Isolation and characterization of novel marine-derived actinomycete taxa rich in bioactive metabolites - PubMed (original) (raw)

Isolation and characterization of novel marine-derived actinomycete taxa rich in bioactive metabolites

Nathan A Magarvey et al. Appl Environ Microbiol. 2004 Dec.

Abstract

A unique selective enrichment procedure has resulted in the isolation and identification of two new genera of marine-derived actinobacteria. Approximately 90% of the microorganisms cultured by using the presented method were from the prospective new genera, a result indicative of its high selectivity. In this study, 102 actinomycetes were isolated from subtidal marine sediments collected from the Bismarck Sea and the Solomon Sea off the coast of Papua New Guinea. A combination of physiological parameters, chemotaxonomic characteristics, distinguishing 16S rRNA gene sequences, and phylogenetic analysis based on 16S rRNA genes provided strong evidence for the two new genera (represented by strains of the PNG1 clade and strain UMM518) within the family Micromonosporaceae. Biological activity testing of fermentation products from the new marine-derived actinomycetes revealed that several had activities against multidrug-resistant gram-positive pathogens, malignant cells, and vaccinia virus replication.

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Figures

FIG. 1.

FIG. 1.

Colony and spore morphologies of marine-derived actinomycetes of group I, group II, and group III. (A) NaST21Cx plate containing different marine actinomycete colonies (arrows) radiating outward from marine sediment particulate matter. (B) Isolation of UMM486 (group I) showing the growth pattern, phenotype characteristics, and concentric ring formation of group I members on NaST21Cx. (C) Isolation of UMM500 (group II) showing the growth pattern and phenotype characteristics of group II members on NaST21Cx. (D) Isolation of UMM518 (group III) showing the growth pattern and phenotype characteristics of group III members on NaST21Cx. (E) Scanning electron micrograph of UMM486 submerged spores (arrows). (F) Scanning electron micrograph of UMM518 submerged spores (arrows).

FIG.2.

FIG.2.

Phylogram depicting the phylogenetic relationships based on the 16S rRNA gene sequences of all described members of the family Micromonosporaceae and selected strains belonging to groups I, II, and III (10, 12-14, 22, 32). Bootstrap values (those calculated to be greater than 50% by both methods) from 1,000 resamplings are shown at their respective nodes with neighbor-joining values on the left and maximum-parsimony values on the right. Actinomadura fulvescens, Streptomyces coelicolor, and Micrococcus luteus were used as outgroups in tree constructions. Marine clades of the family Micromonosporaceae are enclosed in brackets. The closest terrestrial relative of the marine Micromonosporaceae clades, M. nigra, a salt pond isolate (Syracuse, N.Y.) (35), is indicated by an asterisk. The relative distances of branch lengths are a reflection of the substitutions per nucleotide position within the 16S rRNA gene sequences (scale bar).

FIG. 3.

FIG. 3.

Ribotype analysis of PNG1 clade strains (based on PvuII digestion). (Left) Riboprint dendrogram generated by comparing the VCC riboprint patterns of various PNG1 clade strains by using BioNumerics software. (Right) VCC riboprint for PNG1 clade strains.

FIG. 4.

FIG. 4.

Selectivity of the NaST21Cx isolation method. Growth differences of PNG1 clade member UMM553 on NaST21Cx with cellulose (left panel) and NaST21Cx without cellulose (right panel) following 14 days of incubation at 30°C.

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