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Optimizacija biosinteze rekombinantne fitaze v bakteriji Streptomyces rimosus.
Slemc, Lucija (Author), Petković, Hrvoje (Mentor) More about this mentor... This link opens in a new window, Paš, Maja (Co-mentor)

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Abstract
Bakterije iz rodu Streptomyces so poznane predvsem zaradi produkcije antibiotikov. Vse več pa je zanimanja za seve rodu Streptomyces za produkcijo heterolognih proteinov. Cilj magistrske naloge je bil optimizacija produkcijskega gojišča za produkcijo rekombinantne fitaze s pomočjo S. rimosus. Produkcijsko gojišče, ki se trenutno uporablja za kultivacijo S. rimosus je prilagojeno predvsem produkciji tetraciklinov. Pripravili smo različne vrste gojišč in merili fitazno aktivnost. V obsegu magistrske naloge smo izmed dveh sevov S. rimosus izbrali donosnejši sev z oznako e24 za nadaljnja testiranja. Pokazali smo, da ima razmerje med sojino moko in koruznim škrobom vpliv na biosintezo fitaze. Ko je koncentracija sojine moke v gojišču prenizka ali previsoka, se vrednosti fitazne aktivnosti znižajo v primerjavi z osnovnim (nespremenjenim) produkcijskim gojiščem. Vpliv glukoze je odvisen od koncentracije sojine moke in koruznega škroba in časa gojenja, vendar v večini primerov nima pozitivnega vpliva na produkcijo fitaze. Izmed vseh testiranih dodanih virov ogljika in dušika, imajo pozitiven vpliv na produkcijo fitaze: laktoza, dekstrin, sečnina, amonijev sulfat, kvas in kazein. Določili smo optimalno koncentracijo CaCO3, NaCl, cinka, mangana in železa. Dodatek fosforja v produkcijskem gojišču negativno vpliva na produkcijo fitaze. V okviru magistrske naloge smo določili optimalne koncentracije dodatkov in posameznih komponent gojišča za rekombinantno produkcijo fitaze v S. rimosus.

Language:Slovenian
Keywords:biotehnologija, Streptomyces rimosus, rekombinantna produkcija, fitaza, optimizacija gojišča
Work type:Master's thesis/paper (mb22)
Tipology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Year:2017
Publisher:[L. Slemc]
UDC:602.3:579.873.7:604.4:577.15(043.2)
COBISS.SI-ID:8762489 Link is opened in a new window
Views:1034
Downloads:374
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Secondary language

Language:English
Title:Optimisation of biosynthesis of recmobinant phytase in bacteria Streptomyces rimosus
Abstract:
Members of genus Streptomyces are well known antibiotics producers. Streptomyces sp. are also attractive host cells for heterologous protein production. The aim of our thesis was optimization of production medium for recombinant phytase production in S. rimosus. Currently used production medium for cultivation of S. rimosus is adjusted for antibiotics production. We prepared various mediums and measured phytase activity. Within master thesis we selected from two strains S. rimosu more productive strain e24 for further analysis. We have shown that ratio between soybean flour and starch effect on phytase activity. Low or high concentrations of soybean flour in medium decreased phytase activity regard unaltered production medium. Effect of added glucose depends on concentrations of soybean meal and corn starch and time of cultivation: but in most cases glucose repress phytase production. From all tested added carbon and nitrogen sources positive effect on phytase production have: lactose, dextrin, urea, ammonium sulfate, yeast and casein. We determined optimal concentration of CaCO3, NaCl, zinc, manganese and iron. Phosphate supplement has negative effect on phytase production. In context of the master's thesis we determined the optimal concentration of the individual components of the medium for the recombinant production of phytase.

Keywords:biotechnology, Streptomyces rimosus, recombinant production, phytase, medium optimization

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