Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / Bacterial cellulose (BC) is a naturally nanoscale biomaterial with high purity and excellent chemical and mechanical properties. There are several sugar-rich renewable sources and wastes that can be used as alternative media for bacterial cellulose production.The aim of this study was to evaluate the suitability of an extract of mesquite pods as an alternative carbohydrate source for the production of BC through fermentation by Gluconacetobacter hansenii. The amount of sugars, soluble solids, and pH of the extract was characterized. The influence of the initial sugar concentration, pH, and the source of supplementary nitrogen on the fermentation were evaluated. The best production of bacterial cellulose was achieved in the medium with a sugar concentration of 30 g /L, pH 4.0, and supplemented with 10 g /L of yeast extract. The films from the optimized medium were oven dried and characterized by x-ray diffraction (XRD), Fourier Transform Infrared spectroscopy (FTIR), thermal gravimetric analysis (TGA), differential scanning calorimetry (DSC) and electron microscopy (SEM). The results were similar to typical bacterial cellulose films produced in the reference medium HS. Thus, it was possible to demonstrate the suitability of mesquite pod aqueous extract to produce BC. / A celulose bacteriana (CB) à um biomaterial naturalmente nanomÃtrico, com elevado grau de pureza e excelentes propriedades quÃmicas e mecÃnicas. Existem na natureza fontes renovÃveis e resÃduos ricos em aÃÃcares que despertam um crescente interesse como meios alternativos em substituiÃÃo aos tradicionalmente empregados na produÃÃo de celulose. O objetivo deste estudo foi avaliar o extrato de vagens de algaroba (Prosopis juliflora) como fonte alternativa para produÃÃo de CB atravÃs de fermentaÃÃo por Gluconacetobacter hansenii. O extrato foi caracterizado quanto à quantidade de aÃÃcares, pH e sÃlidos solÃveis. AtravÃs da fermentaÃÃo do extrato foram realizados testes de influÃncia da concentraÃÃo inicial de aÃÃcares, influÃncia do pH e efeito da variaÃÃo da suplementaÃÃo com fonte de nitrogÃnio sobre a produÃÃo de CB. A melhor produÃÃo foi proveniente da fermentaÃÃo do extrato de algaroba diluÃdo a uma concentraÃÃo de aÃÃcares de 30 g/L com pH 4,0 suplementado com 10 g/L de extrato de levedura. As pelÃculas obtidas do extrato com condiÃÃes melhoradas foram secas em estufa e caracterizadas por difraÃÃo de raios x (DRX), espectroscopia de absorÃÃo no infravermelho (FTIR), anÃlise termogravimÃtrica (TGA), calorimetria exploratÃria diferencial (DSC) e microscopia eletrÃnica de varredura (MEV), e apresentaram resultados tÃpicos de celulose bacteriana quando comparados à CB produzida no meio de referÃncia HS. Assim, foi possÃvel mostrar a viabilidade de produÃÃo de CB com extrato aquoso de vagens de algaroba.

Made available in DSpace on 2017-05-12T14:36:23Z (GMT). No. of bitstreams: 1 DISSERTACAO AMANDA ALVES MESTRADO EM CIENCIAS FARMACEUTICAS _UNIOESTE 2015.pdf: 10598736 bytes, checksum: 51f0f3eb83858fee62392b7892930766 (MD5) Previous issue date: 2015-12-07 / Lignocellulosic biomass are the raw material most abundant and promising as a natural and renewable resource. These plant materials are complex carbohydrate polymer composed mainly of cellulose, hemicellulose and lignin, which are linked by covalent bonds and can be transformed into value-added products, such as biofuels. The degradation of lignocellulosic material is made mainly from enzymes produced by microorganisms such as filamentous fungi, yeast and bacteria. Ethanol production from agricultural residues, based on the enzymatic hydrolysis, it takes basically four stages: production of enzymes, pretreatment, enzymatic hydrolysis and fermentation. Pretreatment is a work that will break the lignin cellulose complex, reducing the degree of crystallinity of the cellulose and increase the porosity of the material, by increasing the surface area of the biomass. However, pre-treatment products can generate inhibitors which include phenolic and other aromatic, aliphatic acids, aldehydes, furans, inorganic ions. The fermentation and simultaneous saccharification is an important approach for producing cellulosic or ethanol of second generation, where the enzymatic hydrolysis of cellulose and fermentation are simultaneously carried out in the same reactor, in order to obtain ethanol at a high rate and decrease formation of inhibitor compounds. Enzymatic hydrolysis requires, first, that the lignocellulosic biomass is pretreated to increase access to enzymatic