Submitted by Caroline Perez (lopesperezcaroline@gmail.com) on 2017-03-15T13:18:05Z No. of bitstreams: 1 Caroline_Lopes_Perez_Dissert_Final.pdf: 6948213 bytes, checksum: 88316394d4ffc78684a65d0a022b2c93 (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2017-03-21T14:27:52Z (GMT) No. of bitstreams: 1 perez_cl_me_sjrp.pdf: 6948213 bytes, checksum: 88316394d4ffc78684a65d0a022b2c93 (MD5) / Made available in DSpace on 2017-03-21T14:27:52Z (GMT). No. of bitstreams: 1 perez_cl_me_sjrp.pdf: 6948213 bytes, checksum: 88316394d4ffc78684a65d0a022b2c93 (MD5) Previous issue date: 2017-03-03 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Esta dissertação apresenta um estudo sobre a ampliação de escala de biorreatores de leito empacotado para produção de celulases e hemicelulases por fermentação em estado sólido (FES) empregando o fungo termofílico Myceliophthora thermophila I-1D3b e substrato composto por bagaço de cana de açúcar e farelo de trigo. As etapas de desenvolvimento do projeto consistiram em ensaios em biorreator de diâmetro interno e altura iguais a 7,62 e 10 centímetros, de modo que foram realizados ensaios variando tamanho das partículas de bagaço de cana, densidade bulk do leito, taxas de aeração e modo de distribuição da vazão de ar. Com base nos resultados obtidos, observou-se que partículas de bagaço de tamanho reduzido fornecem bons rendimentos de atividades enzimáticas quando submetidas a altas taxas de aeração em leitos de elevada densidade. Partículas maiores forneceram melhores resultados para leitos de �bulk= 0,395 g/cm3 , bem como a utilização de vazão de ar determinada com base no número de Damköhler modificado igual a 1. Resultados obtidos em ensaios em biorreatores de 0,8 metros, com e sem distribuição de ar, mostraram que inserir ar ao longo do leito é uma estratégia que permite aeração eficiente ao longo do sistema fermentativo, sem que ocorra secagem excessiva do substrato. Atividades relativas de CMCase e xilanase atingiram cerca de 1,2 e 0,7, respectivamente. As etapas finais consistiram em fermentações realizadas em biorreator de diâmetro interno e altura iguais a 20 centímetros. Nessas condições, foram testadas duas formas de distribuição de ar ao longo do leito de quatro módulos fermentativos: utilizando um tubo interno perfurado no módulo central inserindo metade da vazão total, e a outra metade inserida pelo inferior do leito e utilizando um tubo interno central, com quatro orifícios por módulo para espalhamento da vazão. Em termos de produção de celulase, o tubo no módulo central se mostrou a melhor alternativa para o aumento de escala, chegando a apresentar 0,8 de atividades enzimáticas relativas de CMCase, e em termos de atividades relativas de xilanase, a melhor opção foi o tubo interno longitudinal, apresentando valores de 0,55. Diante do exposto, o número de Damköhler modificado é interessante do ponto de vista de aumento de escala, no entanto é preciso ainda entender o mecanismo de atividade respiratória dos microrganismos para que os conceitos possam ser aplicados em sinergia, podendo-se assim esperar bom desempenho do processo em escala industrial. / This work presents a study of scale–up packed bed bioreactors for the production of cellulases and hemicellulases by solid-state fermentation (SSF) using the thermophilic fungus Myceliophthora thermophila I-1D3b and substrate composed by sugar cane bagasse and wheat bran. The development phases of the project consist in tests using bioreactors composed by internal diameter and height equal to 7.62 and 10 centimeters, respectively, so that the assays vary the size of the sugar cane bagasse particles, the bulk density, the airflow rate and how’s distributed along the bed. Based on the results obtained, it was defined that the particles of small size provide good results of enzymatic activity yields when submitted to high aeration rates in high bulk density beds. When testing normal size sugarcane bagasse particles, the best results were obtained for the beds with �bulk= 0,395 g/cm3 as well as airflow rate based on the modified Damköhler number equal to 1. Results obtained in tests in 0,8 meter height bioreactors, with and without airflow distribution, show that inserting air in different bed heights is a strategy to ensure efficient aeration throughout the fermentation system without excessive drying of the substrate. Activities related to CMCase and xylanase reached about 1.2 and 0.7, respectively. The final stages consisted in perform fermentation assays in a bioreactor of internal diameter and height equal to 20 centimeters. Under these conditions, two forms of long-term distribution were tested: using an internal ring to insert half of the total flow, together with the other half inserted in the lower part of the bed and using a central internal pipe with 4 holes per module for airflow distribution. In terms of cellulase production, the internal ring showed a better alternative for scaling-up, reaching a valeu of 0.8 CMCase relative enzymatic activities, and with respect to xylanase relative enzymatic activities, the best option was the central pipe, presenting values of 0,55. On the above, the modified Damköhler number is interesting when it refers to scale-up, although it’s necessary to understand the mechanism of respiratory activity of the microorganisms so that the concepts can be applied in synergy, and it can be expected good process performance in industrial scale. / CNPq: 132957/2016-7

