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Estrat?gias de cultivo para produ??o de biomassa e promotores de crescimento por Herbaspirillum seropedicae BR-11417 / Culture strategies for biomass production and growth promoters by Herbaspirillum seropedicae BR-11417SCHEIDT, Wagner 27 October 2015 (has links)
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Previous issue date: 2015-10-27 / CAPES / The bacteria Herbaspirillum seropedicae is established as diazotrophic, nitrogen fixing, gram-negative, vibrioid, ?-proteobacteria, showing ability to synthesize phytohormones growth promoters in plants. The goal of this work was to improve the fermentation production of H. seropedicae, seeking the maximum production of cells and indole compounds with process cost reduction. The optimization of the culture medium Yeast Dextrose Glucose Sucrose - DYGS ? was conducted through fermentation tests on shaker. The original carbon sources were replaced, and the concentrations of nitrogen and nutrient sources were optimized through the Central Composite Rotational Design - CCRD (2?). The simplified culture medium obtained was composed of glycerol (5.5g L-1), yeast extract (2.8 g L-1), MgSO4?7H2O (0.5 g L-1) e K2HPO4 (0.5 g L-1). This medium provided an increase of 87.5 % in biomass production, and is 49.8 % cheaper than the traditional culture medium DYGS. An experimental design CCRD (2?) was carried with the purpose of improving the operational conditions of the bioreactor and the best growing condition was obtained at 35 ?C, 3 LPM of aeration rate, and 180 RPM of agitation speed. Under these conditions, and using the previously optimized culture medium, the maximum cell production (X= 3.23 g L-1) as well as the maximum metabolic product (P= 9.02mg L-1) were obtained. It was also tested a culture medium with glycerol and yeast extract concentrations increased to 8.0 g L-1 and 4.0 g L-1, respectively. Under these conditions, it was observed that the maximum cell production and metabolic product increased to X = 4.18 g L-1 and P = 11.74 mg L-1, respectively. An appropriate mathematical computer simulation model was developed and used to describe the dynamic behavior of the main variables of fermentation bioreactor. However, there was some difficulty to reproduce appropriately the profiles of all variables, probably due to the specific characteristics of this fermentation process, and the necessity of a mathematical model that takes into account parameters associated to limitations and inhibitions caused by substrate and product. / A bact?ria Herbaspirillum seropedicae ? estabelecida como sendo diazotr?fica fixadora de nitrog?nio, gram-negativa, vibri?ide, membro da classe ? das proteobact?rias, apresentando a capacidade de produ??o de fitohorm?nios promotores do crescimento em plantas. Este trabalho apresentou como objetivos aprimorar o processo fermentativo de produ??o de H. seropedicae, buscando a m?xima produ??o de c?lulas e compostos ind?licos, com redu??o de custos do processo. Realizou-se a otimiza??o do meio de cultura Dextrose Yeast Glucose Sucrose modificado ? DYGS ? atrav?s de ensaios fermentativos em shaker. Substitu?ram-se as fontes de carbono originais e aperfei?oaram-se as concentra??es dos demais componentes atrav?s da matriz experimental 2? obtida atrav?s do Delineamento Composto Central Rotacional ? DCCR. Obteve-se um meio de cultura simplificado, composto por glicerol (5,5 g L-1), extrato de levedura (2,8 g L-1), MgSO4?7H2O (0,5 g L-1) e K2HPO4 (0,5 g L-1) que se apresentou 49,8 % mais barato e com capacidade de produ??o de c?lulas 87,5 % maior quando comparado ao meio DYGS. Atrav?s da DCCR (2?) aprimorou-se tamb?m as condi??es operacionais de biorreator, obtendo-se a melhor condi??o de cultivo a temperatura de 35 ?C, aera??o de 3 LPM e agita??o de 180 RPM. Nestas condi??es, com o meio de cultura otimizado, obteve-se a produ??o celular m?xima de X = 3,23 g L-1 e de produto metab?lico m?ximo de P = 9,02 mg L-1. Testou-se um meio de cultura com concentra??es de glicerol e de extrato de levedura aumentadas para 8,0 g L-1 e 4,0 g L-1, respectivamente, sendo observada uma produ??o celular m?xima de X = 4,18 g L-1 e de produto metab?lico m?ximo de P = 11,74 mg L-1. Avaliou-se atrav?s de simula??o computacional, o modelo matem?tico mais adequado para descrever o comportamento din?mico das principais vari?veis das fermenta??es em biorreator. Contudo, observou-se uma alta dificuldade em descrever adequadamente os perfis de todas as vari?veis avaliadas, provavelmente devido ?s caracter?sticas espec?ficas deste processo fermentativo e ? necessidade de um modelo que leve em considera??o par?metros de limita??es e inibi??es causadas pelo substrato e produto.
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