Abstract: Diagnostic enzymes like aspartate aminotransferase
(AST), alanine aminotransferase (ALT) and alkaline phosphatase
(ALP) were determined as indices of heavy metal pollution in Tilapia
guinensis. Three different sets of fishes treated with lead (Pb), iron
(Fe) and copper (Cu) were used for the study while a fourth group
with no heavy metal served as a control. Fishes in each of the groups
were exposed to 2.65mg/l of Pb, 0.85mg/l of Fe and 0.35 mg/l of Cu
in aerated aquaria for 96 hours. Tissue fractionation of the liver
tissues was carried out and the three diagnostic enzymes (AST, ALT,
and ALP) were estimated. Serum levels of the same diagnostic
enzymes were also measured. The mean values of the serum enzyme
activity for ALP in each experimental group were 19.5±1.62,
29.67±2.17 and 1.15±0.27 IU/L for Pb, Fe and Cu groups compared
with 9.99±1.34 IU/L enzyme activity in the control. This result
showed that Pb and Fe caused increased release of the enzyme into
the blood circulation indicating increased tissue damage while Cu
caused a reduction in the serum level as compared with the level in
the control group. The mean values of enzyme activity obtained in
the liver were 102.14±6.12, 140.17±2.06 and 168.23±3.52 IU/L for
Pb, Fe and Cu groups, respectively compared to 91.20±9.42 IU/L
enzyme activity for the control group. The serum and liver AST and
ALT activities obtained in Pb, Fe, Cu and control groups are
reported. It was generally noted that the presence of the heavy metal
caused liver tissues damage and consequent increased level of the
diagnostic enzymes in the serum.
Abstract: Whey is the lactose rich by-product of the dairy
industry, having good amount of nutrient reservoir. Most abundant
nutrients are lactose, soluble proteins, lipids and mineral salts.
Disposing of whey by most of milk plants which do not have proper
pre-treatment system is the major issue. As a result of which, there
can be significant loss of potential food and energy source. Thus,
whey has been explored as the substrate for the synthesis of different
value added products such as enzymes. β-galactosidase is one of the
important enzymes and has become the major focus of research due
to its ability to catalyze both hydrolytic as well as
transgalactosylation reaction simultaneously. The enzyme is widely
used in dairy industry as it catalyzes the transformation of lactose to
glucose and galactose, making it suitable for the lactose intolerant
people. The enzyme is intracellular in both bacteria and yeast,
whereas for molds, it has an extracellular location. The present work
was carried to utilize the whey for the production of β-galactosidase
enzyme using both yeast and fungal cultures. The yeast isolate
Kluyveromyces marxianus WIG2 and various fungal strains have
been used in the present study. Different disruption techniques have
also been investigated for the extraction of the enzyme produced
intracellularly from yeast cells. Among the different methods tested
for the disruption of yeast cells, SDS-chloroform showed the
maximum β-galactosidase activity. In case of the tested fungal
cultures, Aureobasidium pullulans NCIM 1050 was observed to be
the maximum extracellular enzyme producer.
Abstract: Plasmin plays an important role in the human
circulatory system owing to its catalytic ability of fibrinolysis. The
immediate injection of plasmin in patients of strokes has intrigued
many scientists to design vectors that can transport plasmin to the
desired location in human body. Here we predict the structure of
human plasmin and investigate the interaction of plasmin with the
gold-nanoparticle.
Because the crystal structure of plasminogen has been solved, we
deleted N-terminal domain (Pan-apple domain) of plasminogen and
generate a mimic of the active form of this enzyme (plasmin). We
conducted a simulated annealing process on plasmin and discovered a
very large conformation occurs. Kringle domains 1, 4 and 5 had been
observed to leave its original location relative to the main body of the
enzyme and the original doughnut shape of this enzyme has been
transformed to a V-shaped by opening its two arms. This observation
of conformational change is consistent with the experimental results of
neutron scattering and centrifugation.
We subsequently docked the plasmin on the simulated gold surface
to predict their interaction. The V-shaped plasmin could utilize its
Kringle domain and catalytic domain to contact the gold surface.
