Thesis (M.S.)--Oklahoma State University, 2010. / Vita. Includes bibliographical references.

O presente projeto de pesquisa foi desenvolvido no ano de 2015 in loco em 109 nascentes da Sub Bacia de Posses no município de Extrema, MG que está inserida na Bacia Hidrográfica do Rio Jaguari. Entendeu-se que o Biossistema da Nascente é composto por um conjunto de funções biológicas, ecológicas e geológicas que se interagem entre si e se completam com a ação antrópica e o ciclo hidrológico. Objetivou-se neste trabalho apresentar uma ferramenta prática de gestão, que seja objetiva e eficiente para análise da qualidade do Biossistema das Nascentes, baseado no estudo e análise dos indicadores que envolvem a sua dinâmica, valorizando as interações entre as dimensões ecológica, social e antrópica, organizando os aspectos de forma quantitativa e qualitativa com o objetivo de identificar o nível de conservação em que se encontra o Biossistema da Nascente de forma que esta ferramenta de análise denominada Protocolo de Campo venha a ser utilizada por gestores da qualidade das águas das bacias hidrográficas. Para isso foi utilizado o Protocolo de Campo composto por 11 indicadores, também foi feito uma análise exploratória dos parâmetros de qualidade dos Biossistemas das nascentes da Sub Bacia de Posses em Extrema - MG, para isso foram coletadas amostra de águas das nascentes, medição da vazão e análise do número mais provável de coliformes. Os dados obtidos pelo Protocolo de Campo demonstraram que dos 109 Biossistemas de Nascentes analisados, 64 se enquadraram como Perturbado, 41 como Conservados e apenas 4 atingiram a pontuação para se enquadrar como Preservado, sendo que nenhum dos Biossistemas das Nascentes estudados se enquadraram na categoria Degradada, porém notou-se que o entorno das nascentes é ocupado em sua grande maioria por pastagem sendo 92,66%, seguido por Eucalipto com 7,34%, apresentaram fisionomia herbácea 43,12%, fisionomia arbustiva entre 1 a 5 metros 43,12% e 13,76% fisionomia arbustiva acima de 5 metros, contudo mais de 90% dos Biossistemas das Nascentes analisados apresentaram ausência de lixo, ausência de fogo e ausência de assoreamento. / This research was developed in 2015 at 109 water springs of the sub basin from Posses, Extrema, MG, that are inserted at Jaguari\'s River basin. The result is that the water spring\'s biosystem is composed by a group of biological, ecological and geological functions, and they interact with each one and they are completed with antropic action and the hydrological cycle. The objective of this research was to present a practical tool of management, and that must be objective and efficient to analyze the quality of water spring\'s biosystem, based on studies and analysis of the indicators that involves it\'s dynamic, doing the interaction between ecological, social and anthropical dimensions very important, organizing the aspects in a quantitative and qualitative form and the objective must be identify in each level of conservation is the water spring\'s biosystem analized, after that the analyzis tool named by \"Protocolo de Campo\" will be used by water quality managers. To get this, the \"Protocolo de Campo\" was composed by 11 indicators and a research of the quality parameters of the water spring\'s biosystems from Posses in Extrema - MG was made too, for this were collected water samples at the water springs, flow measurement and analysis of the most probable number of coliforms. The data obtained in the field by the \"Protocolo de Campo\" revealed that of the 109 water spring\'s biosystems analized, 64 are in the \"disturbed group\", 41 are in the \"conserved group\" and only 4 are in the \"preserved group\". None of them is in the \"degraded group\", but the research has noticed that the surroundings of the water springs is ocuppied in the most by pasture, being 92,66%, eucalyptus, being 7,34%, they represent 43,12% of herbaceous physiognomy, 43,12% represent shrub physiognomy between 1 and 5 meters and 13,76% represent shrub physiognomy above 5 meters, even more than 90% of the water spring\'s biosystem analized presented absence of rubbish, absence of fire and absence of silting.

