| Gene analysis and enzymatic properties of thermostable beta-glycosidase from Pyrococcus kodakaraensis KOD1. | |
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MedLine Citation:
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PMID: 16232586 Owner: NLM Status: PubMed-not-MEDLINE |
Abstract/OtherAbstract:
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A beta-glycosidase with broad substrate specificity was identified from a hyperthermophilic archaeon, Pyrococcus kodakaraensis KOD1. The gene encoding beta-glycosidase (Pk-gly) consists of 1449 nucleotides corresponding to a polypeptide of 483 amino acids. The protein showed similarity with other beta-glycosidases from family-1 glycosyl hydrolases, in particular, it showed high identity to beta-mannosidase from P. furiosus (55.7%), beta-glycosidase from Sulfolobus solfataricus (42.7%) and beta-glucosidase from P. furiosus (41.9%). The cloned gene was expressed in Escherichia coli and the recombinant protein was purified. The beta-glycosidase showed optimal activity at pH 6.5 and at an extremely high temperature of 100 degrees C, and had a half-life of 18 h at 90 degrees C. The beta-glycosidase hydrolyzed various pNp-beta-glycopyranosides, with kcat K(m) values in the order of pNp-beta-glucopyranoside = pNp-beta-mannopyranoside > pNp-beta-galactopyranoside > pNp-beta-xylopyranoside. pNp-beta-mannopyranoside was the substrate exhibiting the lowest K(m) value [0.254 mM] with a kcat K(m) ratio comparable to that of pNp-beta-glucopyranoside. This substrate specificity was distinct from previously reported beta-glycosidases. We observed that the region in PK-Gly corresponding to the fifth alpha-helix and beta-strand region of beta-glycosidase from S. solfataricus, which constitutes a large portion of the channel for substrate incorporation, displayed a chimeric structure, with the N-terminal region similar to beta-glycosidases and the C-terminal region similar to beta-mannosidases. An exo-type hydrolytic activity and transglycosylation activity were also observed towards cellooligomers. |
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Authors:
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S Ezaki; K Miyaoku; K Nishi; T Tanaka; S Fujiwara; M Takagi; H Atomi; T Imanaka |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Journal of bioscience and bioengineering Volume: 88 ISSN: 1389-1723 ISO Abbreviation: J. Biosci. Bioeng. Publication Date: 1999 |
Date Detail:
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Created Date: 2005-10-19 Completed Date: 2005-10-31 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 100888800 Medline TA: J Biosci Bioeng Country: Japan |
Other Details:
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Languages: eng Pagination: 130-5 Citation Subset: - |
Affiliation:
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Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Yoshida-Honmachi, Sakyo-ku, Kyoto 606-8501, Japan. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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