P-aminosalicylic acid production by enzymatic kolbeschmitt reaction using salicylic acid decarboxylases improved through site-directed mutagenesis

Saori Ienaga, Sachiyo Kosaka, Yuki Honda, Yoshitaka Ishii, Kotaro Kirimura

    Research output: Contribution to journalArticle

    12 Citations (Scopus)

    Abstract

    A reversible salicylic acid decarboxylase (Sdc) catalyzes the carboxylation of m-aminophenol (m-AP) to paminosalicylic acid (PAS) as an antituberculous agent, through an enzymatic KolbeSchmitt reaction. To develop a highyield PAS production system through such an enzymatic reaction, we generated Sdc mutants by site-directed mutagenesis and succeeded in generating several mutants showing increased carboxylation specific activities. Among them, a Y64T-F195Y-Sdc mutant showed a 12-fold higher carboxylation specific activity toward m-AP than wild-type Sdc. By the whole-cell reaction of recombinant Escherichia coli BL21(DE3) expressing the gene encoding Y64T-F195Y-Sdc, 70mM PAS was produced from 100 mM m-AP within 2 h. This reaction time was shortened to one-twelfth that of the PAS production using E. coli BL21(DE3) expressing the gene encoding wild-type Sdc (24 h). Moreover, 140 mM PAS was produced from 200 mM m-AP within 9 h by the whole-cell reaction of recombinant E. coli BL21(DE3) expressing the gene encoding Y64T-F195Y-Sdc.

    Original languageEnglish
    Pages (from-to)628-634
    Number of pages7
    JournalBulletin of the Chemical Society of Japan
    Volume86
    Issue number5
    DOIs
    Publication statusPublished - 2013

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    Aminosalicylic Acid
    Mutagenesis
    Carboxy-Lyases
    Salicylic Acid
    Carboxylation
    Gene encoding
    Acids
    Escherichia coli
    3-aminophenol

    ASJC Scopus subject areas

    • Chemistry(all)

    Cite this

    P-aminosalicylic acid production by enzymatic kolbeschmitt reaction using salicylic acid decarboxylases improved through site-directed mutagenesis. / Ienaga, Saori; Kosaka, Sachiyo; Honda, Yuki; Ishii, Yoshitaka; Kirimura, Kotaro.

    In: Bulletin of the Chemical Society of Japan, Vol. 86, No. 5, 2013, p. 628-634.

    Research output: Contribution to journalArticle

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    abstract = "A reversible salicylic acid decarboxylase (Sdc) catalyzes the carboxylation of m-aminophenol (m-AP) to paminosalicylic acid (PAS) as an antituberculous agent, through an enzymatic KolbeSchmitt reaction. To develop a highyield PAS production system through such an enzymatic reaction, we generated Sdc mutants by site-directed mutagenesis and succeeded in generating several mutants showing increased carboxylation specific activities. Among them, a Y64T-F195Y-Sdc mutant showed a 12-fold higher carboxylation specific activity toward m-AP than wild-type Sdc. By the whole-cell reaction of recombinant Escherichia coli BL21(DE3) expressing the gene encoding Y64T-F195Y-Sdc, 70mM PAS was produced from 100 mM m-AP within 2 h. This reaction time was shortened to one-twelfth that of the PAS production using E. coli BL21(DE3) expressing the gene encoding wild-type Sdc (24 h). Moreover, 140 mM PAS was produced from 200 mM m-AP within 9 h by the whole-cell reaction of recombinant E. coli BL21(DE3) expressing the gene encoding Y64T-F195Y-Sdc.",
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    AU - Kirimura, Kotaro

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