Oxidation of 6,7-dihydro-5H-benzocycloheptene by bacterial strains expressing naphthalene dioxygenase, biphenyl dioxygenase, and toluene dioxygenase yields homochiral monol or cis-diol enantiomers as major products.
Bacterial strains expressing naphthalene, biphenyl, and toluene dioxygenase were examined for their abilities to oxidize 6,7-dihydro-5H-benzocycloheptene (benzocyclohept-1-ene). The major oxidation products were isolated, and their absolute configurations were determined by chiral 1H nuclear magnetic resonance analysis of diastereomeric boronate esters, chiral stationary-phase high-pressure liquid chromatography, and stereo-chemical correlation. Pseudomonas sp. strain 9816/11 and Sphingomonas yanoikuyae (formerly identified as a Beijerinckia sp.) B8/36 expressing naphthalene and biphenyl dioxygenases, respectively, oxidized benzocyclohept-1-ene to a major product identified as (-)-(1R,2S)-cis-dihydroxybenzocycloheptane (> 98% enantiomeric excess [ee], 50 and 90% yield, respectively). In contrast, Pseudomonas putida F39/D expressing toluene dioxygenase oxidized benzocyclohept-1-ene to (+)-(5R)-hydroxybenzocyclohept-1-ene (> 98% ee, 90% yield) as the major metabolite and to the "opposite" diol, (+)-(1S,2R)-cis-dihydroxybenzocycloheptane (> 98% ee, 10% yield). The results indicate that, for benzocyclohept-1-ene, the major reaction catalyzed by naphthalene and biphenyl dioxygenases is dioxygenation whereas toluene dioxygenase catalyzes mainly R-stereospecific benzylic monooxygenation. Although the type of reaction catalyzed by each organism was not predictable, the absolute configuration of the diol and monol products formed by naphthalene and toluene dioxygenases are consistent with the stereochemistry of the products formed by these enzymes from other benzocycloalkene substrates.
Resnick
S M
SM
Department of Microbiology, University of Iowa, Iowa City 52242, USA. sresink@vaxa.weeg.uiowa.edu
Gibson
D T
DT
D001419:Bacteria / Q000378:metabolism*; D001567:Benzocycloheptenes / Q000737:chemistry / Q000378:metabolism*; D049308:Dioxygenases; D007506:Iron-Sulfur Proteins; D009097:Multienzyme Complexes / Q000378:metabolism; D010084:Oxidation-Reduction; D010105:Oxygenases / Q000378:metabolism; D011549:Pseudomonas / Q000378:metabolism; D016958:Pseudomonas putida / Q000378:metabolism; D013237:Stereoisomerism
D001567:Benzocycloheptenes; D007506:Iron-Sulfur Proteins; D009097:Multienzyme Complexes; D010105:Oxygenases; D049308:Dioxygenases; C038703:naphthalene dioxygenase; C023172:toluene dioxygenase; C074161:biphenyl-2,3-dioxygenase