Project Description
Pueraria lobata var mirifica (P. mirifica) is a plant native to Thailand and other parts of Southeast Asia, with its roots being a century long staple in Thai traditional medicine[1]. This usage is due to its content of puerarin, daidzein, and genistein, which are all phytoestrogens that mimic the estrogen hormone[5]. These isoflavonoids also possess essential antioxidative and neuroprotective properties, making P. mirifica a valuable plant. Its medicinal properties contribute to the widespread selling of P. mirifica herbal supplements across Thailand and Southeast Asia[4]. However, the plant’s tuberous roots have been heavily exploited for commercial purposes, putting the species at risk for eventual extinction[1].
Currently, the means of producing P. mirifica supplements is unsustainable with the entire plant getting uprooted during harvest due to its tuberous nature [3]. These practices of harvesting result in significant physical disruptions to the surrounding soil and natural habitat, leading to erosion, degradation of soil structure, and potential damage to local ecosystems. Moreover, P. mirifica has a long maturation period, with a fully differentiated P. mirifica that provides the optimal yield of isoflavanoids taking 3 years to grow [2]; thereby, consuming a lot of water, the scarcity of which is amplified with increasing frequency of droughts in Thailand. All this makes the continual production of P. mirifica supplements significantly more difficult to uphold, making this herbal medicine less accessible to those who could benefit from it.
Our Solution
The Thailand-RIS team aims to address the issues related to the cultivation of P.mirifica by developing a more sustainable and less resource-intensive way to produce isoflavonoids. Through synthetic biology, the enzymes responsible for the production of phytoestrogen in P.mirifica will be relocated to another plant that is easier to cultivate. Currently, the enzymes required to create puerarin are only found in a few leguminous plants [7]. However, by relocating the production of all the key enzymes involved in the isoflavonoid biosynthetic pathway for puerarin to Nicotiana benthamiana (N. benthamiana), we can produce puerarin more efficiently [6]. Unlike P. mirifica, N. benthamiana is easier to grow as it takes only a mere 2 months to mature and is not a tuber, thereby reducing resource consumption, soil disruption, and habitat destruction. This approach not only minimizes environmental impact, but also boosts the production of beneficial isoflavonoids such as puerarin, daidzein, and genistein, making this plant’s medicinal properties more accessible to those who need it.
Figure 1. Isoflavonoid Biosynthetic Pathways in P.mirifica
As illustrated in Fig 1, P. mirifica’s puerarin production involves many key enzymes. Non-leguminous plants typically only produce metabolites up to cinnamic acid and lack a complete Isoflavanoid pathway[4]. By introducing the production of the enzymes CHS, CHR, CHI, IFS, HID, and C-UGT into N. benthamiana, we can promote more sustainable production and improve access to puerarin supplements.
References
1. Cherdshewasart, W., Kitsamai, Y., & Malaivijitnond, S. (2007). Evaluation of the Estrogenic Activity of the Wild Pueraria mirifica by Vaginal Cornification Assay. Journal of Reproduction and Development, 53(2), 385–393. https://doi.org/10.1262/jrd.18065
2. ---. (2008). Variance of estrogenic activity of the phytoestrogen-rich plant. Maturitas, 61(4), 350–357. https://doi.org/10.1016/j.maturitas.2008.09.017
3. Intharuksa, A., Kitamura, N., Peerakam, N., Charoensup, W., Ando, H., Sasaki, Y., and Sirisa-Ard, P. (2020). Evaluation of white Kwao Krua (Pueraria candollei Grah. ex Benth.) products sold in Thailand by molecular, chemical, and microscopic analyses. Journal of Natural Medicines, 74(1), 106–118. https://doi.org/10.1007/s11418-019-01351-2
4. News, T. M. (n.d.). Medical Benefits of Kwao Krua (Pueraria mirifica) For Women - Thailand Medical News. Medical Benefits of Kwao Krua (Pueraria Mirifica) For Women - Thailand Medical News. Retrieved June 14, 2024, from https://www.thailandmedical.news/news/medical-benefits-of-kwao-krua-pueraria-mirifica-for-women
5. (PDF) Pueraria candollei var. mirifica: A precious source of pharmaceuticals and cosmeceuticals. (n.d.). Retrieved September 21, 2024, from https://www.researchgate.net/publication/359237047_Pueraria_candollei_var_mirifica_A_precious_source_of_pharmaceuticals_and_cosmeceuticals
6. Suntichaikamolkul, N., Tantisuwanichkul, K., Prombutara, P., Kobrakul, K., Zumsteg, J., Wannachart, S., Schaller, H., Yamazaki, M., Saito, K., De-ekanmkul, W., Vimolmangkang, S., & Sirikantaramas, S. (2019). Transcriptome analysis of Pueraria candollei var. mirifica for gene discovery in the biosyntheses of isoflavones and miroestrol. BMC Plant Biology, 19(1), 581. https://doi.org/10.1186/s12870-019-2205-0
7. White Kwao Krua (Pueraria mirifica). (n.d.). Retrieved June 14, 2024, from https://en.nanomed.co.th/15169529/white-kwao-krua-pueraria-mirifica