O presente trabalho avaliou o crescimento da microalga Dunaliella tertiolecta pela biofixação do CO2 liberado pela produção de cerveja, reciclando um dos gases responsáveis pelo efeito estufa, reduzindo custo da matéria-prima CO2 e agregando valor ao produzir lipídeos e carotenoides naturais. Para isso a microalga foi cultivada em sistemas integrados entre fotobiorreatores e fermentadores. A diferença nos cultivos foi o tipo e a quantidade de CO2 produzida pelas fermentações. Inicialmente se fez fermentações com meio YPD (Yeast Peptone Dextrose) em fermentadores de 2 L acoplados a cada 24 h aos fotobiorreatores em 4 condições distintas, sendo o último fermentador colocado sempre em 144 h de cultivo de microalgas: 30 g L-1 de dextrose a partir de 72 h de cultivo de microalgas, 60 g L-1 de dextrose a partir de 72 h de cultivo de microalgas, 30 g L-1 de dextrose a partir de 24 h de cultivo de microalgas e variando a concentração de (10 à 60) g L-1 de dextrose a partir de 24 h de cultivo de microalgas (YPD (10-60)/24). Os maiores valores para biomassa, carotenoides, produtividades e lipídeos foram obtidos na condição YPD (10-60)/24. Para reproduzir a essa condição utilizando mosto de cerveja, foi calculada a conversão de substrato em produto para, então, acoplar diariamente volumes diferentes de mosto de cerveja em cultivos de microalgas. Os valores obtidos para os cultivos com CO2 desprendidos por estas fermentações foram 1,10 ± 0,05 g L-1 de biomassa, 0,18 ± 0,01 g L-1 d-1 de produtividade de biomassa, 0,58 ± 0,06 d-1 foi a velocidade específica de crescimento, 4,74 ± 0,59 mg g-1 de carotenoides por biomassa, 0,86 ± 0,06 mg L-1 d-1 de produtividade de carotenoides e 13,5 ± 0,4 % (em massa) de lipídeos. Estes valores foram praticamente o dobro dos valores obtidos para o cultivo com CO2 do ar atmosférico, demonstrando que a integração entre fermentadores e fotobiorreatores é uma boa alternativa para indústria alimentícia. Todos cultivos com D. tertiolecta apresentaram o mesmo perfil de carotenoides representado por 46,7 ± 2,0 % de luteína, 22,5 ± 1,6 % de β-caroteno, 9,50 ± 0,66 % de zeaxantina, 1,10 ± 0,16 % de α-caroteno e 20,2 ± 3,0 % para outros. / This study evaluated the growth of microalgae Dunaliella tertiolecta for CO2 biofixation released by brewing, recycling one of the greenhouse gases, reducing cost of raw material CO2 and adding value to produce lipids and natural carotenoids. For this, microalgae were cultivated in integrated systems between photobioreactors and fermenters. The difference in the cultures was the culture medium and the amount of CO2 produced. Initially, fermentation with medium YPD (Yeast Peptone Dextrose) in 2 L fermenters were coupled every 24 h to photobioreactors in 4 different conditions: 30 g L-1 of dextrose from 72 h culture of microalgae; 60 g L-1 of dextrose from 72 h culture of microalgae; 30 g L-1 of dextrose from 24 h culture of microalgae; and ranging dextrose concentration of (10 to 60) g L-1 from 24 h culture of microalgae (YPD (10-60)/24). The highest values for biomass, carotenoids, productivities and lipids were obtained in the condition YPD (10-60)/24. To reproduce this condition using beer wort, the substrate to product yield was determined and different volumes of beer wort where daily coupled to microalgae cultivations. The values obtained for cultures with CO2 released from these fermentations were 1.10 ± 0.05 g L-1 of biomass, 0.18 ± 0.01 g L-1 d-1 of biomass productivity, 0.58 ± 0.06 d-1 for the specific growth rate, 4.74 ± 0,59 mg g-1 of carotenoids per biomass, 0.86 ± 0.06 mg L-1 d-1 of carotenoids productivity and 13.5 ± 0.4 % (mass fraction) of lipids. These values were almost twice the values observed in the cultivation with CO2 of atmospheric air, showing that the integration between fermenters and photobioreactors is a good alternative to increase microalgae growth. All cultures with D. tertiolecta showed the same profile of carotenoids represented by 46.7 ± 2.0 % of lutein, 22.5 ± 1.6 % of β-carotene, 9.50 ± 0.66 % of zeaxanthin, 1.10 ± 0.16 % of α-carotene and 20.2 ± 3.0 % for others.

