Optimiranje biosinteze rekombinantnog proteina u bakteriji Escherichia coli BL21+P

master's thesis

• Title (english): Optimization of recombinant protein production in Escherichia coli BL21+P
• Author: Davor Valinger
• Mentor: Želimir Kurtanjek (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Process Engineering); (Laboratory for Measurement, Regulation and Automatisation); Zagreb
• Defense date and country: 2006-12-12, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

U radu je uspoređen šaržni uzgoj s 15 g L-1 glukoze u podlozi s tri šaržna uzgoja s pritokom supstrata u kojima je dodano također ukupno 15 g L-1 glukoze, ali u različitim vremenskim razmacima. Pokusi su rađeni u bioreaktorima radnog volumena 1,25 L a radni mikroorganizam je bio rekombinantna bakterija Escherichia coli BL21+P transformirana s ekspresijskim vektorom pXX7 koji sadrži gen za humani protein. Na taj način ispitan je utjecaj dinamike dodavanja glukoze na rast biomase, prinos rekombinantnog proteina, sintezu acetata, potrošnju kisika, anorganskog fosfora, alfa amino dušika i amonijačnog dušika. Efikasnost šaržnog uzgoja je po prinosu biomase i rekombinantnog proteina usporediva sa šaržnim uzgojem s pritokom supstrata ako je dodavanje glukoze rađeno rjeđe (dva i tri puta). U procesu s najčešćim dodavanjem glukoze (četiri puta) dobiveno je znatno više biomase i rekombinantnog proteina, najbrže se trošila glukoza, a bila je i najveća potrošnja alfa amino dušika, anorganskog fosfora i acetata.

Abstract (english)

The paper describes results of batch process with 15 g L-1 glucose in medium and three fed-batch procesess in which the same amount of glucose of 15 g L-1 was added but in different time intervals. Experiments were performed in 1,25 L bioreactors using recombinant bacteria Escherichia coli BL21+P transformed with expression vector pXX7 coding for recombinant human protein. In this way effect of glucose feeding on yield of biomass and recombinant protein, acetate production and cunsumption of oxygen inorganic phosphorus, alpha amino nitrogen and ammonium nitrogen was studied. Efficacy of batch process meassured in yield of biomass and recombinant protein was comparable with fed-batch procesess when glucose feeding was less frequent (two and three times). In the fed-batch process with most frequent glucose feeding (four times) considerable higher yield of biomass and recombinant protein was achieved, glucose was fastest consumpted, and the consumption of alpha amino nitrogen, inorganic phosporus and acetate was highest.

• Keywords: bakterija Escherichia coli; šaržni uzgoj; šaržni uzgoj s pritokom supstrata; glukoza; rekombinantni protein
• Keywords (english): Escherichia coli; batch process; fed-batch process; glucose; recombinant protein
• Language: croatian
• URN:NBN: urn:nbn:hr:159:673715
• Study programme: Title: Biotechnology; specializations in: Biochemical Engineering; Course: Biochemical Engineering; Study programme type: university; Study level: graduate; Academic / professional title: diplomirani inženjer biotehnologije; smjer biokemijsko inženjerstvo (diplomirani inženjer biotehnologije; smjer biokemijsko inženjerstvo)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-09-07 13:52:58