BioModels

application/xml

https://www.ebi.ac.uk/biomodels/model/download/MODEL2310300002?filename=NothaGEM%20iSM1809.xmlhttps://www.ebi.ac.uk/biomodels/model/download/MODEL2310300002?filename=NothaGEM%20iSM1809.zipprimaryOK200Sarayu MuraliNon-curatedconstraint-based modelL3V1FROGhttps://www.ebi.ac.uk/biomodels/MODEL2310300002falseBioModelsModelsSBMLNothaGEM iSM18092023MODEL2310300002Murali SMurali S, Ibrahim M, Rajendran H, Shagun S, Masakapalli SK, Raman K, Srivastava S10.3389/fpls.2023.1207218, Camptothecin (CPT) is a vital monoterpene indole alkaloid used in anti-cancer therapeutics. It is primarily derived from Camptotheca acuminata and Nothapodytes nimmoniana plants that are indigenous to Southeast Asia. Plants have intricate metabolic networks and use them to produce secondary metabolites such as CPT, which is a prerequisite for rational metabolic engineering design to optimize their production. By reconstructing metabolic models, we can predict plant metabolic behavior, facilitating the selection of suitable approaches and saving time, cost, and energy, over traditional hit and trial experimental approaches. In this study, we reconstructed a genome-scale metabolic model for N. nimmoniana (NothaGEM iSM1809) and curated it using experimentally obtained biochemical data. We also used in silico tools to identify and rank suitable enzyme targets for overexpression and knockout to maximize camptothecin production. The predicted over-expression targets encompass enzymes involved in the camptothecin biosynthesis pathway, including strictosidine synthase and geraniol 10-hydroxylase, as well as targets related to plant metabolism, such as amino acid biosynthesis and the tricarboxylic acid cycle. The top-ranked knockout targets included reactions responsible for the formation of folates and serine, as well as the conversion of acetyl CoA and oxaloacetate to malate and citrate. One of the top-ranked overexpression targets, strictosidine synthase, was chosen to generate metabolically engineered cell lines of N. nimmoniana using Agrobacterium tumefaciens-mediated transformation. The transformed cell line showed a 5-fold increase in camptothecin production, with a yield of up to 5 µg g-1.. null, 14. Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai, India.msarayu.1993@gmail.com10.3389/fpls.2023.1207218Indian Institute of Technology MadrasfalseNothaGEM iSM1809No 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