Characterization of a Thermostable β-glucosidase from the Hyperthermophilic Archaeon Thermococcus pacificus

Title
Characterization of a Thermostable β-glucosidase from the Hyperthermophilic Archaeon Thermococcus pacificus
Author(s)
이재은; 김윤재; 이현숙; 강성균; 이정현; 권개경
KIOST Author(s)
Lee, Hyun Sook(이현숙)Kang, Sung Gyun(강성균)Lee, Jung Hyun(이정현)Kwon, Kae Kyoung(권개경)
Publication Year
2014-05-02
Abstract
Hyperthermophilc archaeon, Thermococcus pacificus (Topt of 80-88 °C) could grow on various substrates such as peptides, starch, and so on. Based on the genomic analysis of the strain, we identified a novel GH1 β-glucosidase encoding gene (PAC_orf01364), which was located in glycoside hydrolase gene clusters consisting of cellulose-, laminarin-, and agarose-degrading enzymes. The gene revealed an ORF of 1,464 bp encoding 487 amino acid residues, and the deduced amino acid sequence showed 77% identity with Pyrococcus furiosus β-glucosidase (accession no. NP_577802). The gene was cloned and expressed in Escherchia coli system. The recombinant protein was purified by metal affinity chromatography and characterized. The purified enzyme shows optimum activity at pH 5.5 and 75 °C, and thermostability with a half life of 6 h at 90 °C. Interestingly, the enzyme also exhibits laminarinase activity and β-xylosidase activity.(PAC_orf01364), which was located in glycoside hydrolase gene clusters consisting of cellulose-, laminarin-, and agarose-degrading enzymes. The gene revealed an ORF of 1,464 bp encoding 487 amino acid residues, and the deduced amino acid sequence showed 77% identity with Pyrococcus furiosus β-glucosidase (accession no. NP_577802). The gene was cloned and expressed in Escherchia coli system. The recombinant protein was purified by metal affinity chromatography and characterized. The purified enzyme shows optimum activity at pH 5.5 and 75 °C, and thermostability with a half life of 6 h at 90 °C. Interestingly, the enzyme also exhibits laminarinase activity and β-xylosidase activity.
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/26295
Bibliographic Citation
한국미생물학회 춘계 학술대회, pp.252, 2014
Publisher
한국미생물학회
Type
Conference
Language
English
Publisher
한국미생물학회
Related Researcher
Research Interests

marine biotechnology,molecular microbiology,해양생명공학,분자미생물학

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