A bacterial strain (AD26) capable of utilizing atrazine as a sole nitrogen source for growth was isolated from an industrial wastewater sample by enrichment culture. The 16S rRNA gene sequencing identified AD26 as an ...A bacterial strain (AD26) capable of utilizing atrazine as a sole nitrogen source for growth was isolated from an industrial wastewater sample by enrichment culture. The 16S rRNA gene sequencing identified AD26 as an Arthrobacter sp. PCR assays indicated that AD26 contained atrazine-degrading genes trzN and atzBC. The trzN gene of AD26 only differs from the trzN of Arthrobacter aurescens TC1 by one base (A→T at 907) and one amino acid (Met→Leu at 303). The specific activity of trzN of AD26 in crude cell extract was 0.28 U/mg, which was 1.2 times that of TC1. This strain has shown faster growth and atrazine-degradation rates in atrazine-containing minimal media than two well characterized atrazine-degrading bacteria, Pseudomonas sp. ADP and Arthrobacter aurescens TC1. After incubating for 48 h at 30°C, the OD600 of AD26 reached 2.6 compared with 1.33 of ADP. AD26 was capable of degrading 500 mg/L of atrazine in minimal medium at 95% in 72 h, while the degradative rates by TC1 and ADP were only 90% and 86%, respectively. A bioremediation trial of contaminated soil has indicated that AD26 can degrade as high as 98% of atrazine contained in soil (300 mg/kg) after incubating for 20 d at 26°C, nominating this strain as a good candidate for use in bioremediation programs.展开更多
A novel salicylaldehyde dehydrogenase involved in catabolism of naphthalene from Pseudomonas putida ND6, NahV, has been identified. NahV exhibited lower identity in amino acid sequence with the classic salicylaldehyde...A novel salicylaldehyde dehydrogenase involved in catabolism of naphthalene from Pseudomonas putida ND6, NahV, has been identified. NahV exhibited lower identity in amino acid sequence with the classic salicylaldehyde dehydrogenase, NahF, from P. putida ND6. This is the first report of an isofunctional enzyme of bacterial salicylaldehyde dehydrogenase. Comparison of Km and Vmax values of NahV and NahF demonstrated that NahF has a more efficient catalytic reaction than NahV, while NahV has much higher affinity for salicylaldehyde and NAD+. Both enzymes exhibited broad substrate speci- ficities and catalyzed the oxidation of salicylaldehyde, 5-chlorosalicylaldehyde, formaldehyde, m-nitrobenzaldehyde, o-nitrobenzaldehyde, o-methoxybenxaldehyde, glutaraldehyde, caprylic aldehyde, and glyoxal. However, the relative rates at which the substituted analogs are transformed differ considerably. NahV activity could be enhanced by Fe2+, Cu2+ and Zn2+; whereas NahF activity could only be stimulated by Fe2+. NahF is more stable than NahV at elevated temperatures. Dot-blot hybridization analyses showed that nahF-like genes occurred in all naphthalene-degradation bacteria isolated in this study, whereas nahV-like genes were present in only some naphthalene-degrading bacteria.展开更多
文摘A bacterial strain (AD26) capable of utilizing atrazine as a sole nitrogen source for growth was isolated from an industrial wastewater sample by enrichment culture. The 16S rRNA gene sequencing identified AD26 as an Arthrobacter sp. PCR assays indicated that AD26 contained atrazine-degrading genes trzN and atzBC. The trzN gene of AD26 only differs from the trzN of Arthrobacter aurescens TC1 by one base (A→T at 907) and one amino acid (Met→Leu at 303). The specific activity of trzN of AD26 in crude cell extract was 0.28 U/mg, which was 1.2 times that of TC1. This strain has shown faster growth and atrazine-degradation rates in atrazine-containing minimal media than two well characterized atrazine-degrading bacteria, Pseudomonas sp. ADP and Arthrobacter aurescens TC1. After incubating for 48 h at 30°C, the OD600 of AD26 reached 2.6 compared with 1.33 of ADP. AD26 was capable of degrading 500 mg/L of atrazine in minimal medium at 95% in 72 h, while the degradative rates by TC1 and ADP were only 90% and 86%, respectively. A bioremediation trial of contaminated soil has indicated that AD26 can degrade as high as 98% of atrazine contained in soil (300 mg/kg) after incubating for 20 d at 26°C, nominating this strain as a good candidate for use in bioremediation programs.
基金Supported by the National Natural Science Foundation of China (Grant No. 30270274)Postdoctoral Science Foundation of China (Grant No. 2005038461)
文摘A novel salicylaldehyde dehydrogenase involved in catabolism of naphthalene from Pseudomonas putida ND6, NahV, has been identified. NahV exhibited lower identity in amino acid sequence with the classic salicylaldehyde dehydrogenase, NahF, from P. putida ND6. This is the first report of an isofunctional enzyme of bacterial salicylaldehyde dehydrogenase. Comparison of Km and Vmax values of NahV and NahF demonstrated that NahF has a more efficient catalytic reaction than NahV, while NahV has much higher affinity for salicylaldehyde and NAD+. Both enzymes exhibited broad substrate speci- ficities and catalyzed the oxidation of salicylaldehyde, 5-chlorosalicylaldehyde, formaldehyde, m-nitrobenzaldehyde, o-nitrobenzaldehyde, o-methoxybenxaldehyde, glutaraldehyde, caprylic aldehyde, and glyoxal. However, the relative rates at which the substituted analogs are transformed differ considerably. NahV activity could be enhanced by Fe2+, Cu2+ and Zn2+; whereas NahF activity could only be stimulated by Fe2+. NahF is more stable than NahV at elevated temperatures. Dot-blot hybridization analyses showed that nahF-like genes occurred in all naphthalene-degradation bacteria isolated in this study, whereas nahV-like genes were present in only some naphthalene-degrading bacteria.