attack, so that later the cellulose is broken down by cellulase action. Xylanases include the group of enzymes responsible for the hydrolysis of xylan, the major constituent of hemicellulose. The key enzymes involved in this process are β-1,4-endoxylanase and β-D-xylosidase. Endoxylanase cleave glycosidic linkages of the main chain of xylan releasing xylo-oligosaccharides, which are used by β-xylosidase to produce monomers of xylose. The alfaproteobacteria Caulobacter crescentus is non pathogenic, Gram negative, mainly found in aquatic environments and on many types of soils. This bacterium has about seven genes directly associated with xylan degradation and five of them encoding β-xylosidases. To date, there are only three studies on the β-xylosidase II from C. crescentus. The first characterization of this enzyme showed that it is capable of hydrolyzing substrates such as xylobiose, xylotriose and xilopentose whose optimum pH is 6 and optimum temperature is 55°C, although it is stable at 50°C, which shows a thermotolerance, indicating strong enough to be used in different biotechnological applications. The stability and reusability of enzymes are of fundamental importance, since they reflect significantly on the cost of the final product, and one way to achieve this is with the immobilization of enzymes, consisting of confinement thereof in a matrix or support, which can be inert polymers or inorganic materials, so that its catalytic activity is retained and the enzyme can be used repeatedly and continuously. In the present report, it was found that the β-xylosidase II (CcXynB2) of Caulobacter crescentus increased by 62% of its activity in 5 mM KCl probably as a consequence of a positive role of K+ ions. CCxynB2 was measured against various compounds described as inhibitors of hydrolysis and fermentation of lignocellulosic biomass and showed 61% more tolerant incubation with ethanol (200 mM) at 37 °C for 48 h in the absence of alcohol. The specific activities of CcXynB2 were evaluated in the presence of 10mM phenol or galacturonic acid, 100 mM hydroxymethylfurfural or ferulic acid, 1 mM acetic acid, 200 mM arabinose, glucose or xylose and it was found that were equal (100%) or much higher than the values obtained in the total absence of these compounds after 48 h. When the inhibitors were used in combination, the CcXynB2 retained 67% of its initial activity after testing at 37°C during 48 h. The enzymatic hydrolysis of hemicellulose from corncob was conducted with CcXynB2 alone or in synergism with xylanase and commercial β-glycosidase, which were more efficient in performed the saccharification of hemicellulose from 37-50 °C. The immobilized CcXynB2 in mobile phase resin led to a protective effect of specific activity, which was proportionally parallel to decreased temperatures (60 to -20°C). The data presented here indicate that CcXynB2 is promising and has potential to work in simultaneous saccharification and fermentation processes for cellulosic ethanol production. To our knowledge, is the first time that similar results are reported in the literature to bacterial β-xylosidases. Thus, this work contribute positively by providing essential information to improve the use of β-xylosidase II of Caulobacter crescentus. / Biomassas lignocelulósicas constituem a matéria-prima mais abundante e promissora como recurso natural e renovável. Esses materiais vegetais são polímeros de carboidratos complexos compostos basicamente por celulose, hemicelulose e lignina, que estão unidos entre si por ligações covalentes e podem ser convertidos em produtos de valor agregado, como os biocombustíveis. A degradação dos materiais lignocelulósicos é feita a partir de enzimas produzidas principalmente por micro-organismos como fungos filamentosos, leveduras e bactérias. Para obter etanol a partir de resíduos agroindustriais, baseando-se na hidrólise enzimática, são necessárias, basicamente, quatro etapas: produção de enzimas, pré-tratamento, hidrólise enzimática e fermentação. O pré-tratamento é o processo que irá dissociar o complexo lignina-celulose, reduzir o grau de cristalinidade da celulose e aumentar a porosidade dos materiais, através do aumento da área superficial da biomassa. No entanto, o pré-tratamento pode gerar produtos inibidores, que incluem compostos fenólicos e outros aromáticos, ácidos alifáticos, aldeídos, furanos, íons inorgânicos. A fermentação e sacarificação simultânea é uma estratégia importante para a produção de etanol celulósico ou de segunda geração, onde a hidrólise enzimática da celulose e a fermentação são