Catalytic cryogel columns for the synthesis of 6-APA which is a key ingredient in the production of antibiotics

Penicillin G acylase (PA) (E.C 3.5.1.11) is an industrially important enzyme for the production of 6-aminopenicillanic acid from penicillin G during the manufacturing of semi synthetic antibiotics. PA is an immobilized enzyme which is extensivelyused in the world. Cryogel column sare support materials having high surface area which cause an increase in immobilization efficeincy and letting enhanced mass transport due to the their special structure. In this study, high efficiency catalytic cryogel columns, which are reusable and crosslinked with PA and will be an alternative to the current commercial systems for the production of 6-APA, have improved and reusability of this column has investigated. For this purpose, firstly, the hydroxy ethylmethacrylate (HEMA) based enzymemimicking cryogel column has been synthesized indroducing PA as a co-monomer by photosensitive cross-linking using ANADOLUCA method pantented by Say et al., and characterization studies have been performed. Then, the optimum conditions for these columns have been determined in continuos system. In the last step of the study, purified PA from microbial sources has used in the synthesis of catalytic cryogel column and production performance of 6-APA has been investigated. The swelling ratio of catalytic columns has found to be 583.41%. Optimum pH and temperature have determined as 7,2 and 30 oC, respectively, for free and immobilized PA. It has observed that the column activity has decresed only 26 % after 16 use of the same column. After 150 days of storage (in sodiumazide solution at -20oC), while 80% of catalytic activity of cryogels has conserved, free enzyme has conserved only 42% activity. 31,85μg 6-APA has produced in 60 minute optimum conditions using catalytic cyrogel crosslinked with PA which has purified from Penicillium chryosogenum 807.