| Cloning, expression, purification and activation by Na ion of halophilic alkaline phosphatase from moderate halophile Halomonas sp. 593. | |
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MedLine Citation:
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PMID: 20883791 Owner: NLM Status: In-Data-Review |
Abstract/OtherAbstract:
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We have succeeded in the cloning of alkaline phosphatase gene, haalp, from moderate halophile Halomonas sp. 593. A deduced amino acid sequence showed a high ratio of acidic to basic amino acids, characteristic of halophilic proteins. The gene product was efficiently expressed in Escherichia coli BL21 Star (DE3) pLysS, but in an inactive form. The purified recombinant HaALP was separated into four fractions by gel filtration. When they were dialyzed against 50mM Tris-HCl (pH 8.0)/2mM MgCl(2) buffer containing 3M NaCl, one of these four fractions was activated to almost full activity. This fraction contained a folding intermediate that was converted to the native structure by the salt. Among the additional salts tested, i.e., KCl, KBr, LiCl, MgCl(2), (NH(4))(2)SO(4), and Na(2)SO(4), only Na(2)SO(4) was effective, suggesting the importance of Na ion. |
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Authors:
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Matsujiro Ishibashi; Kazuki Oda; Tsutomu Arakawa; Masao Tokunaga |
Publication Detail:
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Type: Journal Article Date: 2010-09-29 |
Journal Detail:
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Title: Protein expression and purification Volume: 76 ISSN: 1096-0279 ISO Abbreviation: Protein Expr. Purif. Publication Date: 2011 Mar |
Date Detail:
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Created Date: 2010-12-14 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9101496 Medline TA: Protein Expr Purif Country: United States |
Other Details:
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Languages: eng Pagination: 97-102 Citation Subset: IM |
Copyright Information:
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Copyright © 2010 Elsevier Inc. All rights reserved. |
Affiliation:
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Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan. |
Export Citation:
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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