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Study on the Application of Reversed Micelles System in Enzymatic Determination and Bioseparation

Author: HeJinXing
Tutor: LiuChangXian
School: Central South University for Nationalities
Course: Analytical Chemistry
Keywords: Reverse micelles Immobilization Enzymatic determination Alcohol dehydrogenase Lactate dehydrogenase Enzymatic kinetics Extraction Bioseparations
CLC: O648
Type: Master's thesis
Year: 2009
Downloads: 51
Quote: 0
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Abstract


Is a surfactant reverse micelles dispersed in the continuous organic phase with spontaneous formation of the normal structure of the reverse micelles with an aqueous polymer. Since reverse micelles can simulate the natural environment of the enzyme, and thus most of the enzyme activity and stability can be maintained. Reverse micelles enzyme system in biological research on catalysis has made great progress in many laboratories the positioning of the system and the structure of enzymes, enzyme kinetics, catalytic activity and stability of other aspects of theoretical research, further broaden the field of reverse micelles technology has laid a good foundation. When the reverse micelles containing organic solvents and aqueous protein solution in contact, the protein in several forces (electrostatic, affinity, hydrophobic) the synergistic effect of the nucleus into the polar and then adjust the appropriate conditions, the protein from the loaded organic phase re-extracted into the aqueous phase, in order to achieve the purpose of purification. Is a kind of reverse micelles extraction solvent extraction, with a large amount of sample handling, easy to zoom in and continuous operation characteristics of the environment within the reverse micelles close to the intracellular environment, protein denaturation is not easy. Extraction of reverse micelles also has a low cost, a solvent can be used repeatedly, the extraction rate is high and the phase ratio advantages. Therefore, the reverse micelles extraction separation and purification of a protein with industrial development has opened up a promising new approach. Chapter gives an overview of the enzyme reverse micelles technical analysis and the basic theory and applications. Chapter II of the reverse micelle solution of lactate dehydrogenase (LDH) catalytic kinetic properties tested water content, surfactant and co-solvent dosage on the impact of enzymes into reverse micelles. Discussion of temperature, pH and other factors on enzyme activities and LDH investigated the fluorescence spectra. Chapter III studies cetyl trimethyl ammonium bromide (CTAB) - octane - hexanol reverse micelles immobilized alcohol dehydrogenase (ADH) Preparation and application. The effects of moisture, CTAB and hexanol dosage for immobilization of ADH. On the free enzyme and immobilized enzyme kinetic properties showed that: the enzymatic reaction optimum pH values ??were 8.2 and 8.8, the optimum temperature was 31 ℃ and 20 ℃, Michaelis constant (Km) of ethanol were 12 mmol / L and 7.4 mmol / L. 30 ℃, storage After 150 min, the free enzyme inactivated 90%, 50% inactivation of the immobilized enzyme, indicating that the reverse micelle immobilized ADH have better thermal stability. And the application of this system for the determination of the concentration of ethanol in the sample. Chapter IV discusses the reverse micelles extraction of yeast ADH, in this extraction system to CTAB as a surfactant, isooctane and amyl alcohol as solvent and cosolvent. Test surfactants and solubilizing concentration, initial pH of the aqueous phase of the extraction. Detailed discussion of ionic strength aqueous stripping phase, the concentration of isopropyl alcohol stripping effect, established the optimum extraction conditions.

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CLC: > Mathematical sciences and chemical > Chemistry > Physical Chemistry ( theoretical chemistry ),chemical physics > Colloid chemistry ( physical and chemical dispersion system )
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