Orientador: Eleni Gomes / Coorientador: Gustavo O. Bonilla-Rodriguez / Banca: Heizir Ferreira de Castro / Banca: Ernandes Benedito Pereira / Banca: Humberto Márcio Santos Milagre / Banca: Teresa Cristina Zangirolami / Resumo: As enzimas são catalisadores muito eficientes e de grande interesse na aplicação industrial. As lipases (E.C. 3.1.1.3) são serina-hidrolases que agem na hidrólise, esterificação e transesterificação de acilgliceróis de cadeia longa. Lipases microbianas têm sido amplamente usadas devido à sua especificidade. Na transesterificação moléculas de triacilglicerol reagem com um álcool na presença de um catalisador formando uma mistura de glicerol e ésteres de ácidos graxos. O biodiesel, definido como ésteres metílicos ou etílicos, tem atraído crescente interesse como uma fonte de energia renovável, substituindo o diesel produzido a partir de combustíveis fósseis. O presente trabalho tem como objetivo principal efetuar a prospecção de fungos termofílicos que apresentem produção significativa de lipase e, concomitantemente, atividade transesterificante. As linhagens foram selecionadas por detecção de atividade lipolítica em placas de ágar contendo Rodamina B, por fermentação submersa (FSm) e fermentação em estado sólido (FES). Foram testadas linhagens de fungos termofílicos da coleção do laboratório de Bioquímica e Microbiologia Aplicada, sendo Thermomucor indicae-seudaticae N31, Rhizomucor pusillus Myceliophtora sp F2.1.4, Myceliophtora sp M7.7, F2.1.1, F2.1.3 e Thermomyces lanuginosus TO-05 os que mostraram um maior potencial hidrolítico. Estudos adicionais avaliaram a produção de lipase através da modificação da fonte de componentes nutricionais e algumas propriedades físicas na produção de lipase por T. indicae-seudaricae N31 em FSm, e por R. pusillus e T. indicae-seudaticae N31 em FES. Os estudos dos processos fermentativos foram bem sucedidos, havendo um aumento de 16 vezes na produção de lipase de R. pusillus e de 36 vezes na lipase de T indicae-seudaticae... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Enzymes are efficient catalysts and interesting for industrial applications. Lipases (EC 3.1.1.3) constitute a group of serine hydrolases that catalyze the hydrolysis, esterification and transesterification reactions of long chain acylglycerols. Microbial lipases have been widely used for biotechnological applications due to their specificity. In transesterification, molecule of a tryacylglicerol react with an alcohol in the presence of catalyst, producing a mixture of fatty esters and glycerol, Biodiesel, defined as methyl or ethyl fatty esters and has attracted considerable attention as a renewable source of energy, in substitution of fossil fuels. The main goal of the present work is the screening of thermophilic fungi that present outstanding lipase production and in parallel able to perform transesterification reaction. Strains were screened for lipase activity on agar plates containing Rhodamine B, for submerged fermentation (SmF) and solid state fermentation (SSF). The tested thermophilic strains were from the collections of the Biochemistry and Applied Microbiology Laboratory, where Thermomucor indicae-seudancae N31, Rhizomucor pusillus Myceliophtora sp F2.1.4, Myceliophtora sp M7.7, F2.1.1, F2.1.3 and Thermomyces lanuginosus TO-O5 had the highest lipase production. Additional studies attempted to improve lipase production by nutrient source modifications and physicals conditions in FmS by T. indicae-seudaticae N31 and in FSS by T. indicae-seudaticae N31 and R. pusillus. The fermentations studies were successful, with a 16 fold enhancement in lipase yield compared to the initial medim from lipase R. pusillus and 36 fold for the lipase T. indicae-seudaticae N31, both in FES. The lipase from T. indicae-seudaticae N31 cultured in SSF and SmF, exhibited maximum lipolytic activity at 40°C and stability for the pH range from 4 to 8. The enzyme produced by FmS presented maximum activity... (Complete abstract click electronic access below) / Doutor

