关键词: 生物化学工程, 人参发根R9923, 液体培养, 人参皂苷Rb₁, 分离纯化, 硅胶柱层析

Abstract: Hairy roots of ginseng were directly inoculated from the root explants of two-year-old ginseng by Agro bacterium rhizogene A₄. The specific growth rate and the ginsenoside content of sixteen kinds of hairy roots were measured, from which three high-yielding clones were selected. The highestyielding clone was R9923 whose biomass increased by 28.8 times after four weeks culturing. The growth of the hairy root and the development of ginsenoside of clone R9923 were investigated and it was found that the ginsenoside content increases with the biomass of the hairy root. The contents of monomer ginsenoside Rg₁, Re and Rb₁in clone R9923 were determined by HPLC method. The total content of ginsenoside in the hairy root after four weeks culturing came up to 15.2 mg/g, in which 68.3% was Rb₁. The content of Rb₁in the hairy root was 2.3 times of that of three-year-old ginseng. The ginsenoside Rb₁was separated and purified from the hairy root by silica gel column chromatography, in which the optimal solvent ratio of chloroform to methanol was 7.5∶2.5, the purity of Rb₁was 89.95% and the yield was 72.7%.

Key words: biochemistry engineering, ginseng hairy root R9923, liquid culture, ginsenoside Rb₁, separation and purification, silica gel column chromatography