Isolation, Identification, and Characterization of Lambda-Cyhalothrin Pesticide Degrading Bacterium ZC-5

2016 ◽  
Vol 723 ◽  
pp. 628-632
Author(s):  
Rong Hu Zhang ◽  
Zhen Hua Zhou ◽  
Jian Cheng Feng
Keyword(s):  
Sole Source ◽  
Achromobacter Xylosoxidans ◽  
Logarithmic Phase ◽  
Aeration Tank ◽  
16S Rdna Sequence ◽  
Carbon And Nitrogen ◽  
Degrading Bacterium ◽  
Lambda Cyhalothrin ◽  
Novel Strategy

A highly efficient lambda-cyhalothrin-degrading bacterium, designated as strain ZC-5, was isolated from the activated sludge of a sewage aeration tank in a pesticide factory by enrichment acclimation and the streak plate method. Strain ZC-5 can grow on minimal medium with lambda-cyhalothrin as the sole source of carbon and nitrogen. After cultivation for 6 h to 24 h, the biomass of the bacterial strain significantly increased at the logarithmic phase. By contrast, the concentration of lambda-cyhalothrin rapidly decreased. The residual lambda-cyhalothrin presented a concentration of 250 mg/L and a degradation rate of 50%. Gas chromatography revealed that this strain can degrade 87.1% lambda-cyhalothrin (500 mg/L) in the culture within 2 days. Morphological analysis showed the Gram-negative strain as short rods. Physiological and biochemical characterizations, as well as phylogenetic analysis of the 16S rDNA sequence identified the bacterium to be an Achromobacter xylosoxidans strain. Results showed that this strain can provide a novel strategy to biodegrade the pesticide lambda-cyhalothrin.

scholarly journals Identification and Characterization of Cotinine Hydroxylase CotA that Initiates biodegradation of Wastewater Micropollutant Cotinine in Nocardioides sp. Strain JQ2195

2021 ◽  
Author(s):  
Lingling Zhao ◽  
Zhenyang Zhao ◽  
Kaiyun Zhang ◽  
Xuan Zhang ◽  
Siqiong Xu ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Tobacco Consumption ◽  
Hydroxyl Group ◽  
Carbon And Nitrogen ◽  
Enzymatic Transformation ◽  
Degrading Bacterium ◽  
Hydroxylase Gene ◽  
Urine Cotinine ◽  
Genetic Mechanisms

Cotinine is a stable toxic contaminant, produced as a byproduct of smoking. It is of emerging concern due to its global distribution in aquatic environments. Microorganisms have the potential to degrade cotinine, however, the genetic mechanisms of this process are unknown. Nocardioides sp. strain JQ2195 is a pure culture strain that has been reported to degrade cotinine at micropollutant concentrations. This strain utilizes cotinine as its sole carbon and nitrogen source. In this study, a 50 kb gene cluster (designated as cot ) involved in cotinine degradation, was predicted based on genomic and transcriptomic analyses. A novel three-component cotinine hydroxylase gene (designated as cotA1A2A3 ), which initiated cotinine catabolism was identified and characterized. CotA from Shinella sp. HZN7 was heterologously expressed and purified, and shown to convert cotinine into 6-hydroxycotinine. H 2 18 O-labelling and ESI-MS analysis confirmed that the hydroxyl group incorporated into 6-hydroxycotinine was derived from water. This study provides new molecular insights into the microbial metabolism of heterocyclic chemical pollutants. IMPORTANCE In the human body, cotinine is the major metabolite of nicotine, and 10–15% of generated cotinine is excreted in urine. Cotinine is a structural analogue of nicotine and is much more stable than nicotine. Increased tobacco consumption has led to high environmental concentrations of cotinine, which may have detrimental effects on aquatic ecosystems and human health. Nocardioides sp. strain JQ2195 is a unique cotinine-degrading bacterium. However, the underlying genetic and biochemical foundations of cotinine degradation are still unknown. In this study, a 50 kb gene cluster (designated cot ) was identified by genomic and transcriptomic analyses as being involved in the degradation of cotinine. A novel three-component cotinine hydroxylase gene (designated cotA1A2A3 ) catalyzed cotinine to 6-hydroxy-cotinine. This study provides new molecular insights into the microbial degradation and enzymatic transformation of cotinine.

