Abstract: The overall objective of this research is a strain
improvement technology for efficient pectinase production. A novel
cells cultivation technology by immobilization of fungal cells has
been studied in long time continuous fermentations. Immobilization
was achieved by using of new material for absorption of stores of
immobilized cultures which was for the first time used for
immobilization of microorganisms. Effects of various conditions of
nitrogen and carbon nutrition on the biosynthesis of pectolytic
enzymes in Aspergillus awamori 1-8 strain were studied. Proposed
cultivation technology along with optimization of media components
for pectinase overproduction led to increased pectinase productivity
in Aspergillus awamori 1-8 from 7 to 8 times. Proposed technology
can be applied successfully for production of major industrial
enzymes such as α-amylase, protease, collagenase etc.
Abstract: A new strategy for oriented immobilization of proteins was proposed. The strategy contains two steps. The first step is to search for a docking site away from the active site on the protein surface. The second step is trying to find a ligand that is able to grasp the targeted site of the protein. To avoid ligand binding to the active site of protein, the targeted docking site is selected to own opposite charges to those near the active site. To enhance the ligand-protein binding, both hydrophobic and electrostatic interactions need to be included. The targeted docking site should therefore contain hydrophobic amino acids. The ligand is then selected through the help of molecular docking simulations. The enzyme α-amylase derived from Aspergillus oryzae (TAKA) was taken as an example for oriented immobilization. The active site of TAKA is surrounded by negatively charged amino acids. All the possible hydrophobic sites on the surface of TAKA were evaluated by the free energy estimation through benzene docking. A hydrophobic site on the opposite side of TAKA-s active site was found to be positive in net charges. A possible ligand, 3,3-,4,4- – Biphenyltetra- carboxylic acid (BPTA), was found to catch TAKA by the designated docking site. Then, the BPTA molecules were grafted onto silica gels and measured the affinity of TAKA adsorption and the specific activity of thereby immobilized enzymes. It was found that TAKA had a dissociation constant as low as 7.0×10-6 M toward the ligand BPTA on silica gel. The increase in ionic strength has little effect on the adsorption of TAKA, which indicated the existence of hydrophobic interaction between ligands and proteins. The specific activity of the immobilized TAKA was compared with the randomly adsorbed TAKA on primary amine containing silica gel. It was found that the orderly immobilized TAKA owns a specific activity twice as high as the one randomly adsorbed by ionic interaction.
Abstract: Nuclear matrix protein 22 (NMP22) is a FDA approved
biomarker for bladder cancer. The objective of this study is to develop
a simple NMP22 immumosensor (NMP22-IMS) for accurate
measurement of NMP22. The NMP22-IMS was constructed with
NMP22 antibody immobilized on screen-printed carbon electrodes.
The construction procedures and antibody immobilization are simple.
Results showed that the NMP22-IMS has an excellent (r2³0.95)
response range (20 – 100 ng/mL). In conclusion, a simple and reliable
NMP22-IMS was developed, capable of precisely determining urine
NMP22 level.
Abstract: Acute kidney injury (AKI) is a new worldwide public
health problem. A diagnosis of this disease using creatinine is still a
problem in clinical practice. Therefore, a measurement of biomarkers
responsible for AKI has received much attention in the past couple
years. Cytokine interleukin-18 (IL-18) was reported as one of the
early biomarkers for AKI. The most commonly used method to
detect this biomarker is an immunoassay. This study used a planar
platform to perform an immunoassay using fluorescence for
detection. In this study, anti-IL-18 antibody was immobilized onto a
microscope slide using a covalent binding method. Make-up samples
were diluted at the concentration between 10 to 1000 pg/ml to create
a calibration curve. The precision of the system was determined
using a coefficient of variability (CV), which was found to be less
than 10%. The performance of this immunoassay system was
compared with the measurement from ELISA.
Abstract: The biomarker for colorectal cancer (CRC) is CEACAM-6 antigen (C6AG). Therefore, this study aims to develop a novel, simple and low-cost CEACAM-6 antigen immumosensor (C6AG-IMS), based on electrical impedance measurement, for precise determination of C6AG. A low-cost screen-printed graphite electrode was constructed and used as the sensor, with CEACAM-6 antibody (C6AB) immobilized on it. The procedures of sensor fabrication and antibody immobilization are simple and low-cost. Measurement of the electrical impedance at a definite frequency ranges (0.43 – 1.26 MHz) showed that the C6AG-IMS has an excellent linear (r2>0.9) response range (8.125 – 65 pg/mL), covering the normal physiological and pathological ranges of blood C6AG levels. Also, the C6AG-IMS has excellent reliability and validity, with the intraclass correlation coefficient being 0.97. In conclusion, a novel, simple, low-cost and reliable C6AG-IMS was designed and developed, being able to accurately determine blood C6AG levels in the range of pathological and normal physiological regions. The C6AG-IMS can provide a point-of-care and immediate screening results to the user at home.