O presente trabalho avaliou o crescimento da microalga Dunaliella tertiolecta pela biofixação do CO2 liberado pela produção de cerveja, reciclando um dos gases responsáveis pelo efeito estufa, reduzindo custo da matéria-prima CO2 e agregando valor ao produzir lipídeos e carotenoides naturais. Para isso a microalga foi cultivada em sistemas integrados entre fotobiorreatores e fermentadores. A diferença nos cultivos foi o tipo e a quantidade de CO2 produzida pelas fermentações. Inicialmente se fez fermentações com meio YPD (Yeast Peptone Dextrose) em fermentadores de 2 L acoplados a cada 24 h aos fotobiorreatores em 4 condições distintas, sendo o último fermentador colocado sempre em 144 h de cultivo de microalgas: 30 g L-1 de dextrose a partir de 72 h de cultivo de microalgas, 60 g L-1 de dextrose a partir de 72 h de cultivo de microalgas, 30 g L-1 de dextrose a partir de 24 h de cultivo de microalgas e variando a concentração de (10 à 60) g L-1 de dextrose a partir de 24 h de cultivo de microalgas (YPD (10-60)/24). Os maiores valores para biomassa, carotenoides, produtividades e lipídeos foram obtidos na condição YPD (10-60)/24. Para reproduzir a essa condição utilizando mosto de cerveja, foi calculada a conversão de substrato em produto para, então, acoplar diariamente volumes diferentes de mosto de cerveja em cultivos de microalgas. Os valores obtidos para os cultivos com CO2 desprendidos por estas fermentações foram 1,10 ± 0,05 g L-1 de biomassa, 0,18 ± 0,01 g L-1 d-1 de produtividade de biomassa, 0,58 ± 0,06 d-1 foi a velocidade específica de crescimento, 4,74 ± 0,59 mg g-1 de carotenoides por biomassa, 0,86 ± 0,06 mg L-1 d-1 de produtividade de carotenoides e 13,5 ± 0,4 % (em massa) de lipídeos. Estes valores foram praticamente o dobro dos valores obtidos para o cultivo com CO2 do ar atmosférico, demonstrando que a integração entre fermentadores e fotobiorreatores é uma boa alternativa para indústria alimentícia. Todos cultivos com D. tertiolecta apresentaram o mesmo perfil de carotenoides representado por 46,7 ± 2,0 % de luteína, 22,5 ± 1,6 % de β-caroteno, 9,50 ± 0,66 % de zeaxantina, 1,10 ± 0,16 % de α-caroteno e 20,2 ± 3,0 % para outros. / This study evaluated the growth of microalgae Dunaliella tertiolecta for CO2 biofixation released by brewing, recycling one of the greenhouse gases, reducing cost of raw material CO2 and adding value to produce lipids and natural carotenoids. For this, microalgae were cultivated in integrated systems between photobioreactors and fermenters. The difference in the cultures was the culture medium and the amount of CO2 produced. Initially, fermentation with medium YPD (Yeast Peptone Dextrose) in 2 L fermenters were coupled every 24 h to photobioreactors in 4 different conditions: 30 g L-1 of dextrose from 72 h culture of microalgae; 60 g L-1 of dextrose from 72 h culture of microalgae; 30 g L-1 of dextrose from 24 h culture of microalgae; and ranging dextrose concentration of (10 to 60) g L-1 from 24 h culture of microalgae (YPD (10-60)/24). The highest values for biomass, carotenoids, productivities and lipids were obtained in the condition YPD (10-60)/24. To reproduce this condition using beer wort, the substrate to product yield was determined and different volumes of beer wort where daily coupled to microalgae cultivations. The values obtained for cultures with CO2 released from these fermentations were 1.10 ± 0.05 g L-1 of biomass, 0.18 ± 0.01 g L-1 d-1 of biomass productivity, 0.58 ± 0.06 d-1 for the specific growth rate, 4.74 ± 0,59 mg g-1 of carotenoids per biomass, 0.86 ± 0.06 mg L-1 d-1 of carotenoids productivity and 13.5 ± 0.4 % (mass fraction) of lipids. These values were almost twice the values observed in the cultivation with CO2 of atmospheric air, showing that the integration between fermenters and photobioreactors is a good alternative to increase microalgae growth. All cultures with D. tertiolecta showed the same profile of carotenoids represented by 46.7 ± 2.0 % of lutein, 22.5 ± 1.6 % of β-carotene, 9.50 ± 0.66 % of zeaxanthin, 1.10 ± 0.16 % of α-carotene and 20.2 ± 3.0 % for others.

