Immobilization of HRP enzyme on polymeric microspheres and its use in decolourisation of organic dyes.

In this study, horseradish peroxidase (HRP) enzyme was immobilized on Pd(II) containing polymeric microspheres by adsorption method and used for the decolourisation of Methyl Orange (MO) and Rhodamine B (RB) dyes. The synthesized microspheres were characterized by Fourier Transform Infrared Spectroscopy (FTIR) and Scanning Electron Microscopy-Energy Dispersive X-ray (SEM/EDX), Thermal Gravimetric Analysis (TGA). The effects of pH, dye concentration, temperature, and H2O2 concentration on the decolourisation of MO and RB were determined. According to the results of various parameters studied, when 2-AEPS-napht-HRP support was used, MO and RB were biodegraded to 69.72% and 80.65%, respectively, within 60 min. When 2-AEPS-napht-Pd-HRP support was used, MO and RB were biodegraded to 58.35% and 90.81%, respectively, under optimum conditions. When the reproducibility results of the immobilized supports were examined, it was observed that they remained efficient during the first five reusability cycles and even reached 65% decolourisation efficiency after the 9th reuse. The immobilized enzyme (2AEPS-npht-HRP and 2AEPS-npht-Pd-HRP) showed remarkable resistance to higher temperatures compared to the free enzyme.

Improvement of catalysis performance of pepsin and lipase enzymes by double enzyme immobilization method.

Ferrocene-coupled chitosan (CS-Fc) support polymer was synthesized for the immobilization of double enzymes and single enzymes. The immobilization study was carried out after a detailed characterization study. The binding of pepsin and lipase enzymes to the polymer was supported by the shift of peaks in the FTIR spectra of free enzymes. The particle sizes of chitosan (CS) and CS-Fc were evaluated using zeta potential measurement. The optimum conditions of the enzymes in the double enzyme system did not affect each other and each showed their activity. In the presence of double enzyme immobilization, the optimum pH remained the same with free enzyme, while the optimum temperature increased differently with an increase of 10 °C. Vmax values were 0.0020 and 0.0038 for free pepsin and lipase enzyme, respectively. However, due to double enzyme immobilization, the Vmax value increased 1.75 fold for pepsin and 3.94 fold for lipase enzyme. In the double enzyme immobilization, the substrate affinity of pepsin increased by 0.4-fold, while the substrate affinity of lipase increased by about 0.3-fold. In the 5th reuse of pepsin and lipase enzymes in the presence of double enzymes (Cs-Fc-Pep) + (Cs-Fc-Lip), it was determined as 18.21% for pepsin enzyme and 90.81% for lipase enzyme.

Qualitative enzymatic detection of organophosphate and carbamate insecticides.

In this study, new polymers containing amides (TrisPS-Ntaa, and TrisPS-Ntaa-Fc) were synthesized by condensation reaction for qualitative identification of insecticides. The synthesized polymers, including amides were investigated by infrared spectroscopy (IR), scanning electron microscopy-energy dispersion X- ray spectrometry (SEM-EDX), and gel permeation chromatography (GPC). Then, acetylcholinesterase enzyme (AChE) was covalently immobilized on these polymers to improve properties (including activity, reusability, and storage stability). Accordingly, organophosphate (malathion, acephate, chlorpyrifos methyl) and carbamate (carbofuran, methiocarb, methomyl), which are used to prevent harmful organisms in some agricultural products were enzymatically determined based on their inhibitory activity on AChE.

Novel polymeric microspheres: Synthesis, enzyme immobilization, antimutagenic activity, and antimicrobial evaluation against pathogenic microorganisms.

New polymeric microspheres containing azomethine (1a-1c and 2a-2c) were synthesized by condensation to compare the enzymatic properties of the enzyme glucose oxidase (GOx) and to investigate antimutagenic and antimicrobial activities. The polymeric microspheres were characterized by elemental analysis, infrared spectra (FT-IR), proton nuclear magnetic resonance spectra, thermal gravimetric analysis, and scanning electron microscopy analysis. The catalytic activity of the glucose oxidase enzyme follows Michaelis-Menten kinetics. Influence of temperature, reusability, and storage capacity of the free and immobilized glucose oxidase enzyme were investigated. It is determined that immobilized enzymes exhibit good storage stability and reusability. After immobilization of GOx in polymeric supports, the thermal stability of the enzyme increased and the maximum reaction rate (Vmax ) decreased. The activity of the immobilized enzymes was preserved even after 5 months. The antibacterial and antifungal activity of the polymeric microspheres were evaluated by well-diffusion method against some selected pathogenic microorganisms. The antimutagenic properties of all compounds were also examined against sodium azide in human lymphocyte cells by micronuclei and sister chromatid exchange tests.

