Studies of Penicillin G Acylase Immobilization Using Highly Porous Cellulose-Based Polymeric Membrane.

Adikane, H. V.; Thakar, D. M.

The different ionic molecules/compounds were used as a ligand for the immobilization of penicillin G acylase on the highly porous cellulose-based polymeric membrane having buffer flux 1,746 LMH (L m-2 h-1) at 0.5 bar pressure. The immobilized enzyme activity around 250 UApp was obtained with the ligand such as proline, tryptophan, casein acid hydrolysate, and brilliant green. Comparatively, proline showed less IMY% (percentage immobilization yield--58) but higher RTA% (percentage of activity retention--71) and specific activity (145 UApp g-1). However, the crosslinked preparation of brilliant green obtained using glutaraldehyde showed 82±2.7% immobilized enzyme activity after the completion of successive five cycles. In comparison with the free enzyme, the enzyme immobilized on the brilliant green coupled membrane showed around 2.4-fold increase in Km value (47.4 mM) as well as similar optimum pH (7.2) and temperature (40°C). The immobilized enzyme retained almost 50% activity after 107 days and 50 cycles of operation. Almost 50% decrease in buffer flux after enzyme immobilization was observed. At the end of the 30 cycles, flux pattern shows around 38% decrease in buffer flux however, after 16 cycles of operation flux moves closer towards the steady state.

PENICILLIN; ENZYMES; TRYPTOPHAN; CASEINS; GLUTARALDEHYDE; PROLINE

Applied Biochemistry & Biotechnology, 2010, Vol 160, Issue 4, p1130

0273-2289

Academic Journal

10.1007/s12010-009-8686-9