O presente trabalho avaliou o crescimento da microalga Dunaliella tertiolecta pela biofixação do CO2 liberado pela produção de cerveja, reciclando um dos gases responsáveis pelo efeito estufa, reduzindo custo da matéria-prima CO2 e agregando valor ao produzir lipídeos e carotenoides naturais. Para isso a microalga foi cultivada em sistemas integrados entre fotobiorreatores e fermentadores. A diferença nos cultivos foi o tipo e a quantidade de CO2 produzida pelas fermentações. Inicialmente se fez fermentações com meio YPD (Yeast Peptone Dextrose) em fermentadores de 2 L acoplados a cada 24 h aos fotobiorreatores em 4 condições distintas, sendo o último fermentador colocado sempre em 144 h de cultivo de microalgas: 30 g L-1 de dextrose a partir de 72 h de cultivo de microalgas, 60 g L-1 de dextrose a partir de 72 h de cultivo de microalgas, 30 g L-1 de dextrose a partir de 24 h de cultivo de microalgas e variando a concentração de (10 à 60) g L-1 de dextrose a partir de 24 h de cultivo de microalgas (YPD (10-60)/24). Os maiores valores para biomassa, carotenoides, produtividades e lipídeos foram obtidos na condição YPD (10-60)/24. Para reproduzir a essa condição utilizando mosto de cerveja, foi calculada a conversão de substrato em produto para, então, acoplar diariamente volumes diferentes de mosto de cerveja em cultivos de microalgas. Os valores obtidos para os cultivos com CO2 desprendidos por estas fermentações foram 1,10 ± 0,05 g L-1 de biomassa, 0,18 ± 0,01 g L-1 d-1 de produtividade de biomassa, 0,58 ± 0,06 d-1 foi a velocidade específica de crescimento, 4,74 ± 0,59 mg g-1 de carotenoides por biomassa, 0,86 ± 0,06 mg L-1 d-1 de produtividade de carotenoides e 13,5 ± 0,4 % (em massa) de lipídeos. Estes valores foram praticamente o dobro dos valores obtidos para o cultivo com CO2 do ar atmosférico, demonstrando que a integração entre fermentadores e fotobiorreatores é uma boa alternativa para indústria alimentícia. Todos cultivos com D. tertiolecta apresentaram o mesmo perfil de carotenoides representado por 46,7 ± 2,0 % de luteína, 22,5 ± 1,6 % de β-caroteno, 9,50 ± 0,66 % de zeaxantina, 1,10 ± 0,16 % de α-caroteno e 20,2 ± 3,0 % para outros. / This study evaluated the growth of microalgae Dunaliella tertiolecta for CO2 biofixation released by brewing, recycling one of the greenhouse gases, reducing cost of raw material CO2 and adding value to produce lipids and natural carotenoids. For this, microalgae were cultivated in integrated systems between photobioreactors and fermenters. The difference in the cultures was the culture medium and the amount of CO2 produced. Initially, fermentation with medium YPD (Yeast Peptone Dextrose) in 2 L fermenters were coupled every 24 h to photobioreactors in 4 different conditions: 30 g L-1 of dextrose from 72 h culture of microalgae; 60 g L-1 of dextrose from 72 h culture of microalgae; 30 g L-1 of dextrose from 24 h culture of microalgae; and ranging dextrose concentration of (10 to 60) g L-1 from 24 h culture of microalgae (YPD (10-60)/24). The highest values for biomass, carotenoids, productivities and lipids were obtained in the condition YPD (10-60)/24. To reproduce this condition using beer wort, the substrate to product yield was determined and different volumes of beer wort where daily coupled to microalgae cultivations. The values obtained for cultures with CO2 released from these fermentations were 1.10 ± 0.05 g L-1 of biomass, 0.18 ± 0.01 g L-1 d-1 of biomass productivity, 0.58 ± 0.06 d-1 for the specific growth rate, 4.74 ± 0,59 mg g-1 of carotenoids per biomass, 0.86 ± 0.06 mg L-1 d-1 of carotenoids productivity and 13.5 ± 0.4 % (mass fraction) of lipids. These values were almost twice the values observed in the cultivation with CO2 of atmospheric air, showing that the integration between fermenters and photobioreactors is a good alternative to increase microalgae growth. All cultures with D. tertiolecta showed the same profile of carotenoids represented by 46.7 ± 2.0 % of lutein, 22.5 ± 1.6 % of β-carotene, 9.50 ± 0.66 % of zeaxanthin, 1.10 ± 0.16 % of α-carotene and 20.2 ± 3.0 % for others.

