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Master's Thesis
Author of thesis: Ing. Katarína Čačková
Acad. year: 2011/2012
Supervisor: prof. RNDr. Ivana Márová, CSc.
Reviewer: Ing. Emília Breierová, Ph.D.
Carotenoids are naturally occurring pigments of plants also produced by microbes. The area of their application concerns mainly food industry; however, they are used in chemical, pharmaceutical, and cosmetics industry as well. Currently, the isolation of carotenoids from plants is markedly regulated by legislation, so the study of their production is greatly emphasised, where the microbiological, instead of the synthetic, production of carotenoids is being prioritized. This work was made as a comparative study of carotenogenic yeasts of the genes Rhodotorula, Sporobolomyces, and Cystofilobasidium. Their ability to use various waste substrates as a carbon and nitrogen source and source of other nutrition factors was tested. In this work, conditions of random mutagenesis were optimized. Particular yeast strains were also subjected to the effect of mutagen ethyl methanesulfonate (EMS) in order to increase the production of biomass and specific metabolites – carotenoids and other lipid-soluble substances. Random mutagenesis and mutant strain selection was performed using waste subtrates as glycerol, pasta and some pasta hydrolyzed by fungal extracellular enzymes. Subsequently, a control of specific DNA sequences in pigments overproducing mutants was analyzed by PCR/DGGE (denaturating gradient gel electrophoresis). Increased production of β-carotene was achieved in a mutant of Sporobolomyces roseus strain growing on glycerol, pasta, and hydrolyzed pasta. Overproduction of carotenoids by mutant strain of Rhodotorula glutinis was observed in glucose medium only. Mutants of Cystofilobasidium capitatum exhibited a decrease of biomass production; on the other hand, the production of carotenoids increased especially in pasta medium hydrolyzed by enzyme preparative from Fusarium solani. In this work it was confirmed that using random mutagenesis strains capable to utilize waste substrates can be selected. In mutant strains increased carotenoids biosynthesis was observed, which enables effective use of cheap substrates and reduction of the negative effects of wastes on the environment.
carotenoids, red yeasts, waste substrates, random mutagenesis
Date of defence
04.06.2012
Result of the defence
Defended (thesis was successfully defended)
Grading
A
Process of defence
1. Diplomantka seznámila členy komise s náplní a cílem diplomové práce. 2. Byly přečteny posudky na diplomovou práci. 3. Diplomantka akceptovala všechny připomínky oponenta a na všechny otázky odpověděla v plné šíři. Diskuse: doc. Španová: Byly pozorovány nějaké nežádoucí faktory (reakce) ?
Language of thesis
Slovak
Faculty
Fakulta chemická
Department
Institute of Food Science and Biotechnology
Study programme
Chemistry and Technology of Foodstuff (NPCP_CHTP)
Field of study
Food Science and Biotechnology (NPCO_CHTP)
Supervisor’s reportprof. RNDr. Ivana Márová, CSc.
Grade proposed by supervisor: A
Reviewer’s reportIng. Emília Breierová, Ph.D.
Grade proposed by reviewer: A
Responsibility: Mgr. et Mgr. Hana Odstrčilová