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

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Abstract

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 K m 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.

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Acknowledgement

We are thankful for the financial support given by the DBT, New Delhi, India, to carry out membrane preparation work (BT/PR2937/PID/06/147/2002) and Director, NCL, Pune, India, for penicillin G acylase immobilization work (MLP008926).

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  1. Chemical Engineering Division, National Chemical Laboratory, Pune, 411 008, India

    H. V. Adikane & D. M. Thakar

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  1. H. V. Adikane
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Correspondence to H. V. Adikane.

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Adikane, H.V., Thakar, D.M. Studies of Penicillin G Acylase Immobilization Using Highly Porous Cellulose-Based Polymeric Membrane. Appl Biochem Biotechnol 160, 1130–1145 (2010). https://doi.org/10.1007/s12010-009-8686-9

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