39 research outputs found
Synthesis and studies of novel difuranone heterocycles for optoelectronic applications
Organická elektronika je progresivní vědní oblast kombinující znalosti mnoha oborů zabývající se studiem a aplikačními možnostmi organických molekul jak z oblasti nízkomolekulárního segmentu, tak oligomerů a polymerů. Funkční organické materiály jsou charakteristické svými vynikajícími vlastnostmi a jejich strukturální variabilitou. V této práci jsou rozebrány vlastnosti, aplikace a historie vývoje benzodifuranonů a diketopyrrolopyrrolů jakožto zástupců „chytrých“ molekul. Dále, jsou zde popsány jednotlivé syntetické přístupy k přípravě symetrických a nesymetrických benzodifuranonů. Experimentální část bakalářské práce se zabývá přípravou výchozího intermediátu hydroxy(thiofen-2-yl)octové kyseliny pro přípravu analogů benzodifuranonů obsahujících thiofenové aromatické jádro.Organic electronics is a progressive field of science that combines knowledge of many disciplines concerned with the study and application possibilities of organic molecules from both, the low-molecular segment as well as the oligomers and polymers. Such molecules are well recognizable by their excellent properties and variability of their structure. This thesis deals with properties, applications and brief research review of benzodifuranones and diketopyrrolopyrroles as the representatives of smart molecules. Furthermore various synthetic approaches to the preparation of both symmetrical and unsymmetrical benzodifuranones are described. Experimental part is focused on the development of hydroxy(thiophene-2-yl) acetic acid initial intermediate for the development of benzodifuranone analogues containing aromatic thiophene core.
Controlled production of lipids and lipidic substances by selected yeasts and microalgae
Karotenoidy jsou přírodní pigmenty vyskytující se v mikroorganismech jako jsou řasy, kvasinky a sinice. Představují nejrozšířenější skupinu antioxidantů s významným biologickým účinkem. V současnosti vzrůstá zájem o karotenoidy vzhledem k jejich příznivým vlivům na lidské zdraví. Chlorofyly jsou zelená fotosyntetická barviva, která nacházejí uplatnění v potravinářství jako intenzivní zelená barviva. Koenzym Q je znám svým pozitivním vlivem pro správnou funkci řady orgánů v lidském těle. Ergosterol je nedílnou součástí membrán kvasinek a hub. Je to provitamin D2, který je důležitou součástí imunitního systému. Mikrobiální lipidy, nebo také ‚‚Single cell oils‘‘ jsou charakteristické vysokým obsahem zdraví prospěšných nenasycených mastných kyselin, které lze využít ve farmacii či kosmetice. Mikrobiální lipidy jsou dále studovány jako alternativa pro výrobu biopaliv. Dizertační práce byla zaměřena na studium a možnosti optimalizace produkce lipidů a lipidických látek vybranými kmeny karotenogenních kvasinek, mikrořas a sinic. V rámci práce byly testované kvasinky rodu Rhodotorula, Rhodosporidium, Cystofilobasidium a Sporidiobolus podrobené kultivacím na sérií médií s různými C/N poměry v rozsahu 13 až 100, obsahujících upravené odpadní substráty z potravinářského průmyslu. Vybrané kmeny byly poté kultivovány v bioreaktorech v médiu obsahujícím kombinaci odpadních substrátů. Kultivace mikrořas rodu Desmodesmus, Scenedesmus, Chlorella, Coccomyxa, Chlamydomonas, Botryococcus se zabývaly optimalizací jednotlivých komponent média a aplikací různých stresů s cílem navýšení produkce studovaných metabolitů. V rámci experimentů s extrémofilní mikrořasou Coccomyxa byly provedeny pilotní velkoobjemové kultivace v otevřených nádržích. V závěrečné části byl provedeny pilotní screeningové a velkoobjemové bioreaktorové experimenty zaměřené na možnosti kokultivace karotenogenních kvasinek a mikrořas. Testované kmeny kvasinek byly s rozdílnou úspěšností schopny utilizovat média obsahující hydrolyzované odpadní substráty. Nejlepším kmenem byl Sporidiobolus pararoseus, který na médiích dosahoval nejvyšších