Our findings not only reveal the flexibility of plasmin structure but
also provide a guide for the design of a plasmin-gold nanoparticle.
Abstract: Lignocellolusic material is a substance that is resistant to be degraded by microorganisms or hydrolysis enzymes. To be used as materials for biofuel production, it needs pretreatment process to improve efficiency of hydrolysis. In this work, chemical pretreatments on rice straw using three diluted organic acids, including acetic acid, citric acid, oxalic acid, were optimized. Using Response Surface Methodology (RSM), the effect of three pretreatment parameters, acid concentration, treatment time, and reaction temperature, on pretreatment efficiency were statistically evaluated. The results indicated that dilute oxalic acid pretreatment led to the highest enhancement of enzymatic saccharification by commercial cellulase and yielded sugar up to 10.67 mg/ml when using 5.04% oxalic acid at 137.11 oC for 30.01 min. Compared to other acid pretreatment by acetic acid, citric acid, and hydrochloric acid, the maximum sugar yields are 7.07, 6.30, and 8.53 mg/ml, respectively. Here, it was demonstrated that organic acids can be used for pretreatment of lignocellulosic materials to enhance of hydrolysis process, which could be integrated to other applications for various biorefinery processes.
Abstract: Rice straw is lignocellulosic biomass which can be utilized as substrate for the biogas production. However, due to the property and composition of rice straw, it is difficult to be degraded by hydrolysis enzymes. One of the pretreatment methods that modify such properties of lignocellulosic biomass is the application of lignocellulose-degrading microbial consortia. The aim of this study is to investigate the effect of microbial consortia to enhance biogas production. To select the high efficient consortium, cellulase enzymes were extracted and their activities were analyzed. The results suggested that microbial consortium culture obtained from cattle manure is the best candidate compared to decomposed wood and horse manure. A microbial consortium isolated from cattle manure was then mixed with anaerobic sludge and used as inoculum for biogas production. The optimal conditions for biogas production were investigated using response surface methodology (RSM). The tested parameters were the ratio of amount of microbial consortium isolated and amount of anaerobic sludge (MI:AS), substrate to inoculum ratio (S:I) and temperature. Here, the value of the regression coefficient R2 = 0.7661 could be explained by the model which is high to advocate the significance of the model. The highest cumulative biogas yield was 104.6 ml/g-rice straw at optimum ratio of MI:AS, ratio of S:I, and temperature of 2.5:1, 15:1 and 44°C respectively.
Abstract: Beta-glucosidase, chitinase, leucine-aminopeptidase, acid phosphomonoesterase and acetate-esterase enzyme activities in
the soils under the impact of metallurgical industrial activity in Lori
marz (district) were investigated. The results of the study showed that
the activities of the investigated enzymes in the soils decreased with increasing distance from the Shamlugh copper mine, the Chochkan
tailings storage facility and the ore transportation road. Statistical
analysis revealed that the activities of the enzymes were positively
correlated (significant) to each other according to the observation
sites which indicated that enzyme activities were affected by the
same anthropogenic factor. The investigations showed that the soils
were polluted with heavy metals (Cu, Pb, As, Co, Ni, Zn) due to
copper mining activity in this territory. The results of Pearson
correlation analysis revealed a significant negative correlation
between heavy metal pollution degree (Nemerow integrated pollution
index) and soil enzyme activity. All of this indicated that copper
mining activity in this territory causing the heavy metal pollution of
the soils resulted in the inhabitation of the activities of the enzymes
which are considered as biological catalysts to decompose organic
materials and facilitate the cycling of nutrients.
