Studies on immobilized trypsin in high concentrations of organic solvents
作者:
Howard H. Weetall,
William P. Vann,
期刊:
Biotechnology and Bioengineering
(WILEY Available online 1976)
卷期:
Volume 18,
issue 1
页码: 105-118
ISSN:0006-3592
年代: 1976
DOI:10.1002/bit.260180109
出版商: Wiley Subscription Services, Inc., A Wiley Company
数据来源: WILEY
摘要:
AbstractTrypsin was covalently immobilized on porous glass in the presence and absence of a specific substrate and reacted in various organic solvents of different dielectric constants. Optimum solvent concentration, pH profile,Km(app),Vmax(app), productivity versus temperature, activity, and reaction rates were determined. Reaction rates of six lysyl dipeptides were compared. Crystalline trypsin was dansylated for studies by nanosecond fluorescence techniques to determine the effects of introducing high concentrations of organic solvents on the molecule. The results indicated that greater reaction rates were observed with dipeptides having more acidic carboxyl terminal groups. The data also indicated that greater reaction rates were observed in higher concentrations of solvents of lower dielectric constants. Nanosecond fluorescence spectroscopy of trypsin in high concentrations of a low dielectric constant solvent indicated major dehydration even though maximal enzyme‐activity was achieved under these condition
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