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Hokkaido University Collection of Scholarly and Academic Papers >
Graduate School of Fisheries Sciences / Faculty of Fisheries Sciences >
Peer-reviewed Journal Articles, etc >
Characterization of a GHF45 cellulase, AkEG21, from the common sea hare Aplysia kurodai.
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| Title: | Characterization of a GHF45 cellulase, AkEG21, from the common sea hare Aplysia kurodai. |
| Authors: | Rahman, Mohammad M Browse this author | | Inoue, Akira Browse this author →KAKEN DB | | Ojima, Takao Browse this author →KAKEN DB |
| Keywords: | Aplysia kurodai | | AkEG21 | | endo-β-1,4-glucanase | | cellulase | | GHF45 | | cDNAcloning | | primarystructure | | phylogenicanalysis |
| Issue Date: | 6-Aug-2014 |
| Publisher: | Frontiers |
| Journal Title: | Frontiers in Chemistry |
| Volume: | 2 |
| Start Page: | 1 |
| End Page: | 13 |
| Publisher DOI: | 10.3389/fchem.2014.00060 |
| Abstract: | ThecommonseahareAplysiakurodaiisknowntobeagoodsourcefortheenzymesdegradingseaweedpolysaccharides.Recentlyfourcellulases,i.e.,95,66,45,and21kDaenzymes,wereisolatedfromA.kurodai(Tsujietal.,2013).Theformerthreecellulaseswereregardedasglycosyl-hydrolase-family9(GHF9)enzymes,whilethe21kDacellulasewassuggestedtobeaGHF45enzyme.The21kDacellulasewassignificantlyheatstable,andappearedtobeadvantageousinperformingheterogeneousexpressionandprotein-engineeringstudy.Inthepresentstudy,wedeterminedsomeenzymaticpropertiesofthe21kDacellulaseandcloneditscDNAtoprovidethebasisfortheproteinengineeringstudyofthiscellulase.Thepurified21kDaenzyme,termedAkEG21inthepresentstudy,hydrolyzedcarboxymethylcellulosewithanoptimalpHandtemperatureat4.5and40◦C,respectively.AkEG21wasconsiderablyheat-stable,i.e.,itwasnotinactivatedbytheincubationat55◦Cfor30min.AkEG21degradedphosphoric-acid-swollencelluloseproducingcellotrioseandcellobioseasmajorendproductsbuthardlydegradedoligosaccharidessmallerthantetrasaccharide.ThisindicatedthatAkEG21isanendolyticβ-1,4-glucanase(EC3.2.1.4).AcDNAof1013bpencodingAkEG21wasamplifiedbyPCRandtheamino-acidsequenceof197residueswasdeduced.ThesequencecomprisedtheinitiationMet,theputativesignalpeptideof16residuesforsecretionandthecatalyticdomainof180residues,whichlinedfromtheN-terminusinthisorder.Thesequenceofthecatalyticdomainshowed47–62%amino-acididentitiestothoseofGHF45cellulasesreportedinothermollusks.BoththecatalyticresiduesandtheN-glycosylationresiduesknowninotherGHF45cellulaseswereconservedinAkEG21.Phylogeneticanalysisfortheamino-acidsequencessuggestedthecloserelationbetweenAkEG21andfungalGHF45cellulases. |
| Rights: | http://creativecommons.org/licenses/by/3.0/ |
| Type: | article |
| URI: | http://hdl.handle.net/2115/58461 |
| Appears in Collections: | 水産科学院・水産科学研究院 (Graduate School of Fisheries Sciences / Faculty of Fisheries Sciences) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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Submitter: 尾島 孝男
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