원문정보
초록
영어
The rare ginsenosides have superior pharmacological activity than the major ginsenosides. Rare ginsenosides are usually an intermediate product of the ginsenoside biosynthesis pathway and do not accumulate in plants. Therefore, its application in medicine and functional foods is inhibited by the production amount. We developed an effective combinatorial biotechnology approach including large-scale production of ginseng adventitious root in plant bioreactor, hydrolysis of glycosidase combinations and immobilization of glycosidase. Firstly, we used 10 liters to 5 tons of plant bioreactor systems of different sizes to cultivate ginseng adventitious roots, and obtained large amounts of pesticide and heavy metal-free root materials with stable ginsenosides content. The biomass could be increased 30-fold within 60 days and the content of ginsenosides was similar to that of 5-years-old cultivated ginseng. Secondly, we screened out more than ten kinds of glycosidases according to the hydrolysis sites and compared the yield and hydrolysis efficiency of ginsenosides by several combinations of enzymes. Afterwards, a variety of effective combinations were obtained, including Bglsk+Bglmm, BglPm+Bgp1, BglSk+Bgp1, BglSk+BglPm+BgpA. 3g total ginseng saponins extracted from adventitious roots in a 10L bioreactor were converted into more than ten kinds of rare ginsenosides after the enzymes treatment, such as PPD, Rh2, Rg3 and CK. The contents are 336.42 mg, 326.61 mg, 138.54 mg and 279.27 mg, respectively. Thirdly, two kinds of β-glycosidases, BglPm and Bgp1, were immobilized on the hollow fiber membrane. After immobilization, the Km of BglPm and Bgp1 were remain unchanged, while the reaction rates were increased 3-fold and 5-fold, respectively. In the immobilized combination of two glycosidases, the yield of Rh2 was further increased to 511.72 mg. After 2 weeks of immobilization, the β-glycosidases activity remained above 80%. Our combinatorial biotechnology provides an efficient, low-cost, pollution-free and reusable method for large-scale production of diverse rare ginsenosides.