Transfer of information between CAD models and downstream manufacturing process planning software typically involves redundant user interaction. Many existing tools are process-centric and unsuited for selection of a "best process" in the context of existing concurrent engineering design tools. A computer based Feature-Recognition (FR) process is developed to extract critical manufacturing features from engineering product CAD models. FR technology is used for automating the extraction of data from CAD product models and uses wire-frame geometry extracted from an IGES neutral file format. Existing hint-based feature recognition techniques have been extended to encompass a broader range of manufacturing domains than typical in the literature, by utilizing a combination of algorithms, each successful at a limited range of features. Use of wire-frame models simplifies product geometry and has the potential to support rapid manufacturing shape evaluation at the conceptual design stage. Native CAD files are converted to IGES neutral files to provide geometry data marshalling to remove variations in user modelling practice, and to provide a consistent starting point for FR operations. Wire-frame models are investigated to reduce computer resources compared to surface and solid models, and provide a means to recover intellectual property in terms of manufacturing design intent from legacy and contemporary product models. Geometric ambiguity in regard to what is ?solid? and what is not has plagued wire-frame FR development in the past. A new application of crossing number theory (CNT) has been developed to solve the wire-frame ambiguity problem for a range of test parts. The CNT approach works satisfactorily for products where all faces of the product can be recovered and is tested using a variety of mechanical engineering parts. Platform independent tools like Extensible Mark-up Language are used to capture data from the FR application and provide a means to separate FR and decision support applications. Separate applications are composed of reusable software modules that may be combined as required. Combining rule-based and case-based reasoning provides decision support to the manufacturing application as a means of rejecting unsuitable processes on functional and economic grounds while retaining verifiable decision pathways to satisfy industry regulators.

Australia is recognising that carbon capture and storage (CCS) may be a feasible pathway for addressing increasing levels of CO2 emissions. This thesis presents a preliminary economic assessment and comparison of the capture costs for different Australian CO2 emission sources. The capture technologies evaluated include solvent absorption, pressure swing adsorption (PSA), gas separation membranes and low temperature separation. The capture cost estimated for hydrogen production, IGCC power plants and natural gas processing is less than A$30/tonne CO2 avoided. CO2 capture cost for iron production ranges from A$30 to A$40 per tonne CO2 avoided. Higher costs of A$40 to over A$80 per tonne CO2 avoided were estimated for flue gas streams from pulverised coal and NGCC power plants, oil refineries and cement facilities, and IDGCC synthesis gas. Based on 2004 and 2005 EU ETS carbon prices (A$30 to A$45 per tonne CO2 avoided), the cost of capture using current commercially available absorption technology may deter wide-scale implementation of CCS, in particular for combustion processes. A sensitivity analysis was undertaken to explore the opportunities for reducing costs. The high cost for capture using solvent absorption is dependent on the energy needed for solvent regeneration and the high capital costs. Cost reductions can be achieved by using new low regeneration energy solvents coupled with recycling the waste heat from the absorption process back to the steam cycle, and using low cost ???fit-for-purpose??? equipment. For membrane and PSA technologies, the capture costs are dominated by the flue gas and post-capture compressors. Operating the permeate or desorption stream under vacuum conditions provides significant cost reductions. Improvements in membrane and adsorbent characteristics such as the adsorbent loading or membrane permeability, CO2 selectivity, and lower prices for the membrane or adsorbent material provide further cost benefits. For low partial pressure CO2 streams, capture using low temperature ???anti-sublimation??? separation can be an alternative option. Low costs could be achieved by operating under low pressures and integrating with external sources of waste heat. Applying the cost reductions achievable with technology and process improvements reduces the capture and CCS costs to a level less than current carbon prices, making CCS an attractive mitigation option.

Thesis (Ph. D.)--Chemistry and Biochemistry, Georgia Institute of Technology, 2009. / Committee Chair: Ragauskas, Arthur; Committee Member: Bunz, Uwe; Committee Member: Cairney, John; Committee Member: Collard, David; Committee Member: Singh, Preet. Part of the SMARTech Electronic Thesis and Dissertation Collection.