desenvolvidas simultaneamente no mesmo reator, com o intuito de obter etanol em altas taxas e diminuir a formação de compostos inibidores. A hidrólise enzimática necessita, primeiramente, que a biomassa lignocelulósica seja pré-tratada para aumentar o acesso ao ataque enzimático, para que posteriormente a celulose seja quebrada pela ação de celulases. As xilanases compreendem o grupo de enzimas responsáveis pela hidrólise do xilano, principal constituinte da hemicelulose. As principais enzimas envolvidas nesse processo são β-1,4-endoxilanase e a β-D-xilosidase. Endoxilanases clivam as ligações glicosídicas da cadeia principal do xilano liberando xilo-oligossacarídeos, que são utilizados pelas β-xilosidases para liberar xilose. A alfaproteobactéria Caulobacter crescentus é não patogênica, Gram negativa, encontrada principalmente em ambientes aquáticos e em muitos tipos de solos. Essa bactéria apresenta cerca de sete genes envolvidos diretamente na degradação do xilano, sendo que cinco deles codificam para β-xilosidases. Até o momento, existem apenas três trabalhos sobre a β-xilosidase II de C. crescentus. A primeira caracterização da enzima mostrou que esta é capaz de hidrolisar substratos como xilobiose, xilotriose e xilopentose, cujo pH ótimo é 6 e temperatura ótima é 55ºC, embora seja mais estável em 50ºC, o que demonstra uma modesta termotolerância, indicando ser suficientemente resistente para diferentes aplicações biotecnológicas. A estabilidade e a possibilidade de reutilização de enzimas são de fundamental importância, pois refletem significativamente no custo do produto final, e uma forma de conseguir isso é com a imobilização de enzimas, que consiste no confinamento da mesma em uma matriz ou suporte, que podem ser polímeros inertes ou materiais inorgânicos, de modo que sua atividade catalítica fique retida e a enzima possa ser usada repetidamente e continuamente. No presente trabalho, verificou-se que a β-xilosidase II (CcXynB2) de Caulobacter crescentus aumentou 62% da sua atividade em 5 mM de KCl provavelmente em consequência de um papel positivo dos íons K+. CcXynB2 foi avaliada frente a diferentes compostos descritos como inibidores do processo de hidrólise e fermentação da biomassa lignocelulósica e mostrou-se 61% mais tolerante a incubação com etanol (200 mM) a atividades específicas da CcXynB2 foram avaliadas na presença de 10 mM fenol ou ácido galacturônico, 100 mM de hidroximetilfurfural ou ácido ferúlico, 1 mM de ácido acético, 200 mM de arabinose, glicose ou xilose, e verificou-se que foram iguais (100%) ou muito superiores aos valores obtidos na ausência total destes compostos após 48 h. Quando os inibidores foram usados em associação, a CcXynB2 reteve 67% da sua atividade inicial após 48 h de ensaio a 37ºC. A hidrólise enzimática da hemicelulose de sabugo de milho foi conduzida com CcXynB2 isoladamente ou em sinergismo com xilanase e β-glicosidase comerciais, as quais foram mais eficientes em sacarificar a hemicelulose entre 37-50ºC. A imobilização da CcXynB2 em resina de fase móvel levou a um efeito protetor da atividade específica, que ocorreu de forma paralela à diminuição de temperatura (60 a -20ºC). Os dados apresentados aqui indicam que a CcXynB2 é promissora e possui potencial para atuar em processos de sacarificação e fermentação simultânea para produção de etanol celulósico. Segundo nosso conhecimento, é a primeira vez que resultados similares são relatados na literatura para β-xilosidases bacterianas. Dessa forma, este trabalho pode contribuir positivamente, fornecendo informações fundamentais para aprimorar o uso da β-xilosidase II de Caulobacter crescentus

Made available in DSpace on 2017-07-10T13:59:26Z (GMT). No. of bitstreams: 1 DISSERTACAO AMANDA ALVES MESTRADO EM CIENCIAS FARMACEUTICAS _UNIOESTE 2015.pdf: 10598736 bytes, checksum: 51f0f3eb83858fee62392b7892930766 (MD5) Previous issue date: 2015-12-07 / SIM(não especificado) / Lignocellulosic biomass are the raw material most abundant and promising as a natural and renewable resource. These plant materials are complex carbohydrate polymer composed mainly of cellulose, hemicellulose and lignin, which are linked by covalent bonds and can be transformed into value-added products, such as biofuels. The degradation of lignocellulosic material is made mainly from enzymes produced by microorganisms such as filamentous fungi, yeast and bacteria. Ethanol production from agricultural residues, based on the enzymatic hydrolysis, it takes basically four stages: production of enzymes, pretreatment, enzymatic hydrolysis and fermentation. Pretreatment