Rice bran was used as the substrate for screening nine strains of Rhizopus sp. for neutral protease production by solid-state fermentation. The best producer, Rhizopus microsporus NRRL 3671, was used for optimizing the process parameters for enzyme production. Fermentation carried out with 44.44 % initial moisture content at a temperature of 30 C for 72 h was found to be the optimum for enzyme secretion by the fermenting organism. While most of the carbon supplements favored enzyme production, addition of casein resulted in a marginal increase in protease yield. Fermentation was then carried out under optimized conditions to obtain the crude extract of the enzyme, which was partially purified by precipitation and dialysis. A 3-fold increase in the enzyme purity was achieved in this manner. The enzyme was found to be a metalloprotease, being activated by Mn2+, with maximal activity at a temperature of 60 C and pH 7.0. Farelo de arroz foi utilizado como substrato para seleção de nove linhagens de Rhizopus sp. com vistas a produção de protease neutra. A linhagem que apresentou maior produtividade da enzima foi Rhizopus microsporus NRRL 3671, sendo utilizada na otimização dos parâmetros do processos e produção da enzima. As condições otimizadas para produção da enzima foram 44% de umidade inicial, temperatura de 30ºC e 72h de fermentação.A suplementação do farelo de arroz com uma fonte de carbono favoreceu a produção da enzima, porém a adição de caseína resultou em um aumento marginal do rendimento em protease. Condições otimizadas foram utilizadas para obtenção do extrato cru da enzima que foi parcialmente purificado por precipitação e diálise. A enzima purificada teve sua atividade incrementada 3 vezes. A enzima foi classificada como metalo-protease, sendo ativada pelo Mn2+ , sendo que sua atividade máxima foi obtida a temperatura de 60ºC e a pH 7.0
The present work deals with isolation of Streptomyces associated with marine sponges and its bioactive potential. Streptomyces sp. were isolated from the marine sponges Callyspongia diffusa, Mycale mytilorum, Tedania anhelans and Dysidea fragilis. From the initial screening, 94 cultures of Streptomyces were obtained and from these 58 cultures exhibited antagonism against bacteria, 36 strains against fungi and 27 strains exhibited broad spectrum activity against both. The submerged culture extracts of the 58 anti-bacterial isolates were analysed and of these 58 strains, 37 strains showed positive inhibition against Bacillus subtilis, 43 against Staphylococcus aureus, 10 against Vibrio cholerae and 10 against Escherichia coli. The antifungal activities of the 36 strains were also evaluated and 27 strains showed positive inhibition against Aspergillus niger, 23 against Saccharomyces cerevisiae and 16 against Candida albicans. The production of polyene substances from the active extracts was confirmed by UV spectral analysis by the absorbance peaks that ranged from 225 to 262 nm and the TLC (R f values) ranging from 0.40 to 0.78. The results indicate that Streptomyces strains isolated from marine sponges produce potential antibacterial, antifungal and broad spectrum antibiotic compounds.
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