This diploma thesis deals with the natural antioxidants present in cereals, especially in barley (Hordeum vulgare). A close attention is paid to the study of carotenoids determination was conducted. In the experimental part, the method of beta-carotene determination was optimized using high-performance liquid chromatography, diode array detector and mass detector (HPLC/DAD/APCI-MS). The method was used for the beta-carotene and lutein determination in the samples of barleycorn, malt and green barley. This method involves the sample saponification, extraction by diethylether, followed by separation on ODS Hypersil 250x4,6 mm, 5m column, using MTBE/MeOH (20:80) as mobile phase and spectrophotometric detection (450 nm). Quantitative analysis was implemented in the HPLC/DAD system. The MS detector was used for identification of analytes. A number of still unpublished data about the content of beta-carotene and lutein in several varieties of malting barley, malt and green barley are stated in this thesis. The highest content of beta-carotene was found in the green barley sample of variety Malz, harvested in first grow phase (8,49 mg/kg of the dry matter). The content of beta-carotene in barleycorn is relatively low (0,07-0,14 mg/kg of the dry matter). The content of beta-carotene is several times higher in the malt produced from barleycorn (0,24-0,56 mg/kg of the dry matter). The diploma thesis was implemented in the Research Institute of Brewing and Malting, Plc. in Brno.

Potato cultivars exhibit a wide variation in skin and flesh colour due to the presence of pigments. This study established that potato cultivars differ greatly with respect to types and concentrations of carotenoids in tubers. A total of 46 cultivars were evaluated for quantitative and qualitative carotenoid composition in different growing seasons, locations, storage conditions and disease symptoms. Factors controlling carotenoid accumulation were also tested by developing an in vitro minituber system as a new high-throughput model system for carotenogenesis in potato tubers. Tuber flesh colour was found to correlate with total carotenoid content in potato cultivars grown in both New Zealand and Netherlands. The main carotenoids identified in 32 potato cultivars in New Zealand were lutein, neoxanthin, violaxanthin and β-carotene. The ratio of these carotenoids varies between cultivars. Neoxanthin was detected in only 13 cultivars (10.59 to 69.21µg/g DW); violaxanthin was found only in 1 cultivar (32.76 µg/g DW). Whereas lutein and β-carotene were found in most of the cultivars but the concentration varied from (0.00 to 160.63 µg/g DW) and (0.00 to 13.62 µg/g DW) respectively. The main carotenoids identified in 12 cultivars grown in the Netherlands were neoxanthin, violaxanthin and lutein, whereas zeaxanthin was not found in any of the cultivars analysed. Marked differences were observed between the same potato cultivars grown in New Zealand and the Netherlands. Therefore cultivars were analysed over a second growing season to assess stability in carotenoids composition. The carotenoid profiles of the potato tubers grown for two different seasons showed highly significant differences between the cultivars, the seasons, the carotenoid pigments, and all combinations of interactions, indicating the complex nature of factors influencing carotenoid composition. Reflectance colorimeter measurement of yellow hue component in this study confirmed that the higher the total carotenoid content, the greater the yellow intensity colour. Eight cultivars were grown at three locations in New Zealand and Agria and Desiree were grown at eight locations in the Netherlands to further investigate the stability of carotenoid composition. Highly significant differences were observed between the cultivars, the locations, the carotenoid pigments, and all combinations of interactions, which emphasises that changes in carotenoid composition are complex and the responses are not consistent across cultivars. Reflectance colorimeter measurement of yellow hue component confirmed the relationship between the yellow colour intensity of tuber flesh, as well as confirming the interaction between colour and locations. Disease and post harvest storage