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Izolacija lipidov iz paradontalnih bakterij ter njihova interakcija z egerolizinom erilizinom A
ID Tori, Tilen (Author), ID Skočaj, Matej (Mentor) More about this mentor... This link opens in a new window, ID Grundner, Maja (Co-mentor)

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Abstract
Parodontoza je ena izmed najpogostejših (vnetnih) bolezni moderne družbe. Za razvoj bolezni so s svojimi virulentnimi dejavniki ključne predvsem parodontalne bakterije, med katere uvršamo tudi bakteriji Tannerella forsythia in Porphyromonas gingivalis. Z namenom iskanja inovativnih načinov diganostike zgodnjih oblik, obvladovanja ali zdravljenja parodontalne bolezni, smo iz bakterij T. forsythia in P. gingivalis izolirali lipide in jim z lipidomskimi analizami določili lastnosti. Rezultati so pokazali prisotnost različnih oblik manj znanega lipida ceramid fosfoetanolamina. Ker je omenjeni lipid v membranah sesalskih celic prisoten v zelo nizkih količinah, bi lahko predstavljal lipidno vrsto, ki jo v ustni votlini najdemo izključno ob prisotnosti parodontalne bolezni in je v korelaciji s povečananim številom parodontalnih bakterij. V magistrski nalogi smo preverjali ali je egerolizin erilizin A primerno molekularno orodje za detekcijo ceramid fosfoetanolamina v umetnih lipidnih sistemih in to tudi potrdili. S kombinacijo erilizina A in njegovega proteinskega partnerja pleurotolizina B smo s testom permeabilizacije veziklov, ki so bili sestavljeni iz celokupnih bakterijskih lipidov, dodatno potrdili vezavo erilizina A na bakterijske lipide ter pokazali, da kombinacija proteinov erilizina A in pleurotolizina B tvori pore v umetnih bakterijskih veziklih in posledično povzroči njihovo lizo. Glede na dobljene rezultate lahko zaključimo, da je (i) erilizin A potencialna molekula za detekcijo ceramid fosfoetanolamina v umetnih lipidnih membranah, sestavljenih iz celokupnega lipidnega ekstrakta parodontalnih bakterij P. gingivalis in T. forsythia in da (ii) erilizin A v kombinaciji s partnerjem pleurotolizinom B izkazuje litični potencial, kar bi lahko izkoristili v zdravljenju parodontalne bolezni.

Language:Slovenian
Keywords:parodontoza, parodontalne bakterije, Tannerrela forsythia, Porphyromonas gingivalis, erilizin A, ceramid fosfoetanolamin, pleurotolizin B
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Place of publishing:Ljubljana
Publisher:[T. Tori]
Year:2021
PID:20.500.12556/RUL-128041 This link opens in a new window
UDC:579.61:616.314-078:577.2.08
COBISS.SI-ID:69307651 This link opens in a new window
Publication date in RUL:02.07.2021
Views:965
Downloads:64
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Secondary language

Language:English
Title:Isolation of lipids from periodontal bacteria and their interaction with aegerolysin erylysin A
Abstract:
Periodontal disease is one of the most common (inflammatory) diseases nowadays. Responsible organisms for the development of the disease are periodontal bacteria, such as Tannerella forsythia and Porphiromonas gingivalis. To investigate new possibilities in diagnosis and treatment of periodontal disease, we extracted lipids from T. forsythia and P. gingivalis and performed lipidomic analysis. The results showed the presence of several isoforms of the lipid ceramide phosphoethanolamine. This lipid is almost absent in mammalian membranes; therefore, it could be used for detecting pariodontal bacteria as their membranes contain ceramide phosphoethanolamine. On the other hand, proteins from the aegerolysin family emerged as suitable molecules for labeling ceramide phosphoethanolamine. Therefore, we have used aegerolysin erylysin A and successfully confirmed the presence of ceramide phosphoethanolamine in lipid extracts of bacteria T. forsythia and P. gingivalis. The binding of erylysin A to vesicles composed of total lipid extracts from T. forsythia and P. gingivalis was also confirmed with the permeabilization test by using erylysin A and its protein partner pleurotolysin B. Based on the obtained results we can conclude that erylysin A is a promising molecule for detection of ceramide phosphoethanolamine in lipid vesicles obtained from the total lipid extracts from periodontal bacteria P. gingivalis and T. forsythia. In combination with pleurotolysin B, erilysin A could also represent a potential solution for the treatment of periodontal disease.

Keywords:periodontitis, periodontal bacteria, Tannerrela forsythia, Porphyromonas gingivalis, erilysin A, ceramide phosphoethanolamine, pleurotolysin B

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