Glucose biosensor based on immobilization of glucose oxidase on a carbon paste electrode modified with microsphere-attached l-glycine.

In the present study, a novel biosensor that is sensitive to glucose was prepared using the microspheres modified with (4-formyl-3-methoxyphenoxymethyl)polystyrene (FMPS) with l-glycine. Polymeric microspheres having Schiff bases were prepared from FMPS using the glycine condensation method. Glucose oxidase enzyme was immobilized onto modified carbon paste electrode by cross-linking with glutaraldehyde. Oxidation of enzymatically produced H2 O2 (+0.5 V vs. Ag/AgCl) was used for determination of glucose. Optimal temperature and pH were found as 50 °C and 8.0, respectively. The glucose biosensor showed a linear working range from 5.0 × 10-4 to 1.0 × 10-2 M, R2 = 0.999. Storage and operational stability of the biosensor were also investigated. The biosensor gave perfect reproducible results after 20 measurements with 3.3% relative standard deviation. It also had good storage stability.

Spectroscopic analysis and the excellent reusability of sphere-capped ferrocene in the oxidation of glucose oxidase.

Sphere-capped ferrocene nanospheres with Schiff base spacers have been prepared using a template, and used as carriers to immobilize glucose oxidase (GOx). GOx immobilized on spheres with one C-spacer (APS-Fc) exhibited high binding affinity to the substrate, which was attributed to appropriate position for the GOx conformation. When glucose oxidase was immobilized with spacers of different lengths, it was found that storage stability decreased with increasing the length of the spacer. It has been found that nanospheres, including capped ferrocene, exhibit good performance as the immobilized supporters of GOx. (APS-EtFc-GOx) retain more than 10% of the initial activity after forty-two successive cycles, which is a remarkable result.

Useful agents against aflatoxin B1 - antibacterial azomethine and Mn(III) complexes involving L-Threonine, L-Serine, and L-Tyrosine.

The present study is focused on evaluating the antimutagenic properties of Schiff bases and Mn(III) complexes with L-Threonine, L-Serine and L-Tyrosine, which have antimicrobial activity. These six compounds were investigated for antimutagenic properties against Aflatoxin Bi (AFBi) by the micronucleus (MN) assay in a human lymphocyte cell culture in vitro. The protective role of these compounds against AFBi-induced MN is probably related to its doses. A mechanism has been proposed to reduce the effect of AFBi.

Nanoplatforms attached Schiff bases by condensation method; Investigation of glucose oxidase enzyme as biocatalysts.

We report a easy approach for the immobilization of glucose oxidase enzyme (GOx) on nanoparticles. Nanoparticles-modified polystyrene-ANH2 (PSA) with some salicylaldehyde derivatives were synthesized by means of condensation and investigated the enzymatic properties of GOx immobilized on there. Modified polystyrenes were characterized using IR spectra, gel permeation chromatography, and scanning electron microscopy. All the characteristics of the immobilized glucose oxidase (PSA-SalH)-GOx, (PSA-SalCH3)-GOx, and (PSA)-GOx enzyme showed except one of them. Immobilized GOx on to (PSA) showed two optimum pH, other immobilized polymers were one optimum.

Preparation of carbon paste electrodes including poly(styrene) attached glycine-Pt(IV) for amperometric detection of glucose.

In this study, a novel carbon paste electrode that is sensitive to glucose was prepared using the nanoparticles modified (4-Formyl-3-methoxyphenoxymethyl) with polystyren (FMPS) with L-Glycine-Pt(IV) complexes. Polymeric nanoparticles having Pt(IV) ion were prepared from (4-Formyl-3-methoxyphenoxymethyl) polystyren, glycine and PtCl4 by template method. Glucose oxidase enzyme was immobilized to a modified carbon paste electrode (MCPE) by cross-linking with glutaraldehyde. Determination of glucose was carried out by oxidation of enzymatically produced H2O2 at 0.5 V vs. Ag/AgCl. Effects of pH and temperature were investigated, and optimum parameters were found to be 8.0 and 55°C, respectively. Linear working range of the electrode was 5.0×10(-6)-1.0×10(-3) M, R(2)=0.997. Storage stability and operational stability of the enzyme electrode were also studied. Glucose biosensor gave perfect reproducible results after 10 measurements with 2.3% relative standard deviation. Also, it had good storage stability (gave 53.57% of the initial amperometric response at the end of 33th day).

