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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
531

Aprovechamiento de subproductos de zumos de frutos rojos para el diseño de alimentos

Díez Sánchez, Elena 02 September 2021 (has links)
Tesis por compendio / [ES] La presente tesis plantea distintas estrategias para el diseño de alimentos a partir del aprovechamiento del bagazo que se genera como subproducto en la industria de elaboración de zumos de frutos rojos. De este modo, se puede aportar un valor añadido al subproducto, al mismo tiempo que se desarrollan alimentos capaces de cubrir las necesidades y exigencias nutritivas que demanda el consumidor actual. Uno de los aspectos más interesantes que tiene el uso de estos subproductos, es su alto contenido en polifenoles y fibra, de gran importancia en la industria alimentaria por su efecto en las propiedades de los alimentos y sobre la salud. Se realizó un primer estudio para conocer el efecto de la sustitución de grasa en bizcochos por diferentes tipos de fibra: soluble, insoluble y una mezcla de ambas. En este trabajo se observó que las características texturales, estructurales y sensoriales se vieron afectadas con la incorporación de fibra, principalmente en el caso de la fibra insoluble. Por todo esto, se establecieron dos estrategias de mejora. Por una parte, se incorporaron diferentes impulsores químicos en combinación con bicarbonato (normal o encapsulado) en bizcochos formulados con bagazo y se caracterizaron desde un punto de vista físico químico, estructural y sensorial. Por otra parte, se estudió cómo afecta a la textura, estructura y a la digestibilidad in vitro del almidón la sustitución parcial de harina de trigo por harina extrusionada en magdalenas formuladas con bagazo. En ambos estudios se obtuvieron resultados satisfactorios en cuanto a la mejora de las propiedades del producto. Además de las estrategias de mejora en productos horneados, se decidió incorporar bagazo en matrices lácteas para desarrollar nuevas formulaciones de batidos. Se estudió el efecto de un tratamiento no térmico, las altas presiones hidrostáticas, sobre el contenido fenólico y la capacidad antioxidante de los batidos, y sobre la capacidad antimicrobiana de los fenoles presentes en el bagazo. Se observó que la mayor retención de compuestos fenólicos y capacidad antioxidante con la mínima supervivencia microbiológica se obtuvo para las presiones y tiempos más altos (500 MPa durante 10 min) en los batidos con las concentraciones más altas de bagazo estudiadas (10%). Por último, se llevó a cabo un estudio con sistemas modelo para conocer las interacciones que tienen los polifenoles, cuando forman parte del bagazo o cuando se encuentran en forma de extracto, con los principales macronutrientes de los alimentos y sus efectos sobre la bioaccesibilidad de los compuestos fenólicos tras la digestión in vitro. La bioaccesibilidad de los polifenoles cuando se incorporaron en los sistemas modelo en forma de bagazo aumentó en comparación con cuando se adicionaron como extracto. Por otro lado, los sistemas modelo formulados con bagazo y un solo nutriente, presentaron una mayor bioaccesibilidad de los compuestos fenólicos que cuando se encontraron todos los nutrientes en el sistema modelo. / [CAT] La present tesi planteja diferents estratègies per al disseny d'aliments a partir de l'aprofitament del bagàs que es genera com a subproducte en la indústria d'elaboració de sucs de fruits rojos. D'aquesta manera, es pot aportar un valor afegit al subproducte, al mateix temps que es desenvolupen aliments capaços de cobrir les necessitats i exigències nutritives que demanda el consumidor actual. Un dels aspectes més interessants que té l'ús d'aquests subproductes, és el seu alt contingut en polifenols i fibra, de gran importància en la indústria alimentària pel seu efecte en les propietats dels aliments i sobre la salut. Es va realitzar un primer estudi per a conéixer l'efecte de la substitució de greix en bescuits per diferents tipus de fibra: soluble, insoluble i una mescla d'ambdues. En aquest treball es va observar que les característiques texturals, estructurals i sensorials es van veure afectades amb la incorporació de fibra, principalment en el cas de la fibra insoluble. Per tot això, es van establir dues estratègies de millora. D'una banda, es van incorporar diferents impulsors químics en combinació amb bicarbonat (normal o encapsulat) en bescuits formulats amb bagàs i es van caracteritzar des d'un punt de vista físic químic, estructural i sensorial. D'altra banda, es va estudiar com afecta a la textura, estructura i a la digestibilitat in vitro del midó la substitució parcial de farina de blat per farina extrusionada en magdalenes formulades amb bagàs. En tots dos estudis es van obtindre resultats satisfactoris quant a la millora de les propietats del producte. A més de les estratègies de millora en productes enfornats, es va decidir incorporar bagàs en matrius làcties per a desenvolupar noves formulacions de batuts. Es va estudiar l'efecte d'un tractament no tèrmic, les altes pressions hidroestàtiques, sobre el contingut fenòlic i la capacitat antioxidant dels batuts, i sobre la capacitat antimicrobiana dels fenols presents en el bagàs. Es va observar que la major retenció de compostos fenòlics i capacitat antioxidant amb la mínima supervivència microbiològica es va obtindre per a les pressions i temps més alts (500 MPa durant 10 min) en els batuts amb les concentracions més altes de bagàs estudiades (10%). Finalment, es va dur a terme un estudi amb sistemes model per a conéixer les interaccions que tenen els polifenols, quan formen part del bagàs o quan es troben en forma d'extracte, amb els principals macronutrients dels aliments i els seus efectes sobre la bioaccessibilitat dels compostos fenòlics després de la digestió in vitro. La bioaccessibilitat dels polifenols quan es van incorporar en els sistemes model en forma de bagàs va augmentar en comparació amb quan es van addicionar com a extracte. D'altra banda, els sistemes model formulats amb bagàs i un sol nutrient, van presentar una major bioaccessibilitat dels compostos fenòlics que quan es van trobar tots els nutrients en el sistema model. / [EN] This thesis proposes different strategies for the design of foods based on the use of pomace generated as a by-product in the berry juice processing industry. In this way, it is possible to add value to the by-product while developing foods capable of meeting the needs and nutritional requirements demanded by today's consumers. One of the most interesting aspects of the use of these by-products is their high content of polyphenols and fiber, of great importance in the food industry due to their effect on food properties and health. A first study was carried out to determine the effect of replacing fat in sponge cakes with different types of fiber: soluble, insoluble and a mixture of both. In this work it was observed that the textural, structural and sensory characteristics were affected by the incorporation of fiber, mainly in the case of insoluble fiber. For all these reasons, two improvement strategies were determined. On the one hand, different chemical leavenings agents were incorporated in combination with bicarbonate (normal or encapsulated) in sponge cakes formulated with pomace and characterized from a physical-chemical, structural and sensory point of view. On the other hand, it was studied how the partial substitution of wheat flour by extruded flour in pomace-formulated muffins affects texture, structure and in vitro starch digestibility. In both studies satisfactory results were obtained in terms of improved product properties. In addition to the improvement strategies in baked products, it was decided to incorporate pomace in dairy matrices to develop new milkshake formulations. The effect of a non-thermal treatment, such as high hydrostatic pressures, on the phenolic content and antioxidant capacity of the milkshakes, and on the antimicrobial capacity of the phenols present in pomace was studied. It was observed that the highest retention of phenolic compounds and antioxidant capacity with the lowest microbiological survival was obtained for the highest pressures and times (500 MPa for 10 min) in the milkshakes with the highest pomace concentrations studied (10%). Finally, a study was carried out with model systems to learn about the interactions that polyphenols have when they are part of the pomace or when they are in extract form, with the main macronutrients of food and their effects on the bioaccessibility of phenolic compounds after in vitro digestion. The bioaccessibility of polyphenols increased when it is incorporated into the model systems as pomace compared to when it is added as extract. On the other hand, model systems formulated with pomace and a single nutrient presented higher bioaccessibility of phenolic compounds than when all nutrients were found in the model system. / The authors are grateful to Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA-Spain) for financial support through the BERRYPOM. Adding value to processing waste: innovative ways to incorporate fibers from berry pomace in baked and extruded cereal-based foods project included in the ERA-NET-SUSFOOD program, to Spanish Ministry of Science, Innovation and Universities through project AGL2017-86840-C2-2-R, and to FEDER funds from EU. / Díez Sánchez, E. (2021). Aprovechamiento de subproductos de zumos de frutos rojos para el diseño de alimentos [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/172372 / Compendio
532

