TY - JOUR

T1 - A translational synthetic biology platform for rapid access to gram-scale quantities of novel drug-like molecules

AU - Reed, James

AU - Stephenson, Michael J.

AU - Miettinen, Karel

AU - Brouwer, Bastiaan

AU - Leveau, Aymeric

AU - Brett, Paul

AU - Goss, Rebecca J.M.

AU - Goossens, Alain

AU - O'Connell, Maria A.

AU - Osbourn, Anne

N1 - Funding Information: This work was supported by a Norwich Research Park Studentship (J.R.), the joint Engineering and Physical Sciences Research Council/ Biotechnological and Biological Sciences Research Council (BBSRC)-funded OpenPlant Synthetic Biology Research Centre grant BB/L014130/1 (M.S., A.O.), European Union grant KBBE-2013-7 (TriForC) (K.M., A.G., A.O.), a John Innes Centre Knowledge Exchange and Commercialization grant (BB/KEC1740/1), the BBSRC Institute Strategic Programme Grant ?Understanding and Exploiting Plant and Microbial Metabolism? (BB/J004561/1) and the John Innes Foundation (A.O.). R.J.M.G. has received funding from the European Research Council under the European Union's Seventh Framework Programme (FP7/2007?2013/ERC grant agreement no 614779). We would like to thank George Lomonossoff for providing the pEAQ vectors, Andrew Davis for photography and Tyler Wooldridge (School of Pharmacy, University of East Anglia, UK) for assistance with the MTS/ELISA assays. Publisher Copyright: © 2017

PY - 2017

Y1 - 2017

N2 - Plants are an excellent source of drug leads. However availability is limited by access to source species, low abundance and recalcitrance to chemical synthesis. Although plant genomics is yielding a wealth of genes for natural product biosynthesis, the translation of this genetic information into small molecules for evaluation as drug leads represents a major bottleneck. For example, the yeast platform for artemisinic acid production is estimated to have taken >150 person years to develop. Here we demonstrate the power of plant transient transfection technology for rapid, scalable biosynthesis and isolation of triterpenes, one of the largest and most structurally diverse families of plant natural products. Using pathway engineering and improved agro-infiltration methodology we are able to generate gram-scale quantities of purified triterpene in just a few weeks. In contrast to heterologous expression in microbes, this system does not depend on re-engineering of the host. We next exploit agro-infection for quick and easy combinatorial biosynthesis without the need for generation of multi-gene constructs, so affording an easy entrée to suites of molecules, some new-to-nature, that are recalcitrant to chemical synthesis. We use this platform to purify a suite of bespoke triterpene analogs and demonstrate differences in anti-proliferative and anti-inflammatory activity in bioassays, providing proof of concept of this system for accessing and evaluating medicinally important bioactives. Together with new genome mining algorithms for plant pathway discovery and advances in plant synthetic biology, this advance provides new routes to synthesize and access previously inaccessible natural products and analogs and has the potential to reinvigorate drug discovery pipelines.

AB - Plants are an excellent source of drug leads. However availability is limited by access to source species, low abundance and recalcitrance to chemical synthesis. Although plant genomics is yielding a wealth of genes for natural product biosynthesis, the translation of this genetic information into small molecules for evaluation as drug leads represents a major bottleneck. For example, the yeast platform for artemisinic acid production is estimated to have taken >150 person years to develop. Here we demonstrate the power of plant transient transfection technology for rapid, scalable biosynthesis and isolation of triterpenes, one of the largest and most structurally diverse families of plant natural products. Using pathway engineering and improved agro-infiltration methodology we are able to generate gram-scale quantities of purified triterpene in just a few weeks. In contrast to heterologous expression in microbes, this system does not depend on re-engineering of the host. We next exploit agro-infection for quick and easy combinatorial biosynthesis without the need for generation of multi-gene constructs, so affording an easy entrée to suites of molecules, some new-to-nature, that are recalcitrant to chemical synthesis. We use this platform to purify a suite of bespoke triterpene analogs and demonstrate differences in anti-proliferative and anti-inflammatory activity in bioassays, providing proof of concept of this system for accessing and evaluating medicinally important bioactives. Together with new genome mining algorithms for plant pathway discovery and advances in plant synthetic biology, this advance provides new routes to synthesize and access previously inaccessible natural products and analogs and has the potential to reinvigorate drug discovery pipelines.

KW - Combinatorial biosynthesis

KW - Drug discovery

KW - Synthetic biology

KW - Terpenes

KW - Transient plant expression technology

KW - Triterpenoids

U2 - 10.1016/j.ymben.2017.06.012

DO - 10.1016/j.ymben.2017.06.012

M3 - Journal article

C2 - 28687337

AN - SCOPUS:85022047056

VL - 42

SP - 185

EP - 193

JO - Metabolic Engineering

JF - Metabolic Engineering

SN - 1096-7176

ER -