Immobilization Of Penicillin G Acylase--Using Layer Material

It is opening a new field of immobile carrier to use layer material-Zirconium phosphate( ZrP) on the immobilization of Penicillin G acylase(PGA). To use ZrP as main carrier, with lysine or 1,6-hexamethylene diamine intercalation to immobilize Penicillin G acylase, and studies on the immobilized acylase. Moreover, to use hydrotalcite as carrier, immobilizing Penicillin G acylase directly.First, to use ZrP as carrier, with absorption and crosslink to immobilized Penicillin G acylase, optimizing the conditions of ihe immobilized acylase, and a good method is ablained: lysine or 1,6-hexamelhylene diamine intercalation, glutaraldehyde activating, acylase immobilizing. The factors of immobilizing including: quantity of PGA , quantity of intercalating carrier, temperature, time, pH, and concentration and pH of glutartaldehyde.Second, studies on the catalysing conditions of immobilized acylase, including: temperature effection, pH effection, stability and reproducibility. Also using XRD to analyse this immobilized acylase.Hence, the higher activity of immobilized acylase is obtained. The activity of the immobilized Penicillin acylase with lysine intercalation is about 521 u/g, 1,6-hexamethylene diamine intercalation is about 360.8 u/g. The yield of the immobilization of Penicillin G acylase are 37.4% and 36.7%, respectively. Those activity have no obvious decrease after 10 times continuous hydrolization of Penicillin GThird, the Michaelis-Menton constant of the immobilized acylase and free enzyme are measured. The values are: 3.109mmol/l(free enzyme),3.916mmol/l(lysine intercalation), 11.24mmol/l( 1,6-hexamethylene diamine intercalation).Finally, the method of using direct immobilizing and glutaraldehyde crosslinking to immobilize acylase of hydratalcite is studied.