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\title{Highly efficient production of diverse rare ginsenosides using combinatorial biotechnology}



\author[1]{Ling-gai Cao}%
\author[1]{Hao Wu}%
\author[1]{He Zhang}%
\author[1]{Quan Zhao}%
\author[1]{Xue Yin}%
\author[1]{Dongran Zheng}%
\author[1]{Chuanwang Li}%
\author[1]{Min-jun Kim}%
\author[1]{Pyol Kim}%
\author[1]{Zheyong Xue}%
\author[1]{Yu Wang}%
\author[1]{Yuhua Li}%
\affil[1]{Northeast Forestry University}%


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  \date{June 18, 2024}


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\begin{abstract}
The rare ginsenosides are recognized as the functionalized molecules
after 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
MeJA-induced culture of adventitious roots in a 10 L bioreactor
associated with enzymatic hydrolysis using six \selectlanguage{greek}β-\selectlanguage{english}glycosidases and their
combination with yields ranging from 5.54-32.66 mg L-1. The yield of Rh2
was furthermore increased 7\% by using immobilized BglPm and Bgp1 in
optimized pH and temperature condition, with the highest yield reaching
51.17 mg L-1 (17.06\% of PPD-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.%
\end{abstract}%



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