The objective is to develop a host strain and expression
system that will allow high level production of both natural
and genetically engineered terpenoids in E. coli. The
terpenoids play a critical role in such important processes
as regulating membrane fluidity, electron transport, and
cellular development. Commonly they must be extracted from
their native source and purified, often making the compounds
extremely expensive. Recent progress shows that many of the
relevant genes may be cloned from plants and expressed in
functional form in microorganisms.
The specific aims of the ONR funded portion of this
effort are:
1) to introduce into E. coli, the genes for specific
classes of terpenoids and optimize production of these
natural products, and
2) to use laboratory evolution of terpene cyclases to
produce novel terpenoids, or to change the distribution of
products made by terpenoid biosynthetic enzymes.
Emphasis is put on demonstrating the utility of the
system for the production of C10. C15, and C20 terpenoids.
Focus is put on those terpenoids for which genes have been
cloned and are expressible in E. coli. Synthases will be
chosen that use reaction mechanisms as distinct as possible
from one another and that produce different products in an
effort to demonstrate the versatility of the system and to
provide a foundation for laboratory evolution studies.
