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dc.contributor.authorChien, H.C.R.en_US
dc.contributor.authorLin, L.L.en_US
dc.contributor.authorChao, S.H.en_US
dc.contributor.authorChen, C.C.en_US
dc.contributor.authorWang, W.C.en_US
dc.contributor.authorShaw, C.Y.en_US
dc.contributor.authorTsai, Y.C.en_US
dc.contributor.authorHu, H.Y.en_US
dc.contributor.authorHsu, W.H.en_US
dc.description.abstractExtracellular leucine aminopeptidase (LAP) from Aspergillus sojae was purified to protein homogeneity by sequential fast protein liquid chromatography steps. LAP had an apparent molecular mass of 37 kDa. of which approximately 3% was contributed by N-glycosylated carbohydrate. The purified enzyme was most active at pH 9 and 70 degreesC for 30 min. The enzyme preferentially hydrolyzed leucine p-nitroanilide followed by Phe, Lys, and Arg derivatives. The LAP activity was strongly inhibited by metal-chelating agents, and was largely restored by divalent cations like Zn2+ and Co2+. The lap gene and its corresponding cDNA fragment of the A. sojae were cloned using degenerated primers derived from internal amino acid sequences of the purified enzyme. lap is interrupted by three introns and is transcribed in a 1.3-kb mRNA that encodes a 377-amino-acid protein with a calculated molecular mass of 41.061 kDa. The mature LAP is preceded by a leader peptide of 77 amino acids, predicted to include an 18-amino-acid signal peptide and an extra sequence of 59 amino acids, Two putative N-glycosylation sites are identified in Asn-87 and Asn-288. Southern blot analysis suggested that lap is a single-copy gene in the A. sojae genome. The deduced amino acid sequence of A. sojae LAP shares only 11-33.1% identity with those of LAPs from IS organisms. (C) 2002 Elsevier Science B.V. All rights reserved.en_US
dc.relationBiochimica Et Biophysica Acta-Gene Structure and Expressionen_US
dc.relation.ispartofseriesBiochimica Et Biophysica Acta-Gene Structure and Expression, Volume 1576, Issue 1-2, Page(s) 119-126.en_US
dc.subjectleucine aminopeptidaseen_US
dc.subjectlap geneen_US
dc.subjectAspergillus sojaeen_US
dc.subjectstreptomyces-griseus aminopeptidaseen_US
dc.subjecttransition-state analogen_US
dc.subjecttranscription factoren_US
dc.subjectangstrom resolutionen_US
dc.subjectambient phen_US
dc.titlePurification, characterization, and genetic analysis of a leucine aminopeptidase from Aspergillus sojaeen_US
dc.typeJournal Articlezh_TW
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