is a work that will break the lignin cellulose complex, reducing the degree of crystallinity of the cellulose and increase the porosity of the material, by increasing the surface area of the biomass. However, pre-treatment products can generate inhibitors which include phenolic and other aromatic, aliphatic acids, aldehydes, furans, inorganic ions. The fermentation and simultaneous saccharification is an important approach for producing cellulosic or ethanol of second generation, where the enzymatic hydrolysis of cellulose and fermentation are simultaneously carried out in the same reactor, in order to obtain ethanol at a high rate and decrease formation of inhibitor compounds. Enzymatic hydrolysis requires, first, that the lignocellulosic biomass is pretreated to increase access to enzymatic attack, so that later the cellulose is broken down by cellulase action. Xylanases include the group of enzymes responsible for the hydrolysis of xylan, the major constituent of hemicellulose. The key enzymes involved in this process are β-1,4-endoxylanase and β-D-xylosidase. Endoxylanase cleave glycosidic linkages of the main chain of xylan releasing xylo-oligosaccharides, which are used by β-xylosidase to produce monomers of xylose. The alfaproteobacteria Caulobacter crescentus is non pathogenic, Gram negative, mainly found in aquatic environments and on many types of soils. This bacterium has about seven genes directly associated with xylan degradation and five of them encoding β-xylosidases. To date, there are only three studies on the β-xylosidase II from C. crescentus. The first characterization of this enzyme showed that it is capable of hydrolyzing substrates such as xylobiose, xylotriose and xilopentose whose optimum pH is 6 and optimum temperature is 55°C, although it is stable at 50°C, which shows a thermotolerance, indicating strong enough to be used in different biotechnological applications. The stability and reusability of enzymes are of fundamental importance, since they reflect significantly on the cost of the final product, and one way to achieve this is with the immobilization of enzymes, consisting of confinement thereof in a matrix or support, which can be inert polymers or inorganic materials, so that its catalytic activity is retained and the enzyme can be used repeatedly and continuously. In the present report, it was found that the β-xylosidase II (CcXynB2) of Caulobacter crescentus increased by 62% of its activity in 5 mM KCl probably as a consequence of a positive role of K+ ions. CCxynB2 was measured against various compounds described as inhibitors of hydrolysis and fermentation of lignocellulosic biomass and showed 61% more tolerant incubation with ethanol (200 mM) at 37 °C for 48 h in the absence of alcohol. The specific activities of CcXynB2 were evaluated in the presence of 10mM phenol or galacturonic acid, 100 mM hydroxymethylfurfural or ferulic acid, 1 mM acetic acid, 200 mM arabinose, glucose or xylose and it was found that were equal (100%) or much higher than the values obtained in the total absence of these compounds after 48 h. When the inhibitors were used in combination, the CcXynB2 retained 67% of its initial activity after testing at 37°C during 48 h. The enzymatic hydrolysis of hemicellulose from corncob was conducted with CcXynB2 alone or in synergism with xylanase and commercial β-glycosidase, which were more efficient in performed the saccharification of hemicellulose from 37-50 °C. The immobilized CcXynB2 in mobile phase resin led to a protective effect of specific activity, which was proportionally parallel to decreased temperatures (60 to -20°C). The data presented here indicate that CcXynB2 is promising and has potential to work in simultaneous saccharification and fermentation processes for cellulosic ethanol production. To our knowledge, is the first time that similar results are reported in the literature to bacterial β-xylosidases. Thus, this work contribute positively by providing essential information to improve the use of β-xylosidase II of Caulobacter crescentus. / Biomassas lignocelulósicas constituem a matéria-prima mais abundante e promissora como recurso natural e renovável. Esses materiais vegetais são polímeros de carboidratos complexos compostos basicamente por celulose, hemicelulose e lignina, que estão unidos entre si por ligações covalentes e podem ser convertidos em produtos de valor agregado, como os biocombustíveis. A degradação dos materiais lignocelulósicos é feita a partir de enzimas produzidas principalmente por micro-organismos como fungos filamentosos, leveduras