produkcí biomasy i sledovaných metabolitů. Z testovaných odpadních substrátů byla nejlepší kombinace odpadního fritovacího oleje a hydrolyzátu kávové sedliny. Úspěšná optimalizace složení hlavních komponent minerálního média vedla k zvýšené produkci studovaných metabolitů. Největší vliv měl optimální poměr P/N a aplikace oxidačního stresu. Nejlepších výsledků dosáhly mikrořasy rodu Desmodesmus a Scenedesmus. Velkoobjemové kultivace Coccomyxy onubensis potvrdily rezistenci kultury proti kontaminaci vnějšími vlivy a schopnost růstu za vysoké teploty a intenzity světelného záření. Kokultivační experimenty potvrdily schopnost symbiotického růstu kvasinek a mikrořas. Nejlepších výsledků dosahovaly všechny testované kvasinky s mikrořasami rodu Demsodesmus a Scenedesmus a v menší míře i rodu Coccomyxa.Carotenoids are natural pigments found in microorganisms such as algae, yeast and cyanobacteria. They represent the most widespread group of antioxidants with a significant biological effect. Currently, there is a growing interest in carotenoids due to their beneficial effects on human health. Chlorophylls are green photosynthetic pigments that are used in the food industry as intensive green dyes. Coenzyme Q is known for its positive effect on the proper functioning of a number of organs in the human body. Ergosterol is an integral part of the membranes of yeasts and fungi. It is a provitamin D2, which is an important part of the immune system. Microbial lipids, or ‚Single cell oils’, are characterised by a high content of healthy unsaturated fatty acids that can be used in pharmacy or cosmetics. Microbial lipids are further studied as an alternative for the production of biofuels. The dissertation focused on the study and possibilities of optimising the production of lipids and lipid substances by selected strains of carotenogenic yeasts, microalgae and cyanobacteria. The tested yeasts of the genera Rhodotorula, Rhodosporidium, Cystofilobasidium and Sporidiobolus were cultivated on a series of different media with different C/N ratios in the range of 13 to 100, containing processed waste substrates of the food industry. Selected strains were then cultivated in bioreactors with medium containing a combination of waste substrates. Cultures of microalgae of the genera Desmodesmus, Scenedesmus, Chlorella, Coccomyxa, Chlamydomonas, Botryococcus dealt with the optimisation of individual components of the medium and the application of various stresses in order to increase the production of the studied metabolites. Pilot large-volume cultures in open tanks were performed as part of experiments with the extremophilic microalgae Cocomyxa. In the final part, pilot screenings and large-volume bioreactor experiments focused on the possibilities of co-cultivation of carotenogenic yeasts and microalgae were performed. The tested yeast strains were able to utilise media containing hydrolysed waste substrates with varying success. The best strain was Sporidiobolus pararoseus, which achieved the highest biomass production and studied metabolites in the media. The tested waste substrates were a combination of waste frying oil and coffee grounds hydrolysate. Successful optimisation of the composition of the mineral medium's main components led to increased production of the studied metabolites. The most important effect exhibited particular P/N ratio and the application of oxidative stress. Desmodesmus and Scenedesmus microalgae achieved the best results. Large-scale cultures of Coccomyxa onubensis confirmed the resistance of the culture to contamination by external influences and the ability to grow at high temperatures and light intensities. Co-cultivation experiments confirmed the ability of symbiotic growth of yeasts and microalgae. The best results were obtained in all tested yeasts co-cultivated with microalgae of the genus Desmodesmus and Scenedesmus and, to a lesser extent, with the genus Coccomyxa.
