title: Optimizing bioconversion pathways through systems analysis and metabolic engineering.
abstract: We demonstrate a general approach for metabolic engineering of biocatalytic systems comprising the uses of a chemostat for strain improvement and radioisotopic tracers for the quantification of pathway fluxes. Flux determination allows the identification of target pathways for modification as validated by subsequent overexpression of the corresponding gene. We demonstrate this method in the indene bioconversion network of Rhodococcus modified for the overproduction of 1,2-indandiol, a key precursor for the AIDS drug Crixivan.
authors.affiliation: Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
authors.forename: Daniel E
authors.initials: DE
authors.lastname: Stafford
authors.forename: Kurt S
authors.initials: KS
authors.lastname: Yanagimachi
authors.forename: Philip A
authors.initials: PA
authors.lastname: Lessard
authors.forename: Sushil K
authors.initials: SK
authors.lastname: Rijhwani
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Sinskey
authors.forename: Gregory
authors.initials: G
authors.lastname: Stephanopoulos
mesh_terms: D001698:Biomedical Engineering / Q000379:methods*; D002384:Catalysis; D002851:Chromatography, High Pressure Liquid; D004345:Drug Industry; D004851:Epoxide Hydrolases / Q000378:metabolism; D005285:Fermentation; D005818:Genetic Engineering; D006868:Hydrolysis; D007189:Indans / Q000737:chemistry / Q000378:metabolism; D007192:Indenes / Q000737:chemistry / Q000378:metabolism; D008660:Metabolism; D008956:Models, Chemical; D010957:Plasmids / Q000378:metabolism; D012240:Rhodococcus / Q000378:metabolism; D013597:Systems Analysis; D013997:Time Factors
chemical_list: D007189:Indans; D007192:Indenes; C093579:1,2-indanediol; D004851:Epoxide Hydrolases