Schiff bases attached L-glutamine and L-asparagine: first investigation on antimutagenic and antimicrobial analyses.

This study was conducted to evaluate the antimutagenic and antimicrobial activities of Schiff bases attached L-glutamine and L-asparagine. Antibacterial activities of the compounds against S. aureus, Sh. dys. typ 7, L. monocytogenes 4b, E. coli, S. typhi H, S. epidermis, Br. abortus, M. luteus, B. cereus, P. putida, and antifungal activity against Candida albicans were studied. These compounds were investigated for antimutagenic properties against Aflatoxin Bi (AFBi) using micronuclei (MN) assay in human lymphocyte cell culture in vitro. The protective role of these compounds against AFBi-induced MN is probably related to its doses.

Ni(II)-tetrahedral complexes: characterization, antimicrobial properties, theoretical studies and a new family of charge-transfer transitions.

A new amine containing selenium and their five imine, (SeSchX)(X: -H, F, Cl, Br, CH3), and Ni (II) complexes, [Ni(SeSchX)(H2O)2]Cl/[Ni(SeSchCl)(H2O)Cl], were synthesized. The compounds were characterized by means of elemental analyses, (13)C and (1)H NMR (for imine), FT-IR, UV-Visible spectroscopy, TGA/DTA and elemental analyses. [Ni(SeSchCl)(H2O)Cl] complex from Ni(II) complexes changes color from yellow to orange in the range pH 5-7. [Ni(SeSchCl)(H2O)Cl] complex has ligand-to-metal charge-transfer (LMCT) transitions in the basic medium. Excitation characteristics and energetic of [Ni(SeSchCl)(H2O)Cl] complex, examined via TD-DFT calculations, reveals transitions of LMCT and π→π(*) character that matches the experimental values. [Ni(SeSchCl)(H2O)Cl] complex showed the highest antibacterial activity when compared to other complexes reported in this work.

Polystyrene attached Pt(IV)-azomethine, synthesis and immmobilization of glucose oxidase enzyme.

Modified polystyrene with Pt(IV)-azomethine (APS-Sch-Pt) was synthesized by means of condensation and demonstrated to be a promising enzyme support by studying the enzymatic properties of glucose oxidase enzyme (GOx) immobilized on it. The characteristics of the immobilized glucose oxidase (APS-Sch-Pt-GOx) enzyme showed two optimum pH values that were pH = 4.0 and pH = 7. The insertion of stable Pt(IV)-azomethine spacers between the polystyrene backbone and the immobilized GOx, (APS-Sch-Pt-GOx), increases the enzymes' activity and improves their affinity towards the substrate even at pH = 4. The influence of temperature, reusability and storage capacity on the free and immobilized glucose oxidase enzyme was investigated. The storage stability of the immobilized glucose oxidase was shown to be eleven months in dry conditions at +4 °C.

Immobilization of ß-galactosidase on novel polymers having Schiff bases.

We have developed a strategy to immobilize ß-galactosidase as a model enzyme by using polymeric supports having Schiff bases, which were prepared from (aminomethyl)polystyrene and 2-phenlyindole-3-carboxaldehyde by condensation. ß-galactosidase was immobilized onto the new polymer supports via covalent bonds. The influence of temperature, pH, reusability, and storage capacity on the free and immobilized ß-galactosidase was investigated. Our results indicate that the (aminomethyl)polystyrene with Schiff bases is most suitable for the immobilization of ß-galactosidase. These kinds of new supports can be used for the immobilization of ß-galactosidase due to their strong storage capacity and reusability.

Antimicrobial activities of N-(2-hydroxy-1-naphthalidene)-amino acid(glycine, alanine, phenylalanine, histidine, tryptophane) Schiff bases and their manganese(III) complexes.

The in vitro antibacterial and antifungal activities of five different amino acid Schiff bases derived from the reaction of 2-hydroxy-1-naphthaldehyde with glycine, L-alanine L-phenylalanine, L-histidine, L-tryptophane and the manganese(III) complexes of these bases were investigated. Structures of the Schiff bases were proven by 1H-NMR. In vitro activities against some Gram-positive (Staphylococcus aureus and Bacillus polymyxa) and Gram-negative (Escherichia coli) bacteria and the fungus Candida albicans were determined. The antimicrobial activities tended to decrease with the increasing size of the amino acid residues.