| Alanine racemase from the acidophile Acetobacter aceti. | |
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MedLine Citation:
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PMID: 16843006 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Acetobacter aceti converts ethanol to acetic acid, and survives acetic acid exposure by tolerating cytoplasmic acidification. Alanine racemase (Alr) is a pyridoxal 5' phosphate (PLP) -dependent enzyme that catalyzes the interconversion of the d- and l-isomers of alanine and has a basic pH optimum. Since d-alanine is essential for peptidoglycan biosynthesis, Alr must somehow function in the acidic cytoplasm of A. aceti. We report the partial purification of native A. aceti Alr (AaAlr) and evidence that it is a rather stable enzyme. The C-terminus of AaAlr has a strong resemblance to the ssrA-encoded protein degradation signal, which thwarted initial protein expression experiments. High-activity AaAlr forms lacking a protease recognition sequence were expressed in Escherichia coli and purified. Biophysical and enzymological experiments confirm that AaAlr is intrinsically acid-resistant, yet has the catalytic properties of an ordinary Alr. |
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Authors:
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Julie A Francois; T Joseph Kappock |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S. Date: 2006-06-06 |
Journal Detail:
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Title: Protein expression and purification Volume: 51 ISSN: 1046-5928 ISO Abbreviation: Protein Expr. Purif. Publication Date: 2007 Jan |
Date Detail:
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Created Date: 2006-12-04 Completed Date: 2007-03-16 Revised Date: 2007-12-03 |
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: 39-48 Citation Subset: IM |
Affiliation:
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Department of Chemistry, Washington University in St. Louis, St. Louis, MO 63130, USA. |
| Data Bank Information | |
Bank Name/Acc. No.:
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GENBANK/DQ344920; DQ344921 |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Acetobacter
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enzymology* Alanine Racemase / antagonists & inhibitors, genetics, isolation & purification*, metabolism Circular Dichroism Electrophoresis, Polyacrylamide Gel Enzyme Stability Hydrogen-Ion Concentration Kinetics Molecular Sequence Data Recombinant Fusion Proteins / antagonists & inhibitors, isolation & purification |
| Grant Support | |
ID/Acronym/Agency:
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P41RR0954/RR/NCRR NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Recombinant Fusion Proteins; EC 5.1.1.1/Alanine Racemase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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