Abstract: We present a dextran modified silicon microring
resonator sensor for high density antibody immobilization. An array
of sensors consisting of three sensor rings and a reference ring was
fabricated and its surface sensitivity and the limit of detection were
obtained using polyelectrolyte multilayers. The mass sensitivity and
the limit of detection of the fabricated sensor ring are 0.35 nm/ng
mm-2 and 42.8 pg/mm2 in air, respectively. Dextran modified sensor
surface was successfully prepared by covalent grafting of oxidized
dextran on 3-aminopropyltriethoxysilane (APTES) modified silicon
sensor surface. The antibody immobilization on hydrogel dextran
matrix improves 40% compared to traditional antibody
immobilization method via APTES and glutaraldehyde linkage.
Abstract: Biochemical Oxygen Demand (BOD) is a measure of
the oxygen used in bacteria mediated oxidation of organic substances
in water and wastewater. Theoretically an infinite time is required for
complete biochemical oxidation of organic matter, but the
measurement is made over 5-days at 20 0C or 3-days at 27 0C test
period with or without dilution. Researchers have worked to further
reduce the time of measurement.
The objective of this paper is to review advancement made in
BOD measurement primarily to minimize the time and negate the
measurement difficulties. Survey of literature review in four such
techniques namely BOD-BARTTM, Biosensors, Ferricyanidemediated
approach, luminous bacterial immobilized chip method.
Basic principle, method of determination, data validation and their
advantage and disadvantages have been incorporated of each of the
methods.
In the BOD-BARTTM method the time lag is calculated for the
system to change from oxidative to reductive state. BIOSENSORS
are the biological sensing element with a transducer which produces
a signal proportional to the analyte concentration. Microbial species
has its metabolic deficiencies. Co-immobilization of bacteria using
sol-gel biosensor increases the range of substrate. In ferricyanidemediated
approach, ferricyanide has been used as e-acceptor instead
of oxygen. In Luminous bacterial cells-immobilized chip method,
bacterial bioluminescence which is caused by lux genes was
observed. Physiological responses is measured and correlated to
BOD due to reduction or emission.
There is a scope to further probe into the rapid estimation of BOD.
Abstract: Prolonged immobilization leads to significant
weakness and atrophy of the skeletal muscle and can also impair the
recovery of muscle strength following injury. Therefore, it is
important to minimize the period under immobilization and accelerate
the return to normal activity. This study examined the effects of heat
treatment and rest-inserted exercise on the muscle activity of the lower
limb during knee flexion/extension. Twelve healthy subjects were
assigned to 4 groups that included: (1) heat treatment + rest-inserted
exercise; (2) heat + continuous exercise; (3) no heat + rest-inserted
exercise; and (4) no heat + continuous exercise. Heat treatment was
applied for 15 mins prior to exercise. Continuous exercise groups
performed knee flexion/extension at 0.5 Hz for 300 cycles without rest
whereas rest-inserted exercise groups performed the same exercise but
with 2 mins rest inserted every 60 cycles of continuous exercise.
Changes in the rectus femoris and hamstring muscle activities were
assessed at 0, 1, and 2 weeks of treatment by measuring the
electromyography signals of isokinetic maximum voluntary
contraction. Significant increases in both the rectus femoris and
hamstring muscles were observed after 2 weeks of treatment only
when both heat treatment and rest-inserted exercise were performed.
These results suggest that combination of various treatment techniques,
such as heat treatment and rest-inserted exercise, may expedite the
recovery of muscle strength following immobilization.
