magistrsko delo
Jernej Rotar (Author), Željko Knez (Mentor), Maja Leitgeb (Co-mentor)

Abstract

Bakterije Streptomyces clavuligerus v procesu fermentacije sintetizirajo klavulansko kislino, katera učinkovito deluje kot inhibitor β-laktamaze in se uporablja v kombinaciji z antibiotiki kot pomoč pri premagovanju bakterijske odpornosti. Pri tem se pojavlja veliko vprašanj glede vpliva substratov na proizvodnost. Zato je namen te študije izboljšati biosintezo klavulanske kisline, pri čemer se usmerimo k preučevanju vpliva aminokislin in mineralov, ki so ključni gradniki medija za fermentacijo klavulanske kisline. V magistrskem delu na laboratorijskem nivoju eksperimentalno iščemo vpliv sestave gojišča na produkcijsko sposobnost seva ter poskušamo optimizirati gojišče s katerim bi dosegli maksimalno produktivnost. Pri eksperimentalnem delu uporabljamo mikrobiološke in analitske metode. S pomočjo HPLC metode analiziramo vsebnost klavulanske kisline. Rezultati kažejo, da so minerali nepogrešljivi pri biosintezi klavulanske kisline. Železo in cink sta najpomembnejša minerala v fazi produkcije klavulanske kisline. Kalcij in magnezij pa negativno vplivata na produkcijo le te. Ugotovili smo, da dodatek aminokislin k že optimiranemu standardnemu mediju dodatno izboljša produkcijo klavulanske kisline. Dodatek arginina, asparagina, lizina in valina pozitivno vpliva na biosintezo produkta. Manjši pozitiven vpliv imajo tudi treonin, histidin in glicin. Aminokislini metionin in cistein pa negativno vplivata na produkcijo. Najboljše rezultate smo dosegli z dodatkom arginina, in sicer smo z dodatkom 10 mM L-arginina vsebnost izboljšali za 24 %. Dokazali smo, da je produktivnost klavulanske kisline odvisna tudi od same produkcijske zmožnosti bakterij S. clavuligerus, ki se od seva do seva razlikuje.

Keywords

klavulanska kislina;fermentacija;biosinteza;vpliv sestave gojišča;magistrske naloge;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UM FKKT - Faculty of Chemistry and Chemical Engineering
Publisher: [J. Rotar]
UDC: 582.23:[602.42:577.152.3](043.2)
COBISS: 21030166 Link will open in a new window
Views: 1270
Downloads: 26
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Other data

Secondary language: English
Secondary title: Influence of media composition to productivity of strain Streptomyces clavuligerus
Secondary abstract: Bacteria Streptomyces clavuligerus synthesizes clavulanic acid in the fermentation process, which effectively acts as a β-lactamase inhibitor, and is used in combination with antibiotics to help the overcome of bacterial resistance. There are many questions about the influence of substrates on production. The purpose of this study is to improve the clavulanic acid biosynthesis, focus on the study of the effects of amino acids and minerals, which are key building bloks for the fermentation of clavulanic acid. In the master's work at the laboratory level, we look experimentally for the influence of media composition to productivity of strain and we try to optimize the medium to achieve the maximum productivity. In the experimental work we use microbiological and analytical methods. With HPLC method we can measure the content of clavulanic acid in broth. The results indicate that the minerals are indispensable in the biosynthesis of clavulanic acid. Iron and zinc are the most important minerals in the production phase of clavulanic acid. Calcium and magnesium have a negative effect on the production. We think that the addition of amino acids to already optimazed standard medium further improve the productivity. The addition of arginine, asparagine, lysine and valine has a positive effect on biosynthesis of the product. Threonine, histidine and glycine also have a lower positive effect. Amino acids methionine and cysteine have a negative effect on productivity. We can achieve the best results with the addition of arginine, when we add the 10 mM concentration L-arginine, we improve the content of clavulanic acid for 24 %. We show that the productivity of clavulanic acid also depends on the production ability of bacteria S. clavuligerus, which varies from strain to strain.
Secondary keywords: clavulanic acid;Streptomyces clavuligerus;fermentation;biosynthesis;influence of media composition;productivity;
URN: URN:SI:UM:
Type (COBISS): Master's thesis/paper
Embargo end date (OpenAIRE): 2020-11-22
Thesis comment: Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Pages: XII, 64 str.
ID: 10859759