Novel substrate specificity of a thermostable β-glucosidase from the hyperthermophilic archaeon, Thermococcus pacificus P-4 SCOPUS KCI

Title
Novel substrate specificity of a thermostable β-glucosidase from the hyperthermophilic archaeon, Thermococcus pacificus P-4
Author(s)
Kim, Y.J.; Lee, J.E.; Lee, H.S.; Kwon, K.K.; Kang, S.G.; Lee, J.-H.
KIOST Author(s)
Kim, Yun Jae(김윤재)Lee, Hyun Sook(이현숙)Kwon, Kae Kyoung(권개경)Kang, Sung Gyun(강성균)Lee, Jung Hyun(이정현)
Alternative Author(s)
김윤재; 이현숙; 권개경; 강성균; 이정현
Publication Year
2015
Abstract
Based on the genomic analysis of Thermococcus pacificus P-4, we identified a putative GH1 β-glucosidase-encoding gene (Tpa-glu). The gene revealed a 1,464 bp encoding 487 amino acid residues, and the deduced amino acid residues exhibited 77% identity with Pyrococcus furiosus β-glucosidase (accession no. NP_577802). The gene was cloned and expressed in Escherichia coli system. The recombinant protein was purified by metal affinity chromatography and characterized. Tpa-Glu showed optimum activity at pH 7.5 and 75°C, and thermostability with a half life of 6 h at 90°C. Tpa-Glu exhibited hydrolyzing activity against various pNP-glycopyranosides, with kcat/Km values in the order of pNP-β-glucopyranoside, pNP-β-galactopyranoside, pNP-β-mannopyranoside, and pNP-β-xylopyranoside. In addition, the enzyme exhibited exo-hydrolyzing activity toward β-1,3-linked polysaccharide (laminarin) and β-1,3-and β-1,4-linked oligosaccharides. This is the first description of an enzyme from hyperthermophilic archaea that displays exo-hydrolyzing activity toward β-1,3-linked polysaccharides and could be applied in combination with β-1,3-endoglucanase for saccharification of laminarin. © 2015, The Microbiological Society of Korea.
ISSN
0440-2413
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/2588
DOI
10.7845/kjm.2015.5003
Bibliographic Citation
Korean Journal of Microbiology, v.51, no.1, pp.68 - 74, 2015
Publisher
Microbiological Society of Korea
Subject
Archaea; Escherichia coli; Pyrococcus furiosus; Thermococcus pacificus
Keywords
Exo-hydrolyzing activity; Laminarinase activity; Thermococcus pacificus p-4; β-1,3-linked polysaccharide; β-glucosidase
Type
Article
Language
Korean
Document Type
Article
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