Synthetic Metabolism in Plants: Elucidating Vinblastine Biosynthesis and Implementing Strategies to Overproduce Complex Plant Metabolites

Lead Research Organisation: John Innes Centre
Department Name: Contracts Office

Abstract

Nature produces a spectacularly diverse array of complex molecules that are exploited for many industrial and pharmaceutical applications. Elucidating the biosynthetic pathways that are used to construct these molecules allows implementation of metabolic engineering or synthetic biology strategies that can dramatically improve production levels of these compounds. Moreover, identifying the biosynthetic genes facilitates study of the unprecedented biochemistry harboured within these specialised metabolic pathways. Unfortunately, research progress in plant specialised metabolism has lagged, in large part due to the complexities of plant systems. This has severely hampered the application of state-of-the-art approaches that can exploit this rich metabolism.

The availability of inexpensive sequence data makes this an outstanding time to revisit difficult questions in plant metabolism. My group has recently obtained RNA-seq data for 24 tissues for Catharanthus roseus, which produces vinblastine, an anti-cancer drug that is arguably one of the most complex natural products found in plants. We now have a clear path toward identifying the ~13 missing biosynthetic enzymes within the ~39,000 genes comprising the transcriptome of C. roseus.

In Objective 1, we outline a plan to identify vinblastine biosynthetic gene candidates using our transcriptome data and then screen these candidates for function in planta. We also propose a series of in vitro assays by which to characterise promising gene candidates biochemically. In Objective 2, we propose to reconstitute portions of the vinblastine pathway in the model plant Nicotiana benthamiana (tobacco) and Saccharomyces cerevisiae (yeast) and to create high-yielding platforms for production of valuable plant metabolites.

Publications

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Kellner F (2015) Genome-guided investigation of plant natural product biosynthesis. in The Plant journal : for cell and molecular biology

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Kries H (2017) Inverted stereocontrol of iridoid synthase in snapdragon. in The Journal of biological chemistry

 
Description New enzymes that create biologically active natural products have been discovered and investigated. Several new enzymes that encode biologicaly active products have been discovered and reported. Crystal structures and the mechanistic basis for these catalysts have also been reported.
Exploitation Route Others can use these enzymes to make new products; also others can use these as a basis to find similar enzymes in other plants
Sectors Chemicals,Manufacturing, including Industrial Biotechology

 
Description I am collaborating with a company, Conagen, to make alkaloids in yeast.
First Year Of Impact 2016
Sector Chemicals,Manufacturing, including Industrial Biotechology
Impact Types Economic

 
Title Development of self organizing map algorithm 
Description This is a program to analyze RNA seq data 
Type Of Material Model of mechanisms or symptoms - non-mammalian in vivo 
Year Produced 2017 
Provided To Others? Yes  
Impact We have identified many new biosynthetic genes using this method 
 
Description Buell Group Bioinformatics 
Organisation Michigan State University
Country United States 
Sector Academic/University 
PI Contribution The Buell group has provided sequencing and bioinformatics support for our plants. We do the functional characterisation and biochemistry.
Collaborator Contribution The Buell group has seqeunced plants for us, assembled genomes/transcriptomes/annotated them, provided extensive bioinformatics support.
Impact Numerous papers (see publications); an NSF grant from the USA; an extensive database of plant sequence data; more papers underway
Start Year 2009
 
Description Collaboration with Vincent Courdavault 
Organisation François Rabelais University or University of Tours
Country France 
Sector Academic/University 
PI Contribution Courdavaults group has done microscopy to show localization of these enzymes. Our role in this project has been to discover the enzymes
Collaborator Contribution Our partners have provided localization data for the enzymes that we have discovered.
Impact See co-authored papers in publications
 
Description Transporter Collaboration 
Organisation University of Copenhagen
Department Department of Plant and Environmental Sciences
Country Denmark 
Sector Academic/University 
PI Contribution Collaboration with University of Copenhagen PIs Barbara Halkier and Hassam Nour Eldin. They perform biochemical assays with plant transporters that we have identified.
Collaborator Contribution They performed assay of transporters in oocytes.
Impact Publication in Nature Plants in 2017
Start Year 2016
 
Description Talk at Kew Gardens 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact A Kew organized event called "State of the World's Plants"
Year(s) Of Engagement Activity 2017