Cloning, expression, and characterization of a methionyl aminopeptidase from a hyperthermophilic Archaeon Thermococcus sp NA1 SCIE SCOPUS

Cited 4 time in WEB OF SCIENCE Cited 5 time in Scopus
Title
Cloning, expression, and characterization of a methionyl aminopeptidase from a hyperthermophilic Archaeon Thermococcus sp NA1
Author(s)
Lee, H. S.; Kim, Y. J.; Bae, S. S.; Jeon, J. H.; Lim, J. K.; Jeong, B. C.; Kang, S. G.; Lee, J. -H.
KIOST Author(s)
Lee, Hyun Sook(이현숙)Kim, Yun Jae(김윤재)Lim, Jae Kyu(임재규)Kang, Sung Gyun(강성균)Lee, Jung Hyun(이정현)
Alternative Author(s)
이현숙; 김윤재; 배승섭; 임재규; 강성균; 이정현
Publication Year
2006-07
Abstract
Genomic analysis of a hyperthermophilic archaeon Thermococcus sp. NA1 revealed the presence of an 885-bp open reading frame encoding a protein of 295 amino acids with a calculated molecular mass of 32,981 Da. Analysis of the deduced amino acid sequence showed that amino acid residues important for catalytic activity and the metal binding ligands conserved in all of methionyl aminopeptidases (Met AP) were also conserved and belonged to type IIa Met AP. The protein, designated TNA1_MetAP (Thermococcus sp. NA1 MetAP), was cloned and expressed in Escherichia coli. The recombinant enzyme was a Mn2+-, Ni2+-, Fe2+-, or Co2+-dependent metallopeptidase. Optimal MetAP activity against (L)-methionine p-nitroanilide (Met-pNA) (K-m = 0.68 mM) occurred at pH 7.0 and 80 to 90 degrees C. The MetAP was very unstable compared to Pyrococcus furiosus MetAP, which was completely inactivated by heating at 80 degrees C for 5 min. It seemed likely that the cysteine residue (Cys53) played a critical role in regulating the thermostability of TNA1(-)MetAP.
ISSN
1436-2228
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/4868
DOI
10.1007/s10126-005-6124-8
Bibliographic Citation
MARINE BIOTECHNOLOGY, v.8, no.4, pp.425 - 432, 2006
Publisher
SPRINGER
Subject
ESCHERICHIA-COLI; PYROCOCCUS-FURIOSUS; SACCHAROMYCES-CEREVISIAE; MOLECULAR-CLONING; CRYSTAL-STRUCTURE; OVEREXPRESSION; GENE; MECHANISM; PROTEINS; SEQUENCE
Keywords
characterization; cloning; expression; genomic analysis; hyperthermophile; methionyl aminopeptidase (MetAP)
Type
Article
Language
English
Document Type
Article
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