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Squalene Synthase Gene from Medicinal Herb Bacopa monniera: Molecular Characterization, Differential Expression, Comparative Modeling, and Docking Studies

Plant Molecular Biology Reporter volume 33, pages 1675–1685 (2015)Cite this article

Abstract

The widespread pharmaceutically important triterpenoid saponins are synthesized via isoprenoid pathway. The formation of squalene is the key regulatory point in triterpene biosynthesis, catalyzed by squalene synthase (SQS). The present study deals with the detailed characterization of SQS by molecular, biochemical, and computational means from Bacopa monniera, an immensely important medicinal plant rich in triterpenoid saponin, bacosides. A full-length SQS gene was isolated from B. monniera, characterized as B. monniera squalene synthase (BmSQS) (1242 bp) encoding 414 amino acids. Deduced amino acid sequence of BmSQS showed highly conserved consensus aspartate-rich motifs (DXXXD) and catalytic site residues. Phylogenetic analysis showed that BmSQS belongs to dicot group having closest relationship with Salvia miltiorrhiza. Semiquantitative and real-time PCR studies showed that the BmSQS messenger RNA (mRNA) expression level was higher in vegetative parts (roots) as compared to floral parts. Methyl jasmonate induces the BmSQS mRNA expression in all tissues tested, while salicylic acid, cold, and salt induce much higher expression in roots. Homology modeling and docking simulations of BmSQS showed the pivotal roles of Asp77, Asp81, Asp213, Asp217, and Tyr168 in catalysis.

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Abbreviations

AMV-RT:

Avian myeloblastosis virus reverse transcriptase

BmSQS:

Bacopa monniera squalene synthase

FPP:

Farnesyl pyrophosphate

MJ:

Methyl jasmonate

qRT-PCR:

Quantitative real-time PCR

RACE:

Rapid amplification of cDNA ends

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Acknowledgments

Authors are thankful to the Council of Scientific and Industrial Research (CSIR), New Delhi, India, for financial support under the grant NWP0009.

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Authors and Affiliations

  1. Plant Tissue Culture Division, CSIR-National Chemical Laboratory, Dr. Homi Bhabha Road, Pune, 411 008, Maharashtra, India

    Rishi K. Vishwakarma, Krunal Patel, Prashant Sonawane, Uma Kumari, Somesh Singh,  Ruby, Shakeel Abbassi, Dinesh C. Agrawal & Bashir M. Khan

  2. Department of Applied Chemistry, Chaoyang University of Technology, Wufeng, Taichung Taiwan, Republic of China

    Rishi K. Vishwakarma, Dinesh C. Agrawal & Hsin-Sheng Tsay

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  1. Rishi K. Vishwakarma
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  2. Krunal Patel
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  3. Prashant Sonawane
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  4. Uma Kumari
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  5. Somesh Singh
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  6. Ruby
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  9. Hsin-Sheng Tsay
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  10. Bashir M. Khan
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Correspondence to Bashir M. Khan.

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Vishwakarma, R.K., Patel, K., Sonawane, P. et al. Squalene Synthase Gene from Medicinal Herb Bacopa monniera: Molecular Characterization, Differential Expression, Comparative Modeling, and Docking Studies. Plant Mol Biol Rep 33, 1675–1685 (2015). https://doi.org/10.1007/s11105-015-0864-z

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