e bactérias. Para obter etanol a partir de resíduos agroindustriais, baseando-se na hidrólise enzimática, são necessárias, basicamente, quatro etapas: produção de enzimas, pré-tratamento, hidrólise enzimática e fermentação. O pré-tratamento é o processo que irá dissociar o complexo lignina-celulose, reduzir o grau de cristalinidade da celulose e aumentar a porosidade dos materiais, através do aumento da área superficial da biomassa. No entanto, o pré-tratamento pode gerar produtos inibidores, que incluem compostos fenólicos e outros aromáticos, ácidos alifáticos, aldeídos, furanos, íons inorgânicos. A fermentação e sacarificação simultânea é uma estratégia importante para a produção de etanol celulósico ou de segunda geração, onde a hidrólise enzimática da celulose e a fermentação são desenvolvidas simultaneamente no mesmo reator, com o intuito de obter etanol em altas taxas e diminuir a formação de compostos inibidores. A hidrólise enzimática necessita, primeiramente, que a biomassa lignocelulósica seja pré-tratada para aumentar o acesso ao ataque enzimático, para que posteriormente a celulose seja quebrada pela ação de celulases. As xilanases compreendem o grupo de enzimas responsáveis pela hidrólise do xilano, principal constituinte da hemicelulose. As principais enzimas envolvidas nesse processo são β-1,4-endoxilanase e a β-D-xilosidase. Endoxilanases clivam as ligações glicosídicas da cadeia principal do xilano liberando xilo-oligossacarídeos, que são utilizados pelas β-xilosidases para liberar xilose. A alfaproteobactéria Caulobacter crescentus é não patogênica, Gram negativa, encontrada principalmente em ambientes aquáticos e em muitos tipos de solos. Essa bactéria apresenta cerca de sete genes envolvidos diretamente na degradação do xilano, sendo que cinco deles codificam para β-xilosidases. Até o momento, existem apenas três trabalhos sobre a β-xilosidase II de C. crescentus. A primeira caracterização da enzima mostrou que esta é capaz de hidrolisar substratos como xilobiose, xilotriose e xilopentose, cujo pH ótimo é 6 e temperatura ótima é 55ºC, embora seja mais estável em 50ºC, o que demonstra uma modesta termotolerância, indicando ser suficientemente resistente para diferentes aplicações biotecnológicas. A estabilidade e a possibilidade de reutilização de enzimas são de fundamental importância, pois refletem significativamente no custo do produto final, e uma forma de conseguir isso é com a imobilização de enzimas, que consiste no confinamento da mesma em uma matriz ou suporte, que podem ser polímeros inertes ou materiais inorgânicos, de modo que sua atividade catalítica fique retida e a enzima possa ser usada repetidamente e continuamente. No presente trabalho, verificou-se que a β-xilosidase II (CcXynB2) de Caulobacter crescentus aumentou 62% da sua atividade em 5 mM de KCl provavelmente em consequência de um papel positivo dos íons K+. CcXynB2 foi avaliada frente a diferentes compostos descritos como inibidores do processo de hidrólise e fermentação da biomassa lignocelulósica e mostrou-se 61% mais tolerante a incubação com etanol (200 mM) a atividades específicas da CcXynB2 foram avaliadas na presença de 10 mM fenol ou ácido galacturônico, 100 mM de hidroximetilfurfural ou ácido ferúlico, 1 mM de ácido acético, 200 mM de arabinose, glicose ou xilose, e verificou-se que foram iguais (100%) ou muito superiores aos valores obtidos na ausência total destes compostos após 48 h. Quando os inibidores foram usados em associação, a CcXynB2 reteve 67% da sua atividade inicial após 48 h de ensaio a 37ºC. A hidrólise enzimática da hemicelulose de sabugo de milho foi conduzida com CcXynB2 isoladamente ou em sinergismo com xilanase e β-glicosidase comerciais, as quais foram mais eficientes em sacarificar a hemicelulose entre 37-50ºC. A imobilização da CcXynB2 em resina de fase móvel levou a um efeito protetor da atividade específica, que ocorreu de forma paralela à diminuição de temperatura (60 a -20ºC). Os dados apresentados aqui indicam que a CcXynB2 é promissora e possui potencial para atuar em processos de sacarificação e fermentação simultânea para produção de etanol celulósico. Segundo nosso conhecimento, é a primeira vez que resultados similares são relatados na literatura para β-xilosidases bacterianas. Dessa forma, este trabalho pode contribuir positivamente, fornecendo informações fundamentais para aprimorar o uso da β-xilosidase II de Caulobacter crescentus

