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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)

Submitter: 尾島 孝男

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