The generic position of two isolates from soils inside a gold mine cave in Kongju, Korea, was determined by 16S rDNA sequencing and chemotaxonomic characteristics. Phylogenetic analysis indicated that both of the isolates formed a clade with Lentzea albidocapillata and members of the genus Saccharothrix of the family Pseudonocardiaceae. The chemical composition of the isolates and of Lentzea albidocapillata was consistent with that of the genus Saccharothrix, which is characterized by a type III cell wall (the mesoisomer of diaminopimelic acid, and galactose and rhamnose as characteristic whole-cell sugars), MK-9(H 4 ) as the major menaquinone, and a phospholipid type PII pattern (phosphatidylethanolamine as a diagnostic phospholipid). The combination of morphological features, chemotaxonomic characters and phylogenetic data supported the proposal that Lentzea albidocapillata, the only and type strain of the genus Lentzea, should be transferred to the genus
Streptomyces blastmyceticus strain 12-6 was isolated from a forest soil sample of Cheonan area on the basis of strong antifungal activities against plant pathogenic fungi. Butanol extracts of the cultural filtrates were active against C. acutatum, C. coccodes, C. gloeosporioides, F. oxysporum, and T. roseum. Active fractions were prepared by thin layer chromatography using silica gel plate; 12-6-2 (Rf 0.36), 12-6-3 (Rf 0.44). Scanning electron microscopy showed that the active fractions caused a change in surface texture of fungal spores from smooth surface to wrinkled surface. The lethal effect on the spores of the active fractions varied from 56% to 100%. It was shown that the spores of C. acutatum were more sensitive to the antifungal fractions than the spores of F. oxysporum. Fluorescence staining using TOTO-1 indicated that the antifungal fractions could make the spores more sensitive to the fluorescence dye. Thus, it was suggested that antifungal agents prepared in this study exhibited the antifungal activity by damaging the plasma membrane of both fungal spores and hyphae. Identification of antifungal agents in the active fraction using GC-MS analysis revealed the presence of cyclo-(Leu-Pro) and 9-octadecenamide as major components that have already been known as antifungal substances.
Marine bacterial isolates were screened for phospholipase C (PLC) activity on PCY agar plates containing phosphatidylcholine (PC) as substrate. The strain that showed the highest activity on a PCY screening agar plate and a thin-layer chromatography was identified as a strain of Pseudoalteromonas and subsequently designated Pseudoalteromonas sp. J937. The extracellular PLC of the strain J937 was purified to a specific activity of 33 U mg(-1) protein by serial ion exchange and gel filtration column chromatography. It had a molecular mass of 32 kDa estimated by SDS-PAGE. The optimal pH and temperature of the enzyme were about pH 8 and 45 degrees C, respectively. The PLC hydrolyzed phosphatidylethanolamine as well as PC but not other glycerophospholipids. Its activity was enhanced by 150% with Ca2+ (200 mM) and by 180% with Na+ (500 mM), suggesting that the purified PLC is a marine-type enzyme.
During antioxidant screening using 1,1-diphenyl-picrylhydrazyl (DPPH) and a lipid peroxidation assay, a streptomycete strain was found to produce herbimycin A and dihydroherbimycin A as antioxidants in the culture filtrate. These molecules were identified by using spectral analyses, including infrared, ultraviolet, mass spectrum, and nuclear magnetic resonance assays. In the DPPH radical-scavenging assay, dihydroherbimycin A exhibited more potent antioxidant activity (IC(50), 1.3 microM) than alpha-tocopherol (IC(50), 2.7 microM) that was used as a reference compound. In the lipid peroxidation assay, both herbimycin A and dihydroherbimycin A demonstrated antioxidant activities of 61% and 72%, respectively, at 100 microg/ml, while alpha-tocopherol exhibited an activity of 93% at the same concentration. Therefore, dihydroherbimycin A might have the potential to be developed into a new therapeutic agent.
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