A comparison of metabolic activity and use of microalgae and pigment-formed yeasts
Karotenoidy jsou přírodními pigmenty vyskytujícími se v rostlinách a mnohých mikroorganismech, jako jsou řasy, kvasinky a bakterie. Představují nejrozšířenější skupinu antioxidantů s významným biologickým účinkem. Lipidy jsou esenciální složkou všech živých organismů. Jsou zdrojem a zásobárnou energie pro organismy, kterou lze využít také v petrochemickém průmyslu pro produkci biopaliv. Tato práce se zabývá studiem produkčních vlastností karotenoidů a lipidů vybranými kmeny kvasinek, řas a sinic, a to s cílem aplikace získaných poznatků a nalezení vhodné levné alternativy pro mikrobiální biotechnologickou produkci těchto metabolitů s využitím odpadních substrátů. Obsah karotenoidů, ergosterolu a koenzymu Q v buňkách byl stanovován kapalinovou chromatografií. Obsah lipidů a profil mastných kyselin byl analyzován plynovou chromatografií. Morfologie buněk a lokalizace vybraných metabolitů byla analyzována technikou časově rozlišené fluorescenční mikroskopie. V této práci byly využity kvasinkové rody Sporobolomyces, Cystofilobasidium a Rhodotorula. Z řas a sinic byl použity rody Desmodesmus, Cyanothece, Chlamydomonas, Synechoccocus a Chlorella. V celkovém porovnání produkčních vlastností, dosahovaly nejvyšší produkce kvasinky Z vybraných kmenů, bylo největších výtěžků karotenoidů dosaženo u kmenů S. pararoseus, C. capitatum a R. mucilaginosa a lipidů u kmenů C. infirmominiatum a S. metaroseus.Carotenoids are natural pigments occurring in plants and many microorganisms, such as algae, yeast and bacteria. They represent the most common group of antioxidants with significant biological effect. Lipids are an essential component of all living organisms. They are the source and the reservoir of energy for organisms, which can also be used in the petrochemical industry for the production of biofuels. This thesis deals with the production properties of carotenoids and lipids by selected strains of yeasts, algae, and canobacteria, in order to apply the acquired knowledge and find cheap suitable alternatives for microbial biotechnological production of these metabolites using waste substrates. Carotenoids, coenzyme Q, and ergosterol in cells were determined by liquid chromatography. The lipid content and fatty acid profile was analyzed by gas chromatography. Cell morphology and localization of selected metabolites were analyzed by fluorescence lifetime imaging microscopy. In this work yeast genera Sporobolomyces, Cystofilobasidium and Rhodotorula were used. As the representatives of the algae and cyanobacteria strains of Desmodesmus, Cyanothece, Chlamydomonas, Synechoccocus and Chlorella strains were studied. In the overall comparison, the yeast strains were more productive than algae and cyanobacteria. The highest carotenoid production was found in S. pararoseus, C. capitatum and R. mucilaginosa cells, while the highest lipid yield was observed in strains of C. infirmominiatum and S. metaroseus.
Rhodotorula kratochvilovae CCY 20-2-26-The Source of Multifunctional Metabolites
Multifunctional biomass is able to provide more than one valuable product, and thus, it is attractive in the field of microbial biotechnology due to its economic feasibility. Carotenogenic yeasts are effective microbial factories for the biosynthesis of a broad spectrum of biomolecules that can be used in the food and feed industry and the pharmaceutical industry, as well as a source of biofuels. In the study, we examined the effect of different nitrogen sources, carbon sources and CN ratios on the co-production of intracellular lipids, carotenoids, beta-glucans and extracellular glycolipids. Yeast strain R. kratochvilovae CCY 20-2-26 was identified as the best co-producer of lipids (66.7 +/- 1.5% of DCW), exoglycolipids (2.42 +/- 0.08 g/L), beta-glucan (11.33 +/- 1.34% of DCW) and carotenoids (1.35 +/- 0.11 mg/g), with a biomass content of 15.2 +/- 0.8 g/L, by using the synthetic medium with potassium nitrate and mannose as a carbon source. It was shown that an increased C/N ratio positively affected the biomass yield and production of lipids and beta-glucans
Raman spectroscopy online monitoring of biomass production, intracellular metabolites and carbon substrates during submerged fermentation of oleaginous and carotenogenic microorganisms
