Rapid purification of the oxygenase component of toluene dioxygenase from a polyol-responsive monoclonal antibody

Article Abstract:

The oxygenase unit, ISP(sub TOL), of toluene dioxygenase is obtained in the active form by extracting it from an immunoaffinity column by polyols and salts that affect the antigen-antibody links. The immunoaffinity column contains the Escherichia coli ISP(sub TOL) bound to the 301-beta monoclonal antibody specific for the Pseudomonas putida F1 ISP(sub TOL). Elution of an 8-ml column with a buffer containing ethylene glycol, ammonium sulfate, dithiothreitol and ferrous ammonium sulfate gives the enzyme in a form that is 300 times more active than that purified by other methods.

Usage, Monoclonal antibodies, Polyols

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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

Article Abstract:

Bacterial strains producing naphthalene dioxygenase or biphenyl dioxygenase oxidize 6,7-dihydro-5H-benzocycloheptene (benzocyclohept-1-ene) by a dioxygenation reaction to produce cis-diol products. Strains producing toluene dioxygenase (TDO) catalyze benzocyclohept-1-ene by a monooxidation reaction to produce monols. The configuration of the small amount of cis-diols formed by the TDO-producing strains is opposite to that of the diols produced by the other strains. The type of reaction that is catalyzed depends on the nature of the substrate and the enzyme specificity.

Bacteria, Oxidases

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Initial reactions in the oxidation of 1,2-dihydronaphthalene by Sphingomonas yanoikuyae strains

Article Abstract:

Sphingomonas yanoikuyae B8/36 cells oxidize 1,2-dihydronaphthalene (DHN) to (-)-(1R,2S)-cis-1,2-dihydroxy-1,2,3,4-tetrahydronaphthalene (THN), (+)-(1R,2S)-cis-1,2-dihydroxy-1,2-DHN and (+)-(R)-2-hydroxy-1,2-DHN, respectively. In contrast, strain B1 oxidizes 1,2-DHN to form (-)-(1R,2S)-cis-1,2-dihydroxy-1,2,3,4-THN as the major product, and (+)-(R)-2-hydroxy-1,2-DHN and alpha-tetralone in small amounts. The enzymes biphenyl-2,3-dioxygenase and cis-biphenyl dihydrodiol dehydrogenase (BDDH) are involved in the reactions. The enzyme BDDH is inactive in strain B8/36.

Analysis, Microbial metabolism

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