Abstract: Microfibrous palygorskite and tubular halloysite clay mineral combined with nanocrystalline TiO2 are incorporating in the preparation of nanocomposite films on glass substrates via sol-gel route at 450oC. The synthesis is employing nonionic surfactant molecule as pore directing agent along with acetic acid-based sol-gel route without addition of water molecules. Drying and thermal treatment of composite films ensure elimination of organic material lead to the formation of TiO2 nanoparticles homogeneously distributed on the palygorskite or halloysite surfaces. Nanocomposite films without cracks of active anatase crystal phase on palygorskite and halloysite surfaces are characterized by microscopy techniques, UV-Vis spectroscopy, and porosimetry methods in order to examine their structural properties.
The composite palygorskite-TiO2 and halloysite-TiO2 films with variable quantities of palygorskite and halloysite were tested as photocatalysts in the photo-oxidation of Basic Blue 41 azo dye in water. These nanocomposite films proved to be most promising photocatalysts and highly effective to dye’s decoloration in spite of small amount of palygorskite-TiO2 or halloysite-TiO2 catalyst immobilized onto glass substrates mainly due to the high surface area and uniform distribution of TiO2 on clay minerals avoiding aggregation.
Abstract: The efficiency of heavy metals removal from sewage
sludge in bioleaching processes with heterotrophic, chemoautotrophic
(sulphur-oxidizing) sludge cenoses and chemical leaching (in
distilled water, weakly acidic or alkaline medium) was compared.
The efficacy of heavy metals removal from sewage sludge varies
from 83 % (Zn) up to 14 % (Cr) and follows the order: Zn > Mn > Cu
> Ni > Co > Pb > Cr. The advantages of metals bioleaching process
at heterotrophic metabolism were shown. A new process for
bioconversation of sewage sludge into fertilizer at middle
temperatures after partial heavy metals removal was developed. This
process is based on enhancing vital ability of heterotrophic
microorganisms by adding easily metabolized nutrients and synthesis
of metabolites by growing sludge cenoses. These metabolites possess
the properties of heavy metals extractants and flocculants which
provide the enhancement of sludge flocks sedimentation. The process
results in biomineral fertilizer of prolonged action with immobilized
sludge bioelements. The fertilizer satisfies the EU limits for the
sewage sludge of agricultural utilization. High efficiency of the
biomineral fertilizer obtained has been demonstrated in vegetation
experiments.
Abstract: The removal of chromium by living yeast biomass immobilized onto pozzolana was studied. The results obtained in batch experiments indicate that the immobilized yeast on to pozzolana is a excellent biosorbent of Cr(V) with a good removal rates of 85–90%. The initial concentration solution and agitation speed affected Cr(V) removal. The batch studies data were described using the Freundlich and Langmuir models, but the best fit was obtained with Langmuir model. The breakthrough curve from the continuous flow studies shows that immobilized yeast in the fixed-bed column is capable of decreasing Cr(VI) concentration from 15mg/l to a adequate level.
Abstract: An optical chemical sensing film based on
immobilizing of 1,1′- diethyl 2,2′-cyanine (pseudocyanine iodide) in
nafion film was developed for the determination of Fe(III). The
sensing film was homogeneous, transparent, and mechanically stable.
Decrease of the absorbance measured at 518 nm was observed when
the sensing film was immersed in a solution of Fe(III). The optimum
response of the sensing film to Fe(III) was obtained in a solution with
pH 4.0. Linear calibration curve over an Fe(III) concentration range
of 1-30 ppm with a limit of detection of 0.71 ppm was obtained.
Cd(II) is the major interference. The sensing film exhibited good
stability for 2 months and high reproducibility. The proposed method
was applied for the determination of Fe(III) in water samples with
satisfactory results.
