Papain kinetics in the presence of a water-miscible organic solvent
Article first published online: 18 FEB 2004
Copyright © 1991 John Wiley & Sons, Inc.
Biotechnology and Bioengineering
Volume 37, Issue 10, pages 967–972, 25 April 1991
How to Cite
Fernandez, M. M., Clark, D. S. and Blanch, H. W. (1991), Papain kinetics in the presence of a water-miscible organic solvent. Biotechnol. Bioeng., 37: 967–972. doi: 10.1002/bit.260371011
- Issue published online: 18 FEB 2004
- Article first published online: 18 FEB 2004
- Manuscript Accepted: 19 NOV 1990
- Manuscript Received: 10 AUG 1990
- organic solvent;
The effects of various concentrations of a water-miscible organic solvent [a 7:3 (v/v) mixture of N, N dimethylformamide and dimethylsulfoxide] on the kinetics of papain have been investigated. The parameters kcat and Km for the amidase and esterase activity of papain using N-α-benzoyl-DL-arginine-p-nitroanilide (BAPNA) and N-α-benzoyl-L-arginine ethyl ester (BAEE) as substrates were determined. For both types of activity, kcat initially increased (up to about 15% solvent), and then decreased with increasing concentrations of organic solvent. In contrast, Km increased sharply with the organic solvent concentration. Active site titration at 0 and 50% solvent indicated no change in the amount of active enzyme. Fluorometric measurements of the emission spectrum of papain did not indicate any major conformational changes with increasing concentrations of organic solvent.