Abstract: Fungal mutant strains have produced cellulase and
xylanase enzymes, and have induced high hydrolysis with enhanced
of rice straw. The mutants were obtained by exposing Penicillium
strain to UV-light treatments. Screening and selection after treatment
with UV-light were carried out using cellulolytic and xylanolytic
clear zones method to select the hypercellulolytic and
hyperxylanolytic mutants. These mutants were evaluated for their
cellulase and xylanase enzyme production as well as their abilities for
biodegradation of rice straw. The mutant 12 UV/1 produced 306.21%
and 209.91% cellulase and xylanase, respectively, as compared with
the original wild type strain. This mutant showed high capacity of
rice straw degradation. The effectiveness of tested mutant strain and
that of wild strain was compared in relation to enhancing the
composting process of rice straw and animal manures mixture. The
results obtained showed that the compost product of inoculated
mixture with mutant strain (12 UV/1) was the best compared to the
wild strain and un-inoculated mixture. Analysis of the composted
materials showed that the characteristics of the produced compost
were close to those of the high quality standard compost. The results
obtained in the present work suggest that the combination between
rice straw and animal manure could be used for enhancing the
composting process of rice straw and particularly when applied with
fungal decomposer accelerating the composting process.
Abstract: Lipases constitute one of the most important groups of
industrial enzymes that catalyze the hydrolysis of triacylglycerol to
glycerol and fatty acids. Muscarinic antagonist relieves smooth
muscle spasm of the gastrointestinal tract and effect on the
cardiovascular system. In this research the effect of a muscarinic
antagonist on the lipase activity of Pseudomonas aeruginosa was
studied. Lineweaver–Burk plot showed that the drug inhibited the
enzyme by competitive inhibition. The IC50 value (0.16 mM) and Ki
(0.03 mM) of the drug revealed the drug bound to enzyme with high
affinity. Determination of enzyme activity in various pH and
temperature showed that the maximum activity of lipase was at pH 8
and 60oC both in presence and absence of the drug.
Abstract: Diminished antioxidant defense or increased
production of reactive oxygen species in the biological system can
result in oxidative stress which may lead to various
neurodegenerative diseases including Alzheimer’s disease (AD).
Microglial activation also contributes to the progression of AD by
producing several proinflammatory cytokines, nitric oxide (NO) and
prostaglandin E2 (PGE2). Oxidative stress and inflammation have
been reported to be possible pathophysiological mechanisms
underlying AD. In addition, the cholinergic hypothesis postulates that
memory impairment in patient with AD is also associated with the
deficit of cholinergic function in the brain. Although a number of
drugs have been approved for the treatment of AD, most of these
synthetic drugs have diverse side effects and yield relatively modest
benefits. Marine algae have great potential in pharmaceutical and
biomedical applications as they are valuable sources of bioactive
properties such as anticoagulation, antimicrobial, antioxidative,
anticancer and anti-inflammatory. Hence, this study aimed to provide
an overview of the properties of Malaysian seaweeds (Padina
australis, Sargassum polycystum and Caulerpa racemosa) in
inhibiting oxidative stress, neuroinflammation and cholinesterase
enzymes. These seaweeds significantly exhibited potent DPPH and
moderate superoxide anion radical scavenging ability (P
Abstract: A quartz crystal microbalance (QCM) nanosensor was developed to detect lysozyme enzyme by functionalizing its gold surface with the attachment of poly(methacroyl-L-phenylalanine) (PMAPA) nanoparticles. PMAPA was chosen as a hydrophobic matrix. The hydrophobic nanoparticles were synthesized by micro-emulsion polymerization method. Hydrophobic QCM nanosensor was tested for real time detection of lysozyme enzyme from aqueous solution. The kinetic and affinity studies were determined by using lysozyme solutions with different concentrations. The responses related with mass (Δm) and frequency (Δf) shifts were used to evaluate adsorption properties.
Abstract: In order to detect and quantify the phenolic contents
of a wastewater with biosensors, two working electrodes based on
modified Poly(Pyrrole) films were fabricated. Enzyme horseradish
peroxidase was used as biomolecule of the prepared electrodes.
Various phenolics were tested at the biosensor. Phenol detection was
realized by electrochemical reduction of quinones produced by
enzymatic activity. Analytical parameters were calculated and the
results were compared with each other.