Thesis (D.Tech.: Biotechnology)-Dept. of Biotechnology, Durban University of Technology, 2007 xi, 209 leaves / Protein engineering technologies, such as directed evolution and DNA recombination, are often used to modify enzymes on a genetic level for the creation of useful industrial catalysts. Pre-treatment of paper pulps with xylanases have been shown to decrease the amounts of toxic chlorine dioxide used to bleach pulp. This study was undertaken to improve the thermal and alkaline stabilities of the xylanase from the fungus Thermomyces lanuginosus using ep-PCR and DNA shuffling.

Thesis (MSc)--Stellenbosch University, 2000. / ENGLISH ABSTRACT: The extractive materials in wood often cause pitch problems in pulp mills. During pulping and bleaching extractives are released from the wood and pulp and later stick to ceramic and metal parts, forming pitch deposits. Pitch deposits impair both product quality and production rates. It decreases the efficiency of pulp washing, screening, centrifugal cleaning, and refining, and can disrupt many paper machine operations. The deposits also break loose from equipment and cause spots in the final product. There are a few triggering mechanisms that induce pitch deposition. Hydrodynamic or mechanical shear can destabilise the colloidal pitch emulsion, causing pitch to agglomerate and deposits to form. Similarly, sudden temperature drops and/or pH shocks and/or the introduction of water hardness ions from fresh water inlets or showers can also cause pitch deposits by destabilising the colloidal pitch emulsion. Inorganic salts, such as calcium carbonate, can catalyse pitch deposition by acting as the building blocks for the sticky pitch. Calcium ions in the white water can react with fatty acids, forming insoluble, sticky calcium soaps. Triglycerides have also been shown to be a major contributor to pitch deposition in kraft pulping and bleaching mills. It forms a sticky deposit to which less sticky particles attach. To attain an improved understanding of pitch problems associated with the kraft pulping and bleaching of Eucalyptus spp., various analyses were done on wood- and pulp extractives and pitch from a South African kraft pulp mill. High molecular weight compounds (involatile) constituted a large portion of the extracts and pitch. Approximately 40% of volatile Eucalyptus grandis extract was f3-sitosterol, with fatty acids (22.8%) and triglycerides (15.5%) also making a substantial contribution. Fatty acid amides were a prominent fraction of pulp extracts from the latter stages of bleaching. The amides constituted 38.3% and triglycerides 10.1% to total volatile pitch deposits. Lipases hydrolyse triglycerides and could therefore help to reduce pitch problems. Consequently 381 filamentous fungi isolated from indigenous and commercial forests in South Africa were screened for lipase activity on tributyrin and Tween 80. Eight strains were selected and the tributyrin and Tween 80 assays were repeated by monitoring lipase activity over a seven-day period. The selected strains were also assayed for their activity toward p-nitrophenyl palmitate. Ophiostoma piliferum Cartapip 58™ and Phanerochaete chrysosporium BKM-F-1767, two strains known for respectively their biodepitching and biopulping ability, were' used as controls. A few of the strains compared well and even outperformed the control strains, indicating their potential for use in pitch control. The effect of pretreatment with the eight selected fungal strains on E. grandis wood- and pulp extractives was determined. Cartapip 58™ and P. chrysosporium BKM-F-1767 were used as control strains. Several of the strains compared well to the control strains in their ability to reduce the triglyceride content of wood extract. The South African isolate, white-rot fungus Phanerochaete psuedomagnoliae nom. prov., reduced triglyceride content significantly. Consequently it can act as an agent for both biopulping and biodepitching. The treated wood samples had a lower triglyceride content than the sterile controls. Consequently more triglycerides would be released into process waters by the sterile controls than the treated samples. The effect