Genetically encoded system for quantifying rubisco activity in catalyzing carbon fixation
AssigneeThe Scripps Research Institute
InventorAhmed H. Badran
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Abstract
The present invention provides assay systems and related methods for monitoring RuBISCO catalyzed carbon fixation in a manner that is insulated from host metabolism. The invention also provides methods for evolving specific RuBISCO enzymes to identify variants with improved activities.
Claims22
An engineered bacterial cell or cell lysate for quantitatively monitoring RuBisCO-dependent CO2 fixation, comprising (a) mutations in the genome that result in bypassing canonical glycolysis pathway and no production or accumulation of 3-phosphoglycerate (3PG) from glycolysis, thereby insulating host metabolism from RuBisCO-produced products, (b) an artificial biosynthetic pathway that produces RuBisCO substrate ribulose 1,5-bisphosphate (RuBP) with reduced toxicity, and (c) a transcriptional biosensor for monitoring glycerate generated from RuBisCO-produced 3PG.
An engineered E. coli cell that bypasses canonical glycolysis pathway and does not produce or accumulate 3-phosphoglycerate (3PG) from glycolysis, comprising mutations that disrupt expression of one or more functional proteins encoded by serA, eda-edd, garDPLRK, glcCDEFGB, gudPXD, allS-allD gene cassette, cdaR, ghrA, ghrB, and ytjC.
An engineered E. coli cell, comprising an artificial biosynthetic pathway that produces RuBisCO substrate ribulose 1,5-bisphosphate (RuBP) with reduced toxicity.
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Dependencies are inferred from the claim text.