Abstract: Newly synthesized Polypropylene-g-Polyethylene
glycol polymer was first time used for a compartment-less enzymatic
fuel cell. Working electrodes based on Polypropylene-g-Polyethylene
glycol were operated as unmediated and mediated system (with
ferrocene and gold/cobalt oxide nanoparticles). Glucose oxidase and
bilirubin oxidase was selected as anodic and cathodic enzyme,
respectively. Glucose was used as fuel in a single-compartment and
membrane-less cell. Maximum power density was obtained as 0.65
nW cm-2, 65 nW cm-2 and 23500 nW cm-2 from the unmediated,
ferrocene and gold/cobalt oxide modified polymeric film,
respectively. Power density was calculated to be ~16000 nW cm-2 for
undiluted wastewater sample with gold/cobalt oxide nanoparticles
including system.
Abstract: Biofuels production has come forth as a future
technology to combat the problem of depleting fossil fuels. Bio-based
ethanol production from enzymatic lignocellulosic biomass
degradation serves an efficient method and catching the eye of
scientific community. High cost of the enzyme is the major obstacle
in preventing the commercialization of this process. Thus main
objective of the present study was to optimize composition of
medium components for enhancing cellulase production by newly
isolated strain of Bacillus tequilensis. Nineteen factors were taken
into account using statistical Plackett-Burman Design. The significant
variables influencing the cellulose production were further employed
in statistical Response Surface Methodology using Central
Composite Design for maximizing cellulase production. The
optimum medium composition for cellulase production was: peptone
(4.94 g/L), ammonium chloride (4.99 g/L), yeast extract (2.00 g/L),
Tween-20 (0.53 g/L), calcium chloride (0.20 g/L) and cobalt chloride
(0.60 g/L) with pH 7, agitation speed 150 rpm and 72 h incubation at
37oC. Analysis of variance (ANOVA) revealed high coefficient of
determination (R2) of 0.99. Maximum cellulase productivity of 11.5
IU/ml was observed against the model predicted value of 13 IU/ml.
This was found to be optimally active at 60oC and pH 5.5.
Abstract: The study examined the effect of Bonny Light whole
crude oil (WC) and its water soluble fraction (WSF) on the activities
of antioxidant enzymes (catalase (CAT) and superoxide dismutase
(SOD)) and crude mitochondria ATPases in the radicle of
germinating bean (Vigna unguiculata). The percentage germination,
level of lipid peroxidation, antioxidant enzyme and mitochondria
Ca2+ and Mg2+ ATPase activities were measured in the radicle of
bean after 7, 14 and 21 days post germination. Viable bean seeds
were planted in soils contaminated with 10ml, 25ml and 50ml of
whole crude oil (WC) and its water soluble fraction (WSF) to obtain
2, 5 and 10% v/w crude oil contamination. There was dose dependent
reduction of the number of bean seeds that germinated in the
contaminated soils compared with control (p
Abstract: This study is concerned with the optimization of
fermentation parameters for the hyper production of mannanase from
Fusarium oxysporum SS-25 employing two step statistical strategy
and kinetic characterization of crude enzyme preparation. The
Plackett-Burman design used to screen out the important factors in
the culture medium revealed 20% (w/w) wheat bran, 2% (w/w) each
of potato peels, soyabean meal and malt extract, 1% tryptone, 0.14%
NH4SO4, 0.2% KH2PO4, 0.0002% ZnSO4, 0.0005% FeSO4, 0.01%
MnSO4, 0.012% SDS, 0.03% NH4Cl, 0.1% NaNO3 in brewer’s spent
grain based medium with 50% moisture content, inoculated with
2.8×107 spores and incubated at 30oC for 6 days to be the main
parameters influencing the enzyme production. Of these factors, four
variables including soyabean meal, FeSO4, MnSO4 and NaNO3 were
chosen to study the interactive effects and their optimum levels in
central composite design of response surface methodology with the
final mannanase yield of 193 IU/gds. The kinetic characterization
revealed the crude enzyme to be active over broader temperature and
pH range. This could result in 26.6% reduction in kappa number with
4.93% higher tear index and 1% increase in brightness when used to
treat the wheat straw based kraft pulp. The hydrolytic potential of
enzyme was also demonstrated on both locust bean gum and guar
gum.
Abstract: Bacterial strains capable of degradation of malathion
from the domestic sewage were isolated by an enrichment culture
technique. Three bacterial strains were screened and identified as
Acinetobacter baumannii (AFA), Pseudomonas aeruginosa (PS1),
and Pseudomonas mendocina (PS2) based on morphological,
biochemical identification and 16S rRNA sequence analysis.
