Indole Alkaloids Production by Catharanthus roseus Protoplasts with Artificial Cell Walls Containing of Guluronic Acid Rich Alginate Gel

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When low molecular weight alginate(guluronic acid(G)-oligomers, M.W.≨2, 000)was added to Catharanthus roseus cell culture, intracellularly stored ajmalicine was released into the broth. Addition of agarose and high molecular weight alginate did not promote the release of ajmalicine. C.roseus protoplasts were immobilized in guluronic acid rich(G-rich)alginate gel beads(artificial cell wall having a kind of elicitor function)and used for ajmalicine production. Immobilized protoplasts could be cultivated in a shake flask at low osmotic pressure, without disruption. The extracellular ajmalicine production by the protoplasts immobilized in alginate was much higher than that by the cells immobilized in alginate and protoplasts immobilized in agarose. On the 7 d of cultivation, cell wall regeneration in the immobilized protoplasts was detected under a fluorescence microscope. This implies that prevention of cell wall regeneration is a prerequiste for long term process with protoplasts. When 30 mM CaCl_2 was added to the broth, active protoplasts were maintained for 15d with neither cell wall regeneration nor inhibition of indole alkaloid production. The specific productivities of various indole alkaloids(ajmalicine, catharanthine and tryptamine)by the immobilized C.roseus protoplasts in the presence of CaCl_2 in the broth were much higher than those of the immobilized cells.
社団法人日本生物工学会の論文
1998-03-25