Carbon dots as markers for plastic degradation assays
Author
Hess, Yan Jurg
Term
4. semester
Education
Publication year
2024
Pages
39
Abstract
Dette speciale undersøger en fluorescensbaseret screeningsmetode, hvor kulstofprikker (carbon dots) indlejret i polyethylenterephthalat (PET) bruges som visuelle markører for polymernedbrydning med henblik på at identificere proteiner med katalytisk aktivitet mod PET. Kulstofprikker blev syntetiseret og integreret i PET enten som film fremstillet ved spin-coating eller som fibre fremstillet ved elektrospinning, og frigivelsen af kulstofprikker ved enzymatisk nedbrydning blev overvåget ved fluorescencemissionsspektroskopi og reversed-phase HPLC. Elektrospundne PET-fibre gav højere overfladeareal og bedre stabilitet end PET-film og var derfor mere egnede til assayet. I proteinforsøg viste cutinase effektiv nedbrydning af PET-fibrene med markant frigivelse af kulstofprikker og et tydeligt fluorescenssignal, mens andre proteiner gav svagere respons. Metoden fremstår lovende som et mere tilgængeligt og tidsbesparende alternativ til traditionel analyse, men uensartet sammenhæng mellem frigivelsen af kulstofprikker og selve PET-nedbrydningen understreger behovet for yderligere optimering, før metoden kan anvendes bredt.
This thesis explores a fluorescence-based screening assay that uses carbon dots embedded in polyethylene terephthalate (PET) as visual markers of polymer degradation to identify proteins with catalytic activity toward PET. Carbon dots were synthesized and incorporated into PET as spin-coated films or electrospun fibers, and their release upon enzymatic degradation of the PET matrix was monitored by fluorescence emission spectroscopy and reversed-phase HPLC. Electrospun PET fibers provided higher surface area and better stability than PET films and were therefore more suitable substrates. In protein assays, cutinase effectively degraded the PET fibers and triggered substantial carbon-dot release with a clear fluorescent signal, whereas other proteins produced weaker responses. The approach shows promise as a more accessible and time-efficient alternative to traditional analyses, but variability in carbon-dot release as a direct proxy for PET degradation indicates that further optimization is needed before broader application.
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