Isolation of a new antibacterial peptide actinokineosin from Actinokineospora spheciospongiae based on genome mining.
Based on genome mining, a new antibacterial peptide named actinokineosin was isolated from a rare actinomycete Actinokineospora spheciospongiae. The amino acid sequence of the C-terminus of actinokineosin was established by TOF-MS/MS experiments. The amino acid sequence in the macrolactam ring was determined by TOF-MS/MS analyses after cleavage with BNPS-skatole and successive trypsin treatment. As a result of an antibacterial assay using a paper disk, actinokineosin showed antibacterial activity against Micrococcus luteus at a dosage of 50 μg per disk. From the genome sequence data of A. spheciospongiae, the biosynthetic gene cluster of actinokineosin was found and was indicated to consist of 10 genes. Among the genes, the gene aknA encoded the precursor of actinokineosin and the genes including aknC, aknB1 and aknB2 were proposed as modification enzymes to give mature actinokineosin.


CONCLUSIONS
Genome mining is a powerful tool to find new bioactive compounds from the genome database. In this report, we succeeded in isolation and structure determination of a new antibacterial peptide named actinokineosin based on genome mining.
Graduate School of Integrated Science and Technology, Shizuoka University, Suruga-ku, Shizuoka, Japan.
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Takasaka
Graduate School of Integrated Science and Technology, Shizuoka University, Suruga-ku, Shizuoka, Japan.
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Kaweewan
Food Research Institute, National Agriculture and Food Research Organization (NARO), Tsukuba, Ibaraki, Japan.
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Ohnishi-Kameyama
Graduate School of Integrated Science and Technology, Shizuoka University, Suruga-ku, Shizuoka, Japan.|College of Agriculture, Academic Institute, Shizuoka University, Suruga-ku, Shizuoka, Japan.|Graduate School of Science and Technology, Shizuoka University, Suruga-ku, Shizuoka, Japan.
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Kodani
D039903:Actinobacteria / Q000737:chemistry* / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology; D001483:Base Sequence; D016680:Genome, Bacterial; D008826:Microbial Sensitivity Tests; D016982:Micrococcus luteus / Q000187:drug effects*; D005810:Multigene Family; D012862:Skatole / Q000031:analogs & derivatives / Q000737:chemistry; D053719:Tandem Mass Spectrometry; D014357:Trypsin / Q000737:chemistry
D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; C000618131:actinokineosin, Actinokineospora spheciospongiae; C005904:BNPS-skatole; D012862:Skatole; D014357:Trypsin
Actinokineospora spheciospongiae; BNPS-skatole; TOF-MS/MS; antibacterial; biosynthesis; lasso peptide