Design theories and methodologies are guidelines to develop design solutions. Among many, the Axiomatic design theory (ADT) and Systematic design procedures (SDP) are two well-known approaches to design. For practical applications, the choice of the design methodology is difficult as there is no study to compare them. To close in such gap in literature, this thesis presents a study on comparison of these two design approaches. To facilitate the comparison, design of a solid state fermenter was taken as a vehicle. The fermenter chosen for this study is was used for detoxification of phorbol esters from Jatropha curcas. Jatopha curcas is a woody plant and is one of the major sources for the production of bio-diesel as it is readily available and has unique composition. Processing Jatopha curcas for biodiesel also yields protein rich Jatopha curcas seed cake. This can be used as animal feedstock, cattle fodder or live feed stock. It is however known that phorbol esters present in the seed cake hinder the utilization of the seed cake as live feed stock. Solid state fermentation by fungi is an effective process to denature phorbol esters, which has been demonstrated at the laboratory scale. Development of an industrial scale solid state fermenter (SSF) is necessary. This study applies SDP and ADT the same deign problem of SSF and compared based on the result of the design. It is noted that in ADT, the evaluation of design alternatives neither considers the cost of the system under design nor the delivery time of the system, but SDP does. To make the comparison on the same ground, an extension of ADT enabling it to consider the cost and delivery time (or time) was developed. Several conclusions can be drawn from this study and they are: (1) ADT and SDP are complementary to each other and the one that integrates both is more effective to design; (2) The essence of Axiom 2 of ADT is to evaluate design alternatives with all factors that lead to difficulty to realize the design, but unfortunately the information content in the current ADT literature only considers the functional or quality aspect; (3) Previous reports suggest the presence of zigzag process only in ADT, However in this study it is evident that SDP exercises the zigzag process as well; (4) the proposed formulation of information content by taking into consideration of the quality, cost, time aspects is more effective in design practice as quite often the cost and time are very important aspects to the customer. The contribution of this thesis study is of two-fold. First, the SSF designed in this study is a pilot one in the field of the biochemical process and it has potential to be implemented. Second, this study concludes several unique findings of ADT and SDP with their relationship, which have further resulted in an integrated ADT and SDP design approach and a more complete formulation of information content capable of evaluating design alternatives from all aspects rather than the functional aspect only.

Submitted by Lucienne Costa (lucienneferreira@ufcg.edu.br) on 2017-12-01T15:43:01Z No. of bitstreams: 1 LAHYANA RAFAELLA DE FREITAS CUNHA - TESE (PPGEP) 2014.pdf: 2398176 bytes, checksum: 737e0981fa2c124c8593a36e031175d1 (MD5) / Made available in DSpace on 2017-12-01T15:43:01Z (GMT). No. of bitstreams: 1 LAHYANA RAFAELLA DE FREITAS CUNHA - TESE (PPGEP) 2014.pdf: 2398176 bytes, checksum: 737e0981fa2c124c8593a36e031175d1 (MD5) Previous issue date: 2014-08-22 / Capes / Uma das formas de remediar poluentes urbanos, como polímeros plásticos, é a utilização de processos biotecnológicos que exploram a capacidade de micro-organismos decompositores, produzirem enzimas na presença de substratos que possuem fontes indutoras adequadas, a exemplo de fungos do gênero Basidiomiceto em substratos como biomassas lignocelulósicas. O presente trabalho tem como objetivo estudar o potencial de substratos como a casca de coco verde, farinha de soja comercial, farelo da palha de milho verde e cana-de-açúcar na produção de lacase e CMCase por meio da fermentação semissólida (FES) utilizando o micro-organismo Psilocybe castanella (CCIBt 2781) como agente da fermentação, bem como, avaliar a degradação de pedaços de copos de polipropileno por esse basidiomiceto, sem e com interferência de mediadores orgânicos e inorgânicos. O fungo foi previamente crescido em ágar extrato de malte (MEA) 2% e inoculado em substratos a base de farelo de palha de milho verde com bagaço de cana-de-açúcar e bagaço da casca de coco verde com farinha de soja comercial nas proporções adequadas para se obter C/N 90 e com os mediadores, óleo vegetal, Tween 80 e azul brilhante de remazol (RBBR). Durante 60 dias de cultivo foram analisados as atividades enzimáticas de lacase e carboximetilcelulase. A degradação causada pela ação microbiana nos polímeros plásticos foi analisada por medidas de perda de massa, microscopia eletrônica de varredura (MEV) e difração de raio X (DRX). A maior atividade da lacase ocorreu em torno do 15º dia de fermentação (95,67 UL-1), contendo bagaço da casca de coco verde como fonte de carbono e soja como fonte de nitrogênio, sob ação dos mediadores tween e RBBR. Para o substrato a base de milho e cana, a maior atividade foi de 69,77 UL-1 em 30 dias, utilizando tween como mediador. A maior produção de CMCase foi de 0,29 U/g, para o substrato a base de milho e cana e de 0,22 U/g no substrato