Tratamientos de secado para la obtención de ingredientes de alto valor nutritivo a partir del destrío postcosecha de caqui

Martínez González, Cristina 17 January 2022 (has links)
Tesis por compendio / [ES] La presente tesis tiene como objetivo ofrecer estrategias que permitan incrementar la rentabilidad del cultivo de caqui "Rojo Brillante" mediante la valorización de los excedentes y destríos generados en los almacenes. La tesis aborda como objetivos principales la obtención de productos e ingredientes deshidratados con alto contenido en compuestos bioactivos y la evaluación de la digestibilidad in vitro de los mismos. La primera estrategia consistió en obtener caquis semisecos mediante secado "natural" evaluando la necesidad de aplicar un tratamiento previo de desastringencia. El tratamiento de secado en los caquis astringentes disminuyó significativamente el contenido en taninos solubles responsables de la astringencia de la fruta, alcanzando valores similares a los presentes en el caqui no astringente. Además, se observó que, el color de la pulpa permaneció con un tono anaranjado y poco pardeado cuando se partió de caqui astringente, por lo que no sería recomendable la aplicación de un tratamiento previo de desastringencia. Una segunda estrategia se centró en la liofilización para obtener snacks de caqui de alta calidad, al ser éste un tratamiento que permite obtener productos o ingredientes con alto valor añadido. Se establecieron las condiciones óptimas de procesado y almacenamiento mediante la obtención de los valores de humedad y actividad del agua críticos que garantizasen el estado vítreo de la matriz, evitando un aumento en la tasa de reacciones de deterioro, cambios de textura y color, y la pérdida de los compuestos bioactivos de la fruta. Como última estrategia se utilizó el tratamiento de secado por aire caliente en caqui astringente y no astringente, en distintos estados de madurez, para obtener snacks de caqui. Los snacks desarrollados presentaron un tono más anaranjado, en comparación con la fruta fresca, y menor contenido en taninos solubles. Se observó una alta correlación entre el nivel de astringencia percibido por los consumidores y la disminución del contenido en taninos solubles. Los snacks obtenidos a partir de caqui astringente en etapas de madurez avanzadas fueron igualmente aceptados por los consumidores que los obtenidos a partir de caqui no astringente. Seguidamente se estudió el efecto del secado con aire caliente y del estado de madurez sobre la fracción de carotenoides mediante diferentes técnicas cuantitativas y cualitativas como la fotoluminiscencia. El secado no afectó al contenido en carotenoides, pero disminuyó la capacidad antioxidante de los snacks. La fotoluminiscencia evidenció la isomerización de carotenoides y la formación de productos de degradación térmica, por lo que ambos hechos podrían explicar la pérdida de capacidad antioxidante en el caqui sometido a tratamientos de secado por aire caliente. Por último, se llevaron a cabo estudios de digestión in vitro. La recuperación de taninos solubles en la fase del intestino delgado fue mayor en los snacks obtenidos mediante secado por aire caliente que en la fruta fresca y en los snacks liofilizados. Los snacks obtenidos a partir de caqui no astringente presentaron un índice de recuperación de taninos solubles mayor que los obtenidos a partir de caqui astringente. Los taninos insolubles llegaron intactos a la fracción no absorbida, por lo que podrían llegar al colon y ejercer su potencial antioxidante. / [CA] La present tesi té com a objectiu oferir estratègies que permeten incrementar la rendibilitat del cultiu de caqui "Roig Brillant" mitjançant la valorització dels excedents i destriaments generats en els magatzems. La tesi aborda com a objectius principals l'obtenció de productes i ingredients deshidratats amb alt contingut en compostos bioactius i l'avaluació de la digestibilitat in vitro d'aquests. La primera estratègia va consistir a obtindre caquis semisecs mitjançant assecat "natural" avaluant la necessitat d'aplicar un tractament previ de desastringència. El tractament d'assecat en els caquis astringents va disminuir significativament el contingut en tanins solubles responsables de la astringència de la fruita, aconseguint valors similars als presents en el caqui no astringent. A més, es va observar que, el color de la polpa va romandre amb un to ataronjat i poc marró quan es va partir de caqui astringent, per la qual cosa no seria recomanable l'aplicació d'un tractament previ de desastringència. Una segona estratègia es va centrar en la liofilització per a obtindre snacks de caqui d'alta qualitat, en ser aquest un tractament que permet obtindre productes o ingredients amb alt valor afegit. Es van establir les condicions òptimes de processament i emmagatzematge mitjançant l'obtenció dels valors d'humitat i activitat de l'aigua crítics que garantiren l'estat vitri de la matriu, evitant un augment en la taxa de reaccions de deterioració, canvis de textura i color, i la pèrdua dels compostos bioactius de la fruita. Com a última estratègia es va utilitzar el tractament d'assecat per aire calent en caqui astringent i no astringent, en diferents estats de maduresa, per a obtindre snacks de caqui. Els snacks desenvolupats van presentar un to més ataronjat, en comparació amb la fruita fresca, i menor contingut en tanins solubles. Es va observar una alta correlació entre el nivell de astringència percebut pels consumidors i la disminució del contingut en tanins solubles. Els snacks obtinguts a partir de caqui astringent en etapes de maduresa avançades van ser igualment acceptats pels consumidors que els obtinguts a partir de caqui no astringent. Seguidament es va estudiar l'efecte de l'assecat amb aire calent i de l'estat de maduresa sobre la fracció de carotenoids mitjançant diferents tècniques quantitatives i qualitatives com la fotoluminiscència. L'assecat no va afectar el contingut en carotenoids, però va disminuir la capacitat antioxidant dels snacks. La fotoluminiscència va evidenciar la isomerització de carotenoids i la formació de productes de degradació tèrmica, per la qual cosa tots dos fets podrien explicar la pèrdua de capacitat antioxidant en el caqui sotmés a tractaments d'assecat per aire calent. Finalment, es van dur a terme estudis de digestió in vitro. La recuperació de tanins solubles en la fase de l'intestí prim va ser major en els snacks obtinguts mitjançant assecat per aire calent que en la fruita fresca i en els snacks liofilitzats. Els