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

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    Mutagenesis
    Carboxy-Lyases
    Salicylic Acid
    Salicylates
    Tunnels
    Carboxylation
    Substrates
    Phenol
    Alanine
    Yeast
    Lysine
    Substitution reactions
    Amino Acids
    2-cresol

    Keywords

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

    ASJC Scopus subject areas

    • Chemistry(all)

    Cite this

    Expanding substrate specificity of salicylate decarboxylase by site-directed mutagenesis for expansion of the entrance region connecting to the substrate access tunnel. / Kirimura, Kotaro; Araki, Masahiro; Ishihara, Mana; Ishii, Yoshitaka.

    In: Chemistry Letters, Vol. 48, No. 1, 01.01.2019, p. 58-61.

    Research output: Contribution to journalArticle

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    AU - Kirimura, Kotaro

    AU - Araki, Masahiro

    AU - Ishihara, Mana

    AU - Ishii, Yoshitaka

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    AB - 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.

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