Gegenstand der vorliegenden Arbeit war die Untersuchungen von Verunreinigungsprofilen von Aminosäuren aus biotechnologischer Herstellung. Dazu sollten die Aminosäuren Arg, His, Ile, Lys, Phe, Pro, Ser und Trp von verschiedenen Herstellern und aus unterschiedlichen Batches genauer unter die Lupe genommen werden. Mit der im Europäischen Arzneibuch beschriebenen dünnschichtchromatographischen Methode (DC-Methode) für „mit Ninhydrin nachweisbare Substanzen“ können ausschließlich Fremdaminosäuren nachgewiesen werden und dies nur, wenn der jeweilige Gehalt relativ hoch ist. Andere Stoffgruppen, die aus der biotechnologischen Herstellung stammen, werden aufgrund der Trennbedingungen sowie der Detektionsart der DC-Methode nicht erfasst. Deshalb mussten neue Methoden entwickelt werden. Laut Vorschriften der International Conference of Harmonisation (ICH) müssen unbekannte Verunreinigungen in Wirkstoffen für orale Therapeutika auf 0,1 % w/w begrenzt werden können. In die Untersuchungen wurden neben den Fremdaminosäuren auch Peptide, Aminozucker und Nucleinsäuren als potentielle Verunreinigungen, die aus der Herstellung bzw. aus dem Reinigungsprozess kommen, einbezogen. Diese zusätzlichen Stoffgruppen können aus den „Nebenaktivitäten“ der Mikroorganismen entstehen. Aminosäuren, Peptide und Aminozucker wurden versucht, kapillarelektrophoretisch zu quantifizieren. Für die Bestimmung von Nucleinsäuren wurde zusätzlich die UV-Spektroskopie eingesetzt. / Aim of the present work was to investigate the impurity profiles of amino acids of biotechnological origin. Eight amino acids were included: Arg, His, Ile, Lys, Phe, Pro, Ser and Trp. The amino acid samples originating from different producers and different batches had to be studied in deeper detail. The thin-layer chromatographic method (TLC-method) of “ninhydrin-positive substances”, as described in the European Pharmacopoeia, is able to detect primarily other amino acids, if their respective content is relatively high. Other groups of substances of biotechnological origin cannot be detected due to the separation conditions and the detection principle of the TLC-method. Therefore new methods had to be developed. In accordance with the guidelines of the International Conference of Harmonisation (ICH), the content of unknown impurities in active ingredients for oral therapeutics should be limited to 0,1 % w/w. Apart from other amino acids, the study included peptides, amino sugars and nucleic acids as potential impurities, originating from production or purification processes. These additional groups of substances are byproducts of the biosynthesis pathways of microorganisms. An attempt was made to quantify the amino acids, peptides and amino sugars by means of capillary electrophoresis. UV-spectrophotometry was additionally used for the determination of nucleic acids.

No description available.

The study presented here was to investigate the traditional production of the Ugandan fermented cereal beverage, Obushera. The effects of germination and malting of sorghum grains under different steeping treatment were first investigated. The traditional preparation of Obushera beverage was carried out and course of fermentation monitored. The viscosity of Obushera was very low throughout the fermentation process. The microflora responsible for the fermentation of Obushera were identified. After considerable research and conduction of tests were carried out, it was found that there was no detectable quantity of alcohol in Obushera. It was also confirmed that that there were no strains of alcohol producing yeasts, such as Saccharomyces sp. found in the Obushera. / Master of Science (Hons) (Food Science)

Commercial flavour-enhancing enzymes were delivered in an encapsulated form to accelerate Cheddar cheese ripening. Polymers such as alginate, chitosan and k- Carrageenan were screened to be used as encapsulant material for microencapsulation of the commercial protease enzyme, Flavourzyme®. Alginate was found to be a suitable polymer for Flavourzyme encapsulation using the Inotech® encapsulator while _-Carrageenan and chitosan were too viscous for extrusion through the encapsulator nozzle. Gelling of alginate-Flavourzyme microcapsules in 0.1M CaCl2 resulted in poor encapsulation efficiency (ranging 17- 18% depending on the alginate concentration). Incorporation of Hi-Maize™ starch or pectin as filler materials into the alginate-Flavourzyme encapsulation matrix to increase encapsulation efficiency by minimising porosity also resulted in poor encapsulation efficiency. An alternative approach to the modification of the cationic gelling solution, by adding chitosan, significantly increased the encapsulation efficiency to 70-88% and produced mostly