XXII. BIOCHEMICKÝ ZJAZD - Jesseniova lekárska fakulta

Posters
15.
PrODUCTION of rECOMBINaNT prOTEINS USING HIGH CELL DENSITY
CULTUrES of ESCHErICHIa COLI IN BIOreaCTOrs
Lucia Pánčiová, Zdenko Levarski, Pavol Utekal, Stanislav Stuchlík and Ján Turňa
Comenius University Faculty of Natural science, Department of Molecular Biology
Mathematical modeling of processes, information techniques and database generation
have received increasing interest of biotech industry in recent years. For the generation
of predictable mathematical models and databases of fermentation processes a vast
amount of data is required to precisely characterize the growth dynamics and microorganism
metabolism. These data are generated using systematic approaches. Systems
of metabolic engineering are an ideal way for the development of bacterial strains
producing bioproducts or aminoacids. Production of recombinant proteins using high
cell density cultures of Escherichia coli became one of the most often used methods in
biotech industry, mainly because of the high volumetric productivity associated with this
method, but also because of the vast amount of data available regarding to host genetics
and physiology. Application of different cultivation techniques results in achievement
of cell densities higher than 100 g of dry cell weight per liter of cultivation media. The
most common problems during cultivation of cells to high densities are limited oxygen
solubility in cultivation medium, formation of growth inhibitory by-products, accumulation
of acetate and CO 2
and generation of heat by the cells. The basic parameter of the
growth kinetics observation is the specific growth rate, which can be different for every
microorganism, type of the cultivation media or cultivation technique used. In this work,
we have focused on the construction of recombinant strains of E.coli, suitable cultivation
media selection, definition of crucial cultivation parameters, optimization of gene
expression, monitoring of growth kinetics and finally determination of cell viability and
biomass concentration.
130 XXII. Biochemistry Congress, Martin