Single-step purification and immobilization of penicillin acylase using hydrophobic ligands

• Published in: Applied biochemistry and biotechnology Vol. 94; no. 2; pp. 127 - 134
• Main Authors: ADIKANE, Harshvardhan V, SINGH, Rajesh K, THAKAR, Dnyaneshwar M, NENE, Sanjay N
• Format: Journal Article
• Language: English
• Published: Heidelberg Springer 01.05.2001; Springer Nature B.V
• Subjects: Adsorption; Biochemistry; Biological and medical sciences; Bioreactors; Biotechnology; Chromatography - methods; Dose-Response Relationship, Drug; Enzyme engineering; Enzymes; Escherichia coli - enzymology; Fundamental and applied biological sciences. Psychology; Immobilization of enzymes and other molecules; Immobilization techniques; Improved methods for extraction and purification of enzymes; Ligands; Methods. Procedures. Technologies; Penicillin; penicillin acylase; Penicillin Amidase - chemistry; Penicillin Amidase - isolation & purification; Proteins; Salts - pharmacology; Sepharose - chemistry; Studies; Temperature; Time Factors; Penicillin amidase; Purification; Stability; Enzymatic activity; Adsorption; Enzyme; Ligand; Immobilization; Hydrolases; Immobilized enzyme
• ISSN: 0273-2289; 1559-0291; 0273-2289
• DOI: 10.1385/ABAB:94:2:127

Five different hydrophobic ligands immobilized on 4% (4XL) and 6% (6XL) crosslinked agarose were used to study the single-step purification of penicillin acylase from cell lysate. The 4XL gels showed relatively higher specific activity and recovery than the 6XL gels. In single-step purification, highly active enzyme (42 U/mg) was obtained using moderately hydrophobic ligand (octyl). The crude enzyme immobilized on octyl gel by adsorption showed significant operational stability over a period of 30 d at room temperature. Reactor studies demonstrated the feasibility of hydrophobic ligands as a medium for immobilization.