Removal of the four c-terminal glycine-rich repeats enhances the thermostability and substrate binding affinity of barley b-amylase

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dc.contributor.authorMa, Y.
dc.contributor.authorEglinton, J.
dc.contributor.authorEvans, D.
dc.contributor.authorLogue, S.
dc.contributor.authorLangridge, P.
dc.date.issued2000
dc.description.abstractBarley beta-amylase undergoes proteolytic cleavage in the C-terminal region after germination. The implication of the cleavage in the enzyme's characteristics is unclear. With purified native beta-amylases from both mature barley grain and germinated barley, we found that the beta-amylase from germinated barley had significantly higher thermostability and substrate binding affinity for starch than that from mature barley grain. To better understand the effect of the proteolytic cleavage on the enzyme's thermostability and substrate binding affinity for starch, recombinant barley beta-amylases with specific deletions at the C-terminal tail were generated. The complete deletion of the four C-terminal glycine-rich repeats significantly increased the enzyme's thermostability, but an incomplete deletion with one repeat remaining did not change the thermostability. Although different C-terminal deletions affect the thermostability differently, they all increased the enzyme's affinity for starch. The possible reasons for the increased thermostability and substrate binding affinity, due to the removal of the four C-terminal glycine-rich repeats, are discussed in terms of the three-dimensional structure of beta-amylase.
dc.description.statementofresponsibilityYue F. Ma, Jason K. Eglinton, D. Evan Evans, Sue J. Logue, and Peter Langridge
dc.identifier.citationBiochemistry, 2000; 39(44):13350-13355
dc.identifier.doi10.1021/bi000688s
dc.identifier.issn0006-2960
dc.identifier.issn1520-4995
dc.identifier.orcidLangridge, P. [0000-0001-9494-400X]
dc.identifier.urihttp://hdl.handle.net/2440/13371
dc.language.isoen
dc.publisherAmer Chemical Soc
dc.source.urihttps://doi.org/10.1021/bi000688s
dc.subjectHordeum
dc.subjectStarch
dc.subjectbeta-Amylase
dc.subjectGlycine
dc.subjectPeptide Fragments
dc.subjectRecombinant Proteins
dc.subjectMutagenesis, Site-Directed
dc.subjectEnzyme Stability
dc.subjectSequence Deletion
dc.subjectBinding Sites
dc.subjectAmino Acid Sequence
dc.subjectRepetitive Sequences, Amino Acid
dc.subjectSubstrate Specificity
dc.subjectHydrolysis
dc.subjectKinetics
dc.subjectMolecular Sequence Data
dc.subjectHot Temperature
dc.titleRemoval of the four c-terminal glycine-rich repeats enhances the thermostability and substrate binding affinity of barley b-amylase
dc.typeJournal article
pubs.publication-statusPublished

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