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. 2001 Aug;67(8):3333-9.
doi: 10.1128/AEM.67.8.3333-3339.2001.

Biotransformation of various substituted aromatic compounds to chiral dihydrodihydroxy derivatives

H Raschke  1 , M Meier, J G Burken, R Hany, M D Müller, J R Van Der Meer, H P Kohler
Affiliations

Biotransformation of various substituted aromatic compounds to chiral dihydrodihydroxy derivatives

H Raschke et al. Appl Environ Microbiol. 2001 Aug.

Abstract

The biotransformation of four different classes of aromatic compounds by the Escherichia coli strain DH5alpha(pTCB 144), which contained the chlorobenzene dioxygenase (CDO) from Pseudomonas sp. strain P51, was examined. CDO oxidized biphenyl as well as monochlorobiphenyls to the corresponding cis-2,3-dihydro-2,3-dihydroxy derivatives, whereby oxidation occurred on the unsubstituted ring. No higher substituted biphenyls were oxidized. The absolute configurations of several monosubstituted cis-benzene dihydrodiols formed by CDO were determined. All had an S configuration at the carbon atom in meta position to the substituent on the benzene nucleus. With one exception, the enantiomeric excess of several 1,4-disubstituted cis-benzene dihydrodiols formed by CDO was higher than that of the products formed by two toluene dioxygenases. Naphthalene was oxidized to enantiomerically pure (+)-cis-(1R,2S)-dihydroxy-1,2-dihydronaphthalene. All absolute configurations were identical to those of the products formed by toluene dioxygenases of Pseudomonas putida UV4 and P. putida F39/D. The formation rate of (+)-cis-(1R,2S)-dihydroxy-1,2-dihydronaphthalene was significantly higher (about 45 to 200%) than those of several monosubstituted cis-benzene dihydrodiols and more than four times higher than the formation rate of cis-benzene dihydrodiol. A new gas chromatographic method was developed to determine the enantiomeric excess of the oxidation products.

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Figures

FIG. 1
FIG. 1
Designation of the protons of mono- and 1,4-disubstituted cis-benzene dihydrodiols, formed by CDO.
FIG. 2
FIG. 2
Determination of the enantiomeric composition of cis-4-bromo-2,3-dihydroxy-1-methyl-cyclohexa-4,6-diene, which is formed by oxidation of 4-bromotoluene by CDO. Enantiomers were separated as their BB derivatives. The mass spectra of the enantiomers were identical. The major enantiomer was formed with an EE of 77%, which was calculated by integration of the peak areas of the major and minor enantiomer. The isomeric structure of the product was determined by 1H NMR analysis.
FIG. 3
FIG. 3
GC-MS analysis and determination of the absolute configuration of cis-1,2-dihydroxy-1,2-dihydronaphthalene. (A) Separation of the BB derivatives of an authentic standard of (±)-cis-1,2-dihydroxy-1,2-dihydronaphthalene. The mass spectra of the two compounds were identical. (B) BB derivative of an authentic standard of (+)-cis-(1R,2S)-dihydroxy-1,2-dihydronaphthalene. (C) BB derivative of the oxidation product formed from naphthalene by CDO. The BB derivatives in panels B and C had identical retention times and identical mass spectra.
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