Background Monitoring and control of both growth media and microbial biomass is extremely important for the development of economical bioprocesses. Unfortunately, process monitoring is still dependent on a limited number of standard parameters (pH, temperature, gasses etc.), while the critical process parameters, such as biomass, product and substrate concentrations, are rarely assessable in-line. Bioprocess optimization and monitoring will greatly benefit from advanced spectroscopy-based sensors that enable real-time monitoring and control. Here, Fourier transform (FT) Raman spectroscopy measurement via flow cell in a recirculatory loop, in combination with predictive data modeling, was assessed as a fast, low-cost, and highly sensitive process analytical technology (PAT) system for online monitoring of critical process parameters. To show the general applicability of the method, submerged fermentation was monitored using two different oleaginous and carotenogenic microorganisms grown on two different carbon substrates: glucose fermentation by yeast Rhodotorula toruloides and glycerol fermentation by marine thraustochytrid Schizochytrium sp. Additionally, the online FT-Raman spectroscopy approach was compared with two at-line spectroscopic methods, namely FT-Raman and FT-infrared spectroscopies in high throughput screening (HTS) setups. Results The system can provide real-time concentration data on carbon substrate (glucose and glycerol) utilization, and production of biomass, carotenoid pigments, and lipids (triglycerides and free fatty acids). Robust multivariate regression models were developed and showed high level of correlation between the online FT-Raman spectral data and reference measurements, with coefficients of determination (R2) in the 0.94–0.99 and 0.89–0.99 range for all concentration parameters of Rhodotorula and Schizochytrium fermentation, respectively. The online FT-Raman spectroscopy approach was superior to the at-line methods since the obtained information was more comprehensive, timely and provided more precise concentration profiles. Conclusions The FT-Raman spectroscopy system with a flow measurement cell in a recirculatory loop, in combination with prediction models, can simultaneously provide real-time concentration data on carbon substrate utilization, and production of biomass, carotenoid pigments, and lipids. This data enables monitoring of dynamic behaviour of oleaginous and carotenogenic microorganisms, and thus can provide critical process parameters for process optimization and control. Overall, this study demonstrated the feasibility of using FT-Raman spectroscopy for online monitoring of fermentation processes.Raman spectroscopy online monitoring of biomass production, intracellular metabolites and carbon substrates during submerged fermentation of oleaginous and carotenogenic microorganismspublishedVersio
Use of Waste Substrates for the Lipid Production by Yeasts of the Genus Metschnikowia—Screening Study
Oleogenic yeasts are characterized by the ability to accumulate increased amounts of lipids under certain conditions. These microbial lipids differ in their fatty acid composition, which allows them to be widely used in the biotechnology industry. The work focuses on the influence of various stress factors in the cultivation process, such as reduced temperature or nutritional stress through the use of various waste substrates, together with manipulating the ratio of carbon and nitrogen sources in the medium. The ability of yeast to produce significant amounts of unsaturated fatty acids was also demonstrated in the work. The most suitable substrate for lipid production was a medium containing glycerol, where the amount of accumulated lipids in the yeast M. pulcherrima 1232 was up to 36%. In our work, the crude animal fat was used for the production of high-value lipids, which to the best of our knowledge is a new result
Production of Enriched Sporidiobolus sp. Yeast Biomass Cultivated on Mixed Coffee Hydrolyzate and Fat/Oil Waste Materials