contendo bagaço da casca de coco verde e farinha de soja comercial. Todas as amostras de PP submetidas à incubação nos dois substratos apresentaram alterações nas superfícies dos polímeros, indicadas por ranhuras, sulcos e furos, quando analisadas por MEV. Esses resultados mostram que a bioconversão dos resíduos utilizados nesse trabalho por meio da fermentação semissólida (FES) é uma forma viável de obtenção das enzimas lignocelulolíticas, além da relação positiva entre a degradação de polímeros sintéticos (PPs) e a atividade enzimática de lacase. / One way to remedy urban pollutants such as plastic polymers, would be the use of biotechnological processes that exploit the ability of decomposing micro-organisms produce enzymes in the presence of substrates that have appropriate inducing sources, like basidiomycete fungi of the genus substrates as lignocellulosic biomass. Therefore, the aim of this work was to study the potential of coconut husk and commercial soy flour, bran straw corn and cane sugar as substrates in the production of laccase and CMCase by semisolid fermentation (SSF) Psilocybe castanella (CCIBt 2781) microorganism as the fermentation agent, as well as to evaluate the degradation of pieces of polypropylene cups by this basidiomycete with and without interference from inorganic and organic mediators. The fungus was previously grown on malt extract agar (MEA) 2% and inoculated in substrates bran straw corn with crushed sugar cane bagasse and coconut husk with commercial soy flour in the proportions adequate to obtain C/N 90 and mediators, vegetable oil, Tween 80 and Remazol brilliant blue R (RBBR). During cultivation for 60 days were analyzed, the enzymatic activities of laccase and carboxymethylcellulase, besides the degradation of plastics by polymer mass loss, MEV and DRX. The highest activity of laccase occurred around the 15th day of fermentation (95.67 UL-1), containing bagasse coconut husk as carbon source and soybean as nitrogen source, under the action of mediators tween and RBBR. To the substrate from corn and sugarcane, the highest activity was 69.77 UL-1 in 30 days, using tween as a mediator. The largest production of CMCase was 0.29 U/g, for the substrate from corn and sugarcane and 0.22 U/g substrate containing bagasse in the coconut husk and commercial soy flour. All samples analyzed by MEV and DRX were subjected to incubation with both substrates showed changes in the surfaces of polymers, indicated by grooves, grooves and holes. These results show that bioconversion of waste materials through this work semisolid fermentation (SSF) is a feasible way of obtaining the enzymes lignocellulolytic way, besides the positive relationship between the degradation of synthetic polymers (PP) and enzymatic activity.

Made available in DSpace on 2014-06-11T19:32:55Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-06-05Bitstream added on 2014-06-13T19:22:45Z : No. of bitstreams: 1 martins_es_dr_rcla.pdf: 4883436 bytes, checksum: 90e9eddebe5cd615b809d1494e86d994 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Pectinases termoestáveis apresentam características interessantes do ponto de vista da sua aplicação industrial, como alta estabilidade ao pH e à temperatura. Além disso, o tipo de processo fermentativo pode influenciar a produção e propriedades físico-químicas destas enzimas. A produção de poligalacturonase (PG) pelo fungo Thermoascus aurantiacus foi realizada em fermentação submersa (FSM) e em estado sólido (FES), usando substratos contendo pectina comercial ou subprodutos agro- industriais como fonte de carbono. A PG bruta obtida em FES apresentou atividade ótima a 65ºC e pH 5,0, com estabilidade na faixa de pH entre 4,0 e 5,0 e entre 7,5 e 8,5 e manteve 85% da atividade original quando incubada a 60ºC, por 1 hora. Em FSM, o melhor meio de cultivo foi a água amarela, com pH inicial de 5,5, após 5 dias de cultivo a 45ºC. A enzima em sua forma bruta apresentou temperatura ótima de 60ºC e pH ótimo de 5,0, maior estabilidade em pH ácido (3,0 a 4,5) e menor termoestabilidade, quando comparada com a obtida em FES, mantendo apenas 13% da atividade original quando incubada a 60ºC, por 1 hora. As enzimas foram purificadas utilizando-se cromatografias de filtração em gel e troca iônica. A PG purificada proveniente da FSM apresentou pH e temperatura ótimos de 5,5 e 60-65ºC, estabilidade em pH 5,0-5,5 e manteve, após 1 hora de incubação, 100% da atividade original até 50ºC. Resultados similares foram obtidos para a PG proveniente da FES. A PG de FES apresentou massa molar de 29,3 kDa, Km de 1,58 mg/mL e Vmáx de 1553,1 ? mol/min/mg, enquanto que a da FSM apresentou massa molar de 30,1 kDa, km de 1,46 mg/mL e Vmáx de 2433,3 ? mol/min/mg. Íons como Fe+3, Ca+2, e K+ praticamente não afetaram a atividade da enzima... / Thermostable pectinases present important characteristics under the view of their industrial application, as their high stability to pH and temperature. Besides, the