snacks obtinguts a partir de caqui no astringent van presentar un índex de recuperació de tanins solubles major que els obtinguts a partir de caqui astringent. Els tanins insolubles van arribar intactes a la fracció no absorbida, per la qual cosa podrien arribar al còlon i exercir el seu potencial antioxidant. / [EN] This thesis proposes different strategies to increase the profitability of "Rojo Brillante" persimmon by valuing the surpluses and discarded fruit generated in the warehouses. This thesis addresses the development of dehydrated products and ingredients with a high content of bioactive compounds and the evaluation of their in vitro digestibility. The first strategy was to obtain semidry persimmons by "natural" drying, evaluating the need to apply a previous desastringency treatment. The drying treatment in the astringent persimmons significantly decreased the soluble tannin content responsible for the astringency of the fruit, reaching values similar to those present in the non-astringent persimmon. In addition, the color of the pulp remained with an orange hue angle and moderate browning, when astringent persimmon was used, thus the application of a previous desastringency treatment would not be recommended. A second strategy focused on freeze-drying to obtain high-quality persimmon snacks, as a treatment that allows obtaining products or ingredients with high added value. Optimum processing and storage conditions were established by obtaining the critical water content and water activity values that would guarantee the glassy state, avoiding an increase in the rate of deterioration reactions, changes in texture and color, and the loss of bioactive compounds in the fruit. As a last strategy, the hot air-drying treatment was used in astringent and non-astringent persimmon, in different ripening stages, to obtain persimmon snacks. The snacks had a more orange hue angle, compared to fresh fruit, and lower soluble tannin content. A high correlation was observed between the level of astringency perceived by consumers and the decrease in soluble tannin content. Snacks obtained from astringent persimmon in advanced ripening stages were equally accepted by consumers as non-astringent ones. Then, the effect of hot air drying and the ripening stage on the carotenoid fraction was studied using different quantitative and qualitative techniques such as photoluminescence. Hot air-drying did not affect the carotenoid content but decreased the antioxidant capacity of the snacks. The photoluminescence evidenced the isomerization of carotenoids and the formation of thermal degradation products. Both facts could explain the loss of antioxidant capacity in persimmon subjected to hot air-drying treatments. Lastly, in vitro digestion studies were carried out. The recovery of soluble tannins in the small intestine phase was higher in the snacks obtained by hot air drying than in the fresh fruit and the freeze-dried snacks. The snacks obtained from non-astringent persimmon had a higher recovery index of soluble tannin than those obtained from astringent persimmon. The insoluble tannins reached the unabsorbed fraction intact, so that they could reach the colon and exert their potential antioxidant capacity. / The authors thank the Ministerio de Ciencia, Innovación y Universidades for the financial support given throughout Project RTA2017-00045-C02-02 / Martínez González, C. (2021). Tratamientos de secado para la obtención de ingredientes de alto valor nutritivo a partir del destrío postcosecha de caqui [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/180348 / Compendio
533

Modelling, Simulation, and Optimisation of Reverse Osmosis Process with Application in Wastewater Treatment and Food Processing

Al-Obaidi, Mudhar A.A.R. January 2018 (has links)
Reverse Osmosis (RO) is a membrane-based separation process applied in several industrial and food processing applications. In this research, performance of RO process is investigated in respect of two applications (a) wastewater treatment (b) concentration fruit juices using model-based techniques. For this purpose, a number of models (both 1 and 2-dimensional steady state and dynamic) for spiral wound RO process are developed based on Solution-Diffusion model and Irreversible Thermodynamic model. The models are validated against actual experimental data reported in the literature before being used in further simulation and optimisation studies for both wastewater treatment and fruit juice concentration. Wastewater effluents of many industrial applications contain a variety of micro-pollutants and highly-toxic compounds, which are released into a variety of water resources. Such pollutants not only disrupt the biological ecosystem, but they also pose a real threat to the water supply for human consumption and to the aquatic ecosystems. The earlier chapters of the thesis evaluate the performance of RO process in terms of removal efficiency of toxic compounds such as chlorophenol, N-nitrosamine, etc. from wastewater. The effect of several operating parameters such as feed pressure, concentration, flow rate and temperature, on the performance of RO process are evaluated. Also, suitability of a number of different RO configurations for efficient removal of toxic compounds are evaluated. For example, (a) two-stage/two-pass RO design synthesis of RO network for the removal of chlorophenol (b) multistage multi-pass RO process with and without energy recovery option for the removal of N-nitrosamine are investigated. The dynamic response of the RO process for step changes in the operating parameters is investigated for the removal of phenolic compounds. Finally, in the context of wastewater treatment, a case study with multi compounds contaminants is suggested where a multi-objective optimisation problem has achieved the optimum rejection of all the compounds and recovery rate. In respect of food processing, RO has been considered as a prominent process in fruit juice concentration due to its ability to effectively retain the flavour, sensory, aroma and nutritional characteristics and concentrate the juice. This research elucidates one example of apple juice concentration process and focuses on highlighting successful modelling and optimisation methodology. This in turn provides an efficient method of RO process for concentrating apple juice by improving the reliability and efficiency of the underlying separation and concentration process. / Ministry of High Education and Scientific Research of Iraq
534

