京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第21805号 / 農博第2318号 / 新制||農||1065(附属図書館) / 学位論文||H31||N5177(農学部図書室) / 京都大学大学院農学研究科農学専攻 / (主査)教授 白岩 立彦, 教授 奥本 裕, 教授 稲村 達也 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

京都大学 / 0048 / 新制・課程博士 / 博士(エネルギー科学) / 甲第22087号 / エネ博第395号 / 新制||エネ||76(附属図書館) / 京都大学大学院エネルギー科学研究科エネルギー基礎科学専攻 / (主査)教授 森井 孝, 教授 木下 正弘, 教授 片平 正人 / 学位規則第4条第1項該当 / Doctor of Energy Science / Kyoto University / DGAM

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京都大学 / 新制・課程博士 / 博士(エネルギー科学) / 甲第25400号 / エネ博第479号 / 新制||エネ||89(附属図書館) / 京都大学大学院エネルギー科学研究科エネルギー基礎科学専攻 / (主査)教授 森井 孝, 教授 片平 正人, 教授 佐川 尚 / 学位規則第4条第1項該当 / Doctor of Energy Science / Kyoto University / DGAM

Les milieux biologiques bruts, provenant des opérations de transformation de biomasse sont souvent caractérisés à la fois par leur caractère polluant et par un potentiel de valorisation important. Le développement de procédés adaptés au traitement de tels milieux complexes présente ainsi beaucoup d’intérêt. Les jus verts générés par la déshydratation de la luzerne (Medicago Sativa) sont caractérisés par une forte teneur en protéines. Outre leur valeur nutritionnelle importante, ces protéines ont des applications potentielles dans plusieurs domaines, notamment environnemental de part leur teneur élevée en Ribulose 1,5 Bisphosphate Carboxylase Oxydase (Rubisco), enzyme responsable de la fixation du CO2 chez les plantes. Dans ce travail la séparation sélective de la Rubisco à partir du jus de luzerne industriel centrifugé par chromatographie d’échange d’ions a été étudiée. Dans un premier temps une méthode d’analyse qualitative et quantitative a été mise au point pour la détection et la quantification de la Rubisco en solution et ainsi le suivi du procédé de séparation. Dans un deuxième temps, le procédé de séparation a été étudié en colonne, en lit fixe et en lit expansé, en utilisant le support échangeur d’anions Q Hyper Z et l’effet de la dilution du milieu sur la capacité dynamique du procédé a été analysé dans les deux cas. Les résultats obtenus ont montré que les deux modes de contact permettent d’avoir des capacités dynamiques de rétention du même ordre de grandeur que celles de la littérature. Après élution, la Rubisco a été concentrée jusqu’à 21 fois et les fractions produites étaient caractérisées par un grand degré de pureté. Par ailleurs, des études d’équilibre et cinétique d’échange ont été initiées dans ce travail et ont démontré que malgré la taille importante de la protéine d’intérêt (560 kDa) les limitations stériques à son transfert ne sont pas plus importantes que dans le cas de protéines plus simples et plus petites et que le support Q Hyper Z présente effectivement une grande affinité pour la protéine. Enfin une première approche théorique a été conduite pour la compréhension des interactions entre la protéine et l’échangeur dans ce milieu complexe. Elle a permis de confirmer l’importance de la prise en compte de la présence d’autres biomolécules dans le milieu sur la rétention de la Rubisco, peut être même plus que celle des sels / Biological raw material derived from bio-refinery processes, is often considered a source of pollution but it seems also to be a promising alternative to potential material recovery. The development of suitable processes for handling such complex biological material has so many concerns. Green juice produced from mechanical dehydration of Alfalfa (Medicago sativa) is an excellent source of protein with high nutritional quality. Ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco) is the most abundant protein in the green juice, with potential applications in many fields, such as human nutrition, pharmaceuticals, environmental… The aim of this study is to isolate and recover Rubisco produced from an industrial alfalfa green juice, by ion exchange chromatography process. First of all, a qualitative and quantitative analytical method was developed to provide reliable information about Rubisco content monitoring in the separation process. In a second step, the separation process was performed in fixed and expanded bed, using the anion exchanger Q Hyper Z. In both cases, the effect of the dilution of the green juice on the dynamic capacity of the columns was studied. The results showed that the dynamic capacity retention was similar in both columns to those reported in literature. After elution step, Rubisco was concentrated 21 times and produced with high level of purity. Furthermore, kinetic of ion exchange study was initiated. Despite the large size of the protein (560 kDa), steric limitations to mass transfer were not very significant when compared to those of conventional small proteins. The support Q Hyper Z showed an excellent affinity for the protein recovery. Finally, a first theoretical investigation has been conducted for understanding the retention mechanism between the protein and the separation column. This study shows the importance of taking into account the presence of other bio-molecules in order to perform the retention of Rubisco, perhaps even more than that of salts

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Human practices have resulted in great damage to the environment. Carbon-depletion, water and air pollution, as well as global warming are examples of the environmental footprints caused by several industries and their related applications. Detergency (cleaning) is widely practiced operation in household, industry and institutional sectors and thus consumes significant amounts of water, energy and chemicals and, therefore, contributes appreciably to the environmental destruction. This process is still not fully understood, on a molecular level, and not acceptably optimised. Therefore, this study is a contribution toward a better fundamental understanding and optimisation of protein stain removal from interfaces, which may ultimately result in the development of environmentally friendly and sustainable cleaning products and technologies. In this research, the cleaning of rubisco, a grassy protein stain, from different surfaces (hydrophobic, hydrophilic, and dyed) using different cleaning agent formulations was investigated. These studies encompassed experimental work and fundamental analysis in terms of mathematical modelling. The results revealed a consistent correlation between enzyme adsorption kinetics and stain cleanability. Higher adsorption and desorption absolute rates ( a k and d k ) resulted in higher enzyme mobility and thus higher stain cleanability regardless of the enzyme adsorbed amount. Surface chemistry underlying the stain has indirectly influenced stain cleanability through the alteration of enzyme adsorption kinetics. Such fundamental findings may aid in screening enzyme candidates for detergent formulations and may also assist in designing easily cleanable surfaces. Another fundamental finding is the cooperative cleaning mechanism of surfactant and enzyme of rubisco stain from different interfaces. The break down of intermolecular physical bonds between interfacial rubisco molecules by surfactants and the break down of the intramolecular covalent bonds by enzyme resulted in a higher protein displacement from interfaces. The overall protein removal by the two different actions of surfactant and enzyme showed a superiority of a biosurfactant-enzyme formulation. This finding may have significant implications on developing sustainable detergents that have superior cleaning performance and no or minimal environmental hazard. Overall, the findings reported in this Ph.D. thesis may form a basis for further comprehensive scientific research, which may ultimately provide detergent market with more efficient and optimum cleaning products and technologies.