrolysis were determined as F, FA, FAM
and FAMP, respectively. Similar trends were found for the contents of
total soluble nitrogen (TSN) and TCA-soluble nitrogen (TCASN). The
contents of SS, TSN and TCASN in the hydrolyzates together with
electrophoretic mobility maps indicates that the high-pressure
treatment of this study accelerated protein hydrolysis compared to
ambient-pressure treatment.
Abstract: As a result of urbanization, the unpredictable growth of industry and transport, production of chemicals, military activities, etc. the concentration of anthropogenic toxicants spread in nature exceeds all the permissible standards. Most dangerous among these contaminants are organic compounds having great persistence, bioaccumulation, and toxicity along with our awareness of their prominent occurrence in the environment and food chain. Among natural ecological tools, plants still occupying above 40% of the world land, until recently, were considered as organisms having only a limited ecological potential, accumulating in plant biomass and partially volatilizing contaminants of different structure. However, analysis of experimental data of the last two decades revealed the essential role of plants in environment remediation due to ability to carry out intracellular degradation processes leading to partial or complete decomposition of carbon skeleton of different structure contaminants. Though, phytoremediation technologies still are in research and development, their various applications have been successfully used. The paper aims to analyze mechanisms of organic contaminants uptake and detoxification in plants, being the less studied issue in evaluation and exploration of plants potential for environment remediation.
Abstract: Influence of octane and benzene on plant cell
ultrastructure and enzymes of basic metabolism, such as nitrogen
assimilation and energy generation have been studied. Different
plants: perennial ryegrass (Lolium perenne) and alfalfa (Medicago
sativa); crops- maize (Zea mays L.) and bean (Phaseolus vulgaris);
shrubs – privet (Ligustrum sempervirens) and trifoliate orange
(Poncirus trifoliate); trees - poplar (Populus deltoides) and white
mulberry (Morus alba L.) were exposed to hydrocarbons of different
concentrations (1, 10 and 100 mM). Destructive changes in bean and
maize leaves cells ultrastructure under the influence of benzene
vapour were revealed at the level of photosynthetic and energy
generation subcellular organells. Different deviations at the level of
subcellular organelles structure and distribution were observed in
alfalfa and ryegrass root cells under the influence of benzene and
octane, absorbed through roots. The level of destructive changes is
concentration dependent. Benzene at low 1 and 10 mM concentration
caused the increase in glutamate dehydrogenase (GDH) activity in
maize roots and leaves and in poplar and mulberry shoots, though to
higher extent in case of lower, 1mM concentration. The induction
was more intensive in plant roots. The highest tested 100mM
concentration of benzene was inhibitory to the enzyme in all plants.
Octane caused induction of GDH in all grassy plants at all tested
concentrations; however the rate of induction decreased parallel to
increase of the hydrocarbon concentration. Octane at concentration 1
mM caused induction of GDH in privet, trifoliate and white mulberry
shoots. The highest, 100mM octane was characterized by inhibitory
effect to GDH activity in all plants. Octane had inductive effect on
malate dehydrogenase in almost all plants and tested concentrations,
indicating the intensification of Trycarboxylic Acid Cycle.
The data could be suggested for elaboration of criteria for plant
selection for phytoremediation of oil hydrocarbons contaminated
soils.