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ID 1
Eprint ID
1
FullText URL
Title Alternative
Genetic Analysis, Expression in Eschericia coli of Aconitase from Chemolithotrophic Acidithiobacillus thiooxidans
Author
Kanahara, Yohei
Tokuda, Chizuka
Nakamura, Atsuo
Matsukawa, Hirokazu
Abstract
An aconitase from Acidithiobacillus thiooxidans was purified and characterized, and its gene was cloned. The cloned aconitase gene (acn) was expressed in Escherichia coli JM 109; aconitase activity was found in the cell extarct. The acn gene encodes a 646-amino acid polypeptide and is located upstream of the isocitrate dehydrogenase gene (icd). A. thiooxidans aconitase showes high sequence similar to pig heart aconitase and E.coli aconitase B. Twenty-five of twenty-seven active site residues assigned in pig heart aconitase are conserved in A. thiooxidans aconitase. The enzyme was purified by DEAE-Toyopearl 650M column chromatogrophy. The purified enzyme had an optimum pH of 7.5 and an optimum temperature of 60 C. Thermal inactivation studies of the purified enzyme revealed the enzyme activity to be uninfluenced after one hour incubation at 40 c. Enzyme activity was retained 100% after incubation of the enzyme at pH 6.0-9.0 for 60min. The A. thiooxidans aconitase was composed of a single polypeptide chain with a molecular mass of 66 kDa.
Keywords
Aconitase
Acidithiobacillus
Isocitrate dehydrogenase
Published Date
2005-02-01
Publication Title
岡山大学農学部学術報告
Publication Title Alternative
Scientific reports of the Faculty of Agriculture, Okayama University
Volume
volume94
Issue
issue1
Publisher
岡山大学農学部
Publisher Alternative
Faculty of Agriculture,Okayama University
Start Page
1
End Page
7
ISSN
0474-0254
NCID
AN00033029
Content Type
Departmental Bulletin Paper
language
日本語
File Version
publisher
Refereed
False
Eprints Journal Name
srfa