of commerciallipases on deposited brown stock pulp extract was also evaluated. The lipases did not reduce the triglyceride content of the deposited extract. The addition of lipases in pulping and bleaching processes would therefore not affect already deposited pitch. / AFRIKAANSE OPSOMMING: Die ekstrakstowwe van hout veroorsaak dikwels 'n neerslag tydens verpulping. Gedurende verpulping en bleiking kom ekstrakstowwe van die hout enpulp vry en kleef aan keramiek- en metaalonderdele. Gevolglik benadeel dié neerslag produkkwaliteit en produksietempo. Dit verlaag die effektiwiteit van pulpwas, sifting, sentrifugale skoonmaakprosesse en suiwering, en kan die werkverrigting van papiermasjiene ontwrig. Die neerslag kan ook later los breek en kolletjies op die finale produk veroorsaak. Verskeie meganismes kan die neerslag veroorsaak. Hidrodinamiese of meganiese wrywing kan die kolloïdale ekstrakstofemulsie destabiliseer en sodoende die ekstrakstof laat konglomereer en neerslaan. Op soortgelyke wyse veroorsaak skielike temperatuurverlaging en/of pH-skokke en/of die toevoeging van ione in varswater om waterhardheid te beheer ook die neerslag deur die kolloïdale ekstrakstofemulsie te destabiliseer. Anorganiese sout soos kalsiumkarbonaat kan neerslagvorming kataliseer omdat dit optree as bousteen vir die klewerige, sementagtige ekstrakstowwe. Kalsiumione in die proseswater kan ook reageer met vetsure om onoplosbare, klewerige kalsiumsepe te vorm. Dit is bewys dat trigliseriede een van die hoofoorsake is in die vorming van die neerslag tydens kraft verpulpingen bleikingprosesse. Om die neerslagreaksie wat met die kraft verpulping en bleiking van Eucalyptus spp. geassosieer word, beter te verstaan, is verskeie analises op hout- en pulpekstrakte asook die neerslag van 'n Suid-Afrikaanse kraft verpulpingsaanleg uitgevoer. Hoë molekulêre massa (nie-vlugtige) stowwe het 'n groot gedeelte van die ekstrakte en neerslag uitgemaak. Ongeveer 40% van die vlugtige Eucalyptus grand is ekstrak bestaan uit ~-sitosterol met vet sure (22.8%) en trigliseriede (15.5%) wat ook aansienlike bydraes lewer. Vetsuuramiede verteenwoordig 'n beduidende komponent van pulpekstrak by die laaste stadiums van bleiking. Die amiede het 38.3% en trigliseriede 10.1%tot die vlugtige fraksie van die neerslag bygedra. Lipases hidroliseer trigliseriede en kan dus help om neerslagprobleme te voorkom. Gevolglik is 381 filamentagtige fungi geïsoleer uit inheemse en kommersiële woude van Suid-Afrika en hul lipase-aktiwiteit op tributyrin en Tween 80 geëvalueer. Agt rasse is geselekteer en die tributyrin en Tween 80 toetse is herhaal deur lipase-aktiwiteit oor 'n sewe-dag periode te monitor. Die geselekteerde rasse is ook getoets vir lipase-aktiwiteit met p-nitrofenielpalmitaat. Ophiostoma piliferum Cartapip 58™ en Phanerochaete chrysosporium BKM-F-1767, twee rasse wat daarvoor bekend staan vir onderskeidelik hul vermoë om houtekstrakstowwe te verminder en te bioverpulp, is as kontroles gebruik. 'n Paar van die geselekteerde rasse het goed vergelyk en selfs beter presteer as die kontrolerasse; 'n aanduiding van hul potensiaal om neerslagreaksies te beheer. Die effek van voorafbehandeling met die agt geselekteerde fungi rasse op E. grandis hout- en pulpekstrak is vasgestel. Cartapip 58™ en P. chrysosporium BKM-F-1767 is gebruik as kontrolerasse. Verskeie rasse het goed vergelyk met die kontrolerasse in hul vermoë om die trigliseriedinhoud van die houtekstrak te verlaag. Die Suid-Afrikaanse isolaat, witverrottingswam Phanerochaete pseudomagnoliae nom. prov., het ook die trigliseried inhoud beduidend verminder. Gevolglik sou dit as 'n middel kon dien vir beide neerslagvoorkoming en bioverpulping. Die trigliseriedinhoud van die behandelde monsters was laer as dié van steriele kontroles. Gevolglik sal meer trigliseriede in proseswater vrygestel word deur die steriele kontroles as die behandelde monsters. Die effek van kommersiële lipases op ongebleikte kraft pulpekstrakneerslag is ook geëvalueer. Omdat lipases nie die trigliseriedinhoud van die neerslag kon verlaag nie sal die gebruik van lipases dus nie die ekstrakstofneerslag in verpulpings- en bleikingsprosesse beïnvloed nie.

