Structure-based modification of D-alanine-D-alanine ligase from Thermotoga maritima ATCC 43589 for depsipeptide synthesis

Tomoki Nakagawa, Ryoko Satake, Masaru Sato, Kuniki Kino

    Research output: Contribution to journalArticle

    Abstract

    Depsipeptides are peptide-like polymers consisting of amino acids and hydroxy acids, and are expected to be new functional materials for drug-delivery systems and polymer science. In our previous study, D-alanyl-D-lactate, a type of depsipeptide, was enzymatically synthesized using D-alanine-D-alanine ligase from Thermotoga maritima ATCC 43589 (TmDdl) by Y207F substitution. Thereafter, in this study, further mutagenesis was introduced, based on structural comparison between TmDdl and a well-characterized D-alanine-D-alanine ligase from Escherichia coli. The S137A/Y207F mutant showed higher D-alanyl-D-lactate and lower D-alanyl-D-alanine synthesizing activity than the Y207F mutant. This suggests that substitution at the S137 residue contributes to product selectivity. Saturated mutagenesis on S137 revealed that the S137G/Y207F mutant showed the highest D-alanyl-D-lactate synthesizing activity. Moreover, the mutant showed broad substrate specificity toward D-amino acid and recognized D-lactate and D,L-isoserine as substrates. On the basis of these characteristics, various depsipeptides can be produced using S137G/Y207F-replaced TmDdl.

    Original languageEnglish
    Pages (from-to)700-704
    Number of pages5
    JournalBioscience, Biotechnology and Biochemistry
    Volume75
    Issue number4
    DOIs
    Publication statusPublished - 2011

    Fingerprint

    Thermotoga maritima
    Depsipeptides
    Mutagenesis
    Polymers
    Substitution reactions
    Amino Acids
    Hydroxy Acids
    Functional materials
    Substrates
    Drug Delivery Systems
    Substrate Specificity
    Alanine
    Escherichia coli
    Lactic Acid
    Peptides
    alanyllactate
    D-alanylalanine synthetase

    Keywords

    • D-alanine-D-alanine ligase
    • Depsipeptide
    • Dipeptide
    • Enzymatic peptide synthesis
    • Thermotoga maritima

    ASJC Scopus subject areas

    • Biotechnology
    • Biochemistry
    • Molecular Biology
    • Applied Microbiology and Biotechnology
    • Analytical Chemistry
    • Organic Chemistry

    Cite this

    Structure-based modification of D-alanine-D-alanine ligase from Thermotoga maritima ATCC 43589 for depsipeptide synthesis. / Nakagawa, Tomoki; Satake, Ryoko; Sato, Masaru; Kino, Kuniki.

    In: Bioscience, Biotechnology and Biochemistry, Vol. 75, No. 4, 2011, p. 700-704.

    Research output: Contribution to journalArticle

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