Characterization of prolyl oligopeptidase from hyperthermophilic archaeon Thermococcus sp NA1 SCIE SCOPUS

Cited 6 time in WEB OF SCIENCE Cited 7 time in Scopus
Title
Characterization of prolyl oligopeptidase from hyperthermophilic archaeon Thermococcus sp NA1
Author(s)
Lee, Hyun Sook; Kim, Yun Jae; Cho, Yona; Kim, Sang-Jin; Lee, Jung-Hyun; Kang, Sung Gyun
KIOST Author(s)
Lee, Hyun Sook(이현숙)Kim, Yun Jae(김윤재)Lee, Jung Hyun(이정현)Kang, Sung Gyun(강성균)
Alternative Author(s)
이현숙; 김윤재; 조요나; 김상진; 이정현; 강성균
Publication Year
2007-03
Abstract
The prolyl oligopeptidase TNA1_POP was found to be encoded in the genome of the hyperthermophilic archaeon Thermococcus sp. NA1 and showed high similarities to its archaeal homologs (76-83%). The enzyme was found to be a single polypeptide composed of 616 amino acids with conserved signature domains. A recombinant TNA1_POP expressed in Escherichia coli was capable of hydrolyzing succinyl-Ala-Pro-p-nitroanilide (Sue-Ala-Pro-pNA) with temperature and pH optimums of 80 degrees C and 7, respectively. TNA1_POP activity appeared to be significantly activated by pre-incubation at 80 degrees C and 90 degrees C with the optimum temperature unchanged. The heat-activated enzyme exhibited a k(cat), approximately twofold higher than that of the unheated enzyme, however, both enzymes showed the same K-m TNA1_POP was thermostable at 80 degrees C retaining 80% of its heat-activated activity even after 23 h, but it lost its enzymatic activity at 90 degrees C with a half-life of 3 h. The loss of the enzymatic activity at 90 degrees C seemed to be caused by the autodegradation of the enzyme, not by thermal denaturation, as supported by circular dichroism spectropolarimetry. Autodegradation fragments ranging from 2 to 18 kDa were mapped by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry.
ISSN
1389-1723
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/4727
DOI
10.1263/jbb.103.221
Bibliographic Citation
JOURNAL OF BIOSCIENCE AND BIOENGINEERING, v.103, no.3, pp.221 - 228, 2007
Publisher
SOC BIOSCIENCE BIOENGINEERING JAPAN
Subject
PYROCOCCUS-FURIOSUS; ACTIVE-SITE; CLEAVING ENZYME; ENDOPEPTIDASE; PEPTIDASES; CLONING; CELLS; SEQUENCE; SPECIFICITY; EXPRESSION
Keywords
prolyl oligopeptidase; hyperthermophile; archaea; Thermococcus; autodegradation
Type
Article
Language
English
Document Type
Article
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