scholarly journals Characterization of S-Triazine Herbicide Metabolism by a Nocardioides sp. Isolated from Agricultural Soils

2000 ◽  
Vol 66 (8) ◽  
pp. 3134-3141 ◽  
Author(s):  
Edward Topp ◽  
Walter M. Mulbry ◽  
Hong Zhu ◽  
Sarah M. Nour ◽  
Diane Cuppels
Keyword(s):  
Agricultural Soils ◽  
Sole Source ◽  
Bacterial Strains ◽  
Corn Production ◽  
Whole Cells ◽  
Carbon And Nitrogen ◽  
Cell Extracts ◽  
Hydrolytic Reaction ◽  
Central Canada

ABSTRACT Atrazine, a herbicide widely used in corn production, is a frequently detected groundwater contaminant. Nine gram-positive bacterial strains able to use this herbicide as a sole source of nitrogen were isolated from four farms in central Canada. The strains were divided into two groups based on repetitive extragenic palindromic (rep)-PCR genomic fingerprinting with ERIC and BOXA1R primers. Based on 16S ribosomal DNA sequence analysis, both groups were identified as Nocardioides sp. strains. None of the isolates mineralized [ring-U-14C]atrazine. There was no hybridization to genomic DNA from these strains usingatzABC cloned from Pseudomonas sp. strain ADP or trzA cloned from Rhodococcus corallinus. S-Triazine degradation was studied in detail inNocardioides sp. strain C190. Oxygen was not required for atrazine degradation by whole cells or cell extracts. Based on high-pressure liquid chromatography and mass spectrometric analyses of products formed from atrazine in incubations of whole cells with H2 18O, sequential hydrolytic reactions converted atrazine to hydroxyatrazine and then to the end productN-ethylammelide. Isopropylamine, the putative product of the second hydrolytic reaction, supported growth as the sole carbon and nitrogen source. The triazine hydrolase from strain C190 was isolated and purified and found to have a Km for atrazine of 25 μM and a V max of 31 μmol/min/mg of protein. The subunit molecular mass of the protein was 52 kDa. Atrazine hydrolysis was not inhibited by 500 μM EDTA but was inhibited by 100 μM Mg, Cu, Co, or Zn. Whole cells and purified triazine hydrolase converted a range of chlorine or methylthio-substituted herbicides to the corresponding hydroxy derivatives. In summary, an atrazine-metabolizingNocardioides sp. widely distributed in agricultural soils degrades a range of s-triazine herbicides by means of a novel s-triazine hydrolase.

scholarly journals Characterization of a New L-Glutaminase Produced by Achromobacter xylosoxidans RSHG1, Isolated from an Expired Hydrolyzed L-Glutamine Sample

Catalysts ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 1262
Author(s):  
Rabia Saleem ◽  
Safia Ahmed
Keyword(s):  
Bacterial Isolate ◽  
Kinetic Studies ◽  
Morphological Characters ◽  
Achromobacter Xylosoxidans ◽  
16S Rdna Sequence ◽  
Potential Applications ◽  
Biochemical Testing ◽  
Ph Range ◽  
Glutaminase Activity