Mesoporous Silica Nanoparticles Enhance the Anticancer Efficacy of Platinum(IV)-Phenolate Conjugates in Breast Cancer Cell Lines

Predarska, Ivana, Saoud, Mohamad, Draca, Dijana, Morgan, Ibrahim, Komazec, Teodora, Eichhorn, Thomas, Mihajlovi, Ekatarina, Dunderovic, Duško, Mijatovic, Sanja, Maksimovic-Ivanic´, Danijela, Hey-Hawkins, Evamarie, Kaluderovic, Goran N. 20 September 2024 (has links)
The main reasons for the limited clinical efficacy of the platinum(II)-based agent cisplatin include drug resistance and significant side effects. Due to their better stability, as well as the possibility to introduce biologically active ligands in their axial positions constructing multifunctional prodrugs, creating platinum(IV) complexes is a tempting strategy for addressing these limitations. Another strategy for developing chemotherapeutics with lower toxicity relies on the ability of nanoparticles to accumulate in greater quantities in tumor tissues through passive targeting. To combine the two approaches, three platinum(IV) conjugates based on a cisplatin scaffold containing in the axial positions derivatives of caffeic and ferulic acid were prepared and loaded into SBA- 15 to produce the corresponding mesoporous silica nanoparticles (MSNs). The free platinum(IV) conjugates demonstrated higher or comparable activity with respect to cisplatin against different human breast cancer cell lines, while upon immobilization, superior antiproliferative activity with markedly increased cytotoxicity (more than 1000-fold lower IC50 values) compared to cisplatin was observed. Mechanistic investigations with the most potent conjugate, cisplatin-diacetyl caffeate (1), and the corresponding MSNs (SBA-15|1) in a 4T1 mouse breast cancer cell line showed that these compounds induce apoptotic cell death causing strong caspase activation. In vivo, in BALB/c mice, 1 and SBA-15|1 inhibited the tumor growth while decreasing the necrotic area and lowering the mitotic rate.
535

Isoleringsmaterialets inverkan på energianvändning och klimatpåverkan : -En fallstudie

Håkans, Alexander, Östlund, Isaac January 2024 (has links)
Inom EU står byggnadssektorn för en hög andel av energianvändningen och medlemsländerna har satt upp mål för att minska energianvändningen. Målen som har fastställts i Sverige innebär att utsläppen av växthusgaser ska vara netto noll senast år 2045. I det här sammanhanget blir det extra viktigt att använda sig av tekniker som minskar CO2e-utsläppen och gynnar klimatet. En byggnads klimatskärm är en primär faktor till energianvändningen och det ställs höga krav på materialen som installeras i klimatskärmen. Det kommer i detta examensarbete utföras en fallstudie på en byggnad i Ljusne, Söderhamn. Här studeras tre olika isoleringsmaterial och två fönstertyper för att analysera deras påverkan på byggnadens energianvändning och för att ta fram det optimalaste alternativet ur ett klimatneutralt perspektiv. De material som studeras är mineralull, fenolskumskiva och termoträ samt vanligt fönster och passivfönster. För att besvara denna frågeställning utförs det energisimuleringar genom datorprogrammet BV2 samt beräkningar med materialens miljövarudeklarationer (EPD) för att analysera deras klimatpåverkan. Byggnadens klimatpåverkan beräknas med hjälp av byggsektorns miljöberäkningsverktyg (BM). I BM beräknas byggnadens utsläpp i koldioxidekvivalenter ut för renovering, energianvändning per år och under en livscykel på 50 år. Energisimuleringar utförda i BV2 visar att tilläggsisolering med fenolskum och fönsterbyte till passivfönster är den effektivaste metoden vid renovering av ytterväggen ur ett energieffektivt användningssätt. Då denna kombination har den lägsta energianvändningen, primärenergital och genomsnittlig värmegenomgångskoefficient. Det påvisar ett resultat med ett primärenergital på 72,5 kWh/m2 och en energianvändning på 116,8 kWh/m2. Men kombination av termoträ och passivfönster är den mest verkningsfulla på byggnadens klimatpåverkan. Detta illustreras i BM där klimatpåverkan för renovering är 2928,9 kg CO2e, energianvändning ger 3980,7 kg CO2e/år och klimatpåverkan för en livscykel på 50 år blir då 201 964 kg CO2e. Användningen av energisimuleringar gör att effektiva och klimatanpassade material kan studeras närmare för att avgöra dess betydelse. För att klimatmålen i Sverige och även EU ska kunna uppnås är behovet av verkningsfulla material väsentliga. Det här examensarbetet är en del i ledet mot ett klimatneutralt samhälle. / Within the European Union (EU), the building sector accounts for a high proportion of energy use and Member States have set targets to reduce energy use. The targets set in Sweden mean that greenhouse gas emissions should be net zero by 2045. In this context, it is particularly important to use technologies that reduce CO2 emissions and benefit the climate. A building’s envelope is a primary factor in energy consumption and high demands are placed on the materials installed in the climate screen. In this thesis, a case study will be carried out on a building in Ljusne, Söderhamn, Sweden. Three different insulation materials and two types of windows will be studied to investigate their impact on the building´s energy consumption and to find the most optimal alternative from a climate neutral perspective.  The materials studied are mineral wool, phenolic foam board and thermal wood, as well as ordinary windows and passive windows. To answer this question, energy simulations are carried out using the computer program BV2 and calculations on the materials environmental product declarations (EPD) to analyze their climate impact. The climate impact of the building is calculated using the building sector´s environmental calculation tool (BM). The BM calculates the building´s emissions in carbon dioxide equivalents for renovation, energy use per year and over a life cycle of 50 years.  Energy simulations performed in BV2 show that additional insulation with phenolic foam and window replacement with passive windows is the most efficient method for renovating the external wall from an energy efficiency point of view. As this combination has the lowest energy use, primary energy figure and average heat transfer coefficient. It demonstrates a result with primary energy figure of 72,2 kWh/m2 and an energy use of 116,8 kWh/m2. But the combination of thermal wood and passive windows is the most effective on the climate impact of the building. This is illustrated in BM where the climate impact for renovation is 2928,9 kg CO2e, energy use gives 3980,7 kg CO2e/year and the climate impact for a life cycle of 50 years is then 201 964 CO2e/kg. The use of energy simulations allows efficient and climate-friendly materials to be studied more closely to determine their importance. For the climate goals in Sweden and EU to be achieved, the need for efficient materials is essential. This thesis is part of the journey towards a climate-neutral society.
536

