|
|
Monitoring of changes of chemical parameters in wine production
Dlapalová, Kristýna ; Vespalcová, Milena (referee) ; Vránová, Dana (advisor)
This bachelor work deals with chemical ganges of the wine must during the fermentation process. We compered the sample must with addend commercial yeasts and indigenous yeasts. The theoretical part describes the vine and its variety. The variety Saugvinon is central of this work and it is more described.. The theoretical part describes the structure of grape and its chemical technology of wine production and wine disease. The production technology is divided into different steps of the production (eg. picking the grapes, pressing, fermentation and wine training). The experimental part describes the methods used in the determination of chemical parameters including the chemicals and equipment. The results are summarized the results of the determination of reducing sugars, titratable acidity, alcohol and pH must. The results are evaluated and commented upon in the discussion.
|
|
|
Use of molecular techniques to characterize yeasts of the genus Metschnikowia
Schneiderwindová, Nicole ; Brázda, Václav (referee) ; Němcová, Andrea (advisor)
This diploma thesis deals with the possibilities of implementation and use of molecular methods for the characterization of yeasts of the genus Metschnikowia and the application of methods in biotechnology or the food industry. The theoretical part focuses on a brief description of yeast, specially selected species that were used during the practical part of the work, the possibilities of their use, and especially on a detailed description of all molecular techniques used. The practical part focuses on the optimization of the molecular methods, namely the method of pulsed gel electrophoresis and the method of denaturing gradient gel electrophoresis. Initially, yeast was cultured under optimal conditions that are specific to this genus. Furthermore, their DNA was isolated using isolation techniques, which were subsequently processed using PFGE and PCR–DGGE methods. The DNA isolation procedure needed to be optimized the most. Several optimizations of the concentration of lysis enzymes, especially the lyticase enzyme, were performed. It was also necessary to determine the correct ratio of low-melting agarose and isolated DNA, which was essential for the correct consistency of the isolated DNA blocks and their further application in PFGE analysis. Finally, the PFGE method was optimized, which brought the correct distribution of chromosomes, and it was possible to describe the individual chromosomes according to their size according to the standard used CHEF of the yeast Hansenula wingei. To properly optimize the DGGE analysis process itself, it was first necessary to isolate the yeast DNA using a kit, then it was used as a template for the PCR reaction. The annealing temperature was also optimized for the individual groups of primers. The amplicons obtained by this reaction were separated by the DGGE method. This technique mainly required the optimization of basic parameters such as the range of the denaturation gradient or the total separation time. According to the measurement results, it can be determined that the process of yeast DNA isolation and their subsequent analysis using molecular methods of pulsed gel electrophoresis and denaturing gradient gel electrophoresis was successful. We were able to describe the genome and determine the number of chromosomes in all used yeast species of the genus Metschnikowia at least partially.
|
|
|
Study of genome of Metschnikowia yeasts by molecular methods
Schneiderwindová, Nicole ; Skoumalová, Petra (referee) ; Němcová, Andrea (advisor)
Yeasts of the genus Metschnikowia belonging to the family Metschnikowiacea are yeasts characterized by vegetative propagation through multilateral budding. These are yeasts widely distributed in nature. More than 35 species occurring have been defined in the wild. They most often occur on flowers, fruits, but also on insects or human skin. They have a wide range of uses due to their antifungal effects in agriculture and the cosmetics industry. This bachelor thesis deals with the study of usage of molecular methods to characterize selected species of yeasts of the genus Metschnikowia. It focuses on a detailed description of the yeast cell structure, karyotype and methods of reproduction in the theoretical part of the work. In the practical part on optimization and description of molecular methods including pulse gel electrophoresis methods used to separate the yeast genome and their subsequent observation of changes in individual parts of genome. First, the yeast was cultured under special conditions that are characteristic of Metschnikowia yeasts, then yeast DNA was isolated using methods suitable for DNA isolation, which was further examined by the PFGE molecular method. The DNA isolation procedure was first optimized for individual yeast strains, as it was necessary to verify the required ratio of low melting agarose to isolated DNA. That was because of it was important for the resulting gel blocks to be suitable for measurement by PFGE analysis. By optimizing the method was possible to create ideal blocks of isolated yeast DNA, which were subsequently subjected to PFGE analysis. Several measurements of PFGE analysis were performed at different time intervals in order to separate small and large yeast chromosomes. The CHEF standard of the yeast Hansenula wingei and the standard of the yeast Schizosaccharomyces pombe were used for the measurements. According to the measurement results, it can be determined that the yeast DNA isolation procedure and subsequent analysis by pulsed gel electrophoresis were successful, as the number of chromosomes of all used yeast species of the genus Metschnikowia was determined.
