RESUMO
Fermented culture of Streptomyces fulvissimus was found to secrete an antibacterial protein inhibitory to Micrococcus luteus, Bacillus subtilis, Bacillus cereus and methicillin resistant Staphylococcus aureus (MRSA) strains. The extracellular protein from the fermented culture on concentration revealed a high molecular weight peptide of 63kDa on SDS-PAGE gel and the region on gel displayed inhibitory activity against methicillin resistant Staphylococcus aureus. Bioactivity of the extra cellular protein was non-sensitive to proteinase K, alpha chymotrypsin, protease, EDTA (ethylene diamine tetra acetic acid), PMSF (phenyl methyl sulfonyl fluoride) and DMSO (dimethyl sulfoxide) but partially susceptible to amylase and heat. Glycoprotein nature of the proteinaceous compound was confirmed by periodic acid schiffs (PAS) staining. The secretary protein of S. fulvissimus demonstrated a significant activity against MRSA strain. It could be an important source for developing new drugs to control multidrug resistant gram positive bacteria.
Assuntos
Antibacterianos/isolamento & purificação , Proteínas de Bactérias/isolamento & purificação , Eletroforese em Gel de Poliacrilamida , Fermentação , Resistência a Meticilina/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Staphylococcus aureus/efeitos dos fármacos , Streptomyces/metabolismoRESUMO
An actinomycin-D producing strain was isolated from soil and characterized as Streptomyces sindenensis. The culture was subjected to UV irradiation and a mutant with 400 percent higher actinomycin-D production was isolated (400 mg/l-1 as compared to 80 mg/l-1 produced by the parent). Production medium was optimized and antibiotic yield with the mutant was enhanced to 850 mg/l-1 which is 963 percent higher as compared with the parent.
Uma cepa produtora de actinomicina-D foi isolada de solo e caracterizada como Streptomyces sindenensis. A cultura foi submetida à radiação UV, e um mutante capaz de produzir 400 por cento mais actinomicina-D foi isolado (400mg/L comparado a 80mg/L produzido pela cepa parental). O meio de produção do antibiótico foi otimizado e o rendimento aumentou para 850 mg/L, ou seja, 963 por cento mais alto que a cepa parental.
Assuntos
Antibacterianos/isolamento & purificação , Dactinomicina/isolamento & purificação , Mutagênicos , Radiação , Streptomyces antibioticus/isolamento & purificação , Métodos , Solo , MétodosRESUMO
Production of actinomycin-D, by an isolate, S. sindenensis, was optimized by statistical methods. Fructose peptone and NaNO3 were found to be critical for antibiotic production. In the second step, their concentrations were optimized with Central Composite Design and Response Surface Methodology. Fructose, peptone and NaNO3 at 2.55, 0.309 and 0.114% respectively gave approximately 261% higher yield (289 mg/l). Cultivation in fermentor at 600 rpm agitation and 1.5 vvm aeration with optimized medium gave 3.56 folds higher yield (365 mg/l) as compared to the yields in shake flasks using normal production medium (80 mg/l).
Assuntos
Biomassa , Reatores Biológicos , Dactinomicina/biossíntese , Indústria Farmacêutica , Modelos Estatísticos , Dados de Sequência Molecular , RNA Ribossômico 16S , Streptomyces/classificaçãoRESUMO
An actinomycin-D producing strain was isolated from soil and characterized as Streptomyces sindenensis. The culture was subjected to UV irradiation and a mutant with 400% higher actinomycin-D production was isolated (400 mg/l-1 as compared to 80 mg/l-1 produced by the parent). Production medium was optimized and antibiotic yield with the mutant was enhanced to 850 mg/l-1 which is 963% higher as compared with the parent.
Uma cepa produtora de actinomicina-D foi isolada de solo e caracterizada como Streptomyces sindenensis. A cultura foi submetida à radiação UV, e um mutante capaz de produzir 400% mais actinomicina-D foi isolado (400mg/L comparado a 80mg/L produzido pela cepa parental). O meio de produção do antibiótico foi otimizado e o rendimento aumentou para 850 mg/L, ou seja, 963% mais alto que a cepa parental.
RESUMO
Biodiversity is an addition sum of the studies on genetic, taxonomic commercial and ecosystem aspects of living systems. All the living individuals of a species contain a distinct combination of genes and the intrinsic interaction among the gene pool influences evolution, survival and phenotypic/genotypic changes of the part of the biodiversity i.e. community. The amount of genetic diversity within population varies tremendously and much of modern conservation biology is concerned with the maintenance of genetic diversity within the population of plants, animals and microbes. Germplasm, obtained with the vast biodiversity, provides a major source of biological material for the development of medicines, vaccines, pharmaceutical products, improved crop and animal varieties and for other environmental applications. Industrialized nations, who have the technology and resources to patent and develop commercial biological products, are having the benefits of biodiversity through the collected and conserved germplasm flowing through the international research centers. In fact a particular genetic contribution usually represents only a small percentage of the total value of the eventual products. In addition, the research and development process required to commercialize a particular product requires enormous technical efforts. The principle of patenting genes is the morally or ethically correct is a matter of intense debate. However, geneticists, having conceived of the technologies with vast and immediate therapeutic, food and environmental values must try to bring to the material to market as soon as possible.
Assuntos
Biodiversidade , Biotecnologia/economia , Genética Microbiana , Genoma , Genômica , Microbiologia Industrial/economiaRESUMO
Optimization of the fermentation medium for maximum alkaline protease production was carried out with a new strain of Pseudomonas aeruginosa (B-2). Replacing the protein source/inducer (albumin in place of casein) brought about significant increase in yield after 48 hr of inoculation. Three most effective medium constituents identified by initial screening method of Plackett-Burman were albumin, (NH4)2SO4 and glucose. Central Composite Design (CCD) and Response Surface Methodology (RSM) were used in the design of the experiment and in the analysis of the results. Optimum levels of the effective medium constituents were albumin (6.586%); (NH4)2SO4, 0.164%; and glucose, 6.72%. The alkaline protease production increased from 533460 to 793492 Ul(-1).