Expanding substrate specificity of salicylate decarboxylase by site-directed mutagenesis for expansion of the entrance region connecting to the substrate access tunnel

Kotaro Kirimura, Masahiro Araki, Mana Ishihara, Yoshitaka Ishii

    Research output: Contribution to journalArticle

    Abstract

    Salicylate decarboxylase (Sdc, EC 4.1.1.91) from the yeast Trichosporon moniliiforme WU-0401 catalyzes the carboxylation of phenol to salicylic acid and is applicable to enzymatic Kolbe-Schmitt reaction. Based on the 3D-modeling of Sdc, we generated a Sdc mutant, K23A-Sdc, by site-directed mutagenesis for expansion of the entrance region connecting to the substrate access tunnel. K23A-Sdc catalyzed the carboxylation of o-cresol to 3-methyl salicylic acid (3-MS), whereas Sdc showed negligible and slight activity toward o-cresol and 3-MS, respectively. Simultaneously, K23A-Sdc showed approximately 5.8-fold more decarboxylation activity toward 3-MS than Sdc. These results clearly indicate that K23A-Sdc acquired novel substrate specificity by substitution of only one amino acid residue (23rd lysine residue to alanine residue), around the substrate entrance region but not at the active center of Sdc.

    Original languageEnglish
    Pages (from-to)58-61
    Number of pages4
    JournalChemistry Letters
    Volume48
    Issue number1
    DOIs
    Publication statusPublished - 2019 Jan 1

    Keywords

    • Enzymatic Kolbe-Schmitt reaction
    • Salicylate decarboxylase
    • Substrate specificity

    ASJC Scopus subject areas

    • Chemistry(all)

    Fingerprint Dive into the research topics of 'Expanding substrate specificity of salicylate decarboxylase by site-directed mutagenesis for expansion of the entrance region connecting to the substrate access tunnel'. Together they form a unique fingerprint.

  • Cite this