type of fermentative process used can affect the ir production and physical-chemical properties. The polygalacturonase (PG) production by the thermophilic fungus Thermoascus aurantiacus was carried out by submerged fermentation (SMF) and solid state fermentation (SSF) using substrates containing commercial pectin or agro- industrial residues as carbon sources. The crude PG from SSF presented optimum activity at 65ºC and pH 5.0, with stability at pH 4.0-5.0 and 7.5-8.5 and maintained 85% of its original activity at 60º C for 1 hour. In SMF the best cultivation medium was the liquid waste from juice extraction, with initial pH of 5.5, after 5 days of cultivation at 45ºC. The crude enzyme showed an optimum activity at 60ºC and pH 5.0, higher stability in acid ic pH (3.0 to 4.5) and was less thermostable when compared to that obtained in SSF, wich maintained only 13% of its original activity at 60ºC, for 1 hour. Purification of enzymes was carried out using filtration and ion-exchange chromatographies. The purified PG, from SMF, showed optimum pH and temperature of 5.5 and 60-65ºC, stability at pH 5.0-5.5 and preserved, after 1 hour incubation, 100% of its original activity at 50ºC. Similar results were obtained to PG from SSF. The PG obtained by SSF presented molar mass of 29.3 kDa, Km of 1.58 mg/ml and Vmáx of 1553.1 ? mol/min/mg, while that the enzyme from SMF presented molar mass of 30.1 kDa, km of 1.46 mg/ml and Vmáx of 2433.3 ? mol/min/mg. Ions such as Fe3+, Ca2+ and K+ practically did not affect the enzyme activity, while Mg2+, Mn2+ and Zn2+ decreased 7%, 75% and 50%... (Complete abstract, click electronic address below)

Lignocellulose, mostly from agricultural and forestry resources, is a potential renewable material for sustainable development of biorefineries. From previous studies, reducing sugar production through biological pretreatment involves two steps: solid-state fermentation (SSF) for delignification, followed by enzymatic hydrolysis by adding celluloytic enzymes (cellulase and xylanase etc.). In the process described in this thesis, the necessary enzymes are produced in-situ and the hydrolysis proceeds directly after the solid-state fermentation. Enzyme hydrolysis releases free amino nitrogen (FAN), reducing sugar and many other potential nutrients from the fermented materials. This method additionally avoids the need for removal of inhibitors compared with conventional chemical pretreatment processes. A range of solid-state fermentations were carried out to investigate the effect of washing procedure, particle size and nitrogen supplement on Trichoderma longibrachiatum growth. From these preliminary studies it was concluded that nitrogen supplementation is a crucial factor to improve significantly the fungi growth and production of feedstock using sugarcane bagasse as raw material. In order to evaluate the influence of environmental humidity on petri dish experiments, moist environments were investigated, with over 75% relative humidity to limit water evaporation from solid-state fermentation. The results showed that moist environments gave approximately 1.85 times the reducing sugar yield than dry environments. The process of simultaneous enzymatic hydrolysis of substrates and fungal autolysis were also studied. The degree of hydrolysis was affected by initial fermented solid to liquid ratio, temperature and pH range. The optimal conditions for subsequent hydrolysis of fermented solids were determined. The optimal solid to liquid ratio, 4% (w/w), temperature 50°C and pH 7 were established. The highest final reducing sugar, 8.9 g/L and FAN, 560 mg/L, were measured after 48 h. The fungal autolysis was identified by image analysis as well as by the consumption of nutrient and the release of free amino nitrogen and phosphorous. Solid state fermentation in a multi-layer tray bioreactor and a packed-bed bioreactor were also developed, with moist air supply for oxygen provision and heat removal. Fermented solids in the multi-layer bioreactor led to the highest subsequent hydrolysis yield on reducing sugar, FAN and Inorganic Phosphorous (IP), 222.85 mg/g, 11.56 mg/g and 19.9 mg/g, respectively. These series of fermentation experiments illustrate the feasibility for the application of consolidated bioprocessing, through simultaneous pretreatment and enzyme production as a more economic and environment-friendly process compared with those reported for chemical pretreatment followed by commercial enzyme process. A growth kinetic model regarding both growth and respiration is also proposed. Ethanol production was studied using the generic feedstock produced from sugarcane bagasse and soybean hulls. Total ethanol yield reached 0.31 mg g-1 (61.4% of theoretical yield) after 30 h of submerged fermentation. The result of subsequent fermentation has already shown the potential of the generic microbial feedstock to be used to produce varied products depending on the microorganism utilised.

