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High-Level Expression in Escherichia coli of Alkaline Phosphatase from Thermus caldophilus GK24 and Purification of the Recombinant Enzyme  

Lee, Jung-Ha (Department of Genetic Engineering, Sungkyunkwan University)
Cho, Yong-Duk (Department of Genetic Engineering, Sungkyunkwan University)
Choi, Jeong-Jin (Department of Genetic Engineering, Sungkyunkwan University)
Lee, Yoon-Jin (Department of Genetic Engineering, Sungkyunkwan University)
Hoe, Hyang-Sook (Department of Genetic Engineering, Sungkyunkwan University)
Kim, Hyun-Kyu (Genetic Resource R&D Institute, Super Bio Co., Ltd.)
Kwon, Suk-Tae (Department of Genetic Engineering, Sungkyunkwan University)
Publication Information
Journal of Microbiology and Biotechnology / v.13, no.5, 2003 , pp. 660-665 More about this Journal
Abstract
High-level expression of Thermus caldophilus GK24 alkaline phosphatase (Tca APase) was achieved in Escherichia coli using the pET-based expression plasmids, pEAP1 and pEAP2. In the case of plasmid pEAP2, the signal peptide region of Tca APase was replaced by the PelB leader peptide of expression vector pET-22b(+). Furthermore, the expression level was somewhat higher than that of plasmid pEAPl. A rapid purification procedure of Tca APase overproduced in E. coli was developed which involved heating to denature E. coli proteins followed by HiTrap Heparin HP column chromatography. Optimal temperature and pH and $Mg^{2+}$ dependence of the recombinant Tca APase were similar to those of native enzyme isolated from T. caldophilus GK24.
Keywords
Alkaline phosphatase; high-level expression; recombinant Tca alkaline phosphatase; Thermus caldophilus GK24; Thermus caldophilus GK24 alkaline phosphatase;
Citations & Related Records
Times Cited By KSCI : 2  (Citation Analysis)
Times Cited By Web Of Science : 7  (Related Records In Web of Science)
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