Thermostable glutamate dehydrogenase from a commensal thermophile, Symbiobacterium toebii; overproduction, characterization, and application

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Title
Thermostable glutamate dehydrogenase from a commensal thermophile, Symbiobacterium toebii; overproduction, characterization, and application
Author(s)
Jae Seok Ha; K Kim; Jae Jun Song; Jin-Woo Bae; Seung Goo LeeSang Chul LeeHaryoung Poo; C S Shin; M H Sung
Bibliographic Citation
Journal of Molecular Catalysis B: Enzymatic, vol. 26, no. 3, pp. 231-240
Publication Year
2003
Abstract
A gene encoding glutamate dehydrogenase (GDH) was found in the genome sequence of a commensal thermophile, Symbiobacterium toebii. The amino acid sequence deduced from the gdh I of S. toebii was well conserved with other thermostable GDHs. The gdh I which encodes GDH consisting of 409 amino acids was cloned and expressed in E. coli DH5α under the control of a highly constitutive expression (HCE) promoter in a pHCE system. The recombinant GDH was expressed without addition of any inducers in a soluble form. The molecular mass of the GDH was estimated to be 263kDa by Superose 6 HR gel filtration chromatography and 44kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) indicating that the GDH was composed of hexameric form. The optimal temperature and pH of the purified enzyme were 60°C and 9.0, respectively, and the purified GDH retained more than 75% of its original activity after an incubation at 70°C for 30min. Although NADP(H) was the preferred cofactor, S. toebii GDH was able to utilize either NADP(H) or NAD(H) as coenzyme.
Keyword
glutamate dehydrogenaseoverproductionregeneration systemsymbiobacterium toebiithermostability
ISSN
1381-1177
Publisher
Elsevier
DOI
http://dx.doi.org/10.1016/j.molcatb.2003.06.008
Type
Article
Appears in Collections:
Division of Bio Technology Innovation > SME Support Center > 1. Journal Articles
Synthetic Biology and Bioengineering Research Institute > 1. Journal Articles
Division of Biomedical Research > Metabolic Regulation Research Center > 1. Journal Articles
Division of Research on National Challenges > Infectious Disease Research Center > 1. Journal Articles
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