As significant biocatalyst, L-glutaminases find potential applications in various fields, from nourishment to the pharmaceutical industry. Anticancer activity and flavor enhancement are the most promising applications of L-glutaminases. In this study, L-glutaminase was isolated and purified from an old glutamine sample. A selected bacterial isolate was characterized taxonomically by morphological characters, biochemical testing and 16S rDNA sequence homology testing. The taxonomical characterization of the isolate identified it as Achromobacter xylosoxidans strain RSHG1. The isolate showed maximum enzyme production at 30 °C, pH 9, with Sorbitol as a carbon source and L-Glutamine as a nitrogen and inducer source. L-Glutaminsae was purified by using column chromatography on a Sephadex G-75. The enzyme has a molecular weight of 40 KDa, pH optimal 7 and is stable in the pH range of 6–8. The optimum temperature for the catalyst was 40 °C and stable at 35–50 °C. The kinetic studies of the purified L-glutaminase exhibited Km and Vmax of 0.236 mM and 443.8 U/mg, respectively. L-Glutaminase activity was increased when incubated with 20 mM CaCl2, BaCl2, ZnSO4, KCl, MgSO4 and NaCl, whereas EDTA, CoCl2, HgCl, ZnSO4 and FeSO4 decreased the activity of the enzyme. The addition of 8% NaCl enhanced the glutaminase activity. L-Glutaminase immobilized on 3.6% agar was stable for up to 3 weeks.

Characterization of an inducible porter required for L-proline catabolism by Escherichia coli K12

1979 ◽  
Vol 57 (10) ◽  
pp. 1191-1199 ◽  
Author(s):  
Janet M. Wood ◽  
David Zadworny
Keyword(s):  
Escherichia Coli ◽  
Catabolite Repression ◽  
Sole Source ◽  
Proline Oxidase ◽  
Transport Reaction ◽  
Carbon And Nitrogen ◽  
Escherichia Coli K12 ◽  
Proline Catabolism ◽  
Uptake Activity

L-Proline can serve as sole source of carbon and nitrogen for the growth of Escherichia coli K12 and other Enterobacteria. L-Proline uptake and L-proline oxidase are suoject both to catabolite repression and to specific induction by L-proline or glycyl-L-proline, although their regulation is not strictly coordinate. A strain defective for L-proline uptake due to a lesion at the locus putP does not show elevated uptake activity either on relief of catabolite repression or when grown on glycyl-L-proline as nitrogen source. The apparent Km for L-proline uptake decreases up to 14-fold as uptake Vm increases when cells are induced for both L-proline uptake and L-proline oxidase; cells with increased uptake activity, alone, do not show an altered Km. Although L-proline is metabolized during the uptake measurements, uptake is always active. The observed variations in uptake Km are unlikely to result from the escape of radioactive L-proline metabolites or from reversal of the transport reaction during the uptake measurements. We conclude that the L-proline porter encoded in putP is responsible for 80 to 90% of the constitutive and for the inducible L-proline uptake activity of wild-type bacteria. Although this porter is amplified in cells induced for L-proline catabolism, the observed values for uptake Vm may not be taken as a direct indicator of porter concentration.

Isolation and characterization of aPseudomonassp. strain PH1 utilizing meta-aminophenol

2000 ◽  
Vol 46 (3) ◽  
pp. 211-217 ◽  
Author(s):  
Razia Kutty ◽  
Hemant J Purohit ◽  
Purushottam Khanna
Keyword(s):  
Nitrogen Source ◽  
16S Rdna ◽  
Sole Source ◽  
Rdna Sequence ◽  
Pseudomonas Sp ◽  
16S Rdna Sequence ◽  
Industry Waste ◽  
Isolation And Characterization ◽  
Dye Industry

Pseudomonas sp. strain PH1 was isolated from soil contaminated with pharmaceutical and dye industry waste. The isolate PH1 could use m-aminophenol as a sole source of carbon, nitrogen, and energy to support the growth. PH1 could degrade up to 0.32 mM m-aminophenol in 120 h, when provided as nitrogen source at 0.4 mM concentration with citrate (0.5 mM) as a carbon source in the growth medium. The presence of ammonium chloride as an additional nitrogen source repressed the degradation of m-aminophenol by PH1. To identify strain PH1, the 16S rDNA sequence was amplified by PCR using conserved eubacterial primers. The FASTA program was used to analyze the 16S rDNA sequence and the resulting homology patterns suggested that PH1 is a Pseudomonas.Key words: m-aminophenol, resorcinol, DNA sequencing.