conditions markedly influenced the levels of total carotenoid, neoxanthin, violaxanthin, zeaxanthin, lutein and β-carotene in potatoes. The magnitude of these effects depends on the cultivar, time of storage, and the intensity of powdery scab symptoms. Results showed that long term storage resulted in the accumulation of neoxanthin, violaxanthin and zeaxanthin with a concomitant decreased of lutein, β-carotene and total carotenoid content. Genotypes infected with disease (lower and higher scab score) resulted in accumulation of violaxanthin, β-carotene and total carotenoid with a concomitant decreased in neoxanthin and lutein. A high-throughput model system for investigating carotenoid biogenesis in potato tubers was developed. This involved in vitro potato minitubers and was validated by assessing the effects of environmental variables, such as drought stress, light intensity and nutrient availability on carotenoid accumulation. Light influenced the presence of zeaxanthin, whereas water stress and nutrient strength influenced the accumulation of neoxanthin and violaxanthin. Although these factors had an effect on the carotenoid content and profile, the most influential factor appeared to be cultivar selection.

Microscopic green algae produce a number of beneficial substances. The commercially used ones include mainly pigments, especially secondary carotenoids. While primary carotenoids are part of the photosynthetic apparatus, secondary carotenoids are produced only under certain specific conditions (e.g. high exposure to light, nitrogen deficiency). Secondary carotenoids have antioxidant properties that protect the cell from adverse effects. The strains currently in commercial use and the strains that are tested for potential use come almost exclusively from algal collections, so this thesis focuses on strains isolated from nature. Two strains of aeroterrestrial green unicellular algae, which showed potential of producing carotenoids, were found and isolated. These strains were identified as Tetracystis pulchra (clade Dunaliellinia) and Tetracystis sp., a strain related to Tetracystis tetraspora SAG 98.80, belonging to the clade Stephanosphaerinia. The culture conditions for T. pulchra were subsequently optimized and then the culture was subjected to a series of experiments, examining the effect of stress factors on carotenoid formation and accumulation. Specifically, the effect of nitrogen starvation, light intensity, temperature and UVA radiation was tested. It has been shown that especially the light...

This diploma thesis deals with the optimization of processes for extraction of bioactive lipophilic compounds from fruits of sea buckthorn (Hippophae Rhamnoides) into various types of plant oil bases using simple maceration. The theoretical part of this thesis deals with the characterization of this herb, its botanical classification, traditional use, chemical composition and medicinal effects. Increased attention is also paid to the characterization of individual types of plant oils used, namely camellia, camellia organic, passionflower, kukui and kiwi oil. The experimental part of the thesis deals with application of theoretical knowledge. The profile of total and free fatty acids for individual plant oil bases was determined by GC/FID, furthet the basic fat numbers were also determined. When optimizing the extraction, emphasis was placed not only on the effect of the extraction agent used, but also on the extraction time (1, 3, 5, 7, 10, 14, 21 and 66 days). The macerates were continuously subjected to the determination of selected parameters (total amount of carotenoids, total amount of phytosterols, lutein, neoxanthin, astaxanthin, stigmasterol, -sitosterol and vitamin E) using UV-VIS spectroscopy and HPLC/PDA. Likewise, the peroxide number was monitored during maceration to assess the degree of oxidative degradation of macerates. The recovery of selected total parameters in individual oils did not differ significantly in most cases. On the contrary, the yield of individual monitored parameters differed significantly. At the same time, static maceration under our conditions was not very suitable for the extraction of vitamin E, stigmasterol and total phytosterols.