20.500.12556/RUL-170161 Karakterizacija inhibicije treonin dehidrogenaze s specifičnimi derivati pirazola Characterisation of the threonine dehydrogenase inhibition with specific derivatives of pyrasole Pirazol in njegovi derivati postajajo vse bolj pomembni v svetu medicine zaradi svojega dokazanega protimikrobnega delovanja. Razvoj različnih derivatov povečuje verjetnost, da bodo nekateri uporabni pri razvoju novih antibiotikov. Derivat pirazola, 4‑(2‑aminoetil)-1-(piridin-2-il)-1H-pirazol-5-ol (I1), zavira rast bakterije Escherichia coli in inhibira aktivnost encima L-treonin dehidrogenaza (TDH). Ta encim je ključnega pomena za metabolizem aminokislin in tvorbo biofilma. TDH v človeku nima funkcionalnega homologa, zato bi lahko spojina I1 bila osnova za razvoj novih antibiotikov. Cilj raziskovalnega dela je bil preučiti, kako različni derivati pirazola vplivajo na inhibicijo aktivnosti TDH in ali vplivajo tudi na tvorbo biofilmov. Za delovanje encima je potreben kofaktor NAD+, ki se ob reakciji pretvori v NADH, kar omogoča fluorimetrično spremljanje aktivnosti. Uporabili smo rekombinantni TDH ter spremljali njegovo aktivnost v prisotnosti različnih derivatov pirazola. Med testiranimi derivati je bil najbolj potenten derivat I1. Podobno inhibitorno aktivnost je imel tudi derivat I2, vendar njegova slaba topnost ni omogočila testiranja pri višjih koncentracijah. Preizkusili smo tudi vpliv derivatov na tvorbo biofilmov, pri čemer smo ugotovili, da vsi derivati zavirajo tvorbo biofilmov. Le I1 in I2 pa sta delovala neposredno na aktivnost TDH. Na podlagi tega ne moremo z gotovostjo potrditi povezave med inhibicijo TDH in zaviranjem tvorbe biofilma. Pyrazole and its derivatives are becoming increasingly prevalent in medicine due to their proven antimicrobial activity. The development of new derivatives increases the likelihood of discovering new effective antibiotics. The pyrazole derivative, 4-(2-aminoethyl)-1-(pyridine-2-yl)-1H-pyrazole-5-ol (I1), inhibits the growth of Escherichia coli and L-threonine dehydrogenase (TDH) activity. The enzyme is a key component in amino acid metabolism and biofilm formation. TDH is not present in humans nor does it have a homologous protein in humans, hence I1 could be a potential base for new drug development. The aim of the research project was to examine the different types of pyrazole derivatives and their effects on TDH activity and whether they affect the formation of biofilms. For the enzyme to work, the cofactor NAD+ is required, where upon the reaction is converted into NADH, which then makes it possible for fluorometric detection of activity. Among the tested derivatives, I1 was the most promising and potent. Derivative I2 also had similar inhibitory activity. However, due to its poor solubility, it proved impossible to test at higher concentrations. Tests were also conducted to see the impact derivatives have on biofilm formation, and the results showed that all derivatives inhibit biofilm formation. However, only I1 and I2 directly affect TDH activity. Based on these findings, further research is required to establish with certainty to prove the correlation between TDH inhibition and biofilm formation suppression. L-treonin dehidrogenaza biofilmi derivati pirazol L-Threonine dehydrogenase biofilms pyrazole derivatives true false false Slovenski jezik Angleški jezik Diplomsko delo/naloga 2025-07-02 13:32:05 2025-07-02 13:32:08 2025-07-18 12:50:33 0000-00-00 00:00:00 2025 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 25521 243067651 Tusek_Marcel_-_Karakterizacija_inhibicije_treonin_dehidrogenaze_s_specificnimi_derivati_pirazola.pdf Tusek_Marcel_-_Karakterizacija_inhibicije_treonin_dehidrogenaze_s_specificnimi_derivati_pirazola.pdf 1 02A2E4F0B8B1C9EE641CB6E562C1087F 5ce9b94abc116acd0a269a7427ad27fd70d32f119d28c9f801ffee0316fe2c3b 4b1d35c6-5736-11f0-b232-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=212096 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0