Purification and Propeties of (+)- cis -Naphthalene Dihydrodiol Dehydrogenase of Pseudomonas putida

Abstract

Cells of Pseudomonas putida, after growth with naphthalene as sole source of carbon and energy, contain an enzyme that oxidizes (+)-cis-1(R), 2(s)-dihydroxy-1, 2-dihydronaphthalene to 1, 2-dihydroxynaphthalene. The purified enzyme has a molecular weight of 102,000 and apparently consists of four 25,500 molecular weight subunits. The enzyme is specific for nicotinamide adenine dinucleotide as an electron acceptor and also oxidizes several other cis-dihydrodiols. However, no enzymatic activity was observed with trans-i, 2-dihydronaphthalene, or the K-region cis-dihydrodiols of carcinogenic polycyclic hydrocarbons.

Cloning and characterization of a novel cis-naphthalene dihydrodiol dehydrogenase gene (narB) from Rhodococcus sp. NCIMB12038

“…Preparation of cell-free extracts was performed as previously described [18]. The assay for cis-NDD was done according to Patel and Gibson [2]. The substrate, (1R,2S)cis-1,2-dihydroxy-1,2-dihydronaphthalene (cisnaphthalene dihydrodiol) was obtained using a biotransformation method [19].…”

“…The ¢rst step of naphthalene degradation is catalyzed by naphthalene 1,2-dioxygenase (NDO) resulting in the formation of cis-1,2-dihydroxy-1,2-dihydronaphthalene (cisnaphthalene dihydrodiol) [1]. The second step is the conversion of cis-naphthalene dihydrodiol to 1,2-dihydroxynaphthalene by cis-1,2-dihydro-1,2-dihydroxynaphthalene dehydrogenase (cis-NDD) that requires NAD as an electron acceptor [2]. This second enzyme has been puri¢ed from Pseudomonas putida NP and shown to consist of four sub-units each of molecular mass 25 500 [2].…”

“…The second step is the conversion of cis-naphthalene dihydrodiol to 1,2-dihydroxynaphthalene by cis-1,2-dihydro-1,2-dihydroxynaphthalene dehydrogenase (cis-NDD) that requires NAD as an electron acceptor [2]. This second enzyme has been puri¢ed from Pseudomonas putida NP and shown to consist of four sub-units each of molecular mass 25 500 [2]. It is capable of oxidizing other cis-dihydrodiols but not trans-1,2-dihydroxy-1,2-dihydronaphthalene [2,3].…”

“…This second enzyme has been puri¢ed from Pseudomonas putida NP and shown to consist of four sub-units each of molecular mass 25 500 [2]. It is capable of oxidizing other cis-dihydrodiols but not trans-1,2-dihydroxy-1,2-dihydronaphthalene [2,3]. The nucleotide sequence of the P. putida strain G7 cis-NDD has been reported recently [4].…”

Isolation of Rhodococcus rhodochrous NCIMB13064 derivatives with new biodegradative abilities

“…Dehalogenation by resting cells was measured using a Corning Chloride Analyser 926 as described earlier [2]. Assays for naphthalene dioxygenase activity were performed by the method of Ensley and Haigler [5] and naphthalene cis-diol dehydrogenase according to Pate1 and Gibson [6]. (lR,2S) cis-1,2dihydroxy-1,2_dihydronaphthalene…”