OPTIMIZATION OF FRUCTANS EXTRACTION FROM in vitro CULTIVATED CHICORY ‘HAIRY’ ROOTS

Dependence of efficiency of fructans extraction on soaking time, temperature and time of high temperature extraction was investigated. Dried and powdered chicory Cichorium intybus L. cv Pala rossa ?hairy? roots obtained by Agrobacterium rhizogenes-mediated transformation with pCB161 vector were used for study. There were used low- and high-temperature extractions without heating at +22 °C during 0.5; 1 and 24 hours and with heating at +70 °C, +80 °C and +90 °C during 10, 20 and 30 minutes. Fructans fractionation was conducted by two ways: separation of high molecular weight fraction by crystallization at +4 °C and low molecular weight separation by extraction with 95% ethanol. To determine fructans concentration in the extracts McRary and Slattery method was used. Based on the experimental data, a mathematical model of fructan extraction process was created. Its adequacy was tested with the Fisher criterion and coefficient of determination. Optimal parameters of the extraction process chosen using the methods of linear programming were determined. Extraction for 30 minutes at 90 °C without soaking identified as the most tech nological one. It allowed to extract fructans general amount from transgenic roots (146 ±8,77 mg/g of root dry weight). Optimal regime of fructan obtaining from chicory ?hairy? roots is extraction at +90 °C for 30 min. Preliminary soaking time does not affect any effectiveness for such extraction. The most effective mode of obtaining of low- and high molecular fractions of fructans from transgenic chicory roots is twostage extraction with 95% ethanol at +80 °C and water at +90 °C with the duration of each stage of 30 minutes.