spherical capsules with an average diameter of 500_m. Encapsulation efficiency increased with an increase in chitosan concentration from 0.1 to 0.3% (w/v) in the cationic gelling solution of 0.1M CaCl2. Though gelling of alginate-Flavourzyme microcapsules in gelling solution of 0.1M CaCl2 containing 0.3% (w/v) chitosan resulted in higher encapsulation efficiency, a chitosan concentration of 0.1% (w/v) was chosen for further work as higher concentrations of chitosan in the gelling solution resulted in aggregation of capsules during formation. Gelling time of 10 min and alginate concentrations in the range 1.6 to 2.0% (w/v) were found to be optimal encapsulation parameters for Flavourzyme encapsulation while 2.0% (w/v) solution of trisodium citrate was found to be optimal for in vitro release of encapsulated enzymes for measurement of enzyme activity. Flavourzyme capsules stored frozen or freeze-dried were shelf stable for at least 10 weeks retaining about 80% of the initial enzyme activity as opposed to retention of 25-34% activity in air-dried capsules. Leakage of encapsulated Flavourzyme prepared from 1.6% (w/v) alginate was slightly higher than those prepared from 1.8 and 2.0% (w/v) alginate in cheese milk. Flavourzyme-alginate capsules prepared from 1.6, 1.8 and 2.0% (w/v) alginate retained over 70% of the initial enzyme activity under simulated cheese-press pressure. Concentration of alginate had no significant effect (p > 0.05) on the retention of encapsulated Flavourzyme when the capsules were pressed for 4h; however when the simulated cheese press duration increased to 8 and 16h the retention of encapsulated Flavourzyme was significantly higher (p [less than] 0.01) in capsules produced from 2.0% (w/v) alginate. Incorporation of encapsulated enzymes into the milk prior to rennetting resulted in an even distribution of capsules in the cheese matrix compared to aggregation of capsules, when added to milled curd prior to salting. All cheeses; control with no added enzymes and experimental cheeses with free and encapsulated Flavourzyme and/or Palatase showed higher levels of moisture and lower levels of fat compared to standard Cheddar cheese due to the variation in the manufacturing protocol. There was no significant difference (p > 0.05) in fat and final pH between control and experimental cheeses and there was no difference in the numbers of coliforms, E.coli, Salmonella, Listeria, coagulase positive staphylococci, Bacillus cereus, yeast and moulds in control or experimental cheeses. Increased and prolonged proteolysis was observed in cheeses with encapsulated Flavourzyme showing increased release of several peptides, also with the formation of new peptides absent in the control cheese with no added enzymes. Accumulation of high molecular weight/hydrophobic peptides was higher in cheeses with free Flavourzyme followed by cheeses with encapsulated Flavourzyme. Concentration of water-soluble peptides increased with the increase in the concentration of encapsulated Flavourzyme in the cheese. Concentration of water-insoluble peptides was higher in control cheese compared to cheeses with encapsulated Flavourzyme even after 180 days ripening. After 30 days of ripening, concentration of most free amino acids was about 3 times greater in cheeses with encapsulated Flavourzyme than in control and about 7 times higher after 90 days ripening. Concentration of total amino acids was consistently higher in cheeses with encapsulated Flavourzyme compared to control. Cheese grading scores for body, texture and appearance of all cheeses with encapsulated enzymes were lower than control and free enzyme treated cheeses during the entire grading period of about 100 days due to crumbly and pasty texture. Control and cheeses with added Flavourzyme received high overall score for flavour. Flavour score of cheese with encapsulated Flavourzyme at a concentration of 0.75 LAPU/g milk protein was higher than all cheeses around 50 days with better overall flavour score until about 94 days ripening with improved flavour and elimination of bitterness. However the flavour of enzyme treated cheeses deteriorated with time and the control cheese scored the highest for flavour. Though increased concentration of free fatty acids was detected in cheeses treated with encapsulated lipase; Palatase, these cheeses developed rancid, unpleasant, strong lipolytic flavours as early as 55 days ripening. / Doctor of Philosophy (PhD)

Thesis (Ph. D.)--Ohio State University, 2006. / Title from first page of PDF file. Includes bibliographical references (p. 104-114).

