Induction of catalytic activity in proteins by lyophilization in the presence of a transition state analogue

• Published in: Biotechnology and bioengineering 1998-01, Vol.57 (2), p.211-215
• Main Authors: Slade, Christopher J., Vulfson, Evgeny N.
• Format: Article
• Language: English
• Subjects: Alkylation; Amines; Animals; Binding Sites; Biological and medical sciences; Biotechnology; bovine serum albumin; Catalysis; Cattle; Conformational dynamics in molecular biology; conformational memory; Conformations; enzyme; Enzyme kinetics; Freeze Drying; Fundamental and applied biological sciences. Psychology; In Vitro Techniques; Molecular biophysics; Molecular dynamics; molecular imprinting; organic solvent; Organic solvents; Oxidation-Reduction; protein; Protein Conformation; Proteins - chemistry; Proteins - isolation & purification; Proteins - metabolism; Serum Albumin, Bovine - chemistry; Serum Albumin, Bovine - isolation & purification; Serum Albumin, Bovine - metabolism; Substrates; transition state analogue; β-lactoglobulin
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/(SICI)1097-0290(19980120)57:2<211::AID-BIT9>3.0.CO;2-Q

The induction of catalytic activity in proteins by lyophilization in the presence of a transition state analogue (biomolecular imprinting) has been attempted. It was shown that proteins which were freeze‐dried with n‐isopropyl‐4‐nitrobenzyl‐amine (a transition state analogue for the reaction of dehydrofluorination of 4‐fluoro‐4‐[p‐nitrophenyl] butan‐2‐one) displayed higher β‐elimination activity as compared to their‐non‐imprinted counterparts. It was also found that native bovine serum albumin has a high dehydrofluorination activity towards the above substrate with kinetic parameters rather similar to those of a catalytic antibody prepared by Shokat et al. (1989). A comparison of the kinetic parameters determined in this study with those obtained for analogous catalytic antibodies and imprinted polymers was made. © 1998 John Wiley & Sons, Inc. Biotechnol. Bioeng. 57: 211–215, 1998.