Made available in DSpace on 2014-06-11T19:27:21Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-03-05Bitstream added on 2014-06-13T19:35:19Z : No. of bitstreams: 1 pirota_rdpb_me_sjrp.pdf: 578843 bytes, checksum: cdb9880602dbf9a622dd6735c9072440 (MD5) / Muitos fungos decompositores de materiais lignocelulósicos vêm sendo utilizados como produtores de enzimas em processos fermentativos nos quais são utilizados substratos de baixo valor comercial, como os resíduos agrícolas e agroindustriais. No entanto, apenas uma pequena parte destes resíduos são utilizados na alimentação de animais de produção. O presente trabalho visou: 1) a obtenção de preparado enzimático termoestável, com alta atividade de xilanase, pectinase e celulases, a partir do cultivo de fungos filamentosos termofílicos por fermentação em estado sólido; 2) uso de suplemento mineral comercial como meio de cultivo para os fungos na fermentação sólida; 3) avaliar a estabilidade das enzimas frente às características do rúmen; 4) hidrólise enzimática. Os fungos Thermomyces lanuginosus, Thermomucor indicae-seudaticae e Rhizomucor pussilus escolhidos para essa finalidade, foram cultivadas a 45°C por 360 horas em fermentação em estado sólido (FES), utilizando-se como substratos, suplemento comercial (B) (Bellpeso SV* - composto de sais minerais, farelo de algodão e polpa cítrica e para efeitos de comparação farelo de algodão e polpa cítrica (35% e 65%) (A). A amostra foi retirada em 24h e todas as outras de 48h em 48h. A produção máxima de pectinase produzida pelo fungo T. indicae-seudaticae foi obtida em 168h (421,31U/g) no substrato A, e a máxima atividade de xilanase produzida pelo T. lanuginosus TO-03 e ROB foram de 644,30Ug e 865,15 U/g em 168h, respectivemente. A amilase foi produzida por todos os fungos, porém CMCase e Avicelase tiveram uma baixa produção e ligninases não foram detectadas. A temperatura e pH ótimo de pectinase produzida pelo fungo T. indicae-seudaticae foi de 55ºC e pH 4,5 e para xilanase produzida pelas duas linhagens de T. lanuginosus, em ambos os substratos foram 70ºC e pH ótimo 6,5, com exceção... / Many rot fungi of lignocellulosics material are being utilized as producers of enzymes in fermentation processes, in which, substrates of very little commercial value such as agricultural and agroindustrial residues are used. However, only a small portion of these residues are utilized in the production of animal feed. The present project proposes: 1) the attainment of prepared enzymatic thermostable, with high activity of xylanase, pectinase and cellulases, from the cultivation of filamentous thermophylic fungi from solid state fermentation; 2) the use of commercial mineral supplementation as medium of cultivation for the fungi in solid fermentation; 3) to evaluate the enzymes stability when facing the characteristics of the rumen; 4) enzymatic hydrolysis. Thermomyces lanuginosus, Thermomucor indicae-seudaticae and Rhizomucor pusillus fungi chosen for this purpose, were cultivated at 45°C, for 360 hours in solid state fermentation (SSF), utilizing as substrates, commercial supplement (Bellpeso SV* - composed of mineral salt, cotton meal and citrus pulp) (B) and for comparison purpose cotton meal and citrus pulp at 35% and 65% (A). The first sample was taken in 24hrs and all the others every 48hrs. The maximum production of pectinase produced by the fungus T. indicae-seudaticae was obtained in 168hrs (421.31U/g) in the commercial substrate A, and to maximum activity of xylanase produced by the T. lanuginosus TO-03 and ROB was of 644.30 U/g and 865.15 U/g in 168hrs, respectively. Amylase was produced by all of the fungi, however CMCase and Avycelase had a low production and ligninases was not detected. The optimun temperature and pH of pectinase produced by the fungus T. indicae-seudaticae were of 55ºC and pH 4.5 and for xylanase produced by the two lineages of T. lanuginosus, in both substrates were 70ºC and optimun pH 6.5, with the exception to ROB in the substrate B that presented... (Complete abstract click electronic access below)

