Properties of soluble α-chymotrypsin in neat glycerol and water

被引：34

作者：

Castro, GR ^{[1
]}

机构：

[1] MIT, Dept Chem, Cambridge, MA 02139 USA

来源：

ENZYME AND MICROBIAL TECHNOLOGY | 2000年 / 27卷 / 1-2期

关键词：

homogeneous biocatalysis; enzymes in organic media; alpha-chymotrypsin; trypsin; glycerol; enzyme thermostability;

D O I：

10.1016/S0141-0229(00)00197-6

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

UV scanning of cr-chymotrypsin dissolved in neat glycerol and water showed no significant differences in its spectra at pH 7.8. Fluorescence scanning revealed a strong dependence on pH values (between 5.9 to 10.5) of the maximum wavelength emission in water and no pH-dependence in 99% glycerol supplemented with 1% of appropriate buffers. The profile of alpha-chymotrypsin activity dissolved in water-glycerol mixtures with phenyl acetate as substrate displayed two maximum: highest peak was found at 100% water, and the second one was observed in 99% glycerol concentration with about 40% of the relative activity. Optimum pH of the soluble alpha-chymotrypsin in glycerol showed a displacement of 1 pH/U towards the alkaline side compared to water at pH 8.0. Kinetic and thermodynamic analysis using kinetic measurements of the thermal stability of alpha-chymotrypsin showed a higher inactivation rate in neat glycerol as compared to water in 30 to 45 degrees C range, however, when temperature increases enzyme stability in glycerol is better than water. Thermostability of trypsin and alpha-chymotrypsin dissolved in glycerol at 100 degrees C showed a half reaction time of approximately 7 and 20 h, respectively, and less than 1 minute in aqueous buffer for both enzymes. (C) 2000 Elsevier Science Inc. All rights reserved.

引用

页码：143 / 150

页数：8

共 22 条

[1]

Baigori MD, 1996, BIOTECHNOL APPL BIOC, V24, P7

[2]

Breccia JD, 1998, J CHEM TECHNOL BIOT, V71, P241, DOI 10.1002/(SICI)1097-4660(199803)71:3<241::AID-JCTB810>3.0.CO

[3]

2-G

[4]

CASTRO GR, 1997, P PEW ANN M, P62