Effects of Immobilization and Permeabilization Procedures on Growth of Chenopodium rubrum Cells and Amaranthin Concentration

Abstract

Cultures of Chenopodium rubrum were subjected to permeabilization and immobilization procedures to examine their potential for pigment production. Amaranthin content of culture media was highest for cultures treated with dimethylsulfoxide (DMSO) or dissolved chitosan. Labelling of the cells with L‐(U‐14C) tyrosine revealed that amaranthin was released from the cells but degraded rapidly. Product degradation in chitosan immobilized cells was delayed by 12–24 hr. Sufficient cell growth was observed in cultures treated with 0.77 mg chitosan per gram fresh biomass, with 0.42 ml.g‐1 DMSO, or when immobilized in a complex Ca‐alginate‐chitosan gel system. All other treatments resulted in inhibition of growth.

Elicitation and permeabilisation affect the accumulation and storage profile of phytoestrogens in high productive suspension cultures of Genista tinctoria

“…6,7,8). Contrary to literature reports where maximum concentrations of the analysed metabolites were observed in media already half an hour after permeabilisation (Bordelius 1988;Knorr and Berlin 1987), in the described experiment this result was only achieved in the 7th hour of the experiment (Table 5 and Figs. 6,7,8).…”

“…The relatively slow release of isoflavonoids from G. tinctoria cells was probably caused by low concentration of DMSO in the culture (2%) and the fact that it was only supplemented in the stationary phase of biomass growth, i.e. prior to the adoption of the experimental procedure which was different from the previously described models (Bordelius 1988;Gontier et al 2002;Knorr and Berlin 1987;Komolpis et al 1998).…”

“…The effect of a DMSO supplement on the growth of G. tinctoria suspension and the accumulation of isoflavonoids Unlike physical permeabilisation, chemical permeabilisation is a relatively inexpensive and simple method allowing for a release of secondary metabolites from plant matrices to growth media (Bordelius 1988;Kanaraiah et al 2005;Knorr and Berlin 1987;Komolpis et al 1998;Teissie et al 2005). The choice of DMSO to permeabilise G. tinctoria suspension was based on literature reports indicating that this compound is one of the most effective factors changing cell permeability and causing not only a considerable release of the analysed metabolites into the growth media, but also, in low concentration (2-10%) allowing the cultivated tissues to remain biochemically active (Bordelius 1988;Knorr and Berlin 1987;Komolpis et al 1998;Teissie et al 2005).…”

“…The choice of DMSO to permeabilise G. tinctoria suspension was based on literature reports indicating that this compound is one of the most effective factors changing cell permeability and causing not only a considerable release of the analysed metabolites into the growth media, but also, in low concentration (2-10%) allowing the cultivated tissues to remain biochemically active (Bordelius 1988;Knorr and Berlin 1987;Komolpis et al 1998;Teissie et al 2005). In order to avoid decay of G. tinctoria biomass when cultivated in the presence of DMSO (Bordelius 1988;Knorr and Berlin 1987), the compound was added to the medium in relatively low concentrations (20 ml l -1 ), and only on day 26 of the experiment, i.e. after the intensive growth of cells was completed and the maximum concentration of isoflavones in the culture was achieved (Table 4 and Fig.…”

“…This would reduce the costs of industrial production of phytoestrogens based on G. tinctoria suspension (Bourgaud et al 2001;Luczkiewicz and Głód 2005;Ramachandra Rao and Ravishankar 2002). The intention was to stimulate a release of isoflavonoids to the experimental medium by chemical permeabilisation of cellular membranes in G. tinctoria suspension (Bordelius 1988;Bourgaud et al 2001;Knorr and Berlin 1987;Komolpis et al 1998;Ramachandra Rao and Ravishankar 2002). The direct permeabilising factor in the described series of experiments was dimethyl sulfoxide (DMSO), which had been successfully used to evoke a release of secondary metabolites from plant matrices grown in vitro (Bordelius 1988;Kanaraiah et al 2005;Komolpis et al 1998).…”