Optimization of penicillin G acylase multipoint immobilization on to glutaraldehyde–chitosan beads

Abstract

The objective of this work was to study the immobilization of penicillin G acylase from Escherichia coli on to chitosan-glutaraldehyde beads by multipoint covalent binding. This process was optimized using a 2(3) experimental design. The parameters selected for the present study were the concentrations of glutaraldehyde, phenylacetic acid and sodium borohydride. Three responses were chosen, namely immobilization yield and stabilization factors of enzyme derivatives at high temperature and at alkaline pH. All the runs at the maximum (+1) and minimum (-1) levels were performed at random. Three experiments were performed at the centre point, coded as zero, for experimental-error estimation. With respect to immobilization yield, the main effectors were the concentrations of glutaraldehyde and phenylacetic acid. For stabilization factors at 50 degrees C and at alkaline pH, the main effectors were the concentrations of glutaraldehyde and sodium borohydride and the interaction between them.

One-Pot, One-Step Production of Dietary Nucleotides by Magnetic Biocatalysts

“…Even if all of the enzyme molecules are immediately incorporated into the support, it is not certain that the enzyme is covalently attached to the support. In fact, when a highly activated support is used, in most cases, an ionic exchange reaction among the enzyme and amino groups in the support takes place at the beginning [29][30][31][32][33][34][35]. In order to discard that the immobilization takes place through ionic exchange, the derivatives were washed four times with 1 M of NaCl solution to promote protein relase, but the presence of TtHGXPRT in the solution was not detected in the solution.…”

Optimization of Penicillin G Acylase Immobilization Process by Surface Response Methodology Using Central Composite Design

“…Penicillin acylase in its soluble form is unstable, cannot be separated from the reaction mixture easily which will add to the production cost of the final product thus such a commercially, industrially important biocatalyst has been immobilized by different techniques employing various supports by many investigators [1][2][3][4][5][6][7]. Each technique or method has its own limitation and disadvantages where immobilization method can alter kinetic properties of immobilized enzyme as compared to its soluble counterpart.…”