Mass production of secondary metabolite in plant cell culture by treatment of saccharide mixture in medium
The present invention relates to a method of producing secondary metabolites with a high yield using plant cell culture, and a medium for the production of secondary metabolites. More specifically, the method of the present invention is characterized in that the plant cell culture is conducted by adding a saccharide mixture to the culture medium as a carbon source, to increase the productivity of the secondary metabolites. For example, the present invention establishes the method of increasing the productivity of secondary metabolites and shortening the culture time by the use of the mixture of glucose and fructose in the plant cell culture, thereby contributing in producing useful secondary metabolites on an industrial scale using plant cell culture.
1. A method of producing secondary metabolites using Taxus genus plant cell culture, by culturing the plant cells in a culture medium for plant cell culture to which a saccharide mixture is added as the sole carbon source, to increase the productivity of secondary metabolites,
wherein the secondary metabolite is paclitaxel,
wherein the cell culture is conducted by adding the saccharide mixture at a cell growth phase and the secondary metabolite production phase,
wherein the contents of the saccharide mixture is 2 to 12% (w/v), and
wherein the saccharide mixture contains glucose and fructose in the mixture ratio by weight of 1:5 to 5:1 or sucrose and fructose in the mixture ratio by weight of 1:4 to 4:1.
2. The method according to claim 1 , wherein
the culture medium is selected from the group consisting of:
Anderson rhododendron medium, CHU(N6) medium, CLC/Ipomoea medium, Chee & Pool (C2D) vitis medium, De greef & jacobs medium, DKW/JUNGLANS medium, Eriksson(er) medium), Gamborg B5 medium, Gresshof & doy (DBM2) medium, Hellers medium, kao michayluk medium, knudson corchid medium, Lindemann Orchid medium, Litvay medium, Linsmaier & Skoog medium, McCowns woody plant medium, Murashige & Skoog medium, murashige & Miller medium, nitsch medium, NLN medium, orchimax medium, quoirin & Lepoivre medium, rugini olive medium, schenk & hildebrandt medium, S-Medium, vacin and went medium, white medium, westvaco WV3 medium, and a modified medium thereof;
the original carbon source is excluded therefrom; and
the saccharide mixture contains glucose and fructose in the mixture ratio by weight of 1:5 to 5:1 or sucrose and fructose in the mixture ratio by weight of 1:4 to 4:1 as a carbon source in the amount of 2 to 12%(w/v) in respect to the media.
3. The method according to claim 1 , wherein the plant cell is derived from a plant selected from the group consisting of Taxus bacata, Taxus brevifolia, Taxus canadensis, Taxus chinensis, Taxus cuspidata, Taxus floridana, Taxus globosa, Taxus media, Taxus wallichiana and Taxus yunnanensis.
4. The method according to claim 1 , wherein the plant cell culture is conducted by the method selected from the group consisting of a batch culture, a continuous culture, a fed-batch culture, a semi-continuous batch process, an immobilized culture, a two-phase culture, and a two stage culture.
5. The method according to claim 1 , wherein the plant cell culture is conducted by:
(i) culturing plant cells into the medium for plant cell culture containing the saccharide mixture at the temperature of 20 to 25° C.; and
(ii) continuously culturing the plant cells by changing the culture temperature into the range from 26° C. to 32° C.