Acinetobacter baumannii AFA was the most efficient malathion
degrading bacterium, so used for further biodegradation study. AFA
was able to grow in mineral salt medium (MSM) supplemented with
malathion (100 mg/l) as a sole carbon source, and within 14 days,
84% of the initial dose was degraded by the isolate measured by high
performance liquid chromatography. Strain AFA could also degrade
other organophosphorus compounds including diazinon, chlorpyrifos
and fenitrothion. The effect of different culture conditions on the
degradation of malathion like inoculum density, other carbon or
nitrogen sources, temperature and shaking were examined.
Degradation of malathion and bacterial cell growth were accelerated
when culture media were supplemented with yeast extract, glucose
and citrate. The optimum conditions for malathion degradation by
strain AFA were; an inoculum density of 1.5x 10^12CFU/ml at 30°C
with shaking. A specific polymerase chain reaction primers were
designed manually using multiple sequence alignment of the
corresponding carboxylesterase enzymes of Acinetobacter species.
Sequencing result of amplified PCR product and phylogenetic
analysis showed low degree of homology with the other
carboxylesterase enzymes of Acinetobacter strains, so we suggested
that this enzyme is a novel esterase enzyme. Isolated bacterial strains
may have potential role for use in bioremediation of malathion
contaminated.
Abstract: Liver disorders are one of the major problems of the
world. Despite its frequent occurrence, high morbidity and high
mortality, its medical management is currently inadequate. This study
was designed to evaluate the hepatoprotective effect of saponin
extract of the root of Garcinia kola on the integrity of the liver of
paracetamol induced wistar albino rats. Twenty five (25) male adult
wistar albino rats were divided into five (5) groups. Group I was the
Control group that received distilled water only, group II was the
negative control that received 2 g/kg of paracetamol on the 13th day,
and group III, IV and V were pre-treated with 100, 200 and
400mg/kg of the saponin extract before inducing the liver damage on
the 13th day with 2 g/kg of paracetamol. Twenty four (24) h after
administration, the rats were sacrificed and blood samples were
collected. The serum Alanine Transaminase (ALT), Aspartate
Transaminase (AST), Alkaline Phosphatase (ALP) activities,
Bilirubin and conjugated bilirubin, glucose and protein
concentrations were evaluated. The liver was fixed immediately in
Formalin and was processed and stained in Haematoxylin and Eosin
(H&E). Administration of saponin extract from the root of Garcinia
kola significantly decreased paracetamol induced elevated enzymes
in the test group. Also histological observations showed that saponin
extract of the root of Garcinia kola exhibited a significant liver
protection against the toxicant as evident by the cells trying to return
to normal. Saponin extract from the root of Garcinia kola indicated a
protection of structural integrity of the hepatocytic cell membrane
and regeneration of the damaged liver.
Abstract: The biosynthesis of nanoparticles by microorganisms,
on the contrary to chemical synthesis, is an environmentally-friendly
process which has low energy requirements. In this investigation, we
used the microorganism Geobacillus wiegelii, strain GWE1, an
aerobic thermophile belonging to genus Geobacillus, isolated from a
drying oven. This microorganism has the ability to reduce selenite
evidenced by the change of color from colorless to red in the culture.
Elemental analysis and composition of the particles were verified
using transmission electron microscopy and energy-dispersive X-ray
analysis. The nanoparticles have a defined spherical shape and a
selenium elemental state. Previous experiments showed that the
presence of the whole microorganism for the reduction of selenite
was not necessary. The results strongly suggested that an intracellular
NADPH/NADH-dependent reductase mediates selenium
nanoparticles synthesis under aerobic conditions. The enzyme was
purified and identified by mass spectroscopy MALDI-TOF TOF
technique. The enzyme is a 1-pyrroline-5-carboxylate dehydrogenase.
