A Simple Method for the Preparation of Crude Gintonin from Ginseng Root, Stem, and Leaf

Abstract

Ginseng has been used as a general tonic agent to invigorate the human body as an adaptogenic agent. In a previous report, we have shown that ginseng contains a novel glycolipoprotein called gintonin. The main function of gintonin is to transiently enhance intracellular free Ca2+ [Ca2+]i levels in animal cells. The previous method for gintonin isolation included multiple steps using organic solvents. In the present report, we developed a simple method for the preparation of crude gintonin from ginseng root as well as stem and leaf, which produced a higher yield of gintonin than the previous one. The yield of gintonin was 0.20%, 0.29%, and 0.81% from ginseng root, stem, and leaf, respectively. The apparent molecular weight of gintonin isolated from stem and leaf through sodium dodecyl sulfate polyacrylamide gel electrophoresis was almost same as that from root but the compositions of amino acids, carbohydrates or lipids differed slightly between them. We also examined the effects of crude gintonin from ginseng root, stem, and leaf on endogenous Ca2+-activated Cl- channel (CaCC) activity of Xenopus oocytes through mobilization of [Ca2+]i. We found that the order of potency for the activation of CaCC was ginseng root > stem > leaf. The ED50 was 1.4±1.4, 4.5±5.9, and 3.9±1.1 μg/mL for root, stem and leaf, respectively. In the present study, we demonstrated for the first time that in addition to ginseng root, ginseng stem and leaf also contain gintonin. Gintonin can be prepared from a simple method with higher yield of gintonin from ginseng root, stem, and leaf. Finally, these results demonstrate the possibility that ginseng stem and leaf could also be utilized for ginstonin preparation after a simple procedure, rather than being discarded.

An Edible Gintonin Preparation from Ginseng

“…In previous studies we demonstrated that gintonin induced CaCC activation in Xenopus oocytes through the mobilization of [Ca 2+ ] i [5,6]. When we compared the potency for endogenous CaCC activation in Xenopus oocytes with the previously acquired extract, we found that ethanol gintonin was not much different from that of previous gintonin for the activation of CaCC.…”

“…We demonstrated that active agents that caused CaCC activation in Xenopus oocytes and transiently elevated the intracellular calcium concentration ([Ca 2+ ] i ) in mammalian cells are not ginsenosides but gintonin [5]. In addition, we found that the ginseng stem and leaf also contain gintonin [6]. Thus, we provided the evidence that ginseng contains novel ingredients and that one novel ingredient, gintonin, could also be the main agent for Ca 2+ -mediated various cellular events.…”

“…Since Ca 2+ is a second messenger and is involved in numerous Ca 2+ -dependent events such as Ca 2+ -dependent enzymes, ion channels, hormones and neurotransmitter release, and more, gintonin might participate a variety of cellular events. In previous reports, we have shown that ginseng can be extracted with various organic solvents such as methanol and butanol as a first step for gintonin preparation from ginseng root, stem, and leaf [5,6]. For use of gintonin as a functional health food and/or drug, a new method of extraction that does not use harmful organic solvents is necessary.…”

“…First, we showed that gintonin can be prepared using ethanol. Second, we also used a one-step extraction instead of previous multiple step extractions using methanol and butanol [5,6]. This simplified method for gintonin preparation has at least three merits compared to previous methods.…”

“…The crude gintonin from ginseng was subjected to 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) [6]. Edible gintonin (100 μg each) from ginseng root was loaded in each lane.…”