Mineralization of Carbofuran by a Soil Bacterium

A bacterium, tentatively identified as an Arthrobacter sp., was isolated from flooded soil that was incubated at 35°C and repeatedly treated with carbofuran (2,3-dihydro-2,2-dimethyl-7-benzofuranyl N-methylcarbamate). This bacterium exhibited an exceptional capacity to completely mineralize the ring-labeled ¹⁴C in carbofuran to ¹⁴CO₂ within 72 to 120 h in a mineral salts medium as a sole source of carbon and nitrogen under aerobic conditions. Mineralization was more rapid at 35°C than at 20°C. No degradation of carbofuran occurred even after prolonged incubation under anaerobic conditions. The predicted metabolites of carbofuran, 7-phenol (2,3-dihydro-2,2-dimethyl-7-benzofuranol) and 3-hydroxycarbofuran, were also metabolized rapidly. 7-Phenol, although formed during carbofuran degradation, never accumulated in large amounts, evidently because of its further metabolism through ring cleavage. The bacterium readily hydrolyzed carbaryl (1-naphthyl N-methylcarbamate), but its hydrolysis product, 1-naphthol, resisted further degradation by this bacterium.     

Self-immobilized bacterial cultures with potential for application as ready-to-use seeds for petroleum bioremediation

Two hydrocarbon-degrading bacterial isolates, an Arthrobacter sp. and a Gram-negative bacillus isolated from Kuwait oil lakes, exhibited considerable cell-surface hydrophobicity without production of exopolysaccharides in complex media. However, the bacteria produced copious amounts of exopolysaccharides in a low nutrient medium. When incubated with sawdust, Styrofoam or wheat bran, as carriers, under low nutrient conditions, stable exopolysaccharide-mediated immobilized cultures were formed. Such immobilized cultures when air-dried at room temperature survived storage for 6 weeks at 45 °C and still retained the ability to degrade hydrocarbons. Viability was retained by the immobilized Arthrobacter sp. and the Gram-negative bacterium at 45 °C storage for up to 6 and 12 months, respectively.     

A comparison of autoclaved and gamma-irradiated soils as media for microbial colonization experiments

Summary The effects on various properties of Lincoln Clay of a sterilizing dose of gamma radiation (3.0 megarads) were compared to the effects resulting from autoclaving the soil (121°C) for 1 hour. Effects of both treatments were much more drastic when moist, rather than air-dry soil was treated but, in general, radiation had less effect on soluble organic matter and on total water-extractable electrolyte than did autoclaving. Radiation caused a greater release of NH₄-N from soil treated moist than did autoclaving but the reverse was true in dry soil. Alcohol-soluble ninhydrin-positive material was increased by both sterilization procedures with irradiation having the greater effect. The aggregate stability of Lincoln clay was decreased by irradiation and increased by autoclaving.Pure cultures of bothArthrobacter sp. andPseudomonas sp. grew better, on the basis of cell yields, in irradiated than in autoclaved soil. Respiration of mixed soil organisms in an artificial soil amended with an extract of irradiated soil was almost identical with that in an extract of air-dried soil but auto-claved soil extract was only metabolized after a prolonged lag period.This work was supported by grant # A 1702 from the National Research Council of Canada, Ottawa, Canada.     

Only C-2 specific glucose oxidase activity is expressed in ligninolytic cultures of the white rot fungus Phanerochaete chrysosporium

Hydroxylamine oxidation was measured in four recently isolated heterotrophic nitrate-reducing bacteria belonging to the generaPseudomonas, Moraxella, Arthrobacter andAeromonas. A hydroxylamine-cytochromec oxidoreductase activity was detected in periplasmic fractions of thePseudomonas andAeromonas spp. and in total soluble fractions of theArthrobacter sp. A monomeric 19-kDa non-haem iron hydroxylamine-cytochromec oxidoreductase was purified from thePseudomonas species and shown to be similar to hydroxylaminecytochromec oxidoreductase ofParacoccus denitrificans.Abbreviations AMO Ammonia monoxygenase - HAO Hydroxylamine-cytochromec oxidoreductase     

