title: Genetic engineering of Escherichia coli for the production of precorrin-3 in vivo and in vitro.
abstract: The construction of a new recombinant strain of Escherichia coli in which two vitamin B12 biosynthetic genes, cobA and cobI, from Pseudomonas denitrificans are simultaneously overexpressed has resulted in the in vivo synthesis and accumulation of Factor III, an isobacteriochlorin not normally synthesized in E. coli. A lysate of the new strain can take the place of two lysates normally required to provide uroporphyrinogen III methyltransferase (cobA) and precorrin-2 methyltransferase (cobI) in an anaerobic five-enzyme synthesis of the early B12 intermediate, precorrin-3 (the reduced form of Factor III) from delta-aminolevulinic acid.
authors.affiliation: Department of Chemistry, Texas A&M University, College Station 77842-3012, USA.
authors.forename: C A
authors.initials: CA
authors.lastname: Roessner
authors.forename: J H
authors.initials: JH
authors.lastname: Park
authors.forename: A I
authors.initials: AI
authors.lastname: Scott
mesh_terms: D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D005818:Genetic Engineering / Q000379:methods*; D008780:Methyltransferases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D011549:Pseudomonas / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014578:Uroporphyrins / Q000096:biosynthesis*
chemical_list: D011994:Recombinant Proteins; D014578:Uroporphyrins; C078447:precorrin 3; D008780:Methyltransferases; C066231:precorrin-2 O-methyltransferase; C060508:uroporphyrin-III C-methyltransferase