Enhanced enzymatic activity and stability of trypsin by reductive alkylation in solid phase

✍ Scribed by Ryohei F. Tsuji

• Publisher: John Wiley and Sons
• Year: 1990
• Tongue: English
• Weight: 408 KB
• Volume: 36
• Category: Article
• ISSN: 0006-3592
• DOI: 10.1002/bit.260361005

✦ Synopsis

Abstract

Amino groups of trypsin (EC 3.4.21.4) were reductively alkylated in solid phase to obtain a surface‐active and biologically active enzyme in an o/w emulsion system. Trypsin adsorbed on a benzamidine–sepharose column was reductively alkylated with n‐octanal in the presence of sodium borohydride, i.e., trypsin‐C8. Activity of trypsin‐C8 against __N__α‐benzoyl‐L‐arginine‐p‐nitroanilide was three times higher than that of native trypsin. Activities of trypsin and trypsin‐C8 against casein were almost the same. After incubating the trypsin solution at 40°C for 1 h, residual activities in the emulsion and solution systems were 64.2 and 57.4%, respectively. On the other hand, residual activities of native trypsin following incubation were 21.8% in the emulsion system and 33.2% in the solution system. Enhancement of trypsin‐C8 stability in the emulsion system may derive from interaction between the hydrophobic areas of trypsin‐C8 molecules and the hydrophobic phase of the emulsion.