Heterologous expression of alkaloid biosynthetic genes--a review

Gene
T M Kutchan

Abstract

Tetrahydrobenzylisoquinoline alkaloids comprise a diverse class of secondary metabolites with many pharmacologically active members. The biosynthesis at the enzyme level of at-least two tetrahydrobenzylisoquinoline alkaloids, the benzophenanthridine alkaloid sanguinarine in the California poppy, Eschscholtzia californica, and the bisbenzylisoquinoline alkaloid berbamunine in barberry, Berberis stolonifera, has been elucidated in detail starting from the aromatic amino acid (aa) L-tyrosine. In an initial attempt to develop alternate systems for the production of medicinally important alkaloids, one enzyme from each pathway (BBE, a covalently flavinylated enzyme of benzophenanthridine alkaloid biosynthesis and CYP80, a phenol coupling cytochrome P-450-dependent oxidase of bisbenzylisoquinoline alkaloid biosynthesis) has been purified to homogeneity, a partial aa sequence determined, and the corresponding cDNAs isolated with aid of synthetic oligos based on the aa sequences. The recombinant enzymes were actively expressed in Spotloptera frugiperda Sf9 cells using a baculovirus vector, purified and then characterized. Insect cell culture has proven to be a powerful system for the overexpression of alkaloid biosynthetic genes.

References

Dec 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·D J YunY Yamada
Apr 1, 1985·Antimicrobial Agents and Chemotherapy·J L Dzink, S S Socransky
May 9, 1995·Proceedings of the National Academy of Sciences of the United States of America·S BlechertM H Zenk
Mar 14, 1995·Proceedings of the National Academy of Sciences of the United States of America·P F Kraus, T M Kutchan

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Citations

Sep 24, 2004·Nature·Anthony G MillgatePhilip J Larkin
Feb 4, 2012·The Arabidopsis Book·Søren BakDanièle Werck-Reichhart
Jan 22, 2002·Metabolic Engineering·Erik H Hughes, Jacqueline V Shanks

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