Submitted by Bruna Rodrigues (bruna92rodrigues@yahoo.com.br) on 2017-10-03T11:55:00Z No. of bitstreams: 1 DissLAS.pdf: 1333707 bytes, checksum: 130c53c4b36b129960094efeec36e4c7 (MD5) / Approved for entry into archive by Ronildo Prado (bco.producao.intelectual@gmail.com) on 2018-01-29T17:16:17Z (GMT) No. of bitstreams: 1 DissLAS.pdf: 1333707 bytes, checksum: 130c53c4b36b129960094efeec36e4c7 (MD5) / Approved for entry into archive by Ronildo Prado (bco.producao.intelectual@gmail.com) on 2018-01-29T17:16:26Z (GMT) No. of bitstreams: 1 DissLAS.pdf: 1333707 bytes, checksum: 130c53c4b36b129960094efeec36e4c7 (MD5) / Made available in DSpace on 2018-01-29T17:20:51Z (GMT). No. of bitstreams: 1 DissLAS.pdf: 1333707 bytes, checksum: 130c53c4b36b129960094efeec36e4c7 (MD5) Previous issue date: 2017-06-27 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A source of great biotechnological potential for the production of enzymes are endophytic microorganisms. These microorganisms are capable of producing a wide variety of enzymes, among them phytase, which is responsible for phytate hydrolysis. The objective of this work was to evaluate the potential of endophytic fungi of the mangroves of the state of São Paulo for phytase production. Initially, a qualitative selection of 33 isolates with PSM (Phytase Screening Medium) and a second quantitative selection was carried out, in which the preselected fungi were submitted to solid state fermentation (SSF) for phytase production. Among the isolates evaluated, the fungus Aspergillus awamori 9(4) was selected, which presented phytase activity of 43.98 U/100g when submitted to SSF culture for 72 h with soybean meal as substrate. In order to improve phytase production, the use of wheat bran as substrate was evaluated, reaching an activity of 82.77 U/100g, under the same conditions of cultivation. The use of KH2PO4 as an inducer in phytase production and the use of dialysis for the removal of interfering ions from the crude extract in phytase activity were also evaluated. For the experiments performed without the inducer, the dialysis of the extract resulted in activities of 85.42 U/100g with soybean meal and 55.44 U/100g with wheat bran. For the experiments of cultivation with the inductor and then dialysed resulted in activities of 132.35 U/100g with soybean meal and 115.52 U/100g with wheat bran, indicating that the effect of the inducer is positive in the production of Phytase and that dialysis is important for enzyme activity. These results indicate that the endophytic fungus of the mangrove of the State of São Paulo is promising for the production of phytase enzyme, being unpublished the use of endophytic micro-organisms for the production of this enzyme. / Uma fonte de grande potencial biotecnológico para a produção de enzimas são os microrganismos endofíticos. Estes microrganismos são capazes de produzir grande variedade de enzimas, dentre elas a fitase, que é responsável pela hidrólise do fitato. O objetivo deste trabalho foi avaliar o potencial de fungos endofíticos dos manguezais do estado de São Paulo para a produção de fitase. Inicialmente foi realizada uma seleção qualitativa de 33 isolados com meio diferencial PSM (Phytase Screening Medium), e uma segunda seleção quantitativa, em que os fungos pré-selecionados foram submetidos à fermentação em estado sólido (FES) para produção de fitase. Dentre os isolados avaliados, selecionou-se o fungo Aspergillus awamori 9(4), tendo este apresentado atividade de fitase de 43,98 U/100g quando submetido a cultivo FES por 72 h com farelo de soja como substrato. A fim de melhorar a produção de fitase, avaliou-se o uso de farelo de trigo como substrato, atingindo assim atividade de 82,77 U/100g, sob as mesmas condições de cultivo. Avaliou-se também o uso de KH2PO4 como indutor na produção de fitase e o uso de diálise para a remoção de íons interferentes do extrato bruto na atividade de fitase. Para os experimentos realizados sem o indutor, a diálise do extrato resultou em atividades de 85,42 U/100g com farelo de soja e 55,44 U/100g com farelo de trigo. Já para os experimentos de cultivo com o indutor e em seguida dialisados resultaram em atividades de 132,35 U/100g com farelo de soja e 115,52 U/100g com farelo de trigo, indicando que o efeito do indutor é positivo na produção de fitases e que a diálise é importante para a atividade enzimática. Esses resultados indicam que o fungo endofítico do manguezal do Estado de São Paulo é promissor para a produção da enzima fitase, sendo inédito o uso de micro-organismos endofíticos para a produção desta enzima.