Biodegradation of 2, 4-Dinitrotoluene by Pseudomonas fluorescens sp. Isolated from a Nitrobenzene Contaminated Soil

2012 ◽  
Vol 610-613 ◽  
pp. 1233-1236 ◽  
Author(s):  
Zhong Hua Huang ◽  
Xue Pu ◽  
Hui Zhang ◽  
Fang Yuan ◽  
Zheng Li Liu
Keyword(s):  
Pseudomonas Fluorescens ◽  
16S Rdna ◽  
Contaminated Soil ◽  
Sole Source ◽  
Contaminated Groundwater ◽  
Mineral Salts ◽  
16S Rdna Sequence ◽  
Degrading Bacterium ◽  
Groundwater Environment ◽  
Initial Ph

A 2,4-Dinitrotoluene degrading bacterium was isolated from sludge contaminated by DNT and identified as a member of Pseudomonas fluorescens sp. based on 16S rDNA sequence. Under growth substrates limited conditions, the strain was enriched with 2,4-DNT as its sole source of carbon and energy in the groundwater environment. Results indicate the isolate grew optimally at 12(±4)°C and initial pH 7.5 in the mineral salts medium supplemented 2,4-DNT with 40mg/L, thus showing potential for the remediation of 2,4-DNT contaminated groundwater.

scholarly journals Characterization of a broad-specificity non-haem iron N-demethylase from Pseudomonas putida CBB5 capable of utilizing several purine alkaloids as sole carbon and nitrogen source

Microbiology ◽  
2011 ◽  
Vol 157 (2) ◽  
pp. 583-592 ◽  
Author(s):  
Ryan M. Summers ◽  
Tai Man Louie ◽  
Chi Li Yu ◽  
Mani Subramanian
Keyword(s):  
Pseudomonas Putida ◽  
Reductase Activity ◽  
Hypothetical Protein ◽  
Sole Source ◽  
Carbon And Nitrogen ◽  
Native Molecular Mass ◽  
Purine Alkaloids ◽  
Naturally Occurring ◽  
Haem Iron

N-Demethylation of many xenobiotics and naturally occurring purine alkaloids such as caffeine and theobromine is primarily catalysed in higher organisms, ranging from fungi to mammals, by the well-studied membrane-associated cytochrome P450s. In contrast, there is no well-characterized enzyme for N-demethylation of purine alkaloids from bacteria, despite several reports on their utilization as sole source of carbon and nitrogen. Here, we provide what we believe to be the first detailed characterization of a purified N-demethylase from Pseudomonas putida CBB5. The soluble N-demethylase holoenzyme is composed of two components, a reductase component with cytochrome c reductase activity (Ccr) and a two-subunit N-demethylase component (Ndm). Ndm, with a native molecular mass of 240 kDa, is composed of NdmA (40 kDa) and NdmB (35 kDa). Ccr transfers reducing equivalents from NAD(P)H to Ndm, which catalyses an oxygen-dependent N-demethylation of methylxanthines to xanthine, formaldehyde and water. Paraxanthine and 7-methylxanthine were determined to be the best substrates, with apparent K m and kcat values of 50.4±6.8 μM and 16.2±0.6 min−1, and 63.8±7.5 μM and 94.8±3.0 min−1, respectively. Ndm also displayed activity towards caffeine, theobromine, theophylline and 3-methylxanthine, all of which are growth substrates for this organism. Ndm was deduced to be a Rieske [2Fe–2S]-domain-containing non-haem iron oxygenase based on (i) its distinct absorption spectrum and (ii) significant identity of the N-terminal sequences of NdmA and NdmB with the gene product of an uncharacterized caffeine demethylase in P. putida IF-3 and a hypothetical protein in Janthinobacterium sp. Marseille, both predicted to be Rieske non-haem iron oxygenases.