A hyper-thermostable, alkaline lipase from Pseudomonas sp. with the property of thermal activation

A hyper-thermostable, alkaline lipase from a newly-isolated, mesophilic Pseudomonas sp. was optimal at pH 11 and at 90 °C. It had a half-life of more than 13 h at 90 °C. It was activated by 30% when heated at 90 °C for 2 h. The enzyme had a greater affinity for mustard oil (K _m=40 mg ml^–1) than for olive oil (K _m=140 mg ml^–1).     

Aerobic biodegradation of nonylphenol by cold-adapted bacteria

Three strains capable of mineralizing nonylphenol as sole carbon source were isolated from a sample of contaminated soil and characterized as two Pseudomonas spp. and a Stenotrophomonas sp. The two Pseudomonas spp. expressed characteristics typical of psychrophiles growing optimally of 10 °C and capable of growing at 0 °C. The Stenotrophomonas sp. was more likely psychrotrophic because it had an optimal temperature between 14 and 22 °C although it was not capable of growing at 4 °C. At 14 °C, one of the Pseudomonas spp. exhibited the highest rate of degradation of nonylphenol (4.4 mg l^–1 d^–1), when compared with axenic or mixed cultures of the isolates. This study represents, to the best of our knowledge, the first reported case of cold-adapted microorganisms capable of mineralizing nonylphenol.     

Simultaneous mineralization of glyphosate and diuron by a consortium of three bacteria as free- and/or immobilized-cells formulations

A bacterial consortium able to mineralize two herbicides, glyphosate (Pseudomonas 4ASW) and diuron (Arthrobacter sp. N4 and Delftia acidovorans), was cultivated in both a synthetic culture medium without phosphate and a sediment extract medium. In the aim at optimizing glyphosate and diuron mineralization, all the combinations, i.e., free and/or immobilized cells in Ca-alginate beads were tested. With the synthetic medium, the simultaneous mineralization of glyphosate and diuron required at least the immobilization of Pseudomonas 4ASW. Conversely, with the sediment extract medium, only the mineralization of diuron was observed, most probably, because of both nutrient deficiency and phosphate in the sediment extract medium.     

Toxicity of Hexavalent Chromium and Its Reduction by Bacteria Isolated from Soil Contaminated with Tannery Waste

An Arthrobacter sp. and a Bacillus sp., isolated from a long-term tannery waste contaminated soil, were examined for their tolerance to hexavalent chromium [Cr(VI)] and their ability to reduce Cr(VI) to Cr(III), a detoxification process in cell suspensions and cell extracts. Both bacteria tolerated Cr(VI) at 100 mg/ml on a minimal salts agar medium supplemented with 0.5% glucose, but only Arthrobacter could grow in liquid medium at this concentration. Arthrobacter sp. could reduce Cr(VI) up to 50 μg/ml, while Bacillus sp. was not able to reduce Cr(VI) beyond 20 μg/ml. Arthrobacter sp. was distinctly superior to the Bacillus sp. in terms of their Cr(VI)-reducing ability and resistance to Cr(VI). Assays with permeabilized (treated with toluene or Triton X 100) cells and crude extracts demonstrated that the Cr(VI) reduction was mainly associated with the soluble protein fraction of the cell. Arthrobacter sp. has a great potential for bioremediation of Cr(VI)-containing waste. Received: 13 June 2002 / Accepted: 13 September 2002     

Metabolic ability and gene characteristics of Arthrobacter sp. strain DNS10, the sole atrazine-degrading strain in a consortium isolated from black soil

