20.500.12556/DKUM-90899 Izboljšanje stabilnosti fermentacijske brozge klavulanske kisline pred izolacijo z optimiranjem procesnih parametrov magistrsko delo Enhancement of clavulanic acid fermentation broth stability before downstream operations by optimization of process parameters Bakterija Streptomyces clavuligerus med fermentacijskim procesom sintetizira klavulansko kislino (KK), ki učinkovito deluje kot inhibitor β-laktamaz in se uporablja v kombinaciji z antibiotiki za premagovanje bakterijske odpornosti. Nestabilnost KK med proizvodnim procesom predstavlja velik izziv, saj hitro razpada, kar zmanjšuje izkoristek in povečuje stroške proizvodnje. Namen te študije je izboljšati biosintezo KK z osredotočanjem na preučevanje vpliva procesnih parametrov na njeno stabilnost in produkcijo. V magistrskem delu smo razvili kvantitativni spektrometrični model za merjenje koncentracij KK in glicerola. Razvit model omogoča hitro, učinkovito in robustno analizo teh dveh spojin med eksperimentalnimi poskusi, kar predstavlja izboljšanje v primerjavi s klasičnimi kromatografskimi tehnikami, saj pospeši pridobivanje rezultatov. Model smo nato uporabili za spremljanje koncentracij KKin glicerola med eksperimentalnimi poskusi, v katerih smo spreminjali različne procesne parametre, kot so mešanje, zračenje, nadtlak, temperatura, redčenje ter kombinirali brozge iz različnih faz produkcijskega cikla S. clavuligerus. Poleg tega smo merili tudi koncentracijo raztopljenega kisika, viskoznost, pH in prisotnost nečistoče A, ki je razgradni produkt klavulanske kisline. Rezultati so pokazali, da lahko pravilna nastavitev procesnih parametrov bistveno izboljša stabilnost in produktivnost klavulanske kisline. Povečanje zračenja in mešanja ter zvišanje tlaka je prineslo do 5 % izboljšanje izkoristka po 6 urah zadrževalnega časa ter 17 % izboljšanje izkoristka po 27 urah procesa, hkrati pa je koncentracija nečistoče A ostala stabilna, kar kaže na manjšo razgradnjo KK. Posamezno povečanje mešanja, zračenja ali tlaka ni dalo enako učinkovitih rezultatov, čeprav je prispevalo k zmanjšanju razgradnje KK od 11 % do 26 %. Ugotovitve prispevajo k optimizaciji industrijskega procesa proizvodnje KK, saj zmanjšujejo razgradnjo produkta in povečujejo izkoristek, kar vodi v bolj učinkovit proces in nižje stroške proizvodnje. During the fermentation process, the bacterium Streptomyces clavuligerus synthesises clavulanic acid (CA), which acts effectively as an inhibitor of β-lactamases and is used in combination with antibiotics to overcome bacterial resistance. The instability of CA during the production process poses a major challenge as it degrades rapidly, reducing yields and increasing production costs. The aim of this study is to improve the biosynthesis of CA by focusing on the study of the influence of process parameters on its stability and production. In this Master thesis, a quantitative spectrometric model was developed to measure the concentrations of CA and glycerol. The developed model allows a fast, efficient and robust analysis of these two compounds during experimental trials, which is an improvement over classical chromatographic techniques as it speeds up the acquisition of results. The model was then used to monitor the concentrations of CA and glycerol during experimental trials in which different process parameters such as agitation, aeration, overpressure, temperature, dilution were varied and broths from different stages of the S. clavuligerus production cycle were combined. In addition, dissolved oxygen concentration, viscosity, pH and the presence of impurity A, a degradation product of CA, were measured. The results showed that the correct adjustment of process parameters can significantly improve the stability and productivity of CA. Increasing aeration and agitation and increasing the pressure resulted in up to a 5 % improvement in yield after 6 hours of residence time and a 17 % improvement in yield after 27 hours of the process, while the concentration of impurity A remained stable, indicating a lower degradation of the CA. Individual increases in agitation, aeration or pressure did not give the same effective results, although they contributed to a reduction in CA degradation of between 11 % and 26 %. The findings contribute to the optimisation of the industrial CA production process by reducing product degradation and increasing recovery, leading to a more efficient process and lower production costs. klavulanska kislina fermentacijski proces stabilnost optimizacija procesnih parametrov spektrometrija clavulanic acid fermentation process stability optimization of process parameters spectroscopy true true false [S. Zadravec] Slovenski jezik Angleški jezik Magistrsko delo/naloga 2024-10-02 21:01:41 2025-02-28 11:45:42 2025-03-15 03:10:04 0000-00-00 00:00:00 2025 0 Maribor 0 Maribor 1 spletni vir (1 datoteka PDF (XI, 72 str.)) 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 66.011:604.4(043.2) 229007107 MAG_Zadravec_Simon_2025.pdf MAG_Zadravec_Simon_2025.pdf 1 7AE713E3286C64513A06F53DA9A6C406 53ff8f042ad8377f5db6cf6ec07aa577587adca0c399a27d5c4b48d91f0b6cb4 491834d5-f536-11ef-8ea5-00155d000105 https://dk.um.si/Dokument.php?lang=slv&id=188887 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0