Bacterial production of transdihydroxycyclohexadiene carboxylates by metabolic pathway engineering.
Homochiral-cis-cyclohexa-3,5-diene-1,2-diols are important synthons. We found a way to produce trans-configured homochiral diols using recombinant Klebsiella pneumoniae 62-1. Transformation of this mutant (Phe- Trp- Tyr-) with plasmids carrying genes involved in chorismic and isochorismic acid metabolism leads to the production of either (+)-trans-(2S,3S)-2,3-dihydroxycyclohexa-4,6-dienecarboxylic acid or (-)-trans-(3R,4R)-3,4-dihydroxycyclohexa-1,5-dienecarboxylic acid, with a yield of 70 or 90 mg (1 culture broth)-1, respectively. The metabolic shift from one diene to the other is caused by a change in activity of isochorismate hydroxymutase and/or isochorismatase which in turn results from growth under iron deficiency or overexpression of genes (entC and/or entB) involved in chrismate metabolism.
Müller
Rolf
R
Institut fur Pharmazeutische Biologie, Nussallee 6, 53115 Bonn, Germany.
Breuer
Michael
M
Institut fur Pharmazeutische Biologie, Nussallee 6, 53115 Bonn, Germany.
Wagener
Andreas
A
Institut fur Pharmazeutische Biologie, Nussallee 6, 53115 Bonn, Germany.
Schmidt
Karsten
K
Institut fur Pharmazeutische Biologie, Nussallee 6, 53115 Bonn, Germany.
Leistner
Eckhard
E
Institut fur Pharmazeutische Biologie, Nussallee 6, 53115 Bonn, Germany.
D000595:Amino Acid Sequence; D001483:Base Sequence; D002264:Carboxylic Acids / Q000737:chemistry / Q000378:metabolism*; D002827:Chorismic Acid / Q000378:metabolism; D003511:Cyclohexanols / Q000737:chemistry / Q000378:metabolism*; D053138:Cyclohexenes; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D005818:Genetic Engineering; D006867:Hydrolases / Q000235:genetics / Q000378:metabolism; D019751:Intramolecular Transferases; D007501:Iron / Q000378:metabolism; D007535:Isomerases / Q000235:genetics / Q000378:metabolism; D007711:Klebsiella pneumoniae / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids / Q000235:genetics; D013237:Stereoisomerism; D014170:Transformation, Genetic
D002264:Carboxylic Acids; D003511:Cyclohexanols; D053138:Cyclohexenes; D017931:DNA Primers; D004269:DNA, Bacterial; C052985:isochorismic acid; D007501:Iron; D006867:Hydrolases; C064012:isochorismatase; D007535:Isomerases; D019751:Intramolecular Transferases; C058486:isochorismate synthase; D002827:Chorismic Acid