Abstract: In order to evaluate the relationship between the sulphur (S), glucose (G), nitrogen (N) and plant residues (st), sulphur immobilization and microbial transformation were monitored in five soil samples from 0-30 cm of Bastam farmers fields of Shahrood area following 11 treatments with different levels of Sulphur (S), glucose (G), N and plant residues (wheat straw) in a randomized block design with three replications and incubated over 20, 45 and 60 days, the immobilization of SO4 -2-S presented as a percentage of that added, was inversely related to its addition rate. Additions of glucose and plant residues increased with the C-to-S ratio of the added amendments, irrespective of their origins (glucose and plant residues). In the presence of C sources (glucose or plant residues). N significantly increased the immobilization of SO4 -2-S, whilst the effect of N was insignificant in the absence of a C amendment. In first few days the amounts of added SO4 -2-S immobilized were linearly correlated with the amounts of added S recovered in the soil microbial biomass. With further incubation the proportions of immobilized SO4 -2-S remaining as biomass-S decreased. Decrease in biomass-S was thought to be due to the conversion of biomass-S into soil organic-S. Glucose addition increased the immobilization (microbial utilization and incorporation into the soil organic matter) of native soil SO4 -2-S. However, N addition enhance the mineralization of soil organic-S, increasing the concentration of SO4 - 2-S in soil.
Abstract: Batch fermentation of 5, 10 and 25 g/L biodiesel
derived crude glycerol was carried out at 30, 37 and 450C by
Clostridium pasteurianum cells immobilized on silica. Maximum
yield of 1,3-propanediol (PDO) (0.60 mol/mol), and ethanol (0.26
mol/mol) were obtained from 10 g/L crude glycerol at 30 and 370C
respectively. Maximum yield of butanol (0.28 mol/mol substrate
added) was obtained at 370C with 25 g/L substrate. None of the three
products were detected at 45oC even after 10 days of fermentation.
Only traces of ethanol (0.01 mol/mol) were detected at 450C with 5
g/L substrate. The results obtained for 25 g/L substrate utilization
were fitted in first order rate equation to obtain the values of rate
constant at three different temperatures for bioconversion of glycerol.
First order rate constants for bioconversion of glycerol at 30, 37 and
45oC were found to be 0.198, 0.294 and 0.029/day respectively.
Activation energy (Ea) for crude glycerol bioconversion was
calculated to be 57.62 kcal/mol.
Abstract: Considering toxicity of heavy metals and their
accumulation in domestic wastes, immobilization of lead and
cadmium is envisaged inside glass-ceramics. We particularly
focused this work on calcium-rich phases embedded in a
glassy matrix.
Glass-ceramics were synthesized from glasses doped with
12 wt% and 16 wt% of PbO or CdO. They were observed and
analyzed by Electron MicroProbe Analysis (EMPA) and
Analytical Scanning Electron Microscopy (ASEM). Structural
characterization of the samples was performed by powder XRay
Diffraction.
Diopside crystals of CaMgSi2O6 composition are shown to
incorporate significant amounts of cadmium (up to 9 wt% of
CdO). Two new crystalline phases are observed with very
high Cd or Pb contents: about 40 wt% CdO for the cadmiumrich
phase and near 60 wt% PbO for the lead-rich phase. We
present complete chemical and structural characterization of
these phases. They represent a promising way for the
immobilization of toxic elements like Cd or Pb since glass
ceramics are known to propose a “double barrier" protection
(metal-rich crystals embedded in a glass matrix) against metal
release in the environment.
Abstract: The leaching behavior and structure of Li2O-CeO2- Fe2O3-P2O5 glasses incorporated with simulated high level nuclear wastes (HLW) were studied. The leach rates of gross and each constituent element were determined from the total weight loss of the specimen and the leachate analyses by inductively coupled argon plasma spectroscopy (ICP). The gross leach rate of the 4.5Li2O- 9.7CeO2-34.7Fe2O3-51.5P2O5 glass waste form containing 45 mass% simulated HLW is of the order of 10
Abstract: Lipases are enzymes particularly amenable for
immobilization by entrapment methods, as they can work equally
well in aqueous or non-conventional media and long-time stability of
enzyme activity and enantioselectivity is needed to elaborate more
efficient bioprocesses. The improvement of Pseudomonas
fluorescens (Amano AK) lipase characteristics was investigated by
optimizing the immobilization procedure in hybrid organic-inorganic
matrices using ionic liquids as additives. Ionic liquids containing a
more hydrophobic alkyl group in the cationic moiety are beneficial
for the activity of immobilized lipase. Silanes with alkyl- or aryl
nonhydrolizable groups used as precursors in combination with
tetramethoxysilane could generate composites with higher
enantioselectivity compared to the native enzyme in acylation
reactions of secondary alcohols. The optimal effect on both activity
and enantioselectivity was achieved for the composite made from
octyltrimethoxysilane and tetramethoxysilane at 1:1 molar ratio (60%
increase of total activity following immobilization and enantiomeric
ratio of 30). Ionic liquids also demonstrated valuable properties as
reaction media for the studied reactions, comparable with the usual
organic solvent, hexane.