Molecular cloning and characterization of Mn-superoxide dismutase from disk abalone (Haliotis discus discus) SCIE SCOPUS

Cited 35 time in WEB OF SCIENCE Cited 36 time in Scopus
Title
Molecular cloning and characterization of Mn-superoxide dismutase from disk abalone (Haliotis discus discus)
Author(s)
Ekanayake, Prashani Mudika; Kang, Hyun-Sil; De Zyosa, Mahanama; Jee, Youngheun; Lee, Youn-Ho; Lee, Jehee
KIOST Author(s)
Lee, Youn Ho(이윤호)
Publication Year
2006-11
Abstract
The mitochondrial enzyme manganese superoxide dismutase (mitMn-SOD) is one of the antioxidant enzymes involved in cellular defense against oxidative stress and catalyzes the conversion of O-2(.-) into the stabler H2O2. In this study, a putative gene encoding Mn-SOD from disk abalone (Haliotis discus discus, aMn-SOD) was cloned, sequenced, expressed in Escherichia coli K12(TB1) and the protein was purified using pMAL protein purification system. Sequencing resulted ORF of 681 bp, which corresponded to 226 amino acids. The protein was expressed in soluble form with molecular weight of 68 kDa including maltose binding protein and pI value of 6.5. The fusion protein had 2781 U/mg activity. The optimum temperature of the enzyme was 37 degrees C and it was active in a range of acidic pH (from 3.5 to 6.5). The enzyme activity was reduced to 50% at 50 degrees C and completely heat inactivated at 80 degrees C. The alignment of aMn-SOD amino acid sequence with Mn-SODs available in NCBI revealed that the enzyme is conserved among animals with higher than 30% identity. In comparison with human mitMn-SOD, all manganese-binding sites are also conserved in aMn-SOD (H28, H100, D185 and H189). aMn-SOD amino acid sequence was closer to that of Biomphalaria glabrata in phylogenetic analysis. (c) 2006 Elsevier Inc. All rights reserved.
ISSN
1096-4959
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/40489
DOI
10.1016/j.cbpb.2006.08.004
Bibliographic Citation
COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY, v.145, no.3-4, pp.318 - 324, 2006
Publisher
ELSEVIER SCIENCE INC
Subject
SEQUENCE ALIGNMENT; GENE-EXPRESSION; SEEDLINGS; INJURY; CELLS; MICE; PURIFICATION; EVOLUTION; SOD2
Keywords
Haliotis discus discus; antioxidant enzyme; manganese superoxide dismutase (Mn-SOD); protein purification; reactive oxygen species (ROS); cDNA; cloning
Type
Article
Language
English
Document Type
Article
Publisher
ELSEVIER SCIENCE INC
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