Made available in DSpace on 2014-06-11T19:23:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-03-05Bitstream added on 2014-06-13T18:50:37Z : No. of bitstreams: 1 silva_bl_me_sjrp_parcial.pdf: 68972 bytes, checksum: 03431955c8d6cea4c3ed5570639d9b21 (MD5) Bitstreams deleted on 2015-03-16T11:30:17Z: silva_bl_me_sjrp_parcial.pdf,Bitstream added on 2015-03-16T11:31:01Z : No. of bitstreams: 1 000713868.pdf: 583338 bytes, checksum: 2ab049d7d1dee609368d97ffcc42cfc4 (MD5) / Proteases constituem uma das mais importantes enzimas industriais e uma de suas principais aplicações é na indústria láctica para produção de queijos. Devido à escassez do coalho tradicional de origem animal, proteases coagulantes microbianas estão sendo pesquisadas como substitutos. O coalho apresenta duas ações hidrolíticas sobre a caseína que caracterizam sua adequação como um bom coagulante, que são a atividade coagulante e a proteolítica. Quanto maior a razão entre coagulante / proteolítica (AC/AP), melhor o coagulante. O presente trabalho objetivou estudar a produção e caracterização da protease coagulante produzida pelo fungo termofilico Thermomucor indicae-seudaticae N31 via fermentação submersa. As condições de produção estudadas foram: natureza e concentração da fonte de carbono e da solução salina, períodos de incubação e velocidade de agitação. Após a produção foi feita a caracterização físico- química que consistiu em determinar as condições de atuações ótimas da enzima. A partir dos resultados, observou-se que a melhor composição no meio fermentativo foi: 4 % de farelo de trigo, 0,3 % de solução salina, 72 horas de incubação a 45 °C e 150 rpm de agitação. Nestas condições os valores da atividade coagulante e da razão AC/AP foram 60,5 U/mL e 510, respectivamente. A enzima coagulante apresentou as características: pH e temperatura ótimos foram 5,5 e 65 °C, respectivamente; pH e temperatura de estabilidade foram 3,5 – 5,0 (retendo cerca de 80 % da atividade depois de 24 horas à temperatura ambiente) e até 60 °C (após uma hora na ausência de substrato). O conjunto dos resultados sugere a conclusão que a protease coagulante é promissora do ponto de vista tecnológico como substituto do coalho animal, principalmente, devido à sua alta especificidade representada pelo elevado valor da razão / Proteases are one of the most important industrial enzymes and one of its main applications is in the dairy industry for the production of cheese. Due to the scarcity of renin bovine, microbial coagulants proteases are being researched as a substitute. The rennet has two hydrolytic action on casein featuring its suitability as a good coagulant, which are the milk-clotting and proteolytic activity. The higher the ratio milk-clotting / proteolytic (MCA/PA), the better the coagulant. The present study investigated the production and characterization of coagulant protease produced by thermophilic fungus Thermomucor indicae-seudaticae N31 on submerged fermentation. The production conditions were: nature and concentration of the carbon source and saline, incubation and agitation speed. After production was performed physicochemical characterization that determined the conditions optimal of the enzyme. From the results, it was observed that the best composition of the fermentative medium was: 4 % wheat bran, 0,3 % saline, 72 hours of incubation at 45 °C and 150 rpm agitation. In these conditions the values of milk-clotting activity and the ratio MCA/PA were 60,5 U/mL and 510, respectively. The clotting enzyme had the characteristics: optimum of pH and temperature were 5,5 and 65 °C, respectively, pH and temperature of stability were 3,5 – 5,0 (retaining approximately 80 % of activity 24 hours at temperature environment) and even 60 °C (after one hour in the absence of substrate). The group of results showed that coagulant protease is promising technological point of view as a substitute for animal rennet, mainly due to their high specificity shown by the high value of the ratio

Orientador: Roberto da Silva / Banca: Heloiza Ferreira Alves do Prado / Banca: Gustavo Orlando Bonilla Rodriguez / Resumo: A ciclomaltodextrina glucanotransferase (EC 2.4.1.19) é uma enzima capaz de formar ciclodextrinas a partir do amido. As ciclodextrinas são oligossacarídeos cíclicos, os quais os principais tipos são α-, β- e γ-CD, que apresentam 6, 7 e 8 unidades de glicose interligadas por ligações α-1,4. Duas bactérias alcalofílicas Bacillus sp subgrupo alcalophilus E16 e Paenibacillus campinasensis H69-3 foram estudadas por apresentarem boa produção de CGTase, ambas foram isoladas de amostras de solo. A atividade enzimática foi estudada utilizando os métodos já pré-determinados em estudos anteriores, como dextrinizante, CD-fenolftaleína e determinação de proteína. Para alcançar a purificação foram utilizados os processos de ultrafiltração e purificação por gel filtração em resina Sephadex G-75. A partir desses processos obteve-se resultados significativos, para o extrato purificado produzido por Bacillus sp subgrupo alcalophilus E16, obteve-se um fator de purificação de 35,4 vezes, um rendimento de 34.1 % e atividade específica de 13.10 U/mg, enquanto o extrato purificado de Paenibacillus campinasensis H69-3 resultou em um fator de purificação de 94.5 vezes, um rendimento de 33.2 % e a atividade específica de 23.64 U/mg. Foram feitos ensaios