20.500.12556/RUL-124054 Priprava rekombinantnih človeških citokromov P450 in njihovo biokatalizno vrednotenje v sintezi metabolitov izbranih učinkovin Production of recombinant human P450 cytochromes and their biocatalytic evaluation in selected drug metabolite synthesis V okviru magistrskega dela smo optimizirali izražanje več človeških paralogov CYP450 v metilotrofni kvasovki Pichia pastoris. Med gojenjem kvasovk smo spreminjali stopnjo aeracije, pH gojišča in hitrost mešanja. Prav tako smo primerjali tudi rast celic med gojenjem v bioreaktorjih in erlenmajericah ter aktivnost encimov v suspenziji celic. Katalitično aktivnost CYP450 smo ovrednotili z inkubacijo modelnih učinkovin testosterona in diklofenaka v kulturah rekombinantnih kvasovk, ekstrakcijo metabolitov in njihovo analizo s HPLC. As part of the master's thesis, we optimized the expression of several human CYP450 paralogs in the methylotrophic yeast Pichia pastoris. During the cultivation of yeasts, the degree of aeration, pH of the growth medium and mixing speed were modified. Also, yeast growth and enzyme activity following fermentation in bioreactors and shake flasks were compared. The catalytic activity of CYP450 was evaluated by incubation of the model drugs testosterone and diclofenac in recombinant yeast cultures, extraction of metabolites and their analysis by HPLC. Pichia pastoris CYP 450 bioreaktorske enote erlenmajerice biotransformacija HPLC diklofenak testosteron Pichia pastoris CYP 450 bioreactor shake flasks biotransformation HPLC diclofenac testosterone true false false Slovenski jezik Angleški jezik Magistrsko delo/naloga 2020-12-23 07:45:38 2020-12-23 07:45:41 2022-09-04 03:57:23 0000-00-00 00:00:00 2020 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 1970-01-01 78339 1552.pdf 1552.pdf 1 0CFA9DB2ADAA35364056426386F435D6 e7df9145afa342bdaa3ed3b759252e6bf793c4bad5ef914dd3745034cbf04439 76f149ea-a1bb-11eb-a523-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=139441 Fakulteta za farmacijo 0 0 0