RESUMO
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. SIGNIFICANCE AND IMPACT OF THE STUDY: 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.
Assuntos
Actinobacteria/química , Antibacterianos/isolamento & purificação , Peptídeos Catiônicos Antimicrobianos/isolamento & purificação , Micrococcus luteus/efeitos dos fármacos , Actinobacteria/genética , Sequência de Aminoácidos , Animais , Antibacterianos/química , Antibacterianos/farmacologia , Peptídeos Catiônicos Antimicrobianos/química , Peptídeos Catiônicos Antimicrobianos/genética , Peptídeos Catiônicos Antimicrobianos/farmacologia , Sequência de Bases , Genoma Bacteriano , Testes de Sensibilidade Microbiana , Família Multigênica , Escatol/análogos & derivados , Escatol/química , Espectrometria de Massas em Tandem , Tripsina/químicaRESUMO
UDP-N-acetylmuramyl-pentapeptide (UDP-MurNAc-pentapeptide) is well known to be a key intermediate of bacterial peptidoglycan biosynthesis. We first detected the occurrence of UDP-MurNAc-pentapeptide in the cyanobacterium Anabaena cylindrica (NIES-19), and identified the structure of this pentapeptide by two-dimensional 1H-1H and 1H-13C NMR correlation experiments and mass spectra.
Assuntos
Anabaena/química , Uridina Difosfato Ácido N-Acetilmurâmico/análogos & derivados , Espectroscopia de Ressonância Magnética , Uridina Difosfato Ácido N-Acetilmurâmico/análise , Uridina Difosfato Ácido N-Acetilmurâmico/químicaRESUMO
Aeruginosin 103-A was isolated from the cultured freshwater cyanobacterium Microcystis viridis(NIES-103). Its structure was elucidated to be 1 on the basis of 2D NMR data. This linear peptide inhibited thrombin, with an IC50 of 9.0 microgram/mL.
Assuntos
Microcystis/química , Oligopeptídeos/isolamento & purificação , Trombina/antagonistas & inibidores , Cromatografia Líquida de Alta Pressão , Oligopeptídeos/farmacologia , Espectrofotometria Infravermelho , Espectrofotometria UltravioletaRESUMO
Dehydroradiosumin, a novel potent trypsin inhibitory dipeptide, was isolated from the freshwater cyanobacterium Anabaena cylindrica (NIES-19). Its structure was elucidated as 1 on the basis of 2D NMR data and chemical degradation. The IC50 of 1 against trypsin was 0.1 microg/mL.
Assuntos
Anabaena/química , Dipeptídeos/isolamento & purificação , Inibidores da Tripsina/isolamento & purificação , Sequência de Carboidratos , Dipeptídeos/farmacologia , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Dados de Sequência Molecular , Espectrofotometria Infravermelho , Espectrofotometria Ultravioleta , Inibidores da Tripsina/farmacologiaRESUMO
Anabaenopeptins I (1) and J (2), two new ureido bond-containing cyclic peptides, were isolated from the cultured cyanobacterium Aphanizomenon flos-aquae (NIES-81) as potent carboxypeptidase-A (CPA) inhibitors. The gross structures of 1 and 2 were established by spectroscopic analysis, including the 2D NMR techniques. The absolute configurations of 1 and 2 were determined by spectral and chemical methods. Anabaenopeptins I and J inhibited CPA with IC(50) values of 5.2 and 7.6 ng/mL, respectively.