APPLICATIONS OF TECHNOLOGY:
- Production of novel, unnatural bio-active molecules
- Enables the uptake of artificial cofactor by a host cell and thereby the production of unnatural bio-actives
Numerous complex biomolecules have been produced by natural or engineered organisms, however, the structures that can be afforded are still limited by the available chemical reactions catalyzed by natural enzymes. Artificial enzymes or the metalloenzymes (ArMs), on the other hand, have been studied in vitro to catalyze the functionalization of complex molecules such as terpenes.
Researchers at Berkeley Lab have developed a method for engineering a genetically modified host cell where a natural biosynthesis pathway was fully coupled with an in vivo expressed and assembled artificial metalloenzyme. This coupling enables the host cell to produce unnatural terpenoids from simple starting materials such as glucose.
The invention provides a platform for the integration of ArMs that catalyze abiotic reactions into metabolic networks broadening the cache of molecules produced biosynthetically by microorganisms. In this case, this is achieved through the assembly of an ArM containing an iridium-porphyrin complex in the cytoplasm of a terpene producing Escherichia coli by a heterologous heme transport machinery, and insertion of this ArM into a natural biosynthetic pathway to produce an unnatural terpenoid. A high stereometric selectivity (72% dr for cyclopropanated limonene) was reached that will be attractive to industry.
DEVELOPMENT STAGE: Proven principle
- Jing Huang
- Zhennan Liu
- Douglas S. Clark
- Jay D. Keasling
- Aindrila Mukhopadhyay
- John F. Hartwig
STATUS: Patent pending. Available for licensing or collaborative research.