Rapid prototyping and engineering of plant-based products Anne Osbourn anne.osbourn@jic.ac.uk
Plants are a rich source of drugs and other high-value compounds
Triterpenes – examples of plant metabolic diversity Sapindus mukorossi Avena sativa
S. Mukorossi saponin
Avenacin A-1
Molluscicidal effects
Antifungal/ Allelopathy
Plant defence Medicago truncatula
Insecticidal properties
Anti-feedant
Soyasaponin βg 3-GlcA-28-Ara-Rha-Xyl-medicagenate
Glycine max
Triterpenes – examples of plant metabolic diversity Cucumis sativus
Glycyrrhiza uralensis
Cucurbitacins
Anti-tumoral
Glycyrrhizin
Anti-inflammatory
Pharmacological properties Many reported uses
Vaccine adjuvant
Panax ginseng
Ginsenosides
Quillajasaponin
Quillaja saponaria
Avenacin A-1 – An antimicrobial defence compound produced by oat (Avena spp.)
Avenacin A-1
Papadopoulou et al. (1999) PNAS 96:12923
The genes for avenacin biosynthesis are organized in a biosynthetic gene cluster
The biosynthetic pathways for many diverse plant specialized metabolites are encoded by co-localized genes
Nutzmann & Osbourn, Curr Opin. Biotechnol. 2014
The biosynthetic pathways for many diverse plant specialized metabolites are encoded by co-localized genes
HyperTrans - A simple system for rapid transient plant expression
Sainsbury F and Lomonossoff G. (2008) Plant Phys. 148:1212
Advantages of a plant-based expression system • Availability of precursors and co-enzymes • mRNA and protein processing • Localisation and compartmentalisation
Improved vacuum infiltration
Pressure reduced by 880 mbar
Gram quantities of purified triterpene
Michael Stephenson
Generation of structural analogs of oxygenated triterpenes in milligram quantities for bioassays
James Reed, Maria O’Connell m/s submitted
Reference compound: Bardoxolone-methyl A semi-synthetic triterpenoid derived from oleanolic acid.
Assembly of the 11-gene cluster for avenacin biosynthesis in N. benthamiana
Avena (oat)
Nicotiana benthamiana
Accumulation of acylated terpene in the vacuole
Mining for new pathways and chemistries
PREDICT
BUILD
TEST
A pilot and production business on the Norwich Research Park with a purpose-built facility employing the HyperTransÂŽ expression system:
Harnessing plant metabolic diversity Identification/validation of bioactives: • Bioactive discovery (e.g. new antimalarials) • Support local knowledge
PARTNERSHIPS WITH DEVELOPING COUNTRIES: KNOWLEDGE SHARING, TRAINING, Commercial production through CO-LEARNING, BUILDING INFRASTRUCTURE, bioengineering: SUSTAINABLE ECONOMIC DEVELOPMENT, PRESERVING BIODIVERSITY, SOCIETAL BENEFIT
Transient plant expression technology
Production in yeast
Production in the field