Molecular cloning and characterization of a novel cold-active beta-1,4-D-mannanase from the Antarctic springtail, Cryptopygus antarcticus SCIE SCOPUS

Cited 39 time in WEB OF SCIENCE Cited 43 time in Scopus
Title
Molecular cloning and characterization of a novel cold-active beta-1,4-D-mannanase from the Antarctic springtail, Cryptopygus antarcticus
Author(s)
Song, Jung Min; Nam, Ki-Woong; Kang, Sung Gyun; Kim, Choong-Gon; Kwon, Suk-Tae; Lee, Youn-Ho
KIOST Author(s)
Kang, Sung Gyun(강성균)Kim, Choong Gon(김충곤)Lee, Youn Ho(이윤호)
Publication Year
2008-09
Abstract
A cDNA encoding a beta-1,4-D-mannanase (CaMan) was identified among the expressed sequence tags (ESTs) of the Antarctic springtail, Ctyptopygus antarcticus. The open reading frame consisted of 1149 bp encoding 382 amino acids with a putative signal peptide. Amino acid sequence comparison with other mannanases indicated that CaMan likely belongs to subfamily 10 of the glycoside hydrolase family 5, together with mollusc beta-mannanases. CaMan shows typical features of a cold-active enzyme: it has a high frequency of polar residues such as Asn, Gln, and Ser, and a low frequency of hydrophobic residues as well as a low ratio of Arg/(Arg+Lys) compared to the mesophilic beta-mannanases. When CaMan was fused with the thioredoxin gene in pET-32a(+), expressed in E. coli Rosetta-gami (DE3), and purified after thrombin treatment, catalytically active enzyme was obtained. CaMan has high specific activity (416.3 U/mg) toward locust bean gum at an optimal temperature of 30 degrees C and an optimal pH of 3.5. Its optimal temperature is the lowest among those of the known mannanases and the optimal pH is also the lowest except those of fungi. Even at 0-5 degrees C, this enzyme retained 20-40% of its maximum activity. Divalent metal ions such as Ca2+, Mg2+, Cu2+, and Zn2+ enhanced the enzyme activity, but Mn2+, Hg2+, and Ag+ inhibited activity. This study represents the first record of a P-mannanase from an arthropod and provides a new source of carbohydrate hydrolysis enzyme with novel characteristics. (C) 2008 Elsevier Inc. All rights reserved.
ISSN
1096-4959
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/4457
DOI
10.1016/j.cbpb.2008.05.005
Bibliographic Citation
COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY, v.151, no.1, pp.32 - 40, 2008
Publisher
ELSEVIER SCIENCE INC
Subject
MUSSEL MYTILUS-EDULIS; BETA-MANNANASES; SUBSTRATE-SPECIFICITY; SEQUENCE ALIGNMENT; CRYSTAL-STRUCTURE; PICHIA-PASTORIS; PURIFICATION; EXPRESSION; ENZYMES; ADAPTATION
Keywords
Antarctic springtail; Cryptopygus antarcticus; beta-mannanase; cold activity; low pH activity; GH5 family
Type
Article
Language
English
Document Type
Article
Publisher
ELSEVIER SCIENCE INC
Related Researcher
Research Interests

Marine Molecular Genetics,Marine Ecology and Evolution,해양분자유전,해양생태및진화

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