Influence of oxygen addition on the phenolic composition of red wine

Geldenhuys, Lorraine 12 1900 (has links)
Thesis (MscAgric (Viticulture and Oenology))--University of Stellenbosch, 2009. / ENGLISH ABSTRACT: Tannins and colour components in red wine are important quality parameters. These factors can be manipulated in the vineyard by grape growing techniques or in the cellar by different winemaking practices. Grape seeds make a significant contribution to tannin concentration in wine when compared to those from the skins and pulp. Tannins contribute to the ageing potential, organoleptic properties and stabilisation of red wine colour. The colour of a red wine is also influenced by malolactic fermentation, the biological process that transforms malic acid into lactic acid which normally leads to an increase in pH. The subsequent change in pH alters the anthocyanin equilibrium, the primary colour components in red wine. Oxygen contributes to the polymerisation of anthocyanins over time to form more stable pigments that are less sensitive to pH fluctuations and sulphur dioxide bleaching. Limited research has been done on the use of oxygen after alcoholic fermentation and the impact it has on the phenolic composition of red wines. Similarly, only a few studies have examined the impact of either the addition or removal of seeds to the phenolic composition of a red wine in combination with oxygen addition. Additionally, little published data seems to exist on the effect of different pHs on red wine’s phenolic and colour development after oxygen addition. In our results we have shown that it is possible to stabilise wine colour by adding supplementary seeds before alcoholic fermentation. This led to an increase in colour intensity in certain cases. Some red wines produced without seeds had significantly lower colour intensities. This clearly suggests that catechins and proanthocyanidins are extracted from seeds and contribute to wine colour as they combine with other pigments to stabilise wine colour. Spectrophotometric and HPLC analyses have shown that the total phenolic content increased with seed concentration. However, we have observed that a wine may possibly become saturated with phenols when supplementary seeds are added. Anthocyanin concentrations often decreased when oxygen was added, while polymeric phenols and polymeric pigments sometimes increased. When applying different oxygen dosages to a red wine on commercial scale with microoxygenation, it was found that monomeric anthocyanins decreased as more oxygen was added and this decrease in anthocyanins led to the formation of stable polymeric pigments. This was reflected in the significant increase in colour intensity for the wines receiving oxygen. Small differences were detected in the total phenol and tannin concentration for the control and oxygenated wines. However, some of these phenolic and colour differences disappeared during subsequent ageing of the wine. When making wines of different initial pHs, we observed that the colour density decreased as the pH increased. The application of oxygen reduced the decrease in colour during MLF, especially at a lower pH. The addition of oxygen did not result in significantly different polymeric pigment concentrations in the various pH treatments, although the results could have been different if the wines were aged. However, pH differences in the range between 3.4 and 4.0 did not significantly influence the phenol composition of the wines under our conditions. This study led to a better understanding on the effect of oxygen additions under different conditions on red wine's phenolic and colour composition. / AFRIKAANSE OPSOMMING: Tanniene en kleur komponente teenwoordig in rooiwyn is belangrike kwaliteit parameters. Hierdie faktore wat wynkwaliteit beïnvloed kan gemanipuleer word in die wingerd deur verskeie verbouingstegnieke toe te pas en in die kelder deur die toepassing van verskillende wynbereidingsmetodes. Die bydrae van sade tot die finale tannienkonsentrasie in rooiwyn is groot in vergelyking met dié van die pulp en doppe. Tanniene dra by tot die verouderingspotensiaal, organoleptiese eienskappe en die stabilisasie van die wynkleur. Die kleur van ‘n rooi wyn word ook beïnvloed deur appelmelksuurgisting (AMG), die biologiese proses wat appelsuur omskakel na melksuur en ‘n gevolglike toename in die pH van die wyn veroorsaak. Hierdie verandering in die pH van die wyn beïnvloed die antosianien ewewig, die primêre kleur komponente teenwoordig in rooiwyn. Suurstof dra by tot die polimerisasie van antosianiene oor tyd om meer stabiele kleur pigmente te vorm met ‘n hoër kleurintensiteit wat minder sensitief is teenoor pH veranderinge en die bleikingseffek van swaweldioksied. Beperkte navorsing is gedoen op die gebruik van suurstof na alkoholiese gisting en die impak daarvan op die fenoliese samestelling van ‘n rooiwyn. Slegs ‘n paar studies het die invloed van die verwydering of byvoeging van sade in kombinasie met suurstoftoediening op ‘n rooiwyn se fenoliese samestelling ondersoek. Dit wil voorkom of beperkte gepubliseerde data beskikbaar is oor die effek wat verskillende pH’s het op rooi wyn se fenoliese en kleurontwikkeling na suurstof byvoeging. Ons resultate het aangedui dat dit wel moontlik is om in sekere gevalle die kleur van ‘n rooiwyn te stabiliseer deur addisionele sade by te voeg voor alkoholiese fermentasie. Hierdie byvoeging het ‘n toename in kleurintensiteit tot gevolg gehad. Sekere wyne wat gemaak is sonder sade het ‘n kenmerkend laer kleur intensiteit gehad. Hierdie bevinding is ‘n duidelike bewys dat katesjiene en prosianidiene geëkstraheer word vanuit die sade en bydra tot wynkleur deurdat hulle met ander pigmente verbind om die kleur sodoende te stabiliseer. Spektrofotometriese en hoë druk vloeistof chromatografie (HDVC) analises het gewys dat die totale fenoliese konsentrasie neem toe met ‘n toename in saad konsentrasie. Daar is egter waargeneem dat ‘n wyn moontlik versadig kan raak met fenole wanneer addisionele sade bygevoeg word. Antosianien konsentrasies het meestal afgeneem wanneer suurstof bygevoeg is, maar polimeriese fenole en polimeriese pigmente het partykeer toegeneem. Met die toediening van verskillende suurstof dosisse tot ‘n rooiwyn op kommersiële skaal het ons bevind dat monomeriese antosianiene afneem wanneer meer suurstof bygevoeg word. Hierdie afname in antosianiene het egter gelei tot die vorming van stabiele polimeriese pigmente. Dié bevinding was gereflekteer in die toename in kleurintensiteit van wyne wat met suurstof behandel is. Klein verskille was waargeneem vir die totale fenol en tannien konsentrasies tussen die kontrole en wyne behandel met suurstof. Sekere van hierdie fenoliese kleur verskille het egter afgeneem tydens die daaropvolgende veroudering van die wyne. Wyne wat gemaak is met verskillende aanvanklike pH’s se kleurintensiteit neem af soos die pH toeneem. Die toediening van suurstof het die kleurverlies tydens AMG verminder, veral by ‘n laer pH. Die toediening van suurstof het nie verskillende polimeriese pigment konsentrasies by verskillende pH’s veroorsaak nie, maar ‘n verskil kon moontlik waargeneem word indien die wyne verouder was. pH verskille tussen 3.4 en 4.0 het egter nie die fenoliese samestelling van die wyne onder ons omstandighede beduidend beïnvloed nie. Hierdie studie het gelei tot meer kennis oor die effek van suurstoftoedienings onder verskillende kondisies op rooiwyn se fenoliese en kleursamestelling.
537

