Patent ID: 6514733
Filing Date: 2003-02-04
Classification: C12N,C12P

Abstract:
A process for the bioproduction of 1,3-propanediol comprising:(a) contacting under suitable conditions (1) a recombinant E. coli comprising (a) a set of exogenous genes consisting of (i) at least one gene encoding a polypeptide having glycerol or diol dehydratase activity; (ii) at least one gene encoding a polypeptide having glycerol-3-phosphate dehydrogenase activity; (iii) at least one gene encoding a polypeptide having glycerol-3-phosphatase activity; and (iv) at least one gene encoding a dehydratase reactivation factor; and (b) at least one endogenous gene encoding a non-specific catalytic activity to convert 3-hydroxypropionaldehyde to 1,3-propanediol, â€ƒwherein no functional dhaT gene encoding a 1,3-propanediol oxidoreductase activity is present in the recombinant E. coli; or (2) a recombinant E. coli comprising (a) a set of exogenous genes consisting of (i) at least one copy of dhaR, nucleotides 2209-4134 of SEQ ID NO:1; (ii) at least one copy of orfY, complementary to nucleotides 6202-6630 of SEQ ID NO:1; (iii) at least one copy of orfX, complementary to nucleotides 4643-4996 of SEQ ID NO:1; (iv) at least one copy of orfW, complementary to nucleotides 4112-4642 of SEQ ID NO:1; (v) at least one copy of dhaB1, dhaB2, and dhaB3, nucleotides 7044-8711 of SEQ ID NO:1, nucleotides 8724-9308 of SEQ ID NO:1, and nucleotides 9311-9736 of SEQ ID NO:1, respectively; and (vi) at least one copy of orfZ, nucleotides 9749-11572 of SEQ ID NO:1; and (b) at least one endogenous gene encoding a non-specific catalytic activity to convert 3-hydroxypropionaldehyde to 1,3-propanediol, â€ƒwherein no functional dhat gene encoding a 1,3-propanediol oxidoreductase activity is present in the recombinant E. coli; or (3) the recombinant E. coli of (1) or (2) further comprising a set of endogenous genes, each having an inactivating mutation, the set of endogenous genes consisting of: (a) a gene encoding a polypeptide having glycerol kinase activity; (b) a gene encoding a polypeptide having glycerol dehydrogenase activity; and (c) a gene encoding a polypeptide having triosephosphate isomerase activity â€ƒwith at least one carbon source selected from the group consisting of monosaccharides, oligosaccharides, polysaccharides, and single-carbon substrates whereby 1,3-propanediol is produced; and (b) optionally recovering the 1,3-propanediol produced in (a).