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 μM 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 μM 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 Rb₁ 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.