Investigation of bacteriocins from lactic acid bacteria and their impact in winemaking

Knoll, Caroline 12 1900 (has links)
Thesis (MSc (Wine Biotechnology))--University of Stellenbosch, 2007. / Bacteriocins are ribosomally synthesized antimicrobial peptides produced by bacteria and are active against other bacteria, either in the same species (narrow spectrum) or across genera (broad spectrum). The application of bacteriocins during the vinification process might help to prevent the production of undesired compounds by inhibiting the indigenous bacterial microflora and allowing malolactic fermentation to be conducted by a selected bacterial strain. Furthermore, the use of bacteriocins might allow reducing the total sulphur dioxide amount in wine. The purpose of this study was the selection of lactic acid bacteria (LAB) belonging to the genera Oenococcus, Lactobacillus and Pediococcus with the ability to produce bacteriocins, with respective biological activity against undesired indigenous wine LAB and the capability to complete malolactic fermentation. The first objective of this study was the screening of LAB isolated from South African red wines for the production of bacteriocins. Only 27 strains out of 330 wine isolates, belonging to the species Lb. plantarum, Lb. paracasei, Lb. hilgardii and O. oeni, showed activity towards various wine-related and non wine-related indicator strains with the colony-overlay method. It is the first time that bacteriocin activity is reported in O. oeni. The second objective was the detection and identification of known structural bacteriocin genes of Lb. plantarum wine strains. Furthermore, the web server BAGEL was used to in silico analyse putative bacteriocin-encoding genes in the genome of O. oeni and primers were designed to amplify four possible bacteriocin-encoding genes. A PCR-based screening revealed the presence of the plantaricin encoding genes plnA, plnEF, plnJ and plnK in five selected Lb. plantarum strains. Moreover, PCR analysis rendered positive results with all four chosen putative bacteriocin-encoding genes in the eight tested O. oeni strains with antimicrobial activity. The latter genes of O. oeni were heterologously expressed in different Escherichia coli host strains, but no antimicrobial activity could be detected. The third objective of this study was the transformation and expression of the heterologous bacteriocin genes nisin A and pediocin PA-1 in two selected Lb. plantarum strains. To enhance their antimicrobial activity a plasmid containing the nisin A gene was successfully cloned into the two strains. Indeed, an enhanced antimicrobial activity could be detected, but the transformed plasmid was not stable. The fourth objective in this project was the evaluation of bacteriocin production in liquid media. A co-culture experiment with a plantaricin producing Lb. plantarum strain and an Enterococcus faecalis strain as indicator was performed. A complete inhibition of cell growth of Ent. faecalis was observed within 72 hours. The last objective was the evaluation of the impacts of phenolic compounds on the activity of nisin and pediocin. The short term influence of two phenolic acids, two flavan-3-ols, grape tannins and oak tannins on the activity of nisin and pediocin PA-1 was investigated. No influence on the activity was detected. Furthermore, synergistic effects on bacterial growth inhibition were observed. This study confirms the potential use of either bacteriocin additives or bacteriocin-producing LAB in order to control the bacterial microflora during the vinification process.
538

The influence of different winemaking techniques on the extraction of grape tannins

