Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/69785
DC FieldValueLanguage
dc.contributor.authorLin, L.L.en_US
dc.contributor.authorChen, M.H.en_US
dc.contributor.authorChien, H.C.R.en_US
dc.contributor.authorKan, S.C.en_US
dc.contributor.authorChen, C.C.en_US
dc.contributor.authorHu, H.Y.en_US
dc.contributor.authorHsu, W.H.en_US
dc.date2007zh_TW
dc.date.accessioned2014-06-11T05:58:54Z-
dc.date.available2014-06-11T05:58:54Z-
dc.identifier.issn0168-1656zh_TW
dc.identifier.urihttp://hdl.handle.net/11455/69785-
dc.description.abstractThe gene encoding a Deinococcus radiodurans R1 bifunctional aminoacylase/carboxypeptidase (DR_ACY/CP) was amplified by polymerase chain reaction and cloned into pQE-30 to generate pQE-DRAC. The cloned gene consists of an open reading frame of 1197 bp encoding a protein with a molecular mass of 42,729 Da. The predicted amino acid sequence shows high homology with those of Geobacillus kaustophilus aminoacylase, Geobacillus stearothermophilus aminoacylase, Pyrococcus horikoshii carboxypeptidase/aminoacylase and Thermoanaerobacter tengcongensis aminoacylase/carboxypeptidase. The expressed enzyme was purified from the crude extract of IPTG-induced Escherichia coli M15 (pQE-DRAC) to homogeneity by nickel-chelate chromatography. The molecular mass of the purified enzyme was determined to be 43 kDa by SDS-PAGE. Maximal aminoacylase activity with N-acetyl-methionine as the substrate occurred at pH 8.0 and 40 degrees C in the sodium phosphate buffer. The aminoacylase activity was strongly inhibited by metal-chelating agents, and was largely restored by divalent cations, such as Co2+, Mn2+ and Ni2+. The purified enzyme had broad specificity toward N-acetylated L-amino acids as well as N-CBZ-peptides. Carboxypeptidase activity of DR_ACY/CP to N-CBZ-Gly-Ala exhibited K-m and k(cat) values of 4.3 mM and 28 s(-1), respectively. The enzyme also had activity toward the cell wall-related substrates, D-Ala-Gly, D-Ala-Gly-Gly and L-Orn-L-Ala. (c) 2006 Elsevier B.V. All rights reserved.en_US
dc.language.isoen_USzh_TW
dc.relationJournal of Biotechnologyen_US
dc.relation.ispartofseriesJournal of Biotechnology, Volume 128, Issue 2, Page(s) 322-334.en_US
dc.relation.urihttp://dx.doi.org/10.1016/j.jbiotec.2006.10.011en_US
dc.subjectaminoacylaseen_US
dc.subjectcarboxypeptidaseen_US
dc.subjectDeinococcus radioduransen_US
dc.subjectgeneen_US
dc.subjectexpressionen_US
dc.subjectbiochemical characterizationen_US
dc.subjectcomplete genome sequenceen_US
dc.subjectarchaebacterium sulfolobus-solfataricusen_US
dc.subjectfineen_US
dc.subjectchemicalsen_US
dc.subjectl-aminoacylaseen_US
dc.subjectamino-acidsen_US
dc.subjectpurificationen_US
dc.subjectcarboxypeptidaseen_US
dc.subjectarchaeonen_US
dc.subjectgeneen_US
dc.subjectpharmaceuticalsen_US
dc.titleCharacterization of a bifunctional aminoacylase/carboxypeptidase from radioresistant bacterium Deinococcus radiodurans R1en_US
dc.typeJournal Articlezh_TW
dc.identifier.doi10.1016/j.jbiotec.2006.10.011zh_TW
item.grantfulltextnone-
item.openairetypeJournal Article-
item.languageiso639-1en_US-
item.fulltextno fulltext-
item.cerifentitytypePublications-
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
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