Multiple nucleophilic elbows leading to multiple active sites in a single module esterase from Sorangium cellulosum

D.B.R.K. Gupta Udatha, Karina Marie Madsen, Gianni Panagiotou, Lisbeth Olsson

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    The catalytic residues in carbohydrate esterase enzyme families constitute a highly conserved triad: serine, histidine and aspartic acid. This catalytic triad is generally located in a very sharp turn of the protein backbone structure, called the nucleophilic elbow and identified by the consensus sequence GXSXG. An esterase from Sorangium cellulosum Soce56 that contains five nucleophilic elbows was cloned and expressed in Escherichia coli and the function of each nucleophilic elbowed site was characterized. In order to elucidate the function of each nucleophilic elbow, site directed mutagenesis was used to generate variants with deactivated nucleophilic elbows and the functional promiscuity was analyzed. In silico analysis together with enzymological characterization interestingly showed that each nucleophilic elbow formed a local active site with varied substrate specificities and affinities. To our knowledge, this is the first report presenting the role of multiple nucleophilic elbows in the catalytic promiscuity of an esterase. Further structural analysis at protein unit level indicates the new evolutionary trajectories in emerging promiscuous esterases.
    Original languageEnglish
    JournalJournal of Structural Biology
    Volume190
    Issue number3
    Pages (from-to)314-327
    Number of pages14
    ISSN1047-8477
    DOIs
    Publication statusPublished - 2015

    Keywords

    • Nucleophilic elbow
    • Esterase
    • Protein modeling
    • Active sites

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