Biodegradation of organophosphate pesticide quinalphos by Ochrobactrum sp. strain HZM.

AIMS: Isolation and identification of bacteria capable of degrading organophosphate pesticide quinalphos and elucidation of its biodegradative pathway. METHODS AND RESULTS: A bacterium capable of degrading organophosphate pesticides was isolated from the pesticide-contaminated soil samples by selective enrichment on quinalphos (QP) as a sole source of carbon and energy. The bacterial strain was identified as Ochrobactrum sp. strain HZM on the basis of its morphological and biochemical characteristics and by phylogenetic analysis based on 16S rRNA gene sequences. The organism utilized various organophosphate pesticides such as quinalphos, profenofos, parathion-methyl and chlorpyrifos as growth substrates. Response surface methodology (RSM) showed optimum conditions for quinalphos degradation at pH 7 and 27°C. 2-Hydroxyquinoxaline and diethyl phosphate were identified as metabolites of quinalphos degradation by HPLC and GC-MS analysis. Cell-free extract of Ochrobactrum sp. strain HZM grown on quinalphos contained the quinalphos hydrolase activity. CONCLUSIONS: A bacterial strain capable of degrading quinalphos was isolated and identified as Ochrobactrum sp. strain HZM. The organism utilized organophosphate pesticides quinalphos, profenofos, parathion-methyl and chlorpyrifos as carbon sources. The organism degraded quinalphos by hydrolysis to yield 2-hydroxyquinoxaline and diethyl phosphate which were further utilized as carbon sources. SIGNIFICANCE AND IMPACT OF THE STUDY: The isolated bacterium Ochrobactrum sp. strain HZM was versatile in degrading various organophosphate pesticides. There was complete mineralization of quinalphos by Ochrobactrum sp. This strain could potentially be useful in the bioremediation of soil and water contaminated with toxic organophosphate pesticides.

Biodegradation of p-cresol by immobilized cells of Bacillus sp. strain PHN 1.

The Bacillus sp. strain PHN 1 capable of degrading p-cresol was immobilized in various matrices namely, polyurethane foam (PUF), polyacrylamide, alginate and agar. The degradation rates of 20 and 40 mM p-cresol by the freely suspended cells and immobilized cells in batches and semi-continuous with shaken cultures were compared. The PUF-immobilized cells achieved higher degradation of 20 and 40 mM p-cresol than freely suspended cells and the cells immobilized in polyacrylamide, alginate and agar. The PUF- immobilized cells could be reused for more than 35 cycles, without losing any degradation capacity and showed more tolerance to pH and temperature changes than free cells. These results revealed that the immobilized cell systems are more efficient than freely suspended cells for degradation of p-cresol.

Biodegradation of p-cresol by Bacillus sp. strain PHN 1.

A bacterium capable of utilizing p-cresol as sole source of carbon and energy was isolated from soil and identified as a Bacillus species. The organism also utilized phenol, o-cresol, m-cresol, 4-hydroxybenzoic acid, and gentisic acid as growth substrates. The organism degraded p-cresol to 4-hydroxybenzoic acid, which was further metabolized by a gentisate pathway, as evidenced by isolation and identification of metabolites and enzyme activities in the cell-free extract. Such a bacterial strain can be used for bioremediation of environments contaminated with phenolic compounds.

Biodegradation of carbaryl by a Micrococcus species.

A bacterium capable of utilizing carbaryl as sole source of carbon was isolated from garden soil and identified as a Micrococcus species. The organism also utilized carbofuran, naphthalene, 1-naphthol, and several other aromatic compounds as growth substrates. The organism degraded carbaryl by hydrolysis to yield 1-naphthol and methylamine. 1-Naphthol was further metabolized via salicylate by a gentisate pathway, as evidenced by oxygen uptake and enzymatic studies.

Biodegradation of phenanthrene by a Bacillus species.

A bacterial strain capable of utilizing phenanthrene as sole source of carbon was isolated from soil and identified as a Bacillus sp. The organism also utilized naphthalene, biphenyl, anthracene, and other aromatic compounds as growth substrates. The organism degraded phenanthrene through the intermediate formation of 1-hydroxy-2-naphthoic acid, which was further metabolized via o-phthalate by a protocatechuate pathway, as evidenced by oxygen uptake and enzymatic studies.

Biodegradation of 4-chlorobiphenyl by Micrococcus species.

A Micrococcus sp., isolated by enrichment culture, grew on 4-chlorobiphenyl at 2 g/l as sole carbon source and produced 4-chlorobenzoic acid in the culture medium as a dead-end metabolite. The organism degraded 4-chlorobiphenyl by 2,3-dihydroxylation followed by meta-ring cleavage to yield 4-chlorobenzoate and carbon fragments for cell growth.

Purification and properties of 2,3-dihydroxy-p-cumate-3,4-dioxygenase from Bacillus species.

2,3-Dihydroxy-p-cumate-3,4-dioxygenase, an enzyme involved in the catabolism of p-cymene, was purified to homogeneity from Bacillus species by affinity chromatography. Purification of the dioxygenase allowed the observation of the immediate ring cleavage product of 2,3-dihydroxy-p-cumate. The enzyme was optimally active at pH 8.2 and at 35 degrees C. The Km value for 2,3-dihydroxy-p-cumate was 32 microM. The enzyme had a broad substrate specificity for 3-substituted catechols. The activity of the enzyme was inhibited by heavy metals, sulphydryl inhibitors, iron-chelating agents, and substrate analogues. Fe2+ was suggested as a cofactor.