Monitoramento de cultivos submersos de linhagens de Streptomyces clavuligerus para a quantificação de distribuição de fluxos metabólicos

• Ano de defesa: 2015
• Autor(a) principal: Baptista, Amanda Salvador [UNESP]
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Estadual Paulista (Unesp)
• Palavras-chave em Português: Biotecnologia; Compostos bioativos; Metabólitos; Modelos matematicos; Microbiologia; Microbiology
• Link de acesso: http://hdl.handle.net/11449/124413

Streptomyces clavuligerus is a species that produces multiple secondary metabolites of therapeutic interest: clavulanic acid, a potent inhibitor of beta-lactamases, and other twenty clavams, several of them with antifungal activity; cephamycin C, a beta-lactam antibiotic used to produce various semi synthetic antibiotics with activity against anaerobic bacteria; tunicamycin, a mixture of homologues nucleoside that inhibits protein glycosylation in prokaryotes and eukaryotes; holomycin, a dithiolopyrrolone with bacteriostatic and cytotoxic activities in mammals. In this project we compared the behavior of strains ATCC 27064 of S. clavuligerus and Hol.80 mutant, non producer of clavulanic acid in submerged cultures. Kinetic parameters and yield coefficients were determined. The experimental data of the cultures were used to simulate the metabolic fluxes distribution for each strain. It was employed a metabolic model previously built up in a Doctoral Thesis and the simulation was carried on with the aid of a computer program designed for this purpose. Batch cultivations with the wild strain in conventional bioreactor using a combination of maltose and lysine resulted in a production of 90 mgL-1 of cephamycin C and 100 mgL-1 of clavulanic acid, without holomycin production. At same conditions, the mutant produced 50 and 60 mgL-1 of cephamycin C and holomycin, respectively. The same combination (maltose and lysine) supplemented with glutamate stimulated the production of holomycin by the wild strain (95 to 110 mgL-1), with a production of cephamycin C and clavulanic acid 65% lower. The mutant produced in the same medium approximately 120 mgL-1 of cephamycin C and 75 mgL-1 of holomycin. These results showed that different nitrogen sources promote the production of various bioactives. The most significant data obtained from the model simulations indicated that the holomycin biosynthesis...