RESUMEN
Three out of 17 Streptomycetes strains - Streptomyces sp. 35 LBG09, Streptomyces sp. 36 LBG09, and Streptomyces sp. 39 LBG09, were selected based on the high production of proteinase inhibitors with trypsin serine proteinase activity. The strains were isolated from soil samples taken from the area around the Bulgarian station on Livingston Island, Antarctica. Biosynthesis of proteinase inhibitors by the promising strains started at different stages of their development but was generally not associated with the growth of the producers. Peak levels were reached in the stationary phase (96-120 h) of their cultivation. Inducing effects on strain development, biomass accumulation, and proteinase inhibitor biosynthesis were based on the composition of the nutrient medium: the polypeptones contained in Taguchi medium and glucose as a carbon source. The most productive out of the three strains was Streptomyces sp. 36 LBG09. Its maximum inhibitory activity was reached at 96 h in culturing media modified by three different carbon sources. The active proteinase inhibitor biosynthesis proceeded at pH values between 6.8 and 8.6 and the dynamics of production depended on the type of carbon source. Peak levels of extracellular protein and dry biomass were reached at 120 h in the stationary growth phase. The residual sugars were minimal at the end of the process when using soluble starch as a carbon source, and maximal when glucose was used.
Asunto(s)
Inhibidores de Proteasas/metabolismo , Streptomyces/metabolismo , Regiones Antárticas , Carbono , Medios de Cultivo , Concentración de Iones de Hidrógeno , Inhibidores de Proteasas/aislamiento & purificación , Microbiología del Suelo , Streptomyces/crecimiento & desarrollo , Inhibidores de Tripsina/metabolismoRESUMEN
The exploration of habitats with unusual environment and poorly explored areas such as Antarctica is one of the strategies for discovery of new biologically active substances and/or new producers. The aim of this study was to identify the actinomycetes isolated from the soils of the island Livingston - Antarctica and to investigate their potential to synthesize antibacterial agents against phytopathogens. Twenty-three actinomycete strains were the object of this study. Using PCR (polymerase chain reaction) amplification all strains were affiliated to genus Streptomyces. The sequencing of the 16S rRNA for three of the strains showed greatest similarity to Streptomyces tendae for one of them, and revealed that the other strains had closest relations to streptomycetes isolated from anthropogenically unaltered regions including Antarctica. The isolates were studied for production of antibacterial substances both by molecular and culture methods. PCR targeting specific biosynthetic genes involved in the production of some groups of antibiotics was performed. The screening showed that all strains possessed the gene for Type-II polyketide synthase, 11 strains - for non-ribosomal peptide synthetase; 6 strains - for polyene antibiotics; and 4 strains - for glycopeptide antibiotics. The production of antibacterial substances by the strains was tested in vitro against phytopathogenic bacteria. The strains differed in the number of inhibited test - bacteria and in their spectrum of action. Four strains showed a wide range of activity against Gram-positive and Gram-negative phytopathogens. The results obtained revealed that the Antarctic soils are potential source for isolation of streptomycetes producing antibiotics from different groups.
RESUMEN
Preliminary differentiating screening of the antibacterial and antifungal activity of a series of diastereomeric cis/trans-3-aryl(heteroaryl)-3,4-dihydroisocoumarin-4-carboxylic acids (3a-i) was performed by the agar diffusion method against twelve microorganism strains of different taxonomic groups. S. aureus and A. niger were the most sensitive strains to the antibiotic effect of the tested compounds, both inhibited by 10 of 12 compounds. The most potent antibacterial agent was cis-3-phenyl-3,4-dihydroisocoumarin-4-carboxylic acid (cis-3a), exhibiting activity against all seven bacterial test strains.