Metabolic engineering for microbial production of terpenoid products
AssigneeManus Bio Inc.
InventorsAjikumar Parayil KUMARAN, Chin-Giaw LIM, Souvik GHOSH, Christopher PIRIE, Jason DONALD, Aaron LOVE, Hong NAN, Hsien-Chung TSENG, Christine Nicole S. SANTOS, Ryan PHILIPPE
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Abstract
In various aspects and embodiments, the invention relates to bacterial strains and methods for making terpene and terpenoid products. The invention provides bacterial strains with improved carbon flux through the MEP pathway, to thereby increase terpene and/or terpenoid product yield by fermentation with carbon sources such as glucose.
Claims52
A method for production of a terpene or terpenoid product, comprising:
- providing a bacterial strain that produces isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) through an upstream methylerythritol pathway (MEP) and converts the IPP and DMAPP to a terpene or terpenoid product through a downstream synthesis pathway;
- and
- culturing the bacterial strain to produce the terpene or terpenoid product, wherein greater than 1% of carbon entering glycolysis becomes MEP carbon.
A bacterial strain having one or more genetic modifications that increase MEP carbon, the bacterial strain producing isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) through the MEP pathway, and converting the IPP and DMAPP to a terpene or terpenoid product through a downstream synthesis pathway;
- said genetic modifications being selected from:
- (1) a modification that decreases expression or activity of IspB;
- (2) a deletion or inactivation of all or part of the iscR gene, and an inducible or constitutive promoter to drive expression of the genes remaining in the isc operon under conditions used for terpene or terpenoid production;
- (3) a ryhB deletion or inactivation;
- (4) MEP enzyme expression or activity is altered or balanced such that DOX and/or ME do not accumulate above about 2 g/L or above about 1 g/L;
- (5) MEP pathway complementation comprising a dxr gene, optionally with ispD, ispE, and/or ispF, and an ispG and/or ispH gene;
- (6) a mutation in pgi (glucose-6-phosphate isomerase) supporting increased terpenoid titer or MEP carbon.
Dependencies are inferred from the claim text.