Bioprocess Optimization of Furanocoumarin Elicitation by Medium Renewal and Re-elicitation: A Perfusion-Based Approach

Abstract

Effect of various abiotic (methyl jasmonate, salicylic acid) and biotic (yeast extract, Aspergillus niger) elicitors on furanocoumarin production and in situ product removal was studied using shoot cultures of Ruta graveolens L. Elicitation by yeast extract (1% w/v) on day 15 was most effective. It led to 7.8-fold higher furanocoumarin production that was attained 24 h after elicitation and 43% of the product was released into the medium. Changes in the relative concentration of furanocoumarins produced depend on the elicitor used. Molar ratio of bergapten increased to 93% in response to yeast extract. With the perspective of developing a commercially feasible process, an approach for preserving viability of biomass and its reuse needs to be developed. For this, medium renewal strategy was investigated. Removal of the spent medium 48 h after elicitation allowed in situ product removal and proved effective in revival of cultures, allowing reuse of biomass. A week after medium renewal, the revived biomass was re-elicited and a second furanocoumarin production peak was obtained. A perfusion-based bioprocess optimization approach, employing elicitation coupled with medium renewal with subsequent re-elicitation, as a new strategy for improved furanocoumarin production, has been suggested.

Enhanced in vitro production of diosgenin in shoot cultures of Dioscorea deltoidea by elicitation and precursor feeding

“…SA is generally produced in plants and accumulated at pathogen attack sites, and then spreads to several plant parts triggering the biosynthetic pathway for accumulation of secondary metabolite production [45]. This is an agreement with Diwan and Malpathak [41] and Coste et al [34] who reported that 200 µM SA increased the furanocoumarin production in Rutagraveolens shoot cultures and hypericin and pseudohypericin content in Hypericum maculatum shoot cultures. However, 50 µM SA signi cantly reduced the diosgenin accumulation in micro-tubers of Chlorophytum borivilianum [46].…”

Elicitation of furanocoumarins in poison hemlock (Conium maculatum L.) cell culture

“…Furthermore, elicitation of C. maculatum plants with copper(II) chloride (10 mM The differential accumulation of furanocoumarins in the medium or the cells after elicitation with various elicitors is unexpected, but not completely unprecedented. Furanocoumarins are also released into the medium after elicitation by Ruta graveolens (Diwan and Malpathak 2011) and Glehnia littoralis (Ishikawa et al 2009). Diwan and Malpathak (2011) reported that in their elicitation system the amount of furanocoumarins is dependent on the elicitor, the most efficient being yeast extract.…”

“…Furanocoumarins are also released into the medium after elicitation by Ruta graveolens (Diwan and Malpathak 2011) and Glehnia littoralis (Ishikawa et al 2009). Diwan and Malpathak (2011) reported that in their elicitation system the amount of furanocoumarins is dependent on the elicitor, the most efficient being yeast extract. In our system biotic compounds (alginic acid, cellulase and chitosan) were also the most efficient.…”

Enhanced in vitro production of diosgenin in shoot cultures of Dioscorea deltoidea by elicitation and precursor feeding

“…This suggested that elicitor used and its concentrations play an important role on the effectiveness of elicitation. In previous studies, it was reported that MeJa has increased furanocoumarin production in Ruta graveleons shoot cultures and asiaticoside production in Centella asiatica whole plant culture [41,36]. Mendoza et al [28] reported that elicitation with 3 µM MeJa increased phenolics and avonoids content in Thevetia peruviana suspension cultures.…”