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. 2024 Apr 23;14(1):9318.
doi: 10.1038/s41598-024-56441-3.

Endophytic fungi of Panax sokpayensis produce bioactive ginsenoside Compound K in flask fermentation

Subecha Rai  1   2 , Laishram Shantikumar Singh  3   4 , Ramanan Uma Shaanker  5 , Kumaraswamy Jeyaram  1 , Tithi Parija  2 , Dinabandhu Sahoo  1   6
Affiliations

Endophytic fungi of Panax sokpayensis produce bioactive ginsenoside Compound K in flask fermentation

Subecha Rai et al. Sci Rep. .

Abstract

Endophytes of Panax have the potential to produce their host plant secondary metabolites, ginsenosides. Panax sokpayensis, an endemic traditional medicinal plant of the Sikkim Himalayas was explored for the isolation of endophytic fungi. In the present study, we have isolated 35 endophytic fungal cultures from the rhizome of P. sokpayensis and screened for ginsenosides production by HPLC by comparing the peak retention time with that of standard ginsenosides. The HPLC analysis revealed that out of 35 isolates, the mycelial extracts of four fungal endophytes (PSRF52, PSRF53, PSRF49 and PSRF58) exhibited peaks with a similar retention time of the standard ginsenoside, Compound K (CK). LC-ESI-MS/MS analysis led to the confirmation of ginsenoside CK production by the four fungal endophytes which showed a compound with m/z 639.6278, similar to that of standard ginsenoside CK with yield in potato dextrose broth flask fermentation ranging from 0.0019 to 0.0386 mg/g of mycelial mass in dry weight basis. The four prospective fungal endophyte isolates were identified as Thermothielavioides terrestris PSRF52, Aspergillus sp. PSRF49, Rutstroemiaceae sp. strain PSRF53, and Phaeosphaeriaceae sp. strain PSRF58 based on ITS sequencing. The present finding highlights the need for further study on growth optimization and other culture parameters to exploit the endophytes as an alternative source for ginsenoside CK production.

Keywords: Panax sokpayensis; Compound K; Endophytes; Fungi; Ginsenoside.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
Location map showing the sampling site at Jorbotay, Sikkim, India. The green colour star indicates the sample collection site (Map was created using the software ArcGIS 9.3 and 10.8.1; https://arcgis.software.informer.com/9.3/).
Figure 2
Figure 2
(A) Panax sokpayensis Shiva K. Sharma & Pandit in natural habitat and (B) rhizome of the plant.
Figure 3
Figure 3
Overlay HPLC chromatogram of Ginsenoside Compound K peak with retention time of 10.8 min (denoted in black), Spiked samples: mycelial extract of endophytic isolate PSRF53+ Compound K (denoted in pink), and the mycelial extract of endophytic isolate PSRF53 (denoted in blue). The arrow indicates the peak of Ginsenoside Compound K.
Figure 4
Figure 4
LC–ESI–MS/MS spectra. (A) The first mass spectra of mycelial extract of endophytic isolate PSRF53, the arrow indicates the molecular ion of ginsenoside Compound K (CK) at m/z 639.6260. (B) The MS/MS spectra of m/z 639.6260 of endophyte isolate PSRF53. (C) The MS/MS spectra of m/z 639.6278 of ginsenoside Compound K standard.
Figure 5
Figure 5
The phylogenetic tree based on ITS rDNA sequences constructed using the Maximum Likelihood method based on the Kimura 2-parameter model shows the taxonomic positions of four endophytic fungal isolates of P. sokpayensis produces ginsenoside Compound K in flask fermentation.
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