Cloning, Analysis, and Expression of the Gene for Thermostable Polyphosphate Kinase of Thermus caldophilus GK24 and Properties of the Recombinant Enzyme

Cloning, Analysis, and Expression of the Gene for Thermostable Polyphosphate Kinase of Thermus caldophilus GK24 and Properties of the Recombinant Enzyme

The gene encoding Thermus caldophilus GK24 polyphosphate kinase was cloned and sequenced. The gene contains an open reading frame encoding 608 amino acids with a calculated molecular mass of 69,850 Da. The deduced amino acid sequence of Tca PPK showed a 40% homology to Escherichia coli PPK, and 39%...

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Veröffentlicht in:Journal of microbiology and biotechnology 2003, 13(1), , pp.139-145
Hauptverfasser: Hoe, H.S, Lee, S.K, Kwon, S.T. (Sungkyunkwan University, Suwon, Republic of Korea), Lee, D.S. (Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea)
Format: Artikel
Sprache:eng
Schlagworte:
Biological and medical sciences
Biology of microorganisms of confirmed or potential industrial interest
Biotechnology
Fundamental and applied biological sciences. Psychology
gene expression
Genetics
Mission oriented research
polyphosphates
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Zusammenfassung:The gene encoding Thermus caldophilus GK24 polyphosphate kinase was cloned and sequenced. The gene contains an open reading frame encoding 608 amino acids with a calculated molecular mass of 69,850 Da. The deduced amino acid sequence of Tca PPK showed a 40% homology to Escherichia coli PPK, and 39% to Klebsiella aerogenes PPK. The Tca ppk gene was expressed under the control of the T7lac promoter on pET-22b(+) in E. coli and its enzyme was purified about 70-fold with 36% yield, following heating and HiTrap chelating HP column chromatography.
ISSN:1017-7825