Synthesis of glucose oxidase-PEG aldehyde conjugates and improvement of enzymatic stability.

In this article, aldehyde derivative of poly(ethylene glycol) (PEG) was synthesized directly with sodium periodate agent. To obtain a conjugate which possesses better stability, PEG aldehyde was bonded to native enzyme with different molar ratios. The conjugation reaction turned out to be efficient and mild. Colorimetric method was applied to evaluate the enzymatic activity of native GOD and its derivatives by introducing another enzyme, horseradish peroxidase. The GOD-PEG aldehyde conjugate with polymeric chains exhibited reduced enzymatic activity towards the catalytical oxidation of glucose, but with significantly increased thermal stability and elongated lifetime. When GOD was modified with PEG aldehyde the enzymatic activity was decreased 40% at 30 °C. However, when incubated at 60 °C the GOD-PEG aldehyde conjugate still retained the enzyme bioactivity of 40% bioactivity left after 4 h, whereas the native GOD lost almost all the activity in 4 h. The polymer chain attached, the more reduction of the enzymatic activity resulted, however, the longer the lifetime and higher thermal stability of the enzyme obtained.

Determination of minimum enzymatic decolorization time of reactive dye solution by spectroscopic & mathematical approach.

Synthetic dyes are very important for textile dyeing, paper printing, color photography and petroleum products. Traditional methods of dye removal include biodegradation, precipitation, adsorption, chemical degradation, photo degradation, and chemical coagulation. Dye decolorization with enzymatic reaction is an important issue for several research field (chemistry, environment) In this study, minimum decolorization time of Remazol Brilliant Blue R dye with Horseradish peroxidase enzyme was calculated using with mathematical equation depending on experimental data. Dye decolorization was determined by monitoring the absorbance decrease at the specific maximum wavelength for dye. All experiments were carried out with different initial dye concentrations of Remazol Brilliant Blue R at 25 degrees C constant temperature for 30 minutes. The development of the least squares estimators for a nonlinear model brings about complications not encountered in the case of the linear model. Decolorization times for completely removal of dye were calculated according to equation. It was shown that mathematical equation was conformed exponential curve for dye degradation.

Measuring calcium, potassium, and nitrate in plant nutrient solutions using ion-selective electrodes in hydroponic greenhouse of some vegetables.

Generally, the life cycle of plants depends on the uptake of essential nutrients in a balanced manner and on toxic elements being under a certain concentration. Lack of control of nutrient levels in nutrient solution can result in reduced plant growth and undesired conditions such as blossom-end rot. In this study, sensitivity and selectivity tests for various polyvinylchloride (PVC)-based ion-selective membranes were conducted to identify those suitable for measuring typical concentration ranges of macronutrients, that is, NO(3-), K(+), and Ca(2+), in hydroponic solutions. The sensitivity and selectivity of PVC-membrane-based ion-selective sensors prepared with tetradodecylammoniumnitrate for NO(3-), valinomycin for K(+), and Ca ionophore IV for Ca(2+) were found to be satisfactory for measuring NO(3-), K(+), and Ca(2+) ions in nutrient solutions over typical ranges of hydroponic concentrations. Potassium, calcium, and nitrate levels that were utilized by cucumber and tomato seedlings in the greenhouse were different. The findings show that tomato plants consumed less amounts of nitrate than cucumber plants over the first 2 months of their growth. We also found that the potassium intake was higher than other nutritional elements tested for all plants.

Enzymatic decolorization of anthraquinone and diazo dyes using horseradish peroxidase enzyme immobilized onto various polysulfone supports.

In this study, covalent immobilization of the horseradish peroxidase (HRP) onto various polysulfone supports was investigated. For this purpose, different polysulfones were methacrylated with methacryloyl chloride, and then, nonwoven fabric samples were coated by using solutions of these methacrylated polysulfones. Finally, support materials were immersed into aquatic solution of HRP enzyme for covalent immobilization. Structural analysis of enzyme immobilization onto various polysulfones was confirmed with Fourier transform infrared spectroscopy, atomic force microscopy, and proton nuclear magnetic resonance spectroscopy. Decolorization of textile diazo (Acid Black 1) and anthraquinone (Reactive Blue 19) dyes was investigated by UV-visible spectrophotometer. Covalently immobilized enzyme has been used seven times in freshly prepared dye solutions through 63 days. Dye decolorization performance of the immobilized systems was observed that still remained high (70%) after reusing three times. Enzyme activities of immobilized systems were determined and compared to free enzyme activity at different conditions (pH, temperature, thermal stability, storage stability). Enzyme activities of immobilized systems and free enzyme were also investigated at the different temperatures and effects of temperature and thermal resistance for different incubation time at 50 °C. In addition, storage activity of free and immobilized enzymes was determined at 4 °C at different incubation days.

Novel creatine biosensors based on all solid-state contact ammonium-selective membrane electrodes.

Novel creatine bienzymatic potentiometric biosensors were prepared by immobilizing urease and creatinase on all solid-state contact PVC-containing palmitic acid and carboxylated PVC matrix membrane ammonium-selective electrodes without inner reference solution. Potentiometric characteristics of biosensors were examined in physiological model solutions at different creatine concentrations. The linear working range and long-term sensitivity of the biosensors were also determined. The creatine biosensors prepared by using the carboxylated PVC membrane electrodes showed more effective performance than those of the PVC containing palmitic acid membrane electrodes. Creatine assay in serum samples was successfully carried out by using the standard addition method.