One of the most addressed topics today is the transfer from a linear model of economics to a model of circular economics. It is a discipline that seeks to eliminate waste produced by various industries. The food industry generates huge amounts of waste worldwide, particularly the coffee industry, and related industries produce millions of tons of waste a year. These wastes have potential utility in biotechnology, and in the production of energy, fuels, fertilizers and nutrients, using green techniques such as anaerobic digestion, co-digestion, composting, enzymatic action, and ultrasonic and hydrothermal carbonization. This work is focused on the biotechnological use of processed spent coffee grounds (SCG) and waste fat/oil materials by some Sporidiobolus sp. carotenogenic yeasts in the model of circular economics. The results show that selected yeast strains are able to grow on SCG hydrolysate and are resistant to antimicrobial compounds present in media. The most productive strain Sporidiobolus pararoseus CCY19-9-6 was chosen for bioreactor cultivation in media with a mixture of coffee lignocellulose fraction and some fat wastes. Sporidiobolus pararoseus CCY19-9-6 was able to produce more than 22 g/L of biomass in mixture of SCG hydrolysate and both coffee oil and frying oil. The combined waste substrates induced the production of lipidic metabolites, whereby the production of carotenoids exceeded 5 mg/g of dry biomass. On media with coffee oil, this strain produced high amounts of ubiquinone (8.265 +/- 1.648 mg/g) and ergosterol (13.485 +/- 1.275 mg/g). Overall, the results prove that a combination of waste substrates is a promising option for the production of carotenoid- and lipid-enriched yeast biomass
Study of Metabolic Adaptation of Red Yeasts to Waste Animal Fat Substrate
Carotenogenic yeasts are non-conventional oleaginous microorganisms capable to utilize various waste substrates. In this work 4 red yeast strains (Rhodotorula, Cystofilobasidium and Sporobolomyces sp.) were cultivated in media containing crude, emulsified and enzymatically hydrolysed animal waste fat, compared with glucose and glycerol as single C-sources. Cell morphology (cryo-SEM, TEM), production of biomass, lipase, biosurfactants, lipids (GC/FID) carotenoids, ubiquinone, ergosterol (HPLC/PDA) in yeast cells was studied depending on medium composition, C-source and C/N ratio. All studied strains are able to utilize solid and processed fat. Biomass production at C/N=13 was higher on emulsified/hydrolysed fat than on glucose/glycerol. Production of lipids and lipidic metabolites was enhanced for several times on fat; the highest yields of carotenoids (24.8 mg/l) and lipids (54.5%/CDW) were found in S.pararoseus. Simultaneous induction of lipase and biosurfactants was observed on crude fat substrate. Increased C/N ratio (13-100) led to higher biomass production in fat media. Production of total lipids increased in all strains to C/N 50. Oppositely, production of carotenoids, ubiquinone and ergosterol dramatically decreased with increased C/N in all strains. Compounds accumulated in stressed red yeasts are having great application potential and can result from valorization of animal waste fat in the biorefinery concept.Karotenogenní kvasinky jsou schopny utilizovat řadu různých substrátů včetně odpadů. V této práci je popsána kultivace vybraných kmenů karotenogenních kvasinek rodů Rhodotorula, Cystofilobasidium, and Sporobolomyces sp. v médiu obsahujícícm surový odpadní živočišný tuk a upravený substrát emulsifikací a enzymovou hydrolýzou. Výsledky byly srovnány s kultivací na glukóze a glycerolu jako jednoduchých zdrojích uhlíku
Revealing the Potential of Lipid and beta-Glucans Coproduction in Basidiomycetes Yeast
Beta (beta)-glucans are polysaccharides composed of D-glucose monomers. Nowadays, beta-glucans are gaining attention due to their attractive immunomodulatory biological activities, which can be utilized in pharmaceutical or food supplementation industries. Some carotenogenicBasidiomycetesyeasts, previously explored for lipid and carotenoid coproduction, could potentially coproduce a significant amount of beta-glucans. In the present study, we screened elevenBasidiomycetesfor the coproduction of lipids and beta-glucans. We examined the effect of four different C/N ratios and eight different osmolarity conditions on the coproduction of lipids and beta-glucans. A high-throughput screening approach employing microcultivation in microtiter plates, Fourier Transform Infrared (FTIR) spectroscopy and reference analysis was utilized in the study. Yeast strainsC. infirmominiatumCCY 17-18-4 andR. kratochvilovaeCCY 20-2-26 were identified as the best coproducers of lipids and beta-glucans. In addition,C. infirmominiatumCCY 17-18-4,R. kratochvilovaeCCY 20-2-26 andP. rhodozymaCCY 77-1-1 were identified as the best alternative producers of beta-glucans. Increased C/N ratio led to increased biomass, lipid and beta-glucans production for several yeast strains. Increased osmolarity had a negative effect on biomass and lipid production while the beta-glucan production was positively affected