1. M ETABOLIC ENGINEERING OF ALKALOIDS PRODUCTION IN PLANT Juan Pan Department of Plant Pathology

2. OUTLINE alkaloids biosynthesis in plant metabolic engineering summary

3. A LKALOIDS - N-containing small molecule - Amino acid derivative - Secondary metabolites - Defense in plant - Pharmaceutical uses

4. Alkaloid biosynthesis In plant - monoterpene indole alkaloid (MIA) Annu. Rev. Plant Biol. 2008. 59:735-69

5. MIA biosynthesis scheme

6. Alkaloid biosynthesis in plant - benzylisoquinoline alkaloids (BIAs) tyrosine (S)-norcoclaurine Nature chemical biology, 2009, 5 (5): 292-300 dopamine 4-hydroxyphenyl acetaldehyde (S)-reticuline

7. Alkaloid biosynthesis in plant - tropane alkaloids

8. Alkaloid biosynthesis in plant - purine alkaloids

9. Metabolic engineering of alkaloid biosynthesis - Overexpressing rate- limiting genes - Gene silencing to block pathway - Regulatory element

10. H6H PMT and H6H overexpression increased scopolamine production Proc Natl Acad Sci USA 101: 6786-6791

11. tyrosine (S)-norcoclaurine dopamine 4-hydroxyphenyl acetaldehyde (S)-reticuline COR Silencing COR blocked morphine biosynthetic pathway Nat Biotechnol 22: 1559-1566

13. Plant Biotechnology Journal (2008) 6, pp. 160–175 Exogenous regulatory factors increased alkaloids production in poppy

14. S UMMARY - Overexpression of rate-limiting enzyme for higher alkaloid production - Gene silencing to block certain pathway - Manipulating regulatory elements to increase alkaloid production - Combination of several methods