|
|
|
Yeasts and wine
Palíková, Petra ; Vadkertiová, Renata (referee) ; Vránová, Dana (advisor)
This thesis deals with isolation and identification wine yeasts from grapes and must. For analysis was used white wine Sauvignon that was grown and producing after needs ecological agriculture. Remove samples were processed in laboratory and by the help of dilution method were obtained pure culture isolated yeasts. In the following step, by the application of commercial kit UltraCleanTM Microbial DNA Isolation Kit we were able to isolated individual DNA that it was used to the next analysis. Isolated DNA was amplification by PCR method with ITS1 and ITS4 primers. PCR products were detected on agarose gel. Amplification samples were chopped five restriction endonucleases: HaeIII, HinfI, TaqaI, AluI and MseI. Chopped DNA was detected by the same way as PCR products and it was compared with restriction patterns of collection yeasts. In the next step it was compared genetic similarity of isolated yeasts by using BioNumerics software. As a criterion it was used Pearson coefficients and UPGMA clastering analysis. The result is dedrogram of genetics similarity isolated yeasts.
|
|
|
Study of the chemical processes during kombucha technology
Chludilová, Markéta ; Obruča, Stanislav (referee) ; Pořízka, Jaromír (advisor)
The kombucha drink is referred to as leavened tea with added sponge called a scobie. In this way, a low-alcohol beverage is obtained, which gains considerable popularity, especially due to the alleged beneficial properties for human health. The microbial composition of this beverage is very rich, consisting of bacteria and yeasts living in a beneficial symbiosis [11]. The microbial composition of the drink and its beneficial effects on human health are very closely related. This work deals with the analysis of the chemical composition of the commodity beverage, especially with regard to the content of major components such as organic acids, carbohydrates and ethanol. These analytes are among the most common and most profoundly found substances in the carbohydrate beverage and are very closely related to the microbial composition of the scoby itself [3]. Combo beverage samples were supplied by RebornFood, one of which is the commercial production of this drink on the Czech market. At the same time, this company bases itself on purely natural fermentation of beverages, and therefore it is not particularly involved in the production process. The results of this diploma work point to the effect of the scoby used and the associated occurrence of analytes in the beverage, as well as the production process associated with the formation of analytes, which is described in several articles. It is expected that this work will provide information both on the production conditions and on the chemical properties of the beverage related to the condition of the scoby, which will be used further, both for RebornFood and the material used to solve the technical problems of this beverage production.
|
|
|
Controlled production of lipids and lipidic substances by selected yeasts and microalgae
Szotkowski, Martin ; Kráčmar, Stanislav (referee) ; Huelva, Ines Garbayo Nores, University (referee) ; Márová, Ivana (advisor)
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.
|
|
|
Brewing industry
Artýszková, Jana ; Illková, Kateřina (referee) ; Omelková, Jiřina (advisor)
The aim of the theoretical part of the thesis is the study and the treatment of the problem of the manufacturing of beer, focused on the czech conditions and classical Czech technology. Within the theoretical part the history of this branch of industry together with the modern methods, enabling greater productivity of beer production, are summarized. Experimental part of the thesis was carried out in a small brewery in Czech republic, producing aproximately 10 000 hl per year. In this brewery the selected parameters were studied not only in the course of cooking and fermentation of beer, but during the output control in racking to the drums and plastic bottles, as well. The studied parameters were as follows: the sugar content of front and wort, the control of microbiological contamination in fermenting cellar and in beer-storage cellar, the content of CO2 by last stage of fermentation and pH- value and color by the end-product.
