Optimizing bioconversion pathways through systems analysis and metabolic engineering.
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.
Stafford
Daniel E
DE
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Yanagimachi
Kurt S
KS
Lessard
Philip A
PA
Rijhwani
Sushil K
SK
Sinskey
Anthony J
AJ
Stephanopoulos
Gregory
G
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
D007189:Indans; D007192:Indenes; C093579:1,2-indanediol; D004851:Epoxide Hydrolases