Enhancement of ginsenoside biosynthesis in high-density cultivation of Panax notoginseng cells by various strategies of methyl jasmonate elicitation

Abstract

A single addition of 200 microM methyl jasmonate (MJA) to high-density cell cultures of Panax notoginseng enhanced ginsenoside production in both shake-flask (250 ml) and airlift bioreactor (ALR; 1 l working volume). Repeated elicitation with two additions of 200 microM MJA during cultivation further induced the ginsenoside biosynthesis in both cultivation vessels. The content of ginsenosides Rg1, Re, Rb1 and Rd in the ALR was increased from, respectively, 0.18+/-0.01, 0.21+/-0.01, 0.21+/-0.02 and 0 mg per100 mg dry cell weight (DW) in untreated cell cultures (control) to 0.32+/-0.02, 0.36+/-0.02, 0.72+/-0.06 and 0.08+/-0.01 mg per100 mg DW with a single addition of MJA and further increased to 0.43+/-0.02, 0.46+/-0.03, 1.09+/-0.07 and 0.14+/-0.02 mg per100 mg DW with two additions of MJA. Interestingly, the activity of the Rb1 biosynthetic enzyme (UDPG-ginsenoside Rd glucosyltransferase), was also increased with a single elicitation by MJA and increased again by a repeated elicitation, which coincided well with the trend in the increase in Rb(1) content. In order to further improve the cell density and ginsenoside production, a strategy of MJA repeated elicitation combined with sucrose feeding was adopted. The final cell density and total ginsenoside content in the ALR reached 27.3+/-1.5 g/l and 2.02+/-0.06 mg per100 mg DW; and the maximum production of ginsenoside Rg1, Re, Rb1 and Rd was 111.8+/-4.7, 117.2+/-4.6, 290.2+/-5.1 and 32.7+/-8.1 mg/l, respectively. The strategies demonstrated and the information obtained in this work are useful for the efficient large-scale production of bioactive ginsenosides by plant cell cultures.