|
|
|
Application of microbial oils in cosmetics
Reichertová, Klára ; Uhlířová, Renata (referee) ; Němcová, Andrea (advisor)
This bachelor thesis is focused on the application of microbial oil in a cosmetic product. Eight strains of yeasts of the genus Metschnikowia were examined, namely Metschnikowia pulcherrima 145, Metschnikowia pulcherrima 147, Metschnikowia pulcherrima 149, Metschnikowia andauensis 129, Metschnikowia andauensis 1241, Metschnikowia sinensis 1244, Metschnikowia zizyphicola 1247 and Metschnikowia shanxiensis 1250 for production of microbial oil. Substrates made of technical hemp (hemp flour, hydrolyzate and a mixture of leaves and flowers) were used for cultivation. The cultivation was performed at reduced temperature with the advantage of lipid production. The composition and ratio of fatty acids were determined by gas chromatography According to these resulst was choosen the best lipid producer and the most abundant substrate. This was the strain Metschnikowia zizyphicola 1247 and the substrate of hemp flour. At the end of the practical part was prepared a cream, into which the microbial oil was mixed. Finally, an evaluation of the oil with regard to its use in cosmetics and a comparison with vegetable oil with the same fatty acid profile are given.
|
|
|
Characterization of selected yeast strains from the environment
Bečková, Alena ; Omelková, Jiřina (referee) ; Vránová, Dana (advisor)
The aim of this bachelor thesis deals with identification type-yeasts, which were obtained from the Culture Collection SAV in Bratislava. The purpose is extension of yeast database in laboratory of Faculty of Chemistry, which is used by identification yeasts in samples taken from food or environment. Identification of yeasts was implemented by method PCR-RFLP. Obtained segment is located in area 5,8S-ITS and is defined by using primers ITS1 and ITS4. Restriction analysis was carried out by restriction endonucleasis HaeIII, HinfI, HhaI and TaqI. Chosen species were also tested for presence of extracellular lipasis. In this part of characterisation was used SpiritBlue Agar. The presence of blue colour is lost by activity of lipasis.
|
|
|
Study of production properties of Metschnikowia yeasts
Chadimová, Markéta ; Szotkowski, Martin (referee) ; Němcová, Andrea (advisor)
Metschnikowia yeasts are able to produce under different conditions different amounts of lipids, which have remarkable uses in biotechnology and industry. This bachelor thesis is focused on the study of cultivation conditions under which yeasts produce the most lipids and also deals with which fatty acids these triacylglycerols are composed of. The influence of cultivation conditions on the amount of biomass is also monitored. Five yeast strains were examined, namely M. pulcherrima 145, M. pulcherrima 147, M. pulcherrima 149, M. andauensis 129 and M. chrysoperlae 1158. Cultivation media with different C/N ratios, containing glucose or cheap waste substrates (glycerol, coffee and waste fat) were used for cultivation. The cultivations were performed at several temperatures and then the amount of grown biomass was determined, and the composition and ratio of fatty acids were determined by gas chromatography. As yeast that produced the most biomass per time was determined the yeast M. chrysoperlae 1158, which after 336 hours of growth at 15 °C was able to produce 10,66 g/l of biomass on a medium with a ratio of C/N = 100. The highest amount of lipids was obtained with yeast M. andauensis 129 at 8 °C on medium with a ratio of C/N = 100 (21,57 % lipids in dry matter). If biomass production is also taken into account, the best results are obtained by the yeast M. chrysoperlae 1158 at a temperature of 11 ° C on a medium with a ratio of C/N = 100 with a biomass production of 10,15 g/l and 19,58 % lipids in dry matter. This yeast was therefore further cultivated on waste substrates.
|
guest ::
