A continuous fluorometric assay for pectin methylesterase

被引：30
|
作者：
Wojciechowski, CL
Fall, R
机构：
[1] UNIV COLORADO,DEPT CHEM & BIOCHEM,BOULDER,CO 80309
[2] COOPERAT INST RES ENVIRONM SCI,BOULDER,CO 80309
来源：
ANALYTICAL BIOCHEMISTRY | 1996年 / 237卷 / 01期
基金：
美国国家科学基金会;
关键词：
D O I：
10.1006/abio.1996.0206
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
A continuous, fluorometric assay for pectin methylesterase (PME) activity is described. Tn this assay, methanol produced by PME hydrolysis of pectin methyl esters is oxidized to formaldehyde by alcohol oxidase, and the formaldehyde is continuously reacted with 4-amino-3-penten-2-one to create a stable, fluorescent product. The increase in fluorescence intensity is linearly proportional to PME activity. The assay can be used in crude plant or fungal extracts with relatively little interference with chemicals or buffers commonly used in PME purification. The fluorescence assay has a useful pH range, from pH 5.0 to 6.5, which overlaps pH optima for many bacterial and fungal PMEs, but which limits its usefulness in assaying plant PMEs with alkaline pH optima. Nevertheless, the method is valuable for rapid assay of plant PMEs during their purification or for comparison of plant tissue PME activities. (C) 1996 Academic Press, Inc.
引用
收藏
页码：103 / 108
页数：6
相关论文
共 50 条
  • [31] Direct continuous fluorometric assay for monoamine oxidase B
    Zhou, JJP
    Zhong, B
    Silverman, RB
    ANALYTICAL BIOCHEMISTRY, 1996, 234 (01) : 9 - 12
  • [32] Synchronous fluorometric method for continuous assay of monophenolase activity
    Zhang, Ling
    Shang, Qi
    Chen, Chan
    Tang, Weikang
    Xu, Yidian
    Liu, Wenbin
    SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY, 2021, 252 (252)
  • [33] A continuous fluorometric assay for the feline immunodeficiency virus protease
    Fitzgerald, MC
    Laco, GS
    Elder, JH
    Kent, SBH
    ANALYTICAL BIOCHEMISTRY, 1997, 254 (02) : 226 - 230
  • [34] Direct continuous fluorometric assay for monoamine oxidase B
    Department of Chemistry, Northwestern University, Evanston, IL 60208-3113, United States
    ANAL. BIOCHEM., 1 (9-12):
  • [35] A continuous fluorometric assay for the assessment of MazF ribonuclease activity
    Wang, Nora R.
    Hergenrother, Paul J.
    ANALYTICAL BIOCHEMISTRY, 2007, 371 (02) : 173 - 183
  • [36] Characterization and transcript profiling of the pectin methylesterase (PME) and pectin methylesterase inhibitor (PMEI) gene families in flax (Linum usitatissimum)
    Pinzon-Latorre, David
    Deyholos, Michael K.
    BMC GENOMICS, 2013, 14
  • [37] Characterization and transcript profiling of the pectin methylesterase (PME) and pectin methylesterase inhibitor (PMEI) gene families in flax (Linum usitatissimum)
    David Pinzón-Latorre
    Michael K Deyholos
    BMC Genomics, 14
  • [38] Thermal inactivation of pectin methylesterase in tomato homogenate
    Sekine, Y
    Wakayama, T
    Barrett, DM
    PROCEEDINGS OF THE EIGHTH INTERNATIONAL ISHS SYMPOSIUM ON THE PROCESSING TOMATO, 2003, (613): : 385 - 390
  • [39] Functional characterization of pectin enzymatically modified with a thermally tolerant pectin methylesterase
    Luzio, Gary
    Cameron, Randall G.
    ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, 2009, 238 : 177 - 177
  • [40] Pectin Methylesterase Action Reduces Sugar Beet Pulp Water Binding in an In Vitro Functional Assay.
    Cease, Megan
    Tovar, Jose C.
    Zhang, Ningning
    Xu, Jianfeng
    Savary, Brett J.
    IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL, 2015, 51 : S55 - S56
12345