Otimização das condições de cultivo de Rhizopus microsporus var. rhizopodiformis para a produção, isolamento e identificação de metabólitos com atividade antimicrobiana / Otimization of the conditions of cultivation of Rhizopus microsporus var. rhizopodiformis for the production, isolation and identification of metabólites with antimicrobial activity

AUTOR(ES)

Ana Silvia Ciscato Camillo

DATA DE PUBLICAÇÃO

2007

RESUMO

The fungus Rhizopus microsporus var. rhizopodiformis was cultivated in different fermentative media aiming to produce secondary metabolites bearing antimicrobial activity. The conidia storage in silica gel was incubated in oat medium for conidia production. After seven days the conidia (4 x 106 conidia/mL) was transferred to a pre-fermentative medium and incubated for 24 h for mycelium production. After that, it was transferred to three fermentative media: Czapek, Jackson and Vogel, and incubated for different periods of time. After fermentation, cultures were filtered and the broth was submitted to partition using ethyl acetate and n-buthanol in sequence, which were, afterwards, recovered under vacuum. The obtained crude extracts were evaluated under bioautography assay aiming to select the most adequate medium for secondary antimicrobial metabolite production. The ethyl acetate extracts obtained from both Jackson and rice media displayed the higher activities. Therefore, they were submitted to different chromatographic means, including silica gel column and HPLC, furnishing four isolated compounds, from which three were identified: two diketopiperazines (cyclos Leu-Pro and Leu-4-OH-Pro) and one aromatic compound containing a nitrogen in the aliphatic moiety. The pure isolated compounds were submitted to microdilution tests against the following bacteria: Kocuria rhizophila (ATCC 9341), Staphylococcus aureus (ATCC 25923) Candida albicans (ATCC 10231), Escherichia coli (ATCC 25922) e a Pseudomonas aeruginosa (ATCC 27853), aiming to determine their Minimum Inhibitory Concentrations (MIC). All the ethyl acetate extracts displayed antimicrobial activity in the bioautography assays against S. aureus and K. rhizophila, as well as the mycelia methanol extracts. Only the n-butanol extracts did not display activity. Regarding the microdilution assay, three of evaluated pure compounds displayed MIC values between 400 and 350 μg/mL. The obtained results indicated that the studied fungus is an interesting source for biologically active secondary metabolites production.

ASSUNTO(S)

antimicrobial activity metabolismo secundário rhizopodiformis cultivation perfil cromatográfico chromatography profile secondary metabolites rhizopus microsporus var rhizopodiformis cultivo mic atividade antimicrobiana rhizopus microsporus var

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