Implantation de la solution technique UREG au CHU de Charleroi
Purification of urease from Pleurotus sajor-caju. Characterisation of GTPase activity of UreG and...
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Login Register Shibboleth Mobile Cart Home Journals Books Authors Librarians Societies About Us Contact Help Français Home > Authors Services o Submissions o Just-IN o OpenArticle o Reprints & permissions o Video abstracts o Plain language summaries Information o Copyright forms o Permission forms o Graphics o Authors' rights o Resources Publishing Toolkit for Authors The following is a list of all the forms and information you need to submit an article to an NRC Research Press journal. 1. Files & Graphics 2. Copyright agreements for publishing 3. Permission forms 4. Your responsibilities as an author 5. General ethical guidelines Purification of Urease from Pleurotus sajor-caju. Characterisation of GTPase Activity of UreG and Biochemical Properties of the enzyme Antik K. Bose ABSTRACT Biochemical properties of urease obtained from fruiting body extract of Pleurotus sajor- caju have been described. The enzyme was purified 3176.623 fold by CM-cellulose, DEAE- cellulose and Sephadex G-200 column chromatography. The molar mass estimated by Sephadex G-200 was 510KD and saccharide content was 4%. 6% SDS-PAGE of purified urease showed subunits UreA/B homodimer (28.8 KD ), UreC (150 KD), UreD (104 KD), UreE (21.5 KD), UreF (102 KD) and UreG (103KD). GTPase activity of UreG was characterized with K M , V max & K cat to be 2.6mM, 1.1μ mol GTP hydrolyzed/ min and 0.27 min -1 . The enzyme hydrolyzed urea with K M and V max to be 1.85mM and 64.51 μ mol NH3 C and 7.4 respectively. The enzyme activity was enhanced by Ca 2+ (2.94%), 5-10 mM, Mn 2+ ( 2.67-52.406%), 20-50mM Ni 2+ (2.6-6.93% ) and 5-10mM Mncl 2 ( 0.58- 8.02% ). The enzyme showed product inhibition by NH 4 + . Competitive inhibition by thiourea and uncompetitive inhibition by NaF were also studied. Amino acids at the active site have been identified to be Histidine and Aspartic acid using DEPC and Diazomethane with EDAC
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Transcript of Purification of urease from Pleurotus sajor-caju. Characterisation of GTPase activity of UreG and...
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About Us
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Français
Home > Authors
Services
o Submissions
o Just-IN
o OpenArticle
o Reprints & permissions
o Video abstracts
o Plain language summaries
Information
o Copyright forms
o Permission forms
o Graphics
o Authors' rights
o Resources
Publishing Toolkit for Authors
The following is a list of all the forms and
information you need to submit an article to
an NRC Research Press journal.
1. Files & Graphics
2. Copyright agreements for
publishing
3. Permission forms
4. Your responsibilities as an
author
5. General ethical guidelines
Purification of Urease from Pleurotus sajor-caju.
Characterisation of GTPase Activity of UreG and
Biochemical Properties of the enzyme
Antik K. Bose
ABSTRACT
Biochemical properties of urease obtained from fruiting body extract of Pleurotus sajor-
caju have been described. The enzyme was purified 3176.623 fold by CM-cellulose, DEAE-
cellulose and Sephadex G-200 column chromatography. The molar mass estimated by
Sephadex G-200 was 510KD and saccharide content was 4%. 6% SDS-PAGE of purified
urease showed subunits UreA/B homodimer (28.8 KD ), UreC (150 KD), UreD (104 KD),
UreE (21.5 KD), UreF (102 KD) and UreG (103KD). GTPase activity of UreG was
characterized with KM, Vmax & Kcat to be 2.6mM, 1.1µ mol GTP hydrolyzed/ min and 0.27
min-1
. The enzyme hydrolyzed urea with KM and Vmax to be 1.85mM and 64.51 µ mol NH3
C and 7.4 respectively. The enzyme
activity was enhanced by Ca2+
(2.94%), 5-10 mM, Mn2+
( 2.67-52.406%), 20-50mM Ni2+
(2.6-6.93% ) and 5-10mM Mncl2 ( 0.58- 8.02% ). The enzyme showed product inhibition by
NH4+. Competitive inhibition by thiourea and uncompetitive inhibition by NaF were also
studied. Amino acids at the active site have been identified to be Histidine and Aspartic
acid using DEPC and Diazomethane with EDAC