quanto a produção de ciclodextrinas utilizando o extrato bruto concentrado da CGTase por ultrafiltração produzida por Paenibacillus campinasensis H69-3, no perfil de hidrólise de diferentes amidos a 1.0%, a enzima atuou convertendo o amido em β-CD de melhor forma nos amidos de arroz e de mandioca, enquanto que na concentração de 2.5 % a melhor conversão se deu nos amidos de milho e mandioca. Quanto à dextrinização dos amidos, as concentrações de 1.0 % e 2.5 % apresentaram o mesmo perfil de hidrólise, sendo os amidos de mandioca, solúvel e batata apresentaram melhor conversão, quando comparadas aos amidos de arroz e milho. A dextrinização do amido foi mais efetiva na ... / Abstract: The cyclomaltodextrin glucanotransferase ( EC 2.4.1.19 ) is an enzyme able to form cyclodextrin from starch . Cyclodextrins are cyclic oligosaccharides , which are the main types α - , β - and γ - CD which have 6, 7 and 8 glucose units connected by α - 1, 4 bonds. Two bacterial alkalophilic Bacillus sp subgroup alcalophilus E16 and Paenibacillus campinasensis H69 -3 was studied due to their good CGTase production, both of which were isolated from soil samples. The enzymatic activity was studied using the methods already pre-determined in previous studies, as dextrinizante, CD-phenolphthalein and protein determination. To achieve the purification and ultrafiltration processes were used for purification gel filtration resin Sephadex G-75. From these processes significant results were obtained for the purified extract produced by Bacillus sp subgroup alcalophilus E16 gave a purification factor of 35.4 times, a yield of 34.1% and a specific activity of 13.10 U / mg , while the purified extract of Paenibacillus campinasensis H69-3 resulted in a purification factor of 94.5 times , a yield of 33.2 % and specific activity of 23.64 U / mg . Tests were made for the production of cyclodextrins using concentrated crude extract by ultrafiltration CGTase produced by Paenibacillus campinasensis H69 3, hydrolysis of starches from different profile than 1.0 % , the enzyme acted converting starch into β- CD best in rice starch and tapioca , whereas a concentration of 2.5 % and better conversion is given cassava and corn starch . As for dextrinizing the starch, the concentrations of 1.0% and 2.5% showed the same profile hydrolysis , and cassava starches , potato and soluble showed better conversion, when compared to starches of rice and corn. Dextrinisation of starch was more effective at 1.0% concentration , because practically all starch was hydrolyzed at a concentration of 2.5% was slower hydrolysis , for 24 hours, the starches in the study were ... / Mestre

Magnetic resonance (MR) is a highly versatile technique with great potential for use in industrial applications; from the in situ study of unit operations to the optimisation of product properties. This thesis, concerned with the latter, is divided into two parts. Firstly, dynamic MR is applied to characterise the flow behaviour, or rheology, of process fluids. Such characterisation is typically performed using conventional rheometry methods operating offline, with an online, or inline, method sought for process control and optimisation. Until recently, MR was an unlikely choice for this application due to the requirement of high-field MR hardware. However, recent developments in low-field MR hardware mean that the potential of MR in such applications can now be realised. Since the implementation of MR flow imaging is challenging on low-field MR hardware, two new approaches to MR rheometry are described using pulsed field gradient (PFG) MR. A cumulant analysis of the PFG MR signal is first used to characterise the rheology of model power-law fluids, namely xanthan gum-in-water solutions, accurate to within 5% of conventional rheometry, the data being acquired in only 6% of the time required when using MR flow imaging. The second approach utilises a Bayesian analysis of the PFG MR signal to characterise the rheology of model Herschel--Bulkley fluids, namely Carbopol 940-in-water solutions; data are acquired in only 12% of the time required for analysis using MR flow imaging. The suitability of the Bayesian MR approach to study process fluids is demonstrated through experimental study on an alumina-in-acetic acid slurry used by Johnson Matthey. Secondly, MR imaging is used to provide insights into the origins and mechanisms of colloidal gel collapse. Many industrial products are colloidal gels, a space-spanning network of attractive particles with a yield stress. Colloidal gels are, however, known to undergo gravitational collapse after a latency period, thus limiting the shelf-life of products. This remains poorly understood, with a more detailed understanding of both fundamental interest and practical importance. To this end, MR imaging is applied offline to investigate the phase behaviour of colloidal gels. In particular, a comparison of the simulated and experimental phase diagrams suggests gravitational gel collapse to be gravity-driven. Furthermore, measurement of the colloid volume fraction using MR imaging indicates the formation of clusters of colloids at the top of the samples. Whether such clusters initiate gravitational gel collapse is yield stress-dependent; the gravitational stress exerted by a cluster must be sufficient to yield the colloidal gel.

