Manipulation of ginsenoside heterogeneity of Panax notoginseng cells in flask and bioreactor cultivations with addition of phenobarbital

Abstract

Structure-similar ginsenosides have different or even totally opposite biological activities, and manipulation of ginsenoside heterogeneity is interesting and significant to biotechnological application. In this work, addition of 1 mM phenobarbital to cell cultures of Panax notoginseng at a relatively high inoculation size of 7.6 g dry cell weight (DW)/L enhanced the production of protopanaxatriol-type (Rg(1) + Re) ginsenosides in both shake flask and airlift bioreactor (ALR, 1 L working volume). The content of Rg(1) + Re in the ALR was increased from 42.5 +/- 4.0 mg per gram DW in untreated cell cultures (control) to 56.4 +/- 4.6 mg per gram DW with addition of 1.0 mM phenobarbital. The maximum productivity of Rg(1) + Re in the ALR reached 5.66 +/- 0.38 mg L(-1) d(-1), which was almost 3.3-fold that of control. The maximum ratio of the detectable ginsenosides protopanaxatriol:protopanaxadiol (Rb(1)) was 7.6, which was about twofold that of control. The response of protopanaxadiol 6-hydroxylase (P6H) activity to phenobarbital addition coincided with the above-mentioned change of ginsenoside heterogeneity (distribution). Phenobarbital addition is considered as a useful strategy for manipulating the ginsenoside heterogeneity in bioreactor with enhanced biosynthesis of protopanaxatriol by P. notoginseng cells.

Ginsenoside Rb3 exerts antidepressant-like effects in several animal models

“…In our research, the purity of Rb3 is 90% and some other types of ginsenoside such as Rb1, Rc are present as impurities. Yue and Zhong (2008) indicated that structurally similar ginsenosides may have different or even totally opposite effects, which may lead to the biphasic dose-related phenomenon shown in our results. Finally, Jiang et al (2007) researched the effects of different concentrations of Rb3 on persistent Na þ currents in ischemic and normal neurons, and found that Rb3 could inhibit persistent Na þ currents in a biphasic dose-related manner.…”

Enhanced production of individual ganoderic acids by integrating Vitreoscilla haemoglobin expression and calcium ion induction in liquid static cultures of Ganoderma lingzhi

“…The transcription levels of cyp512a2 , cyp512v2 and cyp512a13 were upregulated in the VHb strain, suggesting that the increased ratio may be related to higher transcription levels of those cytochrome P450 genes. In P. notoginseng , the induced activity of the cytochrome P450 enzyme protopanaxadiol 6‐hydroxylase increases the ratio of protopanaxatriol‐type to protopanaxadiol‐type ginsenoside (Yue and Zhong, ). The detailed roles of these cytochrome P450s in regulating GA distribution need to be further investigated in the future.…”

“…External environmental factors and genetic manipulation usually play important roles in regulating the heterogeneity of secondary metabolites. For instance, the ratio of the protopanaxatriol‐type to the protopanaxadiol‐type of ginsenosides can be altered by changing oxygen pressure and with the addition of phenobarbital in cell cultures of Panax notoginseng (Han and Zhong, ; Yue and Zhong, ). In Aurantiochytrium sp ., heterologous expression of the VHb gene increases the ratio of astaxanthin to beta‐carotene significantly (Suen et al ., ).…”

Ginsenosides in Panax genus and their biosynthesis

“…Transgenic ginseng plants with stable heredity and high saponin content productivity need to be further investigated. Main achievements of ginseng tissue and cell cultures in recent years are summarized in Table 2 92 , 94 , 169 , 173 , 178 , 184 , 186 , 187 , 188 , 191 , 192 , 193 , 194 , 195 , 196 , 197 , 198 , 199 , 200 , 201 , 202 , 203 , 204 , 205 , 206 , 207 , 208 , 209 , 210 , 211 , 212 , 213 , 214 , 215 .…”