Protective effect of dextrans on glucose oxidase denaturation and inactivation.

In the present study, the stabilizing effect of dextrans as additives on the denaturation and inactivation of glucose oxidase (GOD) was investigated. Three different molecular weighted dextrans (M(w) 17.5, 75, 188 kD) were used with different concentrations. Dramatically increased enzyme activities were measured after one hour of incubation of enzyme with additives between 25-40°C in water bath. Highest activity value was measured with 75 kDa molecular weighted dextran (in concentration 30% w/v) at pH 5. Dextran as an additive supplied a long shelf-life to the enzyme at 4°C. In the presence of the 75 kDa dextran, the enzyme was more stable and its activity was increased 2.7-fold at 30°C. In addition, dextran protected GOD against inactivation by a n-heptane/aqueous buffer-stirred system.

Additive effect of dextrans on the stability of horseradish peroxidase.

The influence of various concentration (10, 20, and 30% w/v) of different molar weighted dextrans as additives on the stability of HRP has been studied in aqueous medium. Native HRP preparations were formulated with different additives for storage stabilization and better performance at high temperature and pH. The results obtained show a stabilizing effect in the presence of an additive (75 kDa dextran). The enzyme with 75 kDa dextran (in concentration 10% w/v) showed the highest thermal resistance and the best performance for long-term storage at pH 5.0. In the presence of the 75 kDa dextran, the enzyme activity was increased threefold at 25 °C and lost only 15% activity in 2 h at 50 °C in comparison to the native enzyme which lost all its activity. In addition, dextran protected HRP against inactivation by air bubbles.

Decolorization of naphthol blue black using the horseradish peroxidase.

This study evaluates the potential of the enzyme horseradish peroxidase in the decolorization of one common industrial azo dye, naphthol blue black. Studies are carried out to understand the process parameters such as pH, temperature and reaction time. The enzymatic decolorization of the dye was examined by UV-Vis spectrophotometer and LC-MS measurements. Temperature and pH conditions were optimized for obtaining high azo-dye decolorization. Azo-dye removal at a pH range 4-6 was found to be the highest for all temperatures. After 5 minutes of treatment, the color removal of dye was ca. 80-90%. The LC-MS and spectrophotometric analyses indicated that the decolorization of the azo dye with enzyme was due to the reduction of the azo bonds. This study verifies the viability of the use of the horseradish peroxidase in the decolorization of naphthol blue black.

Enhanced stability and decolorization of Coomassie Brilliant Blue R-250 by dextran aldehyde-modified horseradish peroxidase.

Horseradish peroxidase (EC 1.11.1.7) was chemically modified by periodate-activated dextran. The activities of free and modified enzyme against organic-aqueous interface and some chemicals were determined. Modified HRP remained fully active in the presence of organic solvent for 4 h. However, the unmodified enzyme lost 50% of its activity within the first 2 h. The effects of possible inhibitors on enzyme activity were investigated. In addition, Coomassie Brilliant Blue R-250 was efficiently decolorized using the free and modified HRP. After 5 minutes of treatment, the color removal of dye was 80-90%. Modified HRP showed effective performance compared to free HRP.

Characterization of water-soluble complexes of polyacrylic acid with alpha-Amylase from Aspergillus oryzae.

Polycomplex formation of alpha-Amylase from Aspergillus oryzae (TAKA) with polyacrylic acid (PAA) was studied by pH titration, fluorescence, and high performance liquid chromatography (HPLC) methods in water solutions. According to the our results, the complex formation and its solubility were depended on nature of enzyme and the pH of solutions. Both of them correlates isoelectric points (PI). The stability of PAA-amylase complexes was negligibly weak at pH 7 [pH > pI (isoelectric pH)]. Stable water-soluble polycomplexes were formed at pH 5 (pI approximately 4.5) and coexisted with free protein molecules. Insoluble complexes has been observed at pH < 4.5. The frozen storage stabilities of the obtained complexes were also studied by measuring the activities at different pH.

Glucose oxidase-dextran conjugates with enhanced stabilities against temperature and pH.

Multipoint covalent bonding of glucose oxidase (EC 1.1.3.4) to hydrophilic natural polymer dextran and optimization of procedures to obtain, with enhanced temperature and pH stabilities, were studied. Purified enzyme was conjugated with various molecular weight dextrans (17.5, 75, and 188 kD) in a ratio of 20:1, 10:1, 1:1, 1:5, 1:10, 1:15, and 1:20. After 1 h of incubation at pH 7, the activities of purified enzyme and conjugates were determined at different temperatures (25 degrees C, 30 degrees C, 35 degrees C, 40 degrees C, 50 degrees C, 60 degrees C, 70 degrees C, and 80 degrees C), and the results were evaluated for thermal resistance. Increases in temperature from 25 degrees C to 50 degrees C did not change the activities of the conjugates. The conjugate, which was prepared with 75 kDa dextran in a molar ratio of 1:5, showed the highest thermal resistance and even the activity still remains at 80 degrees C at pH 7.0. This conjugate also displayed activity in a wide pH range (pH 4.0-7.0) at high temperatures. Conjugate, which was synthesized with 75 kDa dextran in a molar ratio of 1:5, appears to be feasible and useful for biotechnological applications.