O presente projeto de pesquisa foi desenvolvido no ano de 2015 in loco em 109 nascentes da Sub Bacia de Posses no município de Extrema, MG que está inserida na Bacia Hidrográfica do Rio Jaguari. Entendeu-se que o Biossistema da Nascente é composto por um conjunto de funções biológicas, ecológicas e geológicas que se interagem entre si e se completam com a ação antrópica e o ciclo hidrológico. Objetivou-se neste trabalho apresentar uma ferramenta prática de gestão, que seja objetiva e eficiente para análise da qualidade do Biossistema das Nascentes, baseado no estudo e análise dos indicadores que envolvem a sua dinâmica, valorizando as interações entre as dimensões ecológica, social e antrópica, organizando os aspectos de forma quantitativa e qualitativa com o objetivo de identificar o nível de conservação em que se encontra o Biossistema da Nascente de forma que esta ferramenta de análise denominada Protocolo de Campo venha a ser utilizada por gestores da qualidade das águas das bacias hidrográficas. Para isso foi utilizado o Protocolo de Campo composto por 11 indicadores, também foi feito uma análise exploratória dos parâmetros de qualidade dos Biossistemas das nascentes da Sub Bacia de Posses em Extrema - MG, para isso foram coletadas amostra de águas das nascentes, medição da vazão e análise do número mais provável de coliformes. Os dados obtidos pelo Protocolo de Campo demonstraram que dos 109 Biossistemas de Nascentes analisados, 64 se enquadraram como Perturbado, 41 como Conservados e apenas 4 atingiram a pontuação para se enquadrar como Preservado, sendo que nenhum dos Biossistemas das Nascentes estudados se enquadraram na categoria Degradada, porém notou-se que o entorno das nascentes é ocupado em sua grande maioria por pastagem sendo 92,66%, seguido por Eucalipto com 7,34%, apresentaram fisionomia herbácea 43,12%, fisionomia arbustiva entre 1 a 5 metros 43,12% e 13,76% fisionomia arbustiva acima de 5 metros, contudo mais de 90% dos Biossistemas das Nascentes analisados apresentaram ausência de lixo, ausência de fogo e ausência de assoreamento. / This research was developed in 2015 at 109 water springs of the sub basin from Posses, Extrema, MG, that are inserted at Jaguari\'s River basin. The result is that the water spring\'s biosystem is composed by a group of biological, ecological and geological functions, and they interact with each one and they are completed with antropic action and the hydrological cycle. The objective of this research was to present a practical tool of management, and that must be objective and efficient to analyze the quality of water spring\'s biosystem, based on studies and analysis of the indicators that involves it\'s dynamic, doing the interaction between ecological, social and anthropical dimensions very important, organizing the aspects in a quantitative and qualitative form and the objective must be identify in each level of conservation is the water spring\'s biosystem analized, after that the analyzis tool named by \"Protocolo de Campo\" will be used by water quality managers. To get this, the \"Protocolo de Campo\" was composed by 11 indicators and a research of the quality parameters of the water spring\'s biosystems from Posses in Extrema - MG was made too, for this were collected water samples at the water springs, flow measurement and analysis of the most probable number of coliforms. The data obtained in the field by the \"Protocolo de Campo\" revealed that of the 109 water spring\'s biosystems analized, 64 are in the \"disturbed group\", 41 are in the \"conserved group\" and only 4 are in the \"preserved group\". None of them is in the \"degraded group\", but the research has noticed that the surroundings of the water springs is ocuppied in the most by pasture, being 92,66%, eucalyptus, being 7,34%, they represent 43,12% of herbaceous physiognomy, 43,12% represent shrub physiognomy between 1 and 5 meters and 13,76% represent shrub physiognomy above 5 meters, even more than 90% of the water spring\'s biosystem analized presented absence of rubbish, absence of fire and absence of silting.

The production of hydrogen from low-cost abundant renewable biomass would be vital to sustainable development. Cell-free (in vitro) biosystems comprised of synthetic enzymatic pathways would be a promising biomanufacturing platform due to several advantages, such as high product yield, fast reaction rate, easy control and access, and so on. However, it is essential to produce (purified) enzymes at low costs and stabilize them for long periods to decrease biocatalyst costs. Thermophilic recombinant enzymes as building blocks were discovered and developed: fructose 1,6-bisphosphatase (FBP) from Thermotoga maritime, phosphoglucose isomerase (PGI) from Clostridium thermocellum, triose phosphate isomerase (TIM) from Thermus thermophiles and fructose bisphosphate aldolase (ALD) from T. maritima and T. thermophilus. The recombinant proteins were over-expressed in E. coli, purified and characterized. For purification and stabilization of enzymes, one-step, simple, low-cost purification and immobilization methods were developed based on simple adsorption of cellulose-binding module (CBM)-tagged protein on the external surface of high-capacity regenerated amorphous cellulose. Also, a simple, low-cost purification method of thermophilic enzymes was developed utilizing a combination of heat and ammonium sulfate precipitation. For development of cascade enzymes as building modules (biocatalyst modules), it was discovered that the presence of other enzymes/proteins had a strong synergetic effect on the stabilization of the thermolabile enzyme (e.g., PGI) due to the in vitro macromolecular crowding effect. And substrate channeling among CBM-tagged self-assembled three-enzyme complex (synthetic matabolon) immobilized on the easily-recycled cellulose-containing magnetic nanoparticles can not only increase cascade reaction rates greatly, but also decrease enzyme cost in cell-free biosystems. The high product yield and fast reaction rate of dihydrogen from sucrose was validated in a batch reaction containing fifteen enzymes comprising a non-natural synthetic pathway. The yield of dihydrogen production from 2 mM of sucrose was 96.7 % compared to theoretical yield at 37 °C. The maximum rate was increased 3.1 fold when the substrate concentration was increased from 2 to 50 mM in a fed-batch reaction. The research and development of cell-free biosystems for biomanufacturing require more efforts, especially in low-cost recombinant thermostable enzymes as building blocks, efficient cofactor recycling, enzyme and cofactor stabilization, and fast reaction rates. / Ph. D.