Because of the limited supply of imported crude oil and environmental degradation, renewable energy is becoming commercially feasible and environmentally desirable. In this research, biological and thermal (pyrolysis) conversion pathways for biofuel production from lignocellulosic feedstocks were compared. For biological conversions of sorghum, ethanol yield was improved using M81-E variety (0.072 g/g juice) over Umbrella (0.065 g/g juice) for first-generation biomass (sorghum juice), and 0.042 g/g sorghum was obtained from the cellulosic portion of second-generation biomass. When ultrasonication was combined with hot water pretreatment, yields increased by 15% and 7% for cellulose to glucose, and hemicellulose to pentose, respectively. Ethanol yield was 10% higher when this pretreatment was combined with Accellerase 1500+XC for saccharification. Biological conversion yielded 1,600?2,300 L ethanol/ha for first-generation biomass, and 4,300?4,500 L ethanol/ha from lignocellulosic biomass. For thermal (pyrolysis) conversion of lignocellulosic switchgrass at 600 degrees C, product yield was 37% bio-oil, 26% syngas, and 25% bio-char. At 400 degrees C, product yield was 22% bio-oil, 8% syngas, and 56% bio-char. Bio-oil from pyrolysis was highly oxygenated (37 wt%). It required chemical transformation to increase its volatility and thermal stability, and to reduce its viscosity by removing objectionable oxygen, so the product could be used as transportation fuel (gasoline). As a consequence of upgrading bio-oil by catalytic hydrogenation, bio-oil oxygen decreased from 37?2 wt%, carbon increased from 50?83 wt%, hydrogen increased from 9?15 wt% and heating value increased from 36?46 MJ/kg, resulting in a fuel that was comparable to gasoline. The upgraded product passed the thermal stability test when kept under an oxygen-rich environment. The upgraded product consisted of 14.8% parrafins, 21.7% iso-parrafins, 3% napthene, 42.6% aromatics, 4.7% olefin, 4.7% DMF, 8% alcohol, and 0.6% ketone on a mass basis. Comparing the two pathways, biological conversion had 11 wt% ethanol yield from sorghum, and thermal conversion had 13 wt% gasoline yield from switchgrass. For process efficiency, thermal conversion had 35% energy loss versus 45% energy loss for biological conversions. For the biological pathway, ethanol cost was $2.5/gallon ($4/gallon, gasoline equivalent), whereas for the thermal pathway, switchgrass gasoline cost was $3.7/gallon, both with 15% before tax profit.

L'ensilage permet de conserver le fourrage par un processus de fermentation lactique, ce qui limite les pertes de rendement et de valeur nutritive. Le fourrage ainsi acidifié et maintenu en conditions anaérobies peut alors être conservé pour nourrir les animaux durant la période d'hivernage. Théoriquement, lorsque l'ensilabilité des plantes est bonne, la conservation sera elle aussi adéquate. Toutefois, des études récentes ont démontré que la corrélation entre l'ensilabilité et la conservation n'est pas très élevée. L'une des hypothèses est que le pHw (pH de stabilité anaérobique) n'est pas atteint assez rapidement. Le temps de latence (temps avant que le pH commence à descendre) ainsi que le déclin du pH (descente du pH de l'ensilage jusqu'au pHw) seraient trop long. Le nombre de bactéries lactiques, l'efficacité des ces bactéries à produire de l'acide lactique, de même que la spécificité de leur métabolisme fermentaire pour certains sucres pourraient modifier ces paramètres et ainsi améliorer la conservation. Les deux essais réalisés pour ce projet avaient pour objectif d'étudier ces facteurs. Dans le premier essai, la fléole des prés (Phleum pratense L.) au premier cycle de végétation à 35% de MS, a été traitée avec un inoculant lactique contenant du Lactobacillus plantarum (Biomax SI-50). L'inoculant a été appliqué sur le fourrage à trois doses différentes; 105, 106 et 107 UFC .g de FF-1. Dans le deuxième essai, la même plante au deuxième cycle de végétation à 30% de MS, a été traitée avec le Biomax SI-50 et le Powerstart contenant lui aussi un Lactobacillus plantarum (Aber-F1) mais ayant la caractéristique d'utiliser les fructanes. Ces deux inoculants ont soit été réhydratés dans l'eau juste avant leur application sur les fourrages ou activés entre 22 et 24 heures avant leur application dans un bouillon nutritif. Pour chacun des traitements, des silos expérimentaux avec 250 g de fourrage ont été fabriqués. Des silos ont été analysés après 0, 6, 12, 24, 48, 72 heures ainsi que 1, 2, 4 et 15 semaines d'incubation afin d'étudier la cinétique du pH. Le premier essai a démontré que l'addition d'un inoculant permet une diminution plus rapide du pH que le témoin (P < 0,05). Toutefois, les doses d'inoculant ne permettent pas de diminuer le temps de latence comparativement au témoin (P > 0,05). Dans le cas du deuxième essai, il n'y a pas eu de différence ni sur la vitesse de diminution ni sur le temps de latence entre tous les traitements (P > 0,05). Pour les deux essais, le pH de stabilité anaérobie a été atteint et les teneurs en acides butyriques observées sont difficiles à interpréter. La cinétique des différents acides organiques des ensilages nous indiquent certaines particularités qui semblent être reliées aux sucres. ______________________________________________________________________________ MOTS-CLÉS DE L’AUTEUR : Inoculant lactique, Bactérie lactique, Ensilage, Fermentation, Fléole des prés (Phleum pratense L.), Sucres.