WAP-8294A<sub>2</sub>, A Novel Anti-MRSA Antibiotic Produced by <i>Lysobacter</i> sp.

Kato, Azusa; Nakaya, Seigo; Ohashi, and Yoshitami; Hirata, Haruhisa; and, Kiyonaga Fujii; Harada, Kenichi

doi:10.1021/ja970895o

Cited by 55 publications

(64 citation statements)

References 14 publications

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“…10 Originally, the producing strain, Lysobacter sp., produces a WAP-8294 complex that contains at least 20 components. Of these components, the main one, WAP-8294A2, has been structurally characterized mainly by NMR techniques 11 and WAP-8294A1, A4 and Ax13 were isolated in a pure state by chromatographic techniques. In the present study, we tried to determine the structures of WAP-8294A1, A4 and Ax13 based on the fragmentation patterns of WAP-8294A2 in the ESI MS/MS.…”

Section: Discussionmentioning

confidence: 99%

“…As shown in Figure 1, the structure of 1 was determined as the cyclic depsipeptide by 2D NMR experiments including HMBC ( 1 H-detected multiple-bond HMQC) and ROESY techniques. 11 Usually the sequence of the constituent amino acids of the peptide is mainly established with the help of 2D NMR experiments. However, these minor components were obtained in small amounts of the pure compounds.…”

Section: Introductionmentioning

confidence: 99%

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A new anti-MRSA antibiotic complex, WAP-8294A II. Structure characterization of minor components by ESI LCMS and MS/MS

Kato

Hirata

Ohashi³

et al. 2011

J Antibiot

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The anti-MRSA antibiotic, WAP-8294A, was isolated from the fermentation broth of Lysobacter sp. The major component,and 2 mol of L-Ser, D-Orn and D-3-hydroxy-7-Me-octanoic acid. The structure of the WAP-8294A2 was mainly determined as a cyclic depsipeptide by 2D NMR experiments. However, it was difficult to use the NMR experiment to determine the minor components, A1, A4 and Ax13, isolated in small amounts. In the present study, ESI MS/MS was applied to the structure elucidation of these minor components. The structures of these minor components were determined on the basis of the fragmentation pattern of the product ions of WAP-8294A2 in the ESI MS/MS. As a result, it was confirmed that A1 and A4 had the same amino acid sequence as A2, while A1 and A4 had the 3-OH-octanoic acid and 3-OH-8-Me-nonanoic acid, respectively, in the place of the 3-OH-7-Meoctanoic acid in A2. In the structure of Ax13, it was found that Gly of A2 was changed to b-Ala of Ax13.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A new anti-MRSA antibiotic complex, WAP-8294A II. Structure characterization of minor components by ESI LCMS and MS/MS

Kato

Hirata

Ohashi³

et al. 2011

J Antibiot

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“…These traits include production of extracellular degradative enzymes, such as chitinases, glucanases, and proteases, as well as production of antibiotic compounds (5,20,36,37). Mutations in the clp gene had a global effect on a variety of L. enzymogenes phenotypes, including significant re- (44) demonstrated that chitinases purified from culture filtrates enhanced biocontrol of bipolaris leaf spot by strain C3.…”

Section: Discussionmentioning

confidence: 99%

A clp Gene Homologue Belonging to the Crp Gene Family Globally Regulates Lytic Enzyme Production, Antimicrobial Activity, and Biological Control Activity Expressed by Lysobacter enzymogenes Strain C3