Abstract: Commercially available lipases (Candida antarctica lipase B, Novozyme 435, Thermomyces lanuginosus lipase, and Lipozyme TL IM), as well as sol-gel immobilized lipases, have been screened for their ability to acylate regioselectively xylitol, sorbitol, and mannitol with a phenolic ester in a binary mixture of t-butanol and dimethylsulfoxide. HPLC and MALDI-TOF MS analysis revealed the exclusive formation of monoesters for all studied sugar alcohols. The lipases immobilized by the sol-gel entrapment method proved to be efficient catalysts, leading to high conversions (up to 60%) in the investigated acylation reactions. From a sequence of silane precursors with different nonhydrolyzable groups in their structure, the presence of octyl and i-butyl group was most beneficial for the catalytic activity of sol-gel entrapped lipases in the studied process.
Abstract: Langmuir–Blodgett (LB) films of polyaniline (PANI) grown onto ITO coated glass substrates were utilized for the fabrication of Uric acid biosensor for efficient detection of uric acid by immobilizing Uricase via EDC–NHS coupling. The modified electrodes were characterized by atomic force microscopy (AFM). The response characteristics after immobilization of uricase were studied using cyclic voltammetry and electrochemical impedance spectroscopy techniques. The uricase/PANI/ITO/glass bioelectrode studied by CV and EIS techniques revealed detection of uric acid in a wide range of 0.05 mM to 1.0 mM, covering the physiological range in blood. A low Michaelis–Menten constant (Km) of 0.21 mM indicates the higher affinity of immobilized Uricase towards its analyte (uric acid). The fabricated uric acid biosensor based on PANI LB films exhibits excellent sensitivity of 0.21 mA/mM with a response time of 4 s, good reproducibility, long shelf life (8 weeks) and high selectivity.
Abstract: A biosensor based on glucose oxidase (GOx) immobilized onto nanoparticles zirconium oxide with polyethylene nanocomposite for glucose monitoring has been designed. The CTAB/PEG/ZrO2/GOx nanocomposite was deposited onto screen printed carbon paste (SPCE) electrode via spin coating technique. The properties of CTAB/PEG/ZrO2/GOx were study using scanning electron microscopy (SEM). The SPE modified with the CTAB/PEG/ZrO2/GOx showed electrocatalytical response to the oxidation of glucose when ferrocene carboxaldehyde was used as an artificial redox mediator, which was studied by cyclic voltammetry (CV). Several parameters such as working potential, effect of pH and effect of ZrO2/PEG layers that governed the analytical performance of the biosensor, have been studied. The biosensor was applied to detect glucose with a linear range of 0.4 to 2.0 mmol L−1 with good repetability and reproducibility.
Abstract: This work attempts to improve the permselectivity of poly-ortho-phenylenediamine (PPD) coating for glutamate biosensor applications on Pt microelectrode, using constant potential amperometry and cyclic voltammetry. Percentage permeability of the modified PPD microelectrode was carried out towards hydrogen peroxide (H2O2) and ascorbic acid (AA) whereas permselectivity represents the percentage interference by AA in H2O2 detection. The 50-μm diameter Pt disk microelectrode showed a good permeability value toward H2O2 (95%) and selectivity against AA (0.01%) compared to other sizes of electrode studied here. The electrode was further modified with glutamate oxidase (GluOx) that was immobilized and cross linked with glutaraldehyde (GA, 0.125%), resulting in Pt/PPD/GluOx-GA electrode design. The maximum current density Jmax and apparent Michaelis constant, KM, obtained on Pt/PPD/GluOx-GA electrodes were 48 μA cm-2 and 50 μM, respectively. The linear region slope (LRS) was 0.96 μA cm-2 mM-1. The detection limit (LOD) for glutamate was 3.0 ± 0.6 μM. This study shows a promising glutamate microbiosensor for brain glutamate detection.
Abstract: Separation of propylene-propane mixture using
immobilized liquid membrane was investigated. The effect of transmembrane
pressure and carrier concentration on membrane
separation performance was studied. It was observed that for 30:70
(vol. %) propylene-propane mixture, at pressure of 120kPa and
carrier concentration of 20wt. %, a separation factor of 474 was
obtained.
