Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/68891
DC FieldValueLanguage
dc.contributor.authorLin, L.L.en_US
dc.contributor.authorHsu, W.H.en_US
dc.contributor.authorWu, C.P.en_US
dc.contributor.authorChi, M.C.en_US
dc.contributor.authorChou, W.M.en_US
dc.contributor.authorHu, H.Y.en_US
dc.date2004zh_TW
dc.date.accessioned2014-06-11T05:57:28Z-
dc.date.available2014-06-11T05:57:28Z-
dc.identifier.issn1431-0651zh_TW
dc.identifier.urihttp://hdl.handle.net/11455/68891-
dc.description.abstractTwo degenerate primers established from the consensus sequences of bacterial leucine aminopeptidases (LAP) were used to amplify a 360-bp gene fragment from the chromosomal DNA of thermophilic Bacillus kaustophilus CCRC 11223 and the amplified fragment was successfully used as a probe to clone a leucine aminopeptidase (lap) gene from a genomic library of the strain. The gene consists of an open reading frame (ORF) of 1,494 bp and encodes a protein of 497 amino acid residues with a calculated molecular mass of 53.7 kDa. The complete amino acid sequence of the cloned enzyme showed greater than 30% identity with prokaryotic and eukaryotic LAPs. Phylogenetic analysis showed that B. kaustophilus LAP is closely related to the enzyme from Bacillus subtilis and is grouped with the M17 family. His(6)-tagged LAP was generated in Escherichia coli by cloning the coding region into pQE-30 and the recombinant enzyme was purified by nickel-chelate chromatography. The pH and temperature optima for the purified enzyme were 8 and 65degreesC, respectively, and 50% of its activity remained after incubation at 60degreesC for 32 min. The enzyme preferentially hydrolyzed L-leucine-p-nitroanilide (L-Leu-p-NA) followed by Cys derivative.en_US
dc.language.isoen_USzh_TW
dc.relationExtremophilesen_US
dc.relation.ispartofseriesExtremophiles, Volume 8, Issue 1, Page(s) 79-87.en_US
dc.relation.urihttp://dx.doi.org/10.1007/s00792-003-0364-1en_US
dc.subjectBacillus kaustophilusen_US
dc.subjectgene cloningen_US
dc.subjectleucine aminopeptidaseen_US
dc.subjectphylogenyen_US
dc.subjectescherichia-colien_US
dc.subjectbovine lensen_US
dc.subjectcrystal-structureen_US
dc.subjectgenus bacillusen_US
dc.subjectpurificationen_US
dc.subjectcloningen_US
dc.subjectenzymeen_US
dc.subjectidentificationen_US
dc.subjectthermozymesen_US
dc.subjectexpressionen_US
dc.titleA thermostable leucine aminopeptidase from Bacillus kaustophilus CCRC 11223en_US
dc.typeJournal Articlezh_TW
dc.identifier.doi10.1007/s00792-003-0364-1zh_TW
item.fulltextno fulltext-
item.grantfulltextnone-
item.openairetypeJournal Article-
item.languageiso639-1en_US-
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.cerifentitytypePublications-
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