Made available in DSpace on 2016-04-01T17:54:49Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-06-09. Added 1 bitstream(s) on 2016-04-01T18:00:30Z : No. of bitstreams: 1 000859578.pdf: 2407454 bytes, checksum: 8f8cd1ebd5ec7774fbfb7c2dd87ffc81 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Em conjunto com outras enzimas que degradam amido, as α-amilases e CGTases são incluídas na família 13 glicosilhidrolases caracterizada por uma conformação (α/β)8-barril. As amilases são grupo importante de enzimas industriais, representando cerca de 25% do mercado mundial de enzima.. Foram isoladas linhagens bacterianas de áreas do Cerrado que apresentaram alta produção de amilases em estudos anteriores. Devido ao interesse em ampliar os conhecimentos de α-amilases e CGTase com potencial de aplicação industrial, no presente trabalho, propôs-se realizar estudos de análises físico-químicas e do efeito da fonte de carbono, na produção de α-amilase das linhagens microbianas A-1.2 e A-18 e de CGTase produzida por Paenibacillus campinasensis H69-3 utilizando na fermentação submersa os amidos solúvel, trigo, milho, batata e mandioca; farelo de trigo e mandioca; Maizena e Arrozina como diferentes fontes de carbono alternativas Além disso, propôs-se aperfeiçoar os estudos de produção de amilases pela linhagem microbiana A-1.2 por meio da otimização dos constituintes nutricionais do meio de cultivo em que cada um dos ensaios do planejamento fatorial foi executado em triplicata e dessa forma, consideraram-se as médias dos resultados obtidos dessas repetições. Iniciar estudos de biologia molecular com a CGTase produzida por P. campinasensis H69-3 buscando estratégias para a clonagem e expressão da CGTase em hospedeiro heterólogo (Escherichia coli). As linhagens A-1.2 e A-18 tiveram as fontes de carbono amido solúvel, amido de mandioca, amido de milho e farelo de mandioca como os substratos que maximizaram a produção da enzima por ambas as linhagens, e o tempo de fermentação com maiores atividades foi entre 72 e 96 horas. O perfil de produção ao longo do tempo da CGTase de P. campinasensis H69-3 indicou o amido solúvel, amido de mandioca e a Maizena como as fontes de carbono que propiciaram as mais altas... / Together with other enzymes that degrade starch, α-amylases and CGTases are included in the family 13 glycosylhydrolases, characterized by a conformation (α/β)8-barrel. Amylases are an important group of industrial enzymes, accounting for approximately 25% of the world enzyme market. Bacterial strains from the Cerrado areas, which have shown a high production of amylase in the previous studies, have been isolated. Due to the interest in expanding the knowledge of α-amylase and CGTase with potential for industrial application, the present work proposes to conduct studies of physical and chemical analysis and the effect of carbon source on the production of α-amylase of A-1.2 and A-18 microbial strains, as well as CGTase produced by Paenibacillus campinasensis H69-3 in submerged fermentation using the soluble starch, wheat, corn, potato and cassava; wheat bran and cassava; Maizena and Arrozina like various alternative sources of carbon. Furthermore, it proposes enhancing the amylase study of the microbial strain A-1.2 by optimizing the nutritional components of the culture medium in which each of the factorial design experiments has been carried out in triplicate, and thus considering the average results of these repetitions. To initiate molecular biology studies with CGTase produced by P. campinasensis H69-3 searching strategies for the cloning and expression of the CGTase in a heterologous harbourer (E. coli). The A-1.2 and A-18 strains were the carbon sources of soluble, cassava and corn starch, and cassava bran as the substrate that maximized the enzyme production by both strains. And the fermentation time with higher levels of activity was between 72 and 96 hours. The production profile over the CGTase P. campinasensis H69-3 time has indicated the soluble and cassava starch and Maizena as carbon source that provided the highest enzymatic activities. The 72 hours period of time has been more appropriate in enzymatic tests, ...

