This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Lee, H. S.
Right arrow Articles by Lee, J.-H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Lee, H. S.
Right arrow Articles by Lee, J.-H.
Agricola
Right arrow Articles by Lee, H. S.
Right arrow Articles by Lee, J.-H.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, March 2006, p. 1886-1890, Vol. 72, No. 3
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.3.1886-1890.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Cloning, Expression, and Characterization of Aminopeptidase P from the Hyperthermophilic Archaeon Thermococcus sp. Strain NA1

Hyun Sook Lee,1 Yun Jae Kim,1 Seung Seob Bae,1 Jeong Ho Jeon,1 Jae Kyu Lim,1 Byeong Chul Jeong,2 Sung Gyun Kang,1 and Jung-Hyun Lee1*

Korean Ocean Research & Development Institute, Ansan P.O. Box 29, Seoul 425-600, Korea,1 Division of Bioscience and Bioinformatics, Myongji University, Yongin 449-728, Korea2

Received 28 July 2005/ Accepted 7 December 2005

Genomic analysis of a hyperthermophilic archaeon, Thermococcus sp. strain NA1, revealed the presence of a 1,068-bp open reading frame encoding a protein consisting of 356 amino acids with a calculated molecular mass of 39,714 Da (GenBank accession no. DQ144132). Sequence analysis showed that it was similar to the putative aminopeptidase P (APP) of Thermococcus kodakaraensis KOD1. Amino acid residues important for catalytic activity and the metal binding ligands conserved in bacterial, nematode, insect, and mammalian APPs were also conserved in the Thermococcus sp. strain NA1 APP. The archaeal APP, designated TNA1_APP (Thermococcus sp. strain NA1 APP), was cloned and expressed in Escherichia coli. The recombinant enzyme hydrolyzed the amino-terminal Xaa-Pro bond of Lys(N{varepsilon}-Abz)-Pro-Pro-pNA and the dipeptide Met-Pro (Km, 0.96 mM), revealing its functional identity. Further enzyme characterization showed the enzyme to be a Co2+-, Mn2+-, or Zn2+-dependent metallopeptidase. Optimal APP activity with Met-Pro as the substrate occurred at pH 5 and a temperature of 100°C. The APP was thermostable, with a half-life of >100 min at 80°C. This study represents the first characterization of a hyperthermophilic archaeon APP.

* Corresponding author. Mailing address: Korean Ocean Research & Development Institute, Ansan P.O. Box 29, Seoul 425-600, Korea. Phone: 82-31-400-6243. Fax: 82-31-406-2495. E-mail: jlee{at}kordi.re.kr.

Applied and Environmental Microbiology, March 2006, p. 1886-1890, Vol. 72, No. 3
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.3.1886-1890.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»