Continuous spectrophotometric assay for β-glucuronidase

被引:21
作者
Aich, S [1 ]
Delbaere, LTJ [1 ]
Chen, RD [1 ]
机构
[1] Univ Saskatchewan, Saskatoon, SK, Canada
关键词
D O I
10.2144/01304rr02
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A continuous spectrophotometric assay has been developed for detecting beta -glucuronidase activity. In the assay, Para-nitrophenyl beta -D-glucuronide is cleaved to yield a chromophoric product. With the commercial E. coli enzyme, it is demonstrated that the reactions can be continuously monitored by the increase of absorbance at 405 nm. The method is highly sensitive and able to detect less than 1.4 x 10(-4) U/mL of the enzyme activity in solution. Such a new assay offers significant advantages over the existing discontinuous methods and should be useful for both routine enzyme assay and accurate kinetic studies.
引用
收藏
页码:846 / 850
页数:5
相关论文
共 27 条
[1]   ONE-STEP PURIFICATION OF ESCHERICHIA-COLI BETA-GLUCURONIDASE [J].
BLANCO, C ;
NEMOZ, G .
BIOCHIMIE, 1987, 69 (02) :157-161
[2]   PURIFICATION AND PROPERTIES OF BETA-GLUCURONIDASE FROM HUMAN PLACENTA [J].
BROT, FE ;
BELL, CE ;
SLY, WS .
BIOCHEMISTRY, 1978, 17 (03) :385-391
[3]   A CONTINUOUS SPECTROPHOTOMETRIC ASSAY FOR PROTEIN PHOSPHATASES [J].
CHENG, Q ;
WANG, ZX ;
KILLILEA, SD .
ANALYTICAL BIOCHEMISTRY, 1995, 226 (01) :68-73
[4]  
Gallagher S.R., 1992, GUS PROTOCOLS, P47
[5]   BIOCHEMICAL-PROPERTIES OF RECOMBINANT HUMAN BETA-GLUCURONIDASE SYNTHESIZED IN BABY HAMSTER-KIDNEY CELLS [J].
GEHRMANN, MC ;
OPPER, M ;
SEDLACEK, HH ;
BOSSLET, K ;
CZECH, J .
BIOCHEMICAL JOURNAL, 1994, 301 :821-828
[6]  
GLASER JH, 1973, J LAB CLIN MED, V82, P969
[7]   Intracisternal A-particle element transposition into the murine β-glucuronidase gene correlates with loss of enzyme activity:: a new model for β-glucuronidase deficiency in the C3H mouse [J].
Gwynn, B ;
Lueders, K ;
Sands, MS ;
Birkenmeier, EH .
MOLECULAR AND CELLULAR BIOLOGY, 1998, 18 (11) :6474-6481
[8]   A continuous spectrophotometric assay for monoamine oxidase and related enzymes in tissue homogenates [J].
Holt, A ;
Sharman, DF ;
Baker, GB ;
Palcic, MM .
ANALYTICAL BIOCHEMISTRY, 1997, 244 (02) :384-392
[9]   BETA-GLUCURONIDASE FROM ESCHERICHIA-COLI AS A GENE-FUSION MARKER [J].
JEFFERSON, RA ;
BURGESS, SM ;
HIRSH, D .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1986, 83 (22) :8447-8451
[10]  
Jefferson RA., 1987, PLANT MOL BIOL REP, V5, P387, DOI DOI 10.1007/BF02667740