Strain DNS10 was the only member that could utilize atrazine as the sole nitrogen source for growth in an atrazine-degrading consortium which was isolated from black soil previously in our laboratory. It belongs to the genus Arthrobacter according to the sequence of 16S rRNA gene and is designated as Arthrobacter sp. DNS10. 16S rRNA gene phylogenetic analysis showed that strain DNS10 was located in a different evolutionary branch comparing with other Arthrobacter sp. atrazine-degrading strains. The degrading genes such as trzN, atzB and atzC harbored in strain DNS10 revealed high sequence similarity with those in Arthrobacter aurescens TC1 and Pseudomonas sp. ADP. These genes enabled the strain DNS10 to decompose atrazine to cyanuric acid. This was further proved by the results that the strain DNS10 (10⁸ CFU mL⁻¹) could degrade the whole atrazine (100 mg L⁻¹) in the medium within 24 h at 30 °C and there was 66.13 ± 2.11 mg L⁻¹ cyanuric acid accumulated at 24 h. These results imply that the strain DNS10 seems to be an excellent atrazine-degrading strain. Furthermore, this paper helps us in the better understanding of the strain evolution by comparing the metabolic ability and gene characteristics of strain DNS10 with other geographically distinct atrazine-degrading strains.     

Thermostability of an alkaline protease, AprP, is enhanced by replacements of Ser307 and Ser331 at the cleavage sites

The thermostability of an alkaline protease, AprP from Pseudomonas sp. KFCC 10818, was improved by replacing Ser307 and Ser331 at the autoproteolytic cleavage sites with various amino acids. Six mutant enzymes were purified and characterized. Two of these had half-lives four and three times longer than the wild-type protease at 55 °C in the presence of 1 mM CaCl₂. Three mutant enzymes had half-lives twice as long as the wild-type under the same condition.     

A two-enzyme system for the transformation of unsaturated oils to 9(S)-hydroperoxy fatty acids

A two-enzyme system involving a lipase from a Pseudomonassp. and an extract of potato tubers containing lipoxygenase was used to convert triacylglycerols to 9-hydroperoxy fatty acids. Highest yields (up to 25%) were obtained with 1 to 20 g l^–1 trilinolein as substrate after 5 h under O₂ at pH 6 and 25 °C. The product structure was confirmed by ¹H NMR, MS and IR.     

Isolation and characterization of sulfur-utilizing denitrifiers from the sulfur-oxidizing denitrification process

Of 14 potential sulfur-oxidizing strains, Pseudomonas sp. B21 and Agrobacterium sp. B19 were considered as denitrifiers. Under aerobic conditions, with S⁰ as electron donor, maximum cell growth rates were 0.022 (B21) and 0.043 h^–1 (B19). Both grew optimally at pH 7.5 and 28 °C. When NO₃-N was increased from 10 to 200 mg l^–1 the efficiency of nitrate removal of each strain gradually decreased, from 60 to 40%. Addition of suitable organic compounds (C/N < 4.2) increased the nitrate removal efficiencies of both strains, indicating their mixotrophic characters.     

Depolymerisation and biodegradation of a synthetic tanning agent by activated sludges, the bacteria Arthrobacter globiformis and Comamonas testosteroni, and the fungus Cunninghamella polymorpha

