Mode of Action and Partial Purification of the Active Centre of exo-Poly-α-D-galacturonosidase from Selenomonas ruminantium

[img]PDF - Authorized users only
Language: English
814Kb
Title:Mode of Action and Partial Purification of the Active Centre of exo-Poly-α-D-galacturonosidase from Selenomonas ruminantium
Creators:
Heinrichová, Kveta
Heinrich, Július
Dzúrová, Mária
Ziolecki, Alexander
Journal or Publication Title:
Collection of Czechoslovak Chemical Communications, 58, 3, pp. 681-692

Abstract

In the presented paper are summarized results of the study of the mode of action, dimensions and arrangement of the active centre of the exo-poly-α-D-galacturonosidase, (poly(1,4-α-D-galactosiduronate) digalacturonohydrolase, E.C. 3.2.1.82) produced by the bacteria <i>Selenomonas ruminantium</i>. With this aim we determined experimentally values of Michaelis constants and limiting rates for the catalytic hydrolysis of linear oligo(D-galactosiduronates) of the degree of polymeration in the range of 3 to 8, at pH 7.0 and the temperature 30 °C. We calculated molecular activities <i>k</i><sub>0</sub> and parameters <i>k</i><sub>0</sub>/<i>K</i><sub>m</sub> from these values. From the dependence of log<i>k</i><sub>0</sub> and log<i>k</i><sub>0</sub>/<i>K</i><sub>m</sub> on the degree of polymerization five subsites of the active centre were determined, with the catalytic site being situated between the second and third one. Kinetic data were used for the calculation of the affinities of the fourth and fifth subsites <i>A</i><sub>4</sub> and <i>A</i><sub>5</sub> in accordance with the theory of Hiromi. Product analysis of non-labelled oligo(D-galactosiduronates) and compounds labelled with [1-<sup>3</sup>H] at the reducing end anabled to ascertain approximately the value for the first subsite <i>A</i><sub>1</sub> of the active centre and to study the mode of action of the exo-poly-α-D-galacturonosidase from <i>Selenomonas ruminantium</i>.

Title:Mode of Action and Partial Purification of the Active Centre of exo-Poly-α-D-galacturonosidase from Selenomonas ruminantium
Creators:
Heinrichová, Kveta
Heinrich, Július
Dzúrová, Mária
Ziolecki, Alexander
Divisions:Life and Chemical Sciences > Institute of Organic Chemistry and Biochemistry > Collection of Czechoslovak Chemical Communications
Journal or Publication Title:Collection of Czechoslovak Chemical Communications
Volume:58
Number:3
Page Range:pp. 681-692
ISSN:0010-0765
E-ISSN:1212-6950
Publisher:Institute of Organic Chemistry and Biochemistry
Related URLs:
URLURL Type
http://dx.doi.org/10.1135/cccc19930681UNSPECIFIED
ID Code:190
Item Type:Article
Deposited On:06 Feb 2009 16:58
Last Modified:06 Feb 2009 15:59

Citation

Heinrichová, Kveta; Heinrich, Július; Dzúrová, Mária; Ziolecki, Alexander (1993) Mode of Action and Partial Purification of the Active Centre of exo-Poly-α-D-galacturonosidase from Selenomonas ruminantium. Collection of Czechoslovak Chemical Communications, 58 (3). pp. 681-692. ISSN 0010-0765

Repository Staff Only: item control page