Made available in DSpace on 2014-08-27T14:36:44Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-09-24Bitstream added on 2014-08-27T15:57:05Z : No. of bitstreams: 1 000778079.pdf: 2078209 bytes, checksum: f253bd93ea489feb471ed9cad8e18c13 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A conversão enzimática dos polissacarídeos da biomassa é um fator chave no desenvolvimento de bioetanol de segunda geração. A recalcitrância da lignocelulose para a degradação enzimática e o custo elevado de enzimas hidrolíticas necessárias para a despolimerização de polissacarídeos encontrados na parede celular da planta são barreiras significativas para a produção em larga escala e para a comercialização de biocombustíveis e bioprodutos derivados da biomassa vegetal. A fim de aumentar rapidamente a produção de biocombustíveis celulósicos e bioprodutos, existe a necessidade de desenvolver coquetéis enzimáticos mais eficientes e de menor custo para a conversão de biomassa em açúcares fermentáveis. Nesse contexto, o estudo de enzimas degradadoras da parede celular é essencial. No presente trabalho a enzima endo-1,4-?-D-glucanase de Aspergillus terreus (ATEG_09894) foi clonada para expressão em A. nidulans. O teste de expressão mostrou a expressão solúvel da proteína. Sua identidade foi confirmada por espectrometria de massas. O pH ótimo e a temperatura ótima para sua atividade enzimática foram de 5,0 e 55ºC, respectivamente. A desnaturação térmica mostrou que a partir de 60ºC a enzima começa a perder estrutura. Interessantemente a enzima apresenta atividades ?-glucanase e xiloglucanase, tendo preferência por ?-glucano. A caracterização estrutural mostrou que ATEG_09894 foi expressa corretamente e os resultados de SEC e SAXS mostram que a proteína é monomérica em solução e o modelo de sua estrutura tridimensional indica que a superfície eletrostática da molécula contribui para um monômero estável. Os dados apresentados nesse trabalho são importantes pois identificam peculiaridades da enzima ATEG_09894, que atua na degradação da biomassa, sugerem os determinantes de sua seletividade enzimática e apresenta a degradação, não usual, de ?-glucanos... / The enzymatic conversion of polysaccharides from biomass is a key factor in the development of second generation bioethanol. The recalcitrance of lignocellulose to enzymatic degradation and the high cost of hydrolytic enzymes necessary for the depolymerization of polysaccharides found in plant cell wall are significant barriers to large-scale production and commercialization of biofuels and bioproducts derived from plant biomass. In order to rapidly increase the production of biofuel and byproducts cellulosic, a need exists to develop more efficient and lower cost enzymatic cocktails for the conversion of biomass into fermentable sugars. In this context, the study of cell wall degrading enzymes is essential. In this work the enzyme endo-1,4-?-D-glucanase from Aspergillus terreus (ATEG_09894) was cloned for expression in A. nidulans. The expression test showed the expression of a soluble protein. Its identity was confirmed by mass spectrometry. The optimum pH and optimum temperature for the enzyme activity were 5.0 and 55 ºC, respectively. The thermal denaturation showed that above 60 ºC the enzyme starts to lose structure. Interestingly, the enzyme has ?-glucanase and xiloglucanase activities, preferring ?-glucan. Structural characterization showed that ATEG_09894 was expressed correctly folded and the results of SEC and SAXS show that the protein is monomeric in solution and the model of its three dimensional structure indicates that the electrostatic surface molecule contributes to a stable monomer. The data presented in this study are important because they identify peculiarities of ATEG_09894, which acts in the degradation of biomass, suggest the determinants of its selectivity and enzymatic degradation presents, unusual, of ?-glucans and xyloglucans, promising feature for degradation of lignocellulosic material