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: 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: 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: 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: 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: 16-Mercaptohexadecanoic acid (MHDA) and tripeptide glutathione conjugated with gold nanoparticles (Au-NPs) are characterized by Fourier Transform InfaRared (FTIR) spectroscopy combined with Surface-enhanced Raman scattering (SERS) spectroscopy. Surface Plasmon Resonance (SPR) technique based on FTIR spectroscopy has become an important tool in biophysics, which is perspective for the study of organic compounds. FTIR-spectra of MHDA shows the line at 2500 cm-1 attributed to thiol group which is modified by presence of Au-NPs, suggesting the formation of bond between thiol group and gold. We also can observe the peaks originate from characteristic chemical group. A Raman spectrum of the same sample is also promising. Our preliminary experiments confirm that SERS-effect takes place for MHDA connected with Au-NPs and enable us to detected small number (less than 106 cm-2) of MHDA molecules. Combination of spectroscopy methods: FTIR and SERS – enable to study optical properties of Au- NPs and immobilized bio-molecules in context of a bio-nano-sensors.
Abstract: In this paper, naturally immobilized lipase, Carica
papaya lipase, catalyzed biodiesel production from fish oil was
studied. The refined fish oil, extracted from the discarded parts of
fish, was used as a starting material for biodiesel production. The
effects of molar ratio of oil: methanol, lipase dosage, initial water
activity of lipase, temperature and solvent were investigated. It was
found that Carica papaya lipase was suitable for methanolysis of fish
oil to produce methyl ester. The maximum yield of methyl ester
could reach up to 83% with the optimal reaction conditions: oil:
methanol molar ratio of 1: 4, 20% (based on oil) of lipase, initial
water activity of lipase at 0.23 and 20% (based on oil) of tert-butanol
at 40oC after 18 h of reaction time. There was negligible loss in
lipase activity even after repeated use for 30 cycles.
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: The removal of hydrogen sulphide is required for reasons of health, odour problems, safety and corrosivity problems. The means of removing hydrogen sulphide mainly depend on its concentration and kind of medium to be purified. The paper deals with a method of hydrogen sulphide removal from the air by its catalytic oxidation to elemental sulphur with the use of Fe-EDTA complex. The possibility of obtaining fibrous filtering materials able to remove small concentrations of H2S from the air were described. The base of these materials is fibrous ion exchanger with Fe(III)- EDTA complex immobilized on their functional groups. The complex of trivalent iron converts hydrogen sulphide to elemental sulphur. Bivalent iron formed in the reaction is oxidized by the atmospheric oxygen, so complex of trivalent iron is continuously regenerated and the overall process can be accounted as pseudocatalytic. In the present paper properties of several fibrous catalysts based on ion exchangers with different chemical nature (weak acid,weak base and strong base) were described. It was shown that the main parameters affecting the process of catalytic oxidation are:concentration of hydrogen sulphide in the air, relative humidity of the purified air, the process time and the content of Fe-EDTA complex in the fibres. The data presented show that the filtering layers with anion exchange package are much more active in the catalytic processes of hydrogen sulphide removal than cation exchanger and inert materials. In the addition to the nature of the fibres relative air humidity is a critical factor determining efficiency of the material in the air purification from H2S. It was proved that the most promising carrier of the Fe-EDTA catalyst for hydrogen sulphide oxidation are Fiban A-6 and Fiban AK-22 fibres.
Abstract: Nowadays, butyl acetate, a pineapple flavor has been applied widely in food, beverage, cosmetic and pharmaceutical industries. In this study, Butyl acetate, a flavor ester was successfully synthesized via green synthesis of enzymatic reaction route. Commercial immobilized lipase from Rhizomucor miehei (Lipozyme RMIM) was used as biocatalyst in the esterification reaction between acetic acid and butanol. Various reaction parameters such as reaction time (RT), temperature (T) and amount of enzyme (E) were chosen to optimize the reaction synthesis in solvent-free system. The optimum condition to produce butyl acetate was at reaction time (RT), 18 hours; temperature (T), 37°C and amount of enzyme, 25 % (w/w of total substrate). Analysis of yield showed that at optimum condition, >78 % of butyl acetate was produced. The product was confirmed as butyl acetate from FTIR analysis whereby the presence of an ester group was observed at wavenumber of 1742 cm-1.