| Purification, characterization, gene cloning and nucleotide sequencing of D: -stereospecific amino acid amidase from soil bacterium: Delftia acidovorans. | |
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MedLine Citation:
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PMID: 15959727 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The D-amino acid amidase-producing bacterium was isolated from soil samples using an enrichment culture technique in medium broth containing D-phenylalanine amide as a sole source of nitrogen. The strain exhibiting the strongest activity was identified as Delftia acidovorans strain 16. This strain produced intracellular D-amino acid amidase constitutively. The enzyme was purified about 380-fold to homogeneity and its molecular mass was estimated to be about 50 kDa, on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme was active preferentially toward D-amino acid amides rather than their L-counterparts. It exhibited strong amino acid amidase activity toward aromatic amino acid amides including D-phenylalanine amide, D-tryptophan amide and D-tyrosine amide, yet it was not specifically active toward low-molecular-weight D-amino acid amides such as D-alanine amide, L-alanine amide and L-serine amide. Moreover, it was not specifically active toward oligopeptides. The enzyme showed maximum activity at 40 degrees C and pH 8.5 and appeared to be very stable, with 92.5% remaining activity after the reaction was performed at 45 degrees C for 30 min. However, it was mostly inactivated in the presence of phenylmethanesulfonyl fluoride or Cd2+, Ag+, Zn2+, Hg2+ and As3+ . The NH2 terminal and internal amino acid sequences of the enzyme were determined; and the gene was cloned and sequenced. The enzyme gene damA encodes a 466-amino-acid protein (molecular mass 49,860.46 Da); and the deduced amino acid sequence exhibits homology to the D-amino acid amidase from Variovorax paradoxus (67.9% identity), the amidotransferase A subunit from Burkholderia fungorum (50% identity) and other enantioselective amidases. |
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Authors:
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Tipparat Hongpattarakere; Hidenobu Komeda; Yasuhisa Asano |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2005-06-16 |
Journal Detail:
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Title: Journal of industrial microbiology & biotechnology Volume: 32 ISSN: 1367-5435 ISO Abbreviation: J. Ind. Microbiol. Biotechnol. Publication Date: 2005 Dec |
Date Detail:
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Created Date: 2005-12-05 Completed Date: 2006-02-14 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 9705544 Medline TA: J Ind Microbiol Biotechnol Country: England |
Other Details:
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Languages: eng Pagination: 567-76 Citation Subset: IM |
Affiliation:
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Faculty of Agro-Industry, Prince of Songkla University, Hat Yai, 90112, Songkla, Thailand. tipparat.h@psu.ac.th |
| Data Bank Information | |
Bank Name/Acc. No.:
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GENBANK/AB154822 |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Amidohydrolases*
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chemistry,
genetics,
isolation & purification,
metabolism Amino Acid Sequence Cloning, Molecular Delftia acidovorans / enzymology*, genetics, isolation & purification Molecular Sequence Data Sequence Analysis, DNA Soil Microbiology* Stereoisomerism Substrate Specificity |
| Chemical | |
Reg. No./Substance:
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EC 3.5.-/Amidohydrolases; EC 3.5.1.-/D-amino acid amidase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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