Kobayashi

Reedy

Palumbo

et al. 2005

Appl Environ Microbiol

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Lysobacter enzymogenes strain C3, a biological control agent for plant diseases, produces multiple extracellular hydrolytic enzymes and displays antimicrobial activity against various fungal and oomycetous species. However, little is known about the regulation of these enzymes or their roles in antimicrobial activity and biocontrol. A study was undertaken to identify mutants of strain C3 affected in extracellular enzyme production and to evaluate their biocontrol efficacy. A single mini-Tn5-lacZ 1 -cat transposon mutant of L. enzymogenes strain C3 that was globally affected in a variety of phenotypes was isolated. In this mutant, 5E4, the activities of several extracellular lytic enzymes, gliding motility, and in vitro antimicrobial activity were reduced. Characterization of 5E4 indicated that the transposon inserted in a clp gene homologue belonging to the Crp gene family of regulators. Immediately downstream was a second open reading frame similar to that encoding acetyltransferases belonging to the Gcn5-related N-acetyltransferase superfamily, which reverse transcription-PCR confirmed was cotranscribed with clp. Chromosomal deletion mutants with mutations in clp and between clp and the acetyltransferase gene verified the 5E4 mutant phenotype. The clp gene was chromosomally inserted in mutant 5E4, resulting in complemented strain P1. All mutant phenotypes were restored in P1, although the gliding motility was observed to be excessive compared with that of the wild-type strain. clp mutant strains were significantly affected in biological control of pythium damping-off of sugar beet and bipolaris leaf spot of tall fescue, which was partially or fully restored in the complemented strain P1. These results indicate that clp is a global regulatory gene that controls biocontrol traits expressed by L. enzymogenes C3.

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“…already have illuminated their huge potential for the production of antimicrobial agents (47). Among the antibiotics reported are inhibitors of cell wall biosynthesis, such as cephabacins (48) and tripropeptins (49), as well as a number of compounds which target the bacterial membrane (50)(51)(52). The strains used in this study are not yet known as antibiotic producers, although the biosynthesis of such compounds seems likely in light of previous investigations (26).…”

Section: Discussionmentioning

confidence: 95%

Quantitative Analysis of Lysobacter Predation

Seccareccia

Kost

Nett

2015

Appl Environ Microbiol

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bBacteria of the genus Lysobacter are considered to be facultative predators that use a feeding strategy similar to that of myxobacteria. Experimental data supporting this assumption, however, are scarce. Therefore, the predatory activities of three Lysobacter species were tested in the prey spot plate assay and in the lawn predation assay, which are commonly used to analyze myxobacterial predation. Surprisingly, only one of the tested Lysobacter species showed predatory behavior in the two assays. This result suggested that not all Lysobacter strains are predatory or, alternatively, that the assays were not appropriate for determining the predatory potential of this bacterial group. To differentiate between the two scenarios, predation was tested in a CFU-based bioassay. For this purpose, defined numbers of Lysobacter cells were mixed together with potential prey bacteria featuring phenotypic markers, such as distinctive pigmentation or antibiotic resistance. After 24 h, cocultivated cells were streaked out on agar plates and sizes of bacterial populations were individually determined by counting the respective colonies. Using the CFU-based predation assay, we observed that Lysobacter spp. strongly antagonized other bacteria under nutrient-deficient conditions. Simultaneously, the Lysobacter population was increasing, which together with the killing of the cocultured bacteria indicated predation. Variation of the predator/prey ratio revealed that all three Lysobacter species tested needed to outnumber their prey for efficient predation, suggesting that they exclusively practiced group predation. In summary, the CFU-based predation assay not only enabled the quantification of prey killing and consumption by Lysobacter spp. but also provided insights into their mode of predation.

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WAP-8294A₂, A Novel Anti-MRSA Antibiotic Produced by Lysobacter sp.

Cited by 55 publications

References 14 publications

A new anti-MRSA antibiotic complex, WAP-8294A II. Structure characterization of minor components by ESI LCMS and MS/MS

A new anti-MRSA antibiotic complex, WAP-8294A II. Structure characterization of minor components by ESI LCMS and MS/MS

A clp Gene Homologue Belonging to the Crp Gene Family Globally Regulates Lytic Enzyme Production, Antimicrobial Activity, and Biological Control Activity Expressed by Lysobacter enzymogenes Strain C3

Quantitative Analysis of Lysobacter Predation

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WAP-8294A2, A Novel Anti-MRSA Antibiotic Produced by Lysobacter sp.

Cited by 55 publications

References 14 publications

A new anti-MRSA antibiotic complex, WAP-8294A II. Structure characterization of minor components by ESI LCMS and MS/MS

A new anti-MRSA antibiotic complex, WAP-8294A II. Structure characterization of minor components by ESI LCMS and MS/MS

A clp Gene Homologue Belonging to the Crp Gene Family Globally Regulates Lytic Enzyme Production, Antimicrobial Activity, and Biological Control Activity Expressed by Lysobacter enzymogenes Strain C3

Quantitative Analysis of Lysobacter Predation

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WAP-8294A₂, A Novel Anti-MRSA Antibiotic Produced by Lysobacter sp.