scholarly journals Aerobic Degradation of Dinitrotoluenes and Pathway for Bacterial Degradation of 2,6-Dinitrotoluene

2000 ◽  
Vol 66 (5) ◽  
pp. 2139-2147 ◽  
Author(s):  
Shirley F. Nishino ◽  
George C. Paoli ◽  
Jim C. Spain
Keyword(s):  
Burkholderia Cepacia ◽  
Dienoic Acid ◽  
Sole Source ◽  
Bacterial Degradation ◽  
Ring Cleavage ◽  
Bacterial Strains ◽  
Carbon And Nitrogen ◽  
Isolation And Characterization ◽  
Oxidative Pathway

ABSTRACT An oxidative pathway for the mineralization of 2,4-dinitrotoluene (2,4-DNT) by Burkholderia sp. strain DNT has been reported previously. We report here the isolation of additional strains with the ability to mineralize 2,4-DNT by the same pathway and the isolation and characterization of bacterial strains that mineralize 2,6-dinitrotoluene (2,6-DNT) by a different pathway.Burkholderia cepacia strain JS850 andHydrogenophaga palleronii strain JS863 grew on 2,6-DNT as the sole source of carbon and nitrogen. The initial steps in the pathway for degradation of 2,6-DNT were determined by simultaneous induction, enzyme assays, and identification of metabolites through mass spectroscopy and nuclear magnetic resonance. 2,6-DNT was converted to 3-methyl-4-nitrocatechol by a dioxygenation reaction accompanied by the release of nitrite. 3-Methyl-4-nitrocatechol was the substrate for extradiol ring cleavage yielding 2-hydroxy-5-nitro-6-oxohepta-2,4-dienoic acid, which was converted to 2-hydroxy-5-nitropenta-2,4-dienoic acid. 2,4-DNT-degrading strains also converted 2,6-DNT to 3-methyl-4-nitrocatechol but did not metabolize the 3-methyl-4-nitrocatechol. Although 2,6-DNT prevented the degradation of 2,4-DNT by 2,4-DNT-degrading strains, the effect was not the result of inhibition of 2,4-DNT dioxygenase by 2,6-DNT or of 4-methyl-5-nitrocatechol monooxygenase by 3-methyl-4-nitrocatechol.

Purification and characterization of an alkaline serine endopeptidase from a feather-degradingXanthomonas maltophiliastrain

2002 ◽  
Vol 48 (4) ◽  
pp. 342-348 ◽  
Author(s):  
C H De Toni ◽  
M F Richter ◽  
J R Chagas ◽  
J AP Henriques ◽  
C Termignoni
Keyword(s):  
Sole Source ◽  
Peptide Substrates ◽  
Xanthomonas Maltophilia ◽  
Carbon And Nitrogen ◽  
Step Procedure ◽  
Zone Electrophoresis ◽  
Capillary Zone ◽  
Extracellular Peptidases ◽  
Fluorogenic Peptide

A keratinolytic Xanthomonas maltophilia strain (POA-1), cultured on feather meal broth, using keratin as its sole source of carbon and nitrogen, secretes several extracellular peptidases. The major serine peptidase was purified to homogeneity by a five-step procedure. Its purity was evaluated by capillary zone electrophoresis. This enzyme has a molecular mass of 36 kDa, an optimum pH of 9.0, and an optimum temperature of 60°C. The inhibitory profile using protease inhibitors shows that this enzyme is a serine endopeptidase. Besides keratin, the enzyme is active upon the substrates azokeratin, azocasein, and the following fluorogenic peptide substrates: Abz-Leu-Gly-Met-Ile-Ser-Leu-Met-Lys-Arg-Pro-Gln-EDDnp, Abz-Lys-Leu-Cys(SBzl)-Gly-Pro-Lys-Gln-EDDnp, and Abz-Lys-Pro-Cys(SBzl)-Phe-Ser-Lys-Gln-EDDnp.Key words: serine endopeptidase, Xanthomonas maltophilia, keratinase, alkaline endopeptidase.

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