Thesis submitted in fulfilment of the requirements for the degree of Doctor Technologiae: Chemical Engineering, Faculty of Engineering at the Cape Peninsula University of Technology - Cape Town, South Africa / Polycyclic aromatic hydrocarbons (PAHs) are one of the most common and recalcitrant environmental contaminants – known for their potential toxicity, mutagenicity, and carcinogenicity to humans. Biosurfactant application can enhance the biodegradation of PAHs. The main object of this work was to explore the novelty of biosurfactant produced by the isolated strains of Bacillus sp and Pseudomonas aeruginosa grown exclusively on Beta vulgaris, and the modification of the zeolites nanoparticles by the biosurfactant, for enhanced biodegradation of PAHs in soil. Novel biosurfactant-producing strains were isolated from hydrocarbon-contaminated environments, while several agrowaste were screened as primary carbon sources for the expression of biosurfactants, which were quantified using various standardized methods......

Orientador: Hamilton Cabral / Banca: Rodrigo Simões Ribeiro Leite / Banca: Fabiana Fonseca Zanoelo / Resumo: A produção de proteases de origem microbiana depende das condições de cultivo e da diversidade bioquímica de cada espécie. Estudos comparativos entre fermentação em estado sólido (FES) e fermentação submersa (FSm) usando farelo de trigo e meio sintético, respectivamente, foram realizados para a determinação dos parâmetros de produção de proteases pelo fungo Aspergillus fumigatus Fresenius. A melhor produção de protease foi em FES no período de 96 horas utilizando farelo de trigo, temperatura de 30 ºC e 1x106 esporos/5g de substrato com 1.517 U/mL. Em FSm o pico de produção foi em pH 6,0, a 30ºC, 5x105 esporos/mL de meio no período de 72 e 96 horas em meio contendo 0,5 e 0,25% de caseína, respectivamente, ambos com 40 U/mL. Conforme a produtividade dos processos fermentativos, o extrato enzimático da FES foi utilizado para estudos de purificação e caracterização bioquímica. Neste estudo, a protease purificada apresentou atividade ótima em pH 7,5 e 50ºC, sendo inibida por Fenil-metil-sulfonil-fluoreto (PMSF) e mais intensamente por antipaína (1,6 µM). Sobre efeito de íons, foi observado modulação da atividade proteolítica, principalmente com inibição por AlCl3, cuja atividade proteolítica residual foi de 18% após incubação com este íon. Na presença de Ditiotreitol (DTT) e uréia houve diminuição da atividade proteolítica, apresentando atividades residuais de 63% em 200 mM de DTT e 10% com 5 M de uréia. Comparativamente, na concentração de 0,1% de cada surfactante estudado, notou-se redução da atividade proteolítica, sendo 97% em presença de Brometo de cetil-trimetil amônio (CTAB), 79% para 4 - (1,1,3,3 - Tetrametilbutil) fenil- polietileno glicol (Triton X-100), 55% com Polyoxyethylenesorbitan monolaurato (Tween-20) e completa redução da atividade (0%) em... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The microbial protease production depends on growing conditions and the biochemical diversity of each species. Comparative studies between solid-state fermentation (SSF) and submerged fermentation (SmF) using wheat bran and synthetic medium, respectively, were performed to determine the optimum parameters for protease production by the fungus Aspergillus fumigatus Fresenius. The best protease production was in SSF within 96 hours using wheat bran, temperature 30°C and 1x106 spores/5g of substrate, with 1,517 U/mL. In SmF peak production was at pH 6.0 at 30°C, 5x105 spores/mL of media within 72 and 96 hours in medium containing 0.5 and 0.25% casein, respectively, with 40 U/mL. According to the productivity of the fermentative processes, enzymatic extract was used from SSF to study purification and biochemical characterization. In this study, purified protease showed optimum activity at pH 7.5 and 50°C, and inhibited by Phenylmethylsulfonyl fluoride (PMSF) and more intensely for antipain (1,6 µM). Concerning to the effect of ions, we observed modulation of the proteolytic activity, especially with inhibition by AlCl3, which residual activity was of 18 % after incubation with this ion. In the presence of Dithiothreitol (DTT) and ureia, we observed progressive decrease in proteolytic activity, presenting residual activities of 63% with 200 mM DTT, and 10% with 5 M ureia. Comparatively, in the concentration of 0.1% of each surfactant studied, there was a reduction in the proteolytic activity in 97% in presence of Cetyl trimethylammonium bromide (CTAB), 79% with 4-(1,1,3,3-Tetramethylbutyl)phenyl-polyethylene glycol (Triton X-100), 55% with Polyoxyethylenesorbitan monolaurate (Tween-20) and a complete inactivation in the presence of Sodium dodecyl sulfate... (Complete abstract click electronic access below) / Mestre