A strain of Pseudomonas putida that can utilize naphthalene as its sole source of carbon and energy was isolated from soil. A mutant strain of this organism, P. putida 119, when grown on glucose in the presence of naphthalene, accumulates optically pure (+)-cis-1(R),2(S)-dihydroxy-1,2-dihydronaphthalene in the culture medium. The cis relative stereochemistry in this molecule was established by nuclear magnetic resonance spectrometry. Radiochemical trapping experiments established that this cis dihydrodiol is an intermediate in the metabolism of naphthalene by P. Fluorescens (formerly ATCC, 17483), P. putida (ATCC, 17484), and a Pseudomonas species (NCIB 9816), as well as the parent strain of P. putida described in this report. Formation of the cis dihydrodiol is catalyzed by a dioxygenase which requires either NADH or NADPH as an electron donor. A double label procedure is described for determining the origin of oxygen in the cis dihydrodiol under conditions where this metabolite would not normally accumulate. Several aromatic hydrocarbons are oxidized by cell extracts prepared from naphthalene-grown cells of P. putida. The cis dihydrodiol is converted to 1,2-dihydroxynaphthalene by an NAD+-dependent dehydrogenase. This enzyme is specific for the (+) isomer of the dihydrodiol and shows a primary isotope effect when the dihydrodiol is substituted at C-2 with deuterium.

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.

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A psychrotrophic bacterium isolated from polychlorinated biphenyls (PCBs)-contaminated soil grew on biphenyl as sole carbon and energy source, and actively cometabolized PCBs at low temperature. Analysis of cellular fatty acids indicate that the bacterium is most closely related to Hydrogenophaga taeniospiralis. Resting cells incubated with 10 ppm of Aroclor 1221 at 5 or 30 degrees C for 48 h removed all mono-, most di-, and several trichlorobiphenyls. At 5 degrees C, removal of MCBs (monochlorobiphenyls) was between 63 to 89%, DCBs (dichlorobiphenyls) was between 30 to 78%, and TCBs (trichlorobiphenyls) was between 30 to 75%. At 30 degrees C, removal of MCBs was 100%, DCBs was between 30 to 100%, and TCBs was between 27 to 59%. Congeners with two or more ortho chlorine were generally resistant to degradation. However, removal of di-ortho plus para-substituted congeners at 30 degrees C and not at 5 degrees C, suggest that the presence of a para-chlorine enhanced the cometabolism of these congeners at 30 degrees C. Furthermore, after 72 h, resting cells removed 68 and 83% of 500 microM of 2,4'-dichlorobiphenyl (2,4'-DCB) and, 35 and 44% of 500 microM of 2,3-dichlorobiphenyl (2,3-DCB) at 5 and 30 degrees C, respectively. Analysis of metabolites by GC-MS indicates that the cometabolized 2,3-DCB was completely recovered as 2,3-chlorobenzoic acid (2,3-CBA), while the cometabolized 2,4'-DCB was not completely recovered as chlorobenzoic acid. To our knowledge, it is the first strain of Hydrogenophaga taeniospiralis found to degrade an organic pollutant, and also the first psychrotrophic strain of a member of the genus Hydrogenophaga to grow on biphenyl or cometabolize PCBs at low temperature. Results suggest that the bacterium has potential use in the bioremediation of PCB-contaminated sites in cold regions.

Listeria is an environmental contaminant which has been isolated from marine and fresh waters, as well as various seafoods. Furthermore, Listeria, including Listeria monocytogenes, has been isolated from processed seafood products such as smoked fish, cooked and frozen seafoods, marinated fish, surimi products, etc. The pathogen, L. monocytogenes, does have a certain degree of heat resistance. It was found to survive in internally infected shrimp after boiled for up to 5 min. However, the commercial pasteurization process for crab meat was found to be sufficient to inactivate Listeria. The current recovery methodology for L. monocytogenes from seafoods is the Food and Drug Administration Listeria protocol.