Cloning expression and characterization of a thermostable exopolygalacturonase from Thermotoga maritima

被引：21
作者：
Parisot, J [1 ]
Langlois, V [1 ]
Sakanyan, V [1 ]
Rabiller, C [1 ]
机构：
[1] Fac Sci & Technol, Unite Rech & Biocatal, CNRS & U2230, F-44322 Nantes, France
来源：
CARBOHYDRATE RESEARCH | 2003年 / 338卷 / 12期
关键词：
exopolygalacturonase; thermostable enzyme; Thermotoga maritima; inversion of configuration mechanism;
D O I：
10.1016/S0008-6215(03)00165-4
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
A gene encoding for a thermostable exopolygalacturonase (exo-PG) from hyperthermophilic Thermotoga maritima has been cloned into a T7 expression vector and expressed in Escherichia coli. The gene encoded a polypeptide of 454 residues with a molecular mass of 51,304 Da. The recombinant enzyme was purified to homogeneity by heat treatment and nickel affinity chromatography. The thermostable enzyme had maximum of hydrolytic activity for polygalacturonate at 95 degreesC, pH 6.0 and retains 90% of activity after heating at 90 degreesC for 5 h. Study of the catalytic activity of the exopolygalacturonase, investigated by means of H-1 NMR spectroscopy revealed an inversion of configuration during hydrolysis of alpha-(1 --> 4)-galacturonic linkage. (C) 2003 Elsevier Science Ltd. All rights reserved.
引用
收藏
页码：1333 / 1337
页数：5
相关论文
共 24 条
  • [1] Inversion of configuration during hydrolysis of alpha-1,4-galacturonidic linkage by three Aspergillus polygalacturonases
    Biely, P
    Benen, J
    Heinrichova, K
    Kester, HCM
    Visser, J
    [J]. FEBS LETTERS, 1996, 382 (03) : 249 - 255
  • [2] Preliminary characterization of a new exo-beta-(1,4)-galactanase with transferase activity
    Bonnin, E
    Lahaye, M
    Vigouroux, J
    Thibault, JF
    [J]. INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, 1995, 17 (06) : 345 - 351
  • [3] Thermostability, oligomerization and DNA-binding properties of the regulatory protein ArgR from the hyperthermophilic bacterium Thermotoga neapolitana
    Dimova, D
    Weigel, P
    Takahashi, M
    Marc, F
    Van Duyne, GD
    Sakanyan, V
    [J]. MOLECULAR AND GENERAL GENETICS, 2000, 263 (01): : 119 - 130
  • [4] Purification and characterization of an exo-polygalacturonase from the tomato vascular wilt pathogen Fusarium oxysporum f sp lycopersici
    DiPietro, A
    Roncero, MIG
    [J]. FEMS MICROBIOLOGY LETTERS, 1996, 145 (02) : 295 - 299
  • [5] FISHMAN WH, 1957, J BIOL CHEM, P435
  • [6] GEBLER J, 1992, J BIOL CHEM, V267, P12559
  • [7] PURIFICATION AND CHARACTERIZATION OF AN EXTRACELLULAR EXO-D-GALACTURONANASE OF ASPERGILLUS-NIGER
    HEINRICHOVA, K
    REXOVABENKOVA, L
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA, 1976, 422 (02) : 349 - 356
  • [8] Structural and sequence-based classification of glycoside hydrolases
    Henrissat, B
    Davies, G
    [J]. CURRENT OPINION IN STRUCTURAL BIOLOGY, 1997, 7 (05) : 637 - 644
  • [9] Primary structure and characterization of an exopolygalacturonase from Aspergillus tubingensis
    Kester, HCM
    KustersVanSomeren, MA
    Muller, Y
    Visser, J
    [J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 1996, 240 (03): : 738 - 746
  • [10] Purification and characterization of a β-glucuronidase from Aspergillus niger
    Kuroyama, H
    Tsutsui, N
    Hashimoto, Y
    Tsumuraya, Y
    [J]. CARBOHYDRATE RESEARCH, 2001, 333 (01) : 27 - 39
123