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Updated: Jun 24, 2026

OaAEP1-Mediated Enzymatic Synthesis and Immobilization of Polymerized Protein for Single-Molecule Force Spectroscopy
Published on: February 5, 2020
Optimized polymer-enzyme electrostatic interactions significantly improve penicillin G amidase efficiency in charged
Alessandra Basso1, Beatrice A Maltman, Sabine L Flitsch
1Dipartimento di Scienze Farmaceutiche, Università degli Studi, Piazzale Europa 1, 34127 Trieste, Italy.
Abstract:
Hydrolytic yields as high as 80% were obtained by using penicillin G amidase (PGA) on substrates anchored on optimized positively charged PEGA polymers. By increasing the amount of permanent charges inside the polymer, electrostatic interactions between the positively charged PEGA(+) and the negatively charged PGA (pI = 5.2-5.4) were strengthened, thus favouring the accessibility of the bulky enzyme (MW = 88 kDa) inside the pores. The effect of different amounts of charges on polymer swelling and protein retention inside the polymer was investigated and correlated to the enzyme efficiency demonstrating that electrostatic interactions predominate over swelling properties in determining enzyme accessibility.
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