Highly efficient production of diverse rare ginsenosides using combinatorial biotechnology

The rare ginsenosides are recognized as the functionalized molecules after the oral administration of Panax ginseng and its products. The sources of rare ginsenosides are extremely limited because of low ginsenoside contents in wild plants, hindering their application in functional foods and drugs. We developed an effective combinatorial biotechnology approach including tissue culture, immobilization, and hydrolyzation methods. Rh2 and nine other rare ginsenosides were produced by methyl jasmonate‐induced culture of adventitious roots in a 10 L bioreactor associated with enzymatic hydrolysis using six β‐glycosidases and their combination with yields ranging from 5.54 to 32.66 mg L−1. The yield of Rh2 was furthermore increased by 7% by using immobilized BglPm and Bgp1 in optimized pH and temperature conditions, with the highest yield reaching 51.17 mg L−1 (17.06% of protopanaxadiol‐type ginsenosides mixture). Our combinatorial biotechnology method provides a highly efficient approach to acquiring diverse rare ginsenosides, replacing direct extraction from Panax plants, and can also be used to supplement yeast cell factories. The rare ginsenosides of Panax ginseng have been shown to have multiple medical effects such as anti‐cancer, anti‐aging, anti‐inflammatory and other health‐improving activities. The sources of rare ginsenosides are extremely limited, hindering their application in functional foods and drugs. This work provided a set of solutions including culture of ginseng adventitious roots, hydrolysis of major ginsenosides, and the immobilization of β‐glucosidases, producing large amounts of ten types of rare ginsenosides.