| Display of active enzymes on the cell surface of Escherichia coli using PgsA anchor protein and their application to bioconversion. | |
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MedLine Citation:
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PMID: 16133338 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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We have developed a novel Escherichia coli cell surface display system by employing PgsA as an anchoring motif. In our display system, C-terminal fusion to PgsA anchor protein from Bacillus subtilis was used. The enzymes selected for display were alpha-amylase (AmyA) from Streptococcus bovis 148 and lipase B (CALB) from Candida antarctica. The molecular mass values of AmyA and CALB are approximately 77 and 34 kDa, respectively. The enzymes were displayed on the surface as a fusion protein with a FLAG peptide tag at the C terminus. Both the PgsA-AmyA-FLAG and PgsA-CALB-FLAG fusion proteins were shown to be displayed by immunofluorescence labeling using anti-FLAG antibody. The displayed enzymes were active forms, and AmyA and CALB activities reached 990 U/g (dry cell weight) and 4.6 U/g (dry cell weight), respectively. AmyA-displaying E. coli cells grew utilizing cornstarch as the sole carbon source, while CALB-displaying E. coli cells catalyzed enantioselective transesterification, indicating that they are effective whole-cell biocatalysts. Since a target enzyme with a size of 77 kDa and an industrially useful lipase have been successfully displayed on the cell surface of E. coli for the first time, PgsA protein is probably a useful anchoring motif to display various enzymes. |
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Authors:
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Junya Narita; Kenji Okano; Toshihiro Tateno; Takanori Tanino; Tomomitsu Sewaki; Moon-Hee Sung; Hideki Fukuda; Akihiko Kondo |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2005-08-20 |
Journal Detail:
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Title: Applied microbiology and biotechnology Volume: 70 ISSN: 0175-7598 ISO Abbreviation: Appl. Microbiol. Biotechnol. Publication Date: 2006 May |
Date Detail:
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Created Date: 2006-04-07 Completed Date: 2006-08-21 Revised Date: 2008-11-21 |
Medline Journal Info:
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Nlm Unique ID: 8406612 Medline TA: Appl Microbiol Biotechnol Country: Germany |
Other Details:
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Languages: eng Pagination: 564-72 Citation Subset: IM |
Affiliation:
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Division of Molecular Science, Graduate School of Science and Technology, Kobe University, Nada-ku, Kobe, 657-8501, Japan. |
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| MeSH Terms | |
Descriptor/Qualifier:
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Candida
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enzymology Catalysis Cell Membrane / metabolism* Enzymes, Immobilized / genetics, metabolism* Escherichia coli / cytology*, metabolism* Gene Expression Lipase / genetics, metabolism Membrane Proteins / genetics, metabolism* Recombinant Proteins Transferases (Other Substituted Phosphate Groups) / genetics, metabolism* alpha-Amylases / genetics, metabolism |
| Chemical | |
Reg. No./Substance:
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0/Enzymes, Immobilized; 0/Membrane Proteins; 0/Recombinant Proteins; EC 2.7.8.-/Transferases (Other Substituted Phosphate Groups); EC 2.7.8.5/CDP-diacylglycerol-glycerol-3-phosphate 3-phosphatidyltransferase; EC 3.1.1.-/lipase B, Candida antarctica; EC 3.1.1.3/Lipase; EC 3.2.1.1/alpha-Amylases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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