![](/images/graphics-bg.png)
Purification, Characterization, and Effect of Thiol Compounds on Activity of the Erwinia carotovora L-Asparaginase
Joint Authors
Khobragade, Chandrahas N.
Warangkar, Suchita C.
Source
Issue
Vol. 2010, Issue 2010 (31 Dec. 2010), pp.1-10, 10 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2009-11-01
Country of Publication
Egypt
No. of Pages
10
Main Subjects
Abstract EN
L-asparaginase was extracted from Erwinia carotovora and purified by ammonium sulfate fractionation (60–70%), Sephadex G-100, CM cellulose, and DEAE sephadex chromatography.
The apparent Mr of enzyme under nondenaturing and denaturing conditions was 150 kDa and 37±0.5 kDa, respectively.
L-asparaginase activity was studied in presence of thiols, namely, L-cystine (Cys), L-methionine (Met), N-acetyl cysteine (NAC), and reduced glutathione (GSH).
Kinetic parameters in presence of thiols (10–400 μM) showed an increase in Vmax values (2000, 2223, 2380, 2500, and control 1666.7 μmoles mg-1min-1) and a decrease in Km values (0.086, 0.076, 0.062, 0.055 and control 0.098 mM) indicating nonessential mode of activation.
KA values displayed propensity to bind thiols.
A decrease in Vmax/Km ratio in concentration plots showed inverse relationship between free thiol groups (NAC and GSH) and bound thiol group (Cys and Met).
Enzyme activity was enhanced in presence of thiol protecting reagents like dithiothreitol (DTT), 2-mercaptoethanol (2-ME), and GSH, but inhibited by p-chloromercurybenzoate (PCMB) and iodoacetamide (IA).
American Psychological Association (APA)
Warangkar, Suchita C.& Khobragade, Chandrahas N.. 2009. Purification, Characterization, and Effect of Thiol Compounds on Activity of the Erwinia carotovora L-Asparaginase. Enzyme Research،Vol. 2010, no. 2010, pp.1-10.
https://search.emarefa.net/detail/BIM-451156
Modern Language Association (MLA)
Warangkar, Suchita C.& Khobragade, Chandrahas N.. Purification, Characterization, and Effect of Thiol Compounds on Activity of the Erwinia carotovora L-Asparaginase. Enzyme Research No. 2010 (2010), pp.1-10.
https://search.emarefa.net/detail/BIM-451156
American Medical Association (AMA)
Warangkar, Suchita C.& Khobragade, Chandrahas N.. Purification, Characterization, and Effect of Thiol Compounds on Activity of the Erwinia carotovora L-Asparaginase. Enzyme Research. 2009. Vol. 2010, no. 2010, pp.1-10.
https://search.emarefa.net/detail/BIM-451156
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-451156