title: Production of isoleucine by overexpression of ilvA in a Corynebacterium lactofermentum threonine producer.
abstract: Overproduction of isoleucine, an essential amino acid, was achieved by amplification of the gene encoding threonine dehydratase, the first enzyme in the threonine to isoleucine pathway, in a Corynebacterium lactofermentum threonine producer. Threonine overproduction was previously achieved with C. lactofermentum ATCC 21799, a lysine-hyperproducing strain, by introduction of plasmid pGC42 containing the Corynebacterium homdr and thrB genes (encoding homoserine dehydrogenase and homoserine kinase respectively) under separate promoters. The pGC42 derivative, pGC77, also contains ilvA, which encodes threonine dehydratase. In a shake-flask fermentation, strain 21799(pGC77) produced 15 g/l isoleucine, along with small amounts of lysine and glycine. A molar carbon balance indicates that most of the carbon previously converted to threonine, lysine, glycine and isolecine was incorporated into isoleucine by the new strain. Thus, in our system, simple overexpression of wild-type ilvA sufficed to overcome the effects of feedback inhibition of threonine dehydratase by the end-product, isoleucine.
authors.affiliation: Department of Chemical Engineering, MIT, Cambridge, MA 02139, USA.
authors.forename: G E
authors.initials: GE
authors.lastname: Colón
authors.forename: T T
authors.initials: TT
authors.lastname: Nguyen
authors.forename: M S
authors.initials: MS
authors.lastname: Jetten
authors.forename: A J
authors.initials: AJ
authors.lastname: Sinskey
authors.forename: G
authors.initials: G
authors.lastname: Stephanopoulos
mesh_terms: D000095:Acetolactate Synthase / Q000378:metabolism; D003352:Corynebacterium / Q000235:genetics*; D007532:Isoleucine / Q000302:isolation & purification*; D013912:Threonine / Q000378:metabolism; D013913:Threonine Dehydratase / Q000235:genetics* / Q000378:metabolism
chemical_list: D007532:Isoleucine; D013912:Threonine; D000095:Acetolactate Synthase; D013913:Threonine Dehydratase