Histograms of nanoparticles sizes were obtained. Size distribution
ranged from 40-160 nm, where 70% of nanoparticles have less than
100 nm in size. Spectroscopic analysis showed that the nanoparticles
are composed of elemental selenium. To analyse the effect of pH in
size and morphology of nanoparticles, the synthesis of them was
carried out at different pHs (4.0, 5.0, 6.0, 7.0, 8.0). For
thermostability studies samples were incubated at different
temperatures (60, 80 and 100 ºC) for 1 h and 3 h. The size of all
nanoparticles was less than 100 nm at pH 4.0; over 50% of
nanoparticles have less than 100 nm at pH 5.0; at pH 6.0 and 8.0 over
90% of nanoparticles have less than 100 nm in size. At neutral pH
(7.0) nanoparticles reach a size around 120 nm and only 20% of them
were less than 100 nm. When looking at temperature effect,
nanoparticles did not show a significant difference in size when they
were incubated between 0 and 3 h at 60 ºC. Meanwhile at 80 °C the
nanoparticles suspension lost its homogeneity. A change in size was
observed from 0 h of incubation at 80ºC, observing a size range
between 40-160 nm, with 20% of them over 100 nm. Meanwhile
after 3 h of incubation at size range changed to 60-180 nm with 50%
of them over 100 nm. At 100 °C the nanoparticles aggregate forming
nanorod structures. In conclusion, these results indicate that is
possible to modulate size and shape of biologically synthesized
nanoparticles by modulating pH and temperature.
Abstract: Food contamination occurs during post process
handling. This leads to spoilage and growth of pathogenic
microorganisms in the food, thereby reducing its shelf life or
spreading of food borne diseases. Several methods are tried and one
of which is use of antimicrobial packaging. Here, papain, a protease
enzyme, is covalently immobilized with the help of glutarldehyde on
polyurethane and used as a food wrap to protect food from microbial
contamination. Covalent immobilization of papain was achieved at a
pH of 7.4; temperature of 4°C; glutaraldehyde concentration of 0.5%;
incubation time of 24h; and 50mg of papain. The formation of -C=Nobserved
in the Fourier transform infrared spectrum confirmed the
immobilization of the enzyme on the polymer. Immobilized enzyme
retained higher activity than the native free enzyme. The modified
polyurethane showed better reduction of Staphylococcus aureus
biofilm than bare polymer film (eight folds reduction in live colonies,
two times reduction in protein and 6 times reduction in
carbohydrates). The efficacy of this was studied by wrapping it over
S. aureus contaminated cottage cheese (paneer) and cheese and
stored at a temperature of 4°C for 7days. The modified film reduced
the bacterial contamination by eight folds when compared to the bare
film. FTIR also indicated reduction in lipids, sugars and proteins in
the biofilm.
Abstract: Heavy metals are one of the major groups of
contaminants in the environment and many of them are toxic even at
very low concentration in plants and animals. However, some metals
play important roles in the biological function of many enzymes in
living organisms. Metals such as zinc, iron, and cooper are important
for survival and activity of enzymes in plants, however heavy metals
can inhibit enzyme which is responsible for defense system of plants.
Polyphenol oxidase (PPO) is a copper-containing metalloenzyme
which is responsible for enzymatic browning reaction of plants.
Enzymatic browning is a major problem for the handling of
vegetables and fruits in food industry. It can be increased and
effected with many different futures such as metals in the nature and
ground. In the present work, PPO was isolated and characterized
from green leaves of red poppy plant (Papaverr hoeas). Then, the
effect of some known antibrowning agents which can form
complexes with metals and metals were investigated on the red poppy
PPO activity. The results showed that glutathione was the most
potent inhibitory effect on PPO activity. Cu(II) and Fe(II) metals
increased the enzyme activities however, Sn(II) had the maximum
inhibitory effect and Zn(II) and Pb(II) had no significant effect on the
enzyme activity. In order to reduce the effect of heavy metals, the
effects of metal-antibrowning agent complexes on the PPO activity
were determined. EDTA and metal complexes had no significant
effect on the enzyme. L-ascorbic acid and metal complexes decreased
but L-ascorbic acid-Cu(II)-complex had no effect. Glutathione–metal
complexes had the best inhibitory effect on Red poppy leaf PPO
activity.