Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/71627
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dc.contributor.authorLo, H.F.en_US
dc.contributor.authorChen, Y.H.en_US
dc.contributor.authorHsiao, N.W.en_US
dc.contributor.authorChen, H.L.en_US
dc.contributor.authorHu, H.Y.en_US
dc.contributor.authorHsu, W.H.en_US
dc.contributor.authorLin, L.L.en_US
dc.date2005zh_TW
dc.date.accessioned2014-06-11T06:02:08Z-
dc.date.available2014-06-11T06:02:08Z-
dc.identifier.issn0959-3993zh_TW
dc.identifier.urihttp://hdl.handle.net/11455/71627-
dc.description.abstractHistidine-436 of a truncated Bacillus sp. strain TS-23 alpha-amylase (His(6)-tagged Delta NC) has been known to be responsible for thermostability of the enzyme. To understand further the structural role of this residue, site-directed mutagenesis was conducted to replace His-436 of His(6)-tagged Delta NC with aspartate, lysine, tyrosine or threonine. Starch-plate assay showed that all Escherichia coli M15 transformants conferring the mutated amylase genes retained the amylolytic activity. The over-expressed proteins have been purified to near homogeneity by nickel-chelate chromatography and the molecular mass of the purified enzymes was approximately 54 kDa. The specific activity for H436T was decreased by more than 56%, while H436D, H436K, and H436Y showed a higher activity to that of the wild-type enzyme. Although the mutations did not lead to a significant change in the K-m value, more than 66% increase in the value of catalytic efficiency (k(cat)/K-m) was observed in H436D, H436K, and H436Y. At 70 degrees C, H436D exhibited an increased half-life with respect to the wild-type enzyme.en_US
dc.language.isoen_USzh_TW
dc.relationWorld Journal of Microbiology & Biotechnologyen_US
dc.relation.ispartofseriesWorld Journal of Microbiology & Biotechnology, Volume 21, Issue 4, Page(s) 411-416.en_US
dc.relation.urihttp://dx.doi.org/10.1007/s11274-004-1764-9en_US
dc.subjectBacillus sp strain TS-23en_US
dc.subjectalpha-amylaseen_US
dc.subjectsite-directed mutagenesisen_US
dc.subjecthistidineen_US
dc.subjectthermostabilityen_US
dc.subjectmeningosepticum glycerol kinaseen_US
dc.subjectsequence-based classificationen_US
dc.subjectcrystal-structureen_US
dc.subjectescherichia-colien_US
dc.subjectcyclodextrin glycosyltransferaseen_US
dc.subjectenzymatic-propertiesen_US
dc.subjectglycosyl hydrolasesen_US
dc.subjectangstrom resolutionen_US
dc.subjectamylolytic enzymesen_US
dc.subjectprotein stabilityen_US
dc.titleStabilization of a truncated Bacillus sp strain TS-23 alpha-amylase by replacing histidine-436 with aspartateen_US
dc.typeJournal Articlezh_TW
dc.identifier.doi10.1007/s11274-004-1764-9zh_TW
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.openairetypeJournal Article-
item.cerifentitytypePublications-
item.fulltextno fulltext-
item.languageiso639-1en_US-
item.grantfulltextnone-
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