Degradation of a synthetic tanning agent CNSF (a condensation product of 2-naphthalenesulfonic acid (2-NSA) and formaldehyde) by four activated sludges, two previously characterised bacterial strains, Arthrobacter sp. 2AC and Comamonas sp. 4BC, and the fungus Cunninghamella polymorpha, was studied in batch culture at 25°C by determining the changes in the concentrations of CNSF and its component monomers and oligomers (n2–n11). The loss of individual oligomers was correlated with the length of the NSA-CH₂ chain. Approximately 25% of the total CNSF was degraded (i.e. mineralised) by the microbes contained in the four activated sludges and by the two bacterial isolates but with different lag phases and at different overall rates. The decline in CNSF concentration was due almost entirely to the biodegradation of the monomers (34.3% of CNSF) and, in particular, 2-NSA (27% of CNSF). There was no change in the n2–n11 components. The growth of C. polymorpha, on the other hand, arose from extracellular depolymerisation of CNSF oligomers and the biodegradation of the lower molecular mass products. Between 38% and 42% of total CNSF was degraded by C. polymorpha at 25°C. The order of oligomer degradation was inversely related to degree of polymerisation. Eighty percent and 90% of the n4 and n5 and 100% oligomers n6–n11 were degraded after 120h. At a higher temperature (37°C) oligomers n4–n11 were degraded completely after 120h. A combination of biodegradation (75%) and sorption to fungal biomass (25%) accounted for the measured loss of all oligomers from the solution phase. The CNSF degradation rates and the volume of fungal biomass produced (and therefore the extent of biosorption) were dependent on the presence of a second carbon source (both optimum at glucose 5g/l). This is the first report that identifies and distinguishes between depolymerisation, sorption and biodegradation processes in the removal of CNSF and its component oligomers. The use of combinations of the depolymerising fungus C.polymorpha, and the monomer-degrading bacteria, Arthrobacter sp. 2AC and Comamonas sp. 4BC, have potential for wastewater treatment.     

Production of alkali-tolerant cellulase-free xylanase by <Emphasis Type="Italic">Pseudomonas</Emphasis> sp. WLUN024 with wheat bran as the main substrate

An alkali-tolerant cellulase-free xylanase producer, WLI-11, was screened from soil samples collected from a pulp and paper mill in China. It was subsequently identified as a Pseudomonas sp. A mutant, WLUN024, was selected by consecutive mutagenesis by u.v. irradiation and NTG treatment using Pseudomonas sp. WLI-11 as parent strain. Pseudomonas sp. WLUN024 produced xylanase when grown on xylosidic materials, such as hemicellulose, xylan, xylose, and wheat bran. Effects of various nutritional factors on xylanase production by Pseudomonas sp. WLUN024 with wheat bran as the main substrate were investigated. A batch culture of Pseudomonas sp. WLUN024 was conducted under suitable fermentation conditions, where the maximum activity of xylanase reached 1245 U ml⁻¹ after incubating at 37 °C for 24 h. Xylanase produced by Pseudomonas sp. WLUN024 was purified and the molecular weight was estimated as 25.4 kDa. Primary studies on the characteristics of the purified xylanase revealed that this xylanase was alkali-tolerant (optimum pH 7.2–8.0) and cellulase-free. In addition, the xylanase was also capable of producing high quality xylo-oligosaccharides, which indicated its application potential in not only pulp bio-bleaching processes but also in the nutraceutical industry.     

Microbiology of the fermentation ofTelfaria seeds for ogiri production

Summary Ogiri was prepared from seeds of fluted pumpkin in the traditional way. Microorganisms associated with the fermentation were identified asBacillus sp.,Staphylococcus aureus,Pseudomonas sp. andLactobacillus sp. The temperature increased to 40°C during the fermentation, while the pH first increased to 7.8 then dropped to 6.0 by the end of fermentation. The characteristic taste and strong aroma of the ogiri is attributed to the proteolytic and lipolytic activities of the isolates.

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Microbiologie de la fermentation des graines de Telefaria pour la production d'ogiri

Résumé On a préparé l'ogiri à partir de graines de potiron à cannelures à la manière traditionnelle. Les microorganismes associés à la fermentation ont été identifiés commeBacillus sp.,Staphylococcus aureus, Pseudomonas sp. etLactobacillus sp. La température monte à 40°C pendant la fermentation, tandis que le pH augmente d'abord à 7.8 pour décroître ensuite à 6.0 à la fin de la fermentation. Le goût caractéristique et l'arôme fort de l'ogiri sont attribuables aux activités protéolytiques et lipolytiques des isolats.

     

Arbuscular Mycorrhizal Fungi and Rhizospheric Bacteria Diversity Along an Altitudinal Gradient in South American Puna Grassland

Rhizospheric soil samples were taken from Puna native grasses