A novel L-amino acid ligase from Bacillus subtilis NBRC3134, a microorganism producing peptide-antibiotic rhizocticin

Kuniki Kino, Yoichi Kotanaka, Toshinobu Arai, Makoto Yagasaki

    Research output: Contribution to journalArticle

    40 Citations (Scopus)

    Abstract

    L-Amino acid ligase catalyzes the formation of an α-peptide bond from unprotected L-amino acids in an ATP-dependent manner, and this enzyme is very useful in efficient peptide production. We performed enzyme purification to obtain a novel L-amino acid ligase from Bacillus subtilis NBRC3134, a microorganism producing peptide-antibiotic rhizocticin. Rhizocticins are dipep-tide or tripeptide antibiotics and commonly possess L-arginyl-L-2-amino-5- phosphono-3-cis-pentenoic acid. The purification was carried out by detecting L-arginine hydroxamate synthesis activity, and a target enzyme was finally purified 1,280-fold with 0.8% yield. The corresponding gene was then cloned and designated rizA. rizA was 1,242 bp and coded for 413 amino acid residues. Recombinant RizA was prepared, and it was found that the recombinant RizA synthesized dipeptides whose N-terminus was L-arginine in an ATP-dependent manner. RizA had strict substrate specificity toward l-arginine as the N-terminal substrate; on the other hand, the substrate specificity at the C-terminus was relaxed.

    Original languageEnglish
    Pages (from-to)901-907
    Number of pages7
    JournalBioscience, Biotechnology and Biochemistry
    Volume73
    Issue number4
    DOIs
    Publication statusPublished - 2009

    Fingerprint

    Bacilli
    Ligases
    Bacillus subtilis
    Microorganisms
    Anti-Bacterial Agents
    Arginine
    Amino Acids
    Peptides
    NSC 153174
    Substrate Specificity
    Purification
    Substrates
    Enzymes
    Adenosine Triphosphate
    Dipeptides
    Tides
    Genes
    Acids

    Keywords

    • L-amino acid ligase
    • Peptide synthesis
    • Rhizocticin

    ASJC Scopus subject areas

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

    Cite this

    A novel L-amino acid ligase from Bacillus subtilis NBRC3134, a microorganism producing peptide-antibiotic rhizocticin. / Kino, Kuniki; Kotanaka, Yoichi; Arai, Toshinobu; Yagasaki, Makoto.

    In: Bioscience, Biotechnology and Biochemistry, Vol. 73, No. 4, 2009, p. 901-907.

    Research output: Contribution to journalArticle

    @article{e7dd128b066549f480f430a9914ddd2e,
    title = "A novel L-amino acid ligase from Bacillus subtilis NBRC3134, a microorganism producing peptide-antibiotic rhizocticin",
    abstract = "L-Amino acid ligase catalyzes the formation of an α-peptide bond from unprotected L-amino acids in an ATP-dependent manner, and this enzyme is very useful in efficient peptide production. We performed enzyme purification to obtain a novel L-amino acid ligase from Bacillus subtilis NBRC3134, a microorganism producing peptide-antibiotic rhizocticin. Rhizocticins are dipep-tide or tripeptide antibiotics and commonly possess L-arginyl-L-2-amino-5- phosphono-3-cis-pentenoic acid. The purification was carried out by detecting L-arginine hydroxamate synthesis activity, and a target enzyme was finally purified 1,280-fold with 0.8{\%} yield. The corresponding gene was then cloned and designated rizA. rizA was 1,242 bp and coded for 413 amino acid residues. Recombinant RizA was prepared, and it was found that the recombinant RizA synthesized dipeptides whose N-terminus was L-arginine in an ATP-dependent manner. RizA had strict substrate specificity toward l-arginine as the N-terminal substrate; on the other hand, the substrate specificity at the C-terminus was relaxed.",
    keywords = "L-amino acid ligase, Peptide synthesis, Rhizocticin",
    author = "Kuniki Kino and Yoichi Kotanaka and Toshinobu Arai and Makoto Yagasaki",
    year = "2009",
    doi = "10.1271/bbb.80842",
    language = "English",
    volume = "73",
    pages = "901--907",
    journal = "Bioscience, Biotechnology and Biochemistry",
    issn = "0916-8451",
    publisher = "Japan Society for Bioscience Biotechnology and Agrochemistry",
    number = "4",

    }

    TY - JOUR

    T1 - A novel L-amino acid ligase from Bacillus subtilis NBRC3134, a microorganism producing peptide-antibiotic rhizocticin

    AU - Kino, Kuniki

    AU - Kotanaka, Yoichi

    AU - Arai, Toshinobu

    AU - Yagasaki, Makoto

    PY - 2009

    Y1 - 2009

    N2 - L-Amino acid ligase catalyzes the formation of an α-peptide bond from unprotected L-amino acids in an ATP-dependent manner, and this enzyme is very useful in efficient peptide production. We performed enzyme purification to obtain a novel L-amino acid ligase from Bacillus subtilis NBRC3134, a microorganism producing peptide-antibiotic rhizocticin. Rhizocticins are dipep-tide or tripeptide antibiotics and commonly possess L-arginyl-L-2-amino-5- phosphono-3-cis-pentenoic acid. The purification was carried out by detecting L-arginine hydroxamate synthesis activity, and a target enzyme was finally purified 1,280-fold with 0.8% yield. The corresponding gene was then cloned and designated rizA. rizA was 1,242 bp and coded for 413 amino acid residues. Recombinant RizA was prepared, and it was found that the recombinant RizA synthesized dipeptides whose N-terminus was L-arginine in an ATP-dependent manner. RizA had strict substrate specificity toward l-arginine as the N-terminal substrate; on the other hand, the substrate specificity at the C-terminus was relaxed.

    AB - L-Amino acid ligase catalyzes the formation of an α-peptide bond from unprotected L-amino acids in an ATP-dependent manner, and this enzyme is very useful in efficient peptide production. We performed enzyme purification to obtain a novel L-amino acid ligase from Bacillus subtilis NBRC3134, a microorganism producing peptide-antibiotic rhizocticin. Rhizocticins are dipep-tide or tripeptide antibiotics and commonly possess L-arginyl-L-2-amino-5- phosphono-3-cis-pentenoic acid. The purification was carried out by detecting L-arginine hydroxamate synthesis activity, and a target enzyme was finally purified 1,280-fold with 0.8% yield. The corresponding gene was then cloned and designated rizA. rizA was 1,242 bp and coded for 413 amino acid residues. Recombinant RizA was prepared, and it was found that the recombinant RizA synthesized dipeptides whose N-terminus was L-arginine in an ATP-dependent manner. RizA had strict substrate specificity toward l-arginine as the N-terminal substrate; on the other hand, the substrate specificity at the C-terminus was relaxed.

    KW - L-amino acid ligase

    KW - Peptide synthesis

    KW - Rhizocticin

    UR - http://www.scopus.com/inward/record.url?scp=67149125327&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=67149125327&partnerID=8YFLogxK

    U2 - 10.1271/bbb.80842

    DO - 10.1271/bbb.80842

    M3 - Article

    C2 - 19352016

    AN - SCOPUS:67149125327

    VL - 73

    SP - 901

    EP - 907

    JO - Bioscience, Biotechnology and Biochemistry

    JF - Bioscience, Biotechnology and Biochemistry

    SN - 0916-8451

    IS - 4

    ER -