Cloning, expression, and characterization of a hyperalkaline phosphatase from the thermophilic bacterium Thermus sp T351
- Authors
- Choi, JJ[Choi, JJ]; Park, JW[Park, JW]; Shim, H[Shim, H]; Lee, S[Lee, S]; Kwon, M[Kwon, M]; Yang, JS[Yang, JS]; Hwang, H[Hwang, H]; Kwon, ST[Kwon, ST]
- Issue Date
- Feb-2006
- Publisher
- KOREAN SOC MICROBIOLOGY & BIOTECHNOLOGY
- Keywords
- alkaline phosphatase; sequence analysis; thermostable enzyme; Thermus sp T351
- Citation
- JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY, v.16, no.2, pp.272 - 279
- Indexed
- SCIE
SCOPUS
KCI
- Journal Title
- JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY
- Volume
- 16
- Number
- 2
- Start Page
- 272
- End Page
- 279
- URI
- https://scholarworks.bwise.kr/skku/handle/2021.sw.skku/87673
- ISSN
- 1017-7825
- Abstract
- The gene encoding Thermus sp. T351 alkaline phosphatase (T351 APase) was cloned and sequenced. The gene consisted of 1,503 bp coding for a protein with 500 amino acid residues including a signal peptide. The deduced amino acid sequence of T351 APase showed relatively low similarity to other Thermus APases. The T351 APase gene was expressed under the control of the T7lac promoter on the expression vector pET-22b(+) in Escherichia coli BL21(DE3). The expressed enzyme was purified by heat treatment, and UNO (TM) Q and HiTrap (TM) Heparin HP column chromatographies. The purified enzyme exhibited high activity at extremely alkaline pHs, reaching a maximum at pH 12.0. The optimum temperature of the enzyme was 80 degrees C, and the half-life at 85 degrees C was approximately 103 min. The enzyme activity was found to be dependent on metal ions: the addition of Mg2+ and Co2+ increased the activity, whereas EDTA inhibited it. With p-nitrophenyl phosphate as the substrate, T351 APase had a Michaelis constant (K-m) of 3.9 x 10(-5) M. The enzyme catalyzed the hydrolysis of a wide variety of phosphorylated compounds.
- Files in This Item
- There are no files associated with this item.
- Appears in
Collections - Biotechnology and Bioengineering > Department of Bio-mechatronic Engineering > 1. Journal Articles
- Biotechnology and Bioengineering > Integrative Biotechnology > 1. Journal Articles
- Biotechnology and Bioengineering > Department of Genetic Engineering > 1. Journal Articles
Items in ScholarWorks are protected by copyright, with all rights reserved, unless otherwise indicated.