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Optimizacija gojenja bakterije Streptomyces rimosus za pridobivanje rekombinantne serinske proteinaze pernizin
Štefančić, Elena (Author), Petković, Hrvoje (Mentor) More about this mentor... This link opens in a new window, Paš, Maja (Co-mentor)

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Abstract
Cilj magistrske naloge je bila optimizacija produkcijskega gojišča za pridobivanje rekombinantne proteinaze pernizina s pomočjo Streptomyces rimosus. Optimizacijo gojišča smo začeli z gojiščem GOTC, ki je bilo v preteklosti optimizirano za produkcijo antibiotika oksitetraciklina. Preučevali smo, kako različni viri ogljika in dušika v gojišču vplivajo na produkcijo pernizina. Ugotovili smo, da koncentracija sojine moke kot vir dušika zelo vpliva na produkcijo pernizina. Koncentracija škroba nima bistvenega vpliva na produkcijo pernizina, kljub temu da smo pri nižjih koncentracijah škroba v gojišču določili višjo aktivnost pernizina. V prvem delu magistrske naloge smo uporabili dva različna produkcijska seva S. rimosus PER4 in PER5 ter ju primerjali s kontrolnim sevom KON (divji tip, ki ne producira pernizina). Glede na rezultate prvih poskusov smo v nadaljevanju uporabili le produkcijski sev PER5, ker smo pri tem sevu izmerili višjo aktivnost pernizina. V drugem delu magistrske naloge pa smo ugotovili, da dosežemo najvišjo aktivnost pernizina po 5 dneh gojenja. Prav tako smo ugotovili, da je razmerje med ogljikom in dušikom zelo pomembno. Ovrednotili smo tudi učinek različnih komponent gojišča, kot so enostavni sladkorji, anorganski in organski viri dušika in olja. Največji učinek na aktivnost pernizina sta imela ribje in sojino olje v nižjih koncentracijah (do 10 g/L). Visoko aktivnost pernizina smo izmerili tudi v gojiščih z drugimi viri dušika, kot so: sečnina, amonijev sulfat, hidrolizat sojine moke in kvas. Od testiranih virov ogljika, sta imela pozitiven učinek le manitol in dekstrin.

Language:Slovenian
Keywords:biotehnologija, Streptomyces rimosus, rekombinantna produkcija, serinska proteinaza, pernizin, optimizacija gojišča, sekundarni metaboliti
Work type:Master's thesis/paper (mb22)
Tipology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Year:2018
Publisher:[E. Štefančić]
UDC:602.3:579.873.7+604.4:577.15
COBISS.SI-ID:4888184 Link is opened in a new window
Views:463
Downloads:301
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Secondary language

Language:English
Title:Medium optimisation for production of recombinant serine proteinase pernisine in bacteria Streptomyces rimosus
Abstract:
The aim of the master's thesis was to optimize the production medium for obtaining recombinant proteinase pernisine using Streptomyces rimosus. The optimization of the medium was started with the GOTC medium, which has been optimized previously for the production of oxytetracycline antibiotics. We have studied how different sources of carbon and nitrogen in the production medium affect the production of perinsine. We found that the concentration of soy flour as a source of nitrogen greatly influences the production of pernisine. Concentration of starch does not have a significant effect on the production of pernisine, however, at lower concentrations of starch in the medium, higher pernisine activity was measured. In the first part of the experiment, two different S. rimosus production strains were used (PER4 and PER) and compared with the control strain KON (wild type not producing pernisine). Given the results of the first experiment, only the PER5 production strain was used hereafter, since higher pernisine activity was measured in this strain. In the second part of the master's thesis, however, we found that we achieved the highest pernsine activity after 5 days of cultivation. We also found that the ratio between carbon and nitrogen is very important. We evaluated the effect of various media components, such as simple sugars, inorganic and organic sources of nitrogen and oils. The most positive effect on the pernisine production was influenced by fish and soy oil at lower concentrations (up to 10 g/L). High pernisine activity was also measured in media with other sources of nitrogen, such as: urea, ammonium sulphate, soy protein flour and yeast hydrolyzate. Of the tested carbon sources, only mannitol and dextrin had a positive effect.

Keywords:biotechnology, Streptomyces rimosus, recombinant production, serine proteinase, pernisine, medium optimization, secondary metabolites

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