Ispitivanje aktivnosti analoga sintaza N-acetilneuraminske kiseline na neprirodne supstrate

undergraduate thesis

• Title (english): Screening of N-acetylneuraminic acid synthase analogues on unnatural substrates
• Author: Lucija Drandić
• Mentor: Zvjezdana Findrik Blažević (mentor)
• Committee member: Zvjezdana Findrik Blažević (predsjednik povjerenstva)
• Committee member: Martina Sudar (član povjerenstva)
• Committee member: Marija Vuković Domanovac (član povjerenstva)
• Granter: University of Zagreb; Faculty of Chemical Engineering and Technology; Zagreb
• Defense date and country: 2024-09-17, Croatia
• Scientific / art field, discipline and subdiscipline: TECHNICAL SCIENCES; Chemical Engineering

Abstract

Sijalinske kiseline mogu posredovati ili modulirati u širokom spektru fizioloških i patoloških procesa zbog svoje sveprisutne distribucije i strateške lokacije. U posljednjem desetljeću postignut je značajan napredak u proučavanju glikana na ljudskim stanicama, kao i njihove uloge u interakcijama među stanicama. Ugljikohidrati igraju mnogobrojne uloge u različitim organizmima, uključujući razvoj, signalizaciju, interakcije između domaćina i parazita te imunološku funkciju. Iako je provedeno mnogo istraživanja, biologija sijalinske kiseline je još uvijek rastuće područje velikog potencijala čije su primjene tek započele. U ovom radu provedena je kondenzacijska reakcija šećera (L-arabinoza, D-galaktoza, D-liksoza,
D-(+)-manoza) i fosfoenolpiruvata uz prisutnost metalnog kofaktora Co2+. Reakcije su katalizirane analozima sintaze N-acetinneuraminske kiseline (PNH2, PNH5), umjetno dobiveni na temelju sličnosti sa sintazom neuraminske kiseline iz bakterije Neisseria meningitidis. Produkt reakcija su različite sijalinske kiseline. Provedena su ispitivanja s ekstraktom stanica te s pročišćenim enzimom radi potvrde aktivnosti enzima i nastajanja analognih produkata sijalinske kiseline. Uočena je najveća kompatibilnost enzima PNH5 s testiranim šećerima.

Abstract (english)

Sialic acids can mediate or modulate a wide range of physiological and pathological processes due to their omnipresent distribution and strategic location. In the past decade, significant progress has been made in the study of glycans on human cells, as well as their role in cell-cell interactions. Carbohydrates play numerous roles in various organisms, including development, signaling, host-parasite interactions, and immune function. Although much research has been conducted, sialic acid biology is still an emerging field with great potential, while its applications are just beginning. In this study, a condensation reaction of sugars (L-arabinose, D-galactose, D-lyxose, D-(+)-mannose) and phosphoenolpyruvate in the presence of the metal cofactor Co2+. The reactions were catalyzed by analogs of
N-acetylneuraminic acid synthase (PNH2, PNH5), which were artificially derived based on their similarity to the neuraminic acid synthase from the bacterium Neisseria meningitidis.
The product of the reactions are different sialic acids. Assays were conducted on both
cell-free extracts and purified enzyme to confirm the enzyme activity and the formation of sialic acid analogs. The highest compatibility of the enzyme PNH5 with the tested sugars was observed.

• Keywords: enzimi; aktivnost enzima; fosfoenolpiruvat; sijalinska kiselina; N-acetilneuraminska kiselina
• Keywords (english): enzymes; enzyme activity; phosphoenolpyruvate; sialic acid; N-acetylneuraminic acid
• Language: croatian
• URN:NBN: urn:nbn:hr:149:802459
• Project: Number: 956631 — CC-TOP — H2020-MSCA-ITN-2020; Title: C-C Bond Formation Using Top Performing Enzymes; Acronym: C-C Top; Leader: Zvjezdana Findrik Blažević; Jurisdiction: eu; Funder: Europska unija; Funding stream: Horizon 2020
• Study programme: Title: Applied Chemistry - Undergraduate study; specializations in: Environmental chemistry and green technologies, Advanced materials and technologies, Applied organic chemistry; Course: Environmental chemistry and green technologies; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) primijenjene kemije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) primijenjene kemije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Embargoed access; Embargo expiration date: 2026-10-01
• Created on: 2024-11-19 09:10:27