Abstract
Background
Picrorhiza kurroa has been reported as an age-old ayurvedic hepato-protection to treat hepatic disorders due to the presence of iridoids such as picroside-II (P-II), picroside-I, and kutkoside. The acylation of catalpol and vanilloyl coenzyme A by acyltransferases (ATs) is critical step in P-II biosynthesis. Since accumulation of P-II occurs only in roots, rhizomes and stolons in comparison to leaves uprooting of this critically endangered herb has been the only source of this compound. Recently, we reported that P-II acylation likely happen in roots, while stolons serve as the vital P-II storage compartment. Therefore, developing an alternate engineered platform for P-II biosynthesis require identification of P-II specific AT/s.
Methods and results
In that direction, egg-NOG function annotated 815 ATs from de novo RNA sequencing of tissue culture based ‘shoots-only’ system and nursery grown shoots, roots, and stolons varying in P-II content, were cross-compared in silico to arrive at ATs sequences unique and/or common to stolons and roots. Verification for organ and accession-wise upregulation in gene expression of these ATs by qRT-PCR has shortlisted six putative ‘P-II-forming’ ATs. Further, six-frame translation, ab initio protein structure modelling and protein-ligand molecular docking of these ATs signified one MBOAT domain containing AT with preferential binding to the vanillic acid CoA thiol ester as well as with P-II, implying that this could be potential AT decorating final structure of P-II.
Conclusions
Organ-wise comparative transcriptome mining coupled with reverse transcription real time qRT-PCR and protein-ligand docking led to the identification of an acyltransferases, contributing to the final structure of P-II.
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Data Availability
All data generated or analyzed during this study are included in this published article [and its supplementary information files].
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Acknowledgements
The authors thank the Department of Biotechnology, Ministry of Science and Technology, Government of India for providing financial support to RSC in the form of Programme Support on high altitude medicinal herbs. SS thank Bennett University for providing PhD research fellowship.
Funding
This work was financially supported by Department of Biotechnology, Ministry of Science and Technology, Government of India (Grant number: BT/COE/34/SP15268/2015) in the form of a Programme Support on high-value medicinal plants under Centres of Excellence.
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RSC conceived and designed the central flow and objectives to be investigated in the current research. AK, SS, and AS performed more than 90% of the crucial experiments (in silico and wet lab) described in this study and compiled the results section. NN was involved during pursual of primary initiative work for this publication. RP provided the HPLC chromatograms. DB analysed the data and wrote the first complete draft of the manuscript. RSC analysed the data, meticulously edited and finalized the complete draft manuscript for communication.
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Kharb, A., Sharma, S., Sharma, A. et al. Capturing acyltransferase(s) transforming final step in the biosynthesis of a major Iridoid Glycoside, (Picroside-II) in a Himalayan Medicinal Herb, Picrorhiza kurroa. Mol Biol Rep 49, 5567–5576 (2022). https://doi.org/10.1007/s11033-022-07489-9
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DOI: https://doi.org/10.1007/s11033-022-07489-9