Nel, Anton Pieter 03 1900 (has links)
Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosch, 2011. / Includes bibliography. / ENGLISH ABSTRACT: Grape and wine phenols consist of flavanols which is the building blocks for tannins. These building blocks are called monomers which consist of catechins, epicatechins, epigallocatechins and epicatechin-gallate. Tannin is important in wine as it contributes to bitterness, mouth feel (astringency) and maturation potential of the wine. Futhermore it has a health benefit as an antioxidant. Anthocyanins are responsible for the colour of red wine. The anthocyanins combine with tannins to form stable polymeric pigments. Due to the importance of tannins and anthocyanins in wine, it is imperitative that different winemaking techniques are used to extract as much of these components as possible and that the analysis is done quickly and accurately. The aim of this study was to evaluate different winemaking techniques and their extraction of tannins and anthocyanins into the wine. Too much tannin extraction can have a negative effect on the sensory quality of the wine. Therefore a second aim was to evaluate the mouth feel properties of a Shiraz wine. A third aim was to compare the two tannin precipitation methods in terms of time efficiency, repeatability and the ease of practice. To investigate the amount of tannin concentration extracted by different winemaking techniques, two cultivars (Cabernet Sauvignon and Shiraz) were used. These treatments included the addition of an enzyme during fermentation [E], cold maceration [CM], post maceration [PM] and the combination of cold and post maceration [CM+PM]. The grapes were harvested in two different climatic areas during the 2008 and 2009 vintages. The two climatic areas were classified according to the Winkler scale as a III (Morgenster) and a IV (Plaisir de Merle). The grapes were harvested at two different ripeness levels in order to evaluate the effect of the different winemaking processes on the extraction of tannins and anthocyanins. One harvest was before (LB) and the other after (HB) the commercial harvest. The results of this study showed significant differences in the phenolic composition of the wines. It was found that the warmer area showed higher tannin concentrations than the cooler area for both cultivars. In the 2008 Cabernet Sauvignon the CM extracted higher concentrations of tannin from the cooler area at both ripeness levels. In the warmer area, CM extracted the highest tannin concentration HB, but the CM+PM extracted the highest tannin concentration from Cabernet Sauvignon at the LB and CM at the HB of the warmer area. In 2009 the PM extracted the highest concentration of tannin at the lower ripeness level, while the E treatment extracted the highest concentration from the warmer area. In the cooler area the CM+PM extracted the highest concentration of tannin at a lower ripeness level, while there were no siginicant differences between the different treatments at the higher ripeness level. The highest anthocyanin concentration was found in the cooler area. The CM treatment was found to have no effect on anthocyanin extraction. Different methods are available to quantify the tannin concentration in wine. Two of the most popular tannin analytical methods are the bovine serum albumin (BSA) and the methyl cellulose precipitable tannin (MCP) methods. The BSA method is a very complex method which uses at least 3 times more reagents than the MCP method. The MCP method only analyzes tannins, while the BSA method analyzes tannins, monomeric pigments (MP), small polymeric pigments (SPP) and large polymeric pigments (LPP). In this study a good correlation was found between the two tannin precipitation methods (R2 – 0.88). There is controversy regarding the variability of these methods. Some scientists found that the two methods show a good correlation with HPLC, while others found that there was no such correlation between the precipitation methods and the HPLC. The MCP method had a practical advantage as it could be performed in half the time required for the BSA method. This has a significant impact in scenarios where a high sample throughput is required although it only measures total tannin. The phenolic composition and mouth feel of the wine was strongly influenced by the climatic area. In the warmer area the effect of tannin concentration on mouth feel was much less than in the cooler area. The wine made of riper grapes, was more grippy, bitter and numbing than the wines made from greener grapes. The E treatment was especially associated with a dry, grippy sensation. / AFRIKAANSE OPSOMMING: Druif en wyn fenole bestaan uit flavanole wat weer die boublokke is van tanniene. Hierdie boublokke, wat bekend staan as monomere, betsaan uit katesjiene, epikatesjiene, epigallokatesjiene an epikatesjien-gallaat. Tanniene is belangrik in wyn aangesien dit bydra tot bitterheid, mondgevoel (vrankheid) asook die verouderingspotensiaal van wyn. As antioksidante hou dit ook gesondheidsvoordele in. Antosianiene dra by tot die kleur van rooiwyn. Antosianiene kombineer met tanniene om meer stabiele polimeriese pigmente te vorm. As gevolg van die belangrikheid van tanniene en antosianiene is dit van uiterse belang dat verskillende wynmaak tegnieke gebruik word om ekstraksie in die wyn te bevoordeel en dat die analitiese metode so vinnig en akkuraat as moontlik gedoen word. Die eerste doel van hierdie studie was om die ekstraksie van tanniene en antosianiene deur middel van verskillende wynmaak tegnieke te evalueer. Te veel tanniene in die wyn kan negatiewe sensoriese kwaliteit tot gevolg het. Daarom is die tweede doel om die sensoriese kwaliteit van Shiraz wyn te evalueer. Die derde doel van hierdie studie was die twee tannien presipitasie metodes met mekaar te vergelyk in terme van die moeilikheidsgraad van die metode, tyd doeltreffendheid en herhaalbaarheid. Verskillende wynmaak tegnieke (ensiem byvoegings [E], koue maserasie [CM], verlengde dopkontak [PM] en ‘n kombinasie van koue maserasie en verlengde dopkontak [CM+PM]) is vergelyk ten opsigte van tannien en antiosianien ekstraksie. In 2008 en 2009 is twee kultivars (Cabernet Sauvignon en Shiraz) in twee verskillende klimatologiese areas gepars. Hierdie areas is geklassifiseer in die Winklerskaal as ‘n IV (Plaisir de Merle) en ‘n III (Morgenster). Om die effek van die verskillende wynmaak tegnieke op die ekstraksie van antosianiene en tanniene te vergelyk, is hierdie twee kultivars by twee verskillende rypheidsgrade geoes. Die eerste oes was net voor kommersiële oes (LB) en die tweede oes het net na kommersiële oes (HB) plaasgevind. Die 2009 Shiraz wyn is organolepties beoordeel om die effek van die verskillende wynmaak tegnieke op die wyn se mondgevoel te vergelyk. Die resultate van hierdie studie toon beduidende verskille in die fenoliese samestelling van die wyne. Dit is gevind dat die warmer area hoër tannien konsentrasies het as die koeler area. In 2008 het die CM+PM die meeste tanniene uit die Cabernet Sauvignon geëkstraheer by LB en die CM by HB in die warmer area. Die CM het in die koeler area meer tanniene geëkstraheer by beide die LB en HB rypheidsgrade. In 2009 het PM die meeste tanniene geëkstraheer by LB terwyl E die meeste tanniene geëkstraheer in die warmer area. In die koeler area het CM+PM die meeste tanniene geëkstraheer, terwyl geen van die behandelings ‘n effek gehad het by HB. Die meeste antosianien konsentrasie was in die koeler area gevind as in die warmer area. In beide 2008 (LB en HB) en 2009 (LB) het CM die meeste antosianiene geëkstraheer, terwyl geen behandeling ‘n effek gehad het by HB. Twee van die mees populêre tannien analitiese metodes is die BSA (bovine serum albumien) en die MCP (metielsellulose presipitasie) metodes. Die BSA metode is ‘n baie meer ingewikkelde metode waarvoor drie keer meer reagense gebruik word as vir die MCP metode. Maar waar die MCP net tanniene ontleed, ontleed die BSA metode tanniene, monomere (MP), klein polimeriese pigmente (SPP) en groot polimeriese pigmente (LPP). Dit help indien daar gekyk wil word na die evolusie van polimeriese pigmente. In hierdie studie is bevind dat daar ‘n redelike korrelasie (R2 – 0.88) tussen die BSA en MCP metode bestaan. Die herhaalbaarheid van die metodes het redelike kontroversie veroorsaak, waar sommige navorsers bevind het dat die BSA metode nie so herhaalbaar is soos eers bevind is nie. Die MCP metode het ’n praktiese voordeel aangesien dit in die helfde van die tyd van die BSA metode uitgevoer kan word. Dit het ‘n groot impak indien ‘n groot hoeveelheid monsters ontleed moet word. Die fenoliese samestelling en mondgevoel word sterk beïnvloed deur die klimatologiese area. In die warmer area was die effek van tannien konsentrasie op mondgevoel kleiner as in die koeler area. Die wyn van ryper druiwe het meer harder, verdowingseffek en bitter nasmaak gehad as by die wyn van groener druiwe. Die ensiem behandeling was meer geassossieerd met droë mond gevoel.
539

The long-term effects of Sargassum muticum (Yendo) Fensholt invasion on Zostera marina L. and its associated epibiota

DeAmicis, Stacey Lynn January 2012 (has links)
In this thesis I review how Sargassum muticum (Yendo) Fensholt, an invasive alga from Asia, has spread globally due to human activities and describe how this species can affect seagrass ecosystems. Abiotic factors such as nutrient and substratum availability may facilitate the spread of S. muticum into Zostera marina L. meadows, but analyses of seawater nutrients, and sediment particle size and % organic content revealed no significant differences between experimental quadrats in seagrass meadows either with, or without the presence of S. muticum. Phenolic compounds were examined because they form the basis of defensive mechanisms in plants and algae, therefore any change in phenolic content may affect the ability of Z. marina to protect itself from disease, herbivory and invasive species through allelopathic interactions. Results from a four year field study and multiple annual laboratory experiments showed significant reductions (p = 0.034 and p = 0.002, respectively) in the caffeic and tannic acids equivalents content of Z. marina when in the presence of S. muticum. As the abundance of S. muticum increases, other changes in the physiology of Z. marina may occur including variations in growth rates, nutrient partitioning and chlorophyll fluorescence, but data from multiple laboratory experiments illustrated no significant differences in growth. Chlorophyll fluorescence analyses revealed significant differences between treatments with and without S. muticum (p = 0.008), but pairwise comparisons indicated these differences only occurred in 2008 (p < 0.001). Significant differences were also found in nutrient partitioning amongst functional regions of the shoots (p = 0.024), but pairwise comparisons detected these differences between a biomass control treatment (ZZ: Zostera + Zostera) and the ZS (with S. muticum) and ZM (Z. marina on its own at a lower biomass per replicate) treatments (p = 0.013 and p = 0.019, respectively), but not between ZS and ZM. Previous in situ research has found negative effects of S. muticum presence on densities of kelp and other algae. Results from the long-term field study indicated significantly lower mean in situ Z. marina densities within the ZS treatment (p < 0.001). Epibiota found living on the blades of Z. marina provide food for organisms within seagrass ecosystems and also create microhabitats for other species to occupy. Alterations in the abundances of epibiota and microhabitats formed could further modify seagrass ecosystems through shifts in timing of food availability, food preferences and microhabitats created. The long-term field study data revealed significantly lower epibiota abundances within the ZS treatment (p = 0.019), but differences in biomass between treatments were not detected. Changes in the biochemistry, physiology, vegetative physiognomy and epibiota assemblages of Z. marina revealed during experimental manipulations are presented and considered within the context of long-term seagrass survival in light of increasing S. muticum invasion.
540

Production et purification d'acide férulique estérases. Application à la synthèse d'esters phénoliques / Production and purification of ferulic acid esterases. Application to the synthesis of phenolic esters

Kheder, Fadi 25 October 2007 (has links)
L’induction de la synthèse d’une acide férulique estérase (AFE) a été étudiée chez Streptomyces ambofaciens ATCC 23877. L’activité la plus élevée a été détectée en présence de son de blé désamidonné ou de xylane d’avoine (0,22, 0,21 mU/mg protéine, respectivement). Des productions d’AFE en bioréacteur ont également été réalisées en utilisant 1% (p/v) de son de blé comme inducteur. Le niveau de production de l’AFE a été trois fois plus important en bioréacteur qu’en fiole d’Erlenmeyer. L’AFE de Streptomyces ambofaciens ATCC 23877 et celle de Humicola sp., présente dans un mélange enzymatique commercial (DepolTM 740L), ont été partiellement purifiées et caractérisées. A l’issue de la purification, l’activité AFE de Streptomyces ambofaciens ATCC 23877 a été trop faible pour pouvoir être utilisée ultérieurement en synthèse. Par contre, le potentiel de l’AFE de Humicola sp., concentrée par précipitation à l’acétone, pour la synthèse de différents esters phénoliques a été testé. Les meilleurs rendements de conversion ont été observés lors de l’absence de substitutions sur le cycle aromatique de l’acide phénolique ou en présence de groupements hydroxyles. Les synthèses en milieu non aqueux (M2B2) se sont montrées infructueuses en raison, peut-être, d’un effet néfaste du solvant sur l’enzyme / The induction of the ferulic acid esterase (FAE) synthesis was studied with Streptomyces ambofaciens ATCC 23877. The highest activity was detected in the presence of either destarched wheat bran or oat spelt xylan (0,22, 0,21 mU/mg protein, respectively). FAE productions in bioreactor were also carried out using 1% (w/v) of wheat bran as inducer. The FAE production level was three times higher in bioreactor than in Erlenmeyer flask. FAE of Streptomyces ambofaciens ATCC 23877 and that of Humicola sp., present in an enzymatic commercial mixture (DepolTM 740L), were partially purified and characterised. At the end of the purification, FAE activity of Streptomyces ambofaciens ATCC 23877 was too weak to be used later in synthesis. However, the FAE potential of Humicola sp., concentrated by acetone precipitation, for the synthesis of various phenolic esters was tested. The best conversion yields were observed in the absence of substitution on the phenolic acid aromatic cycle or in the presence of hydroxyl groups. The synthesis in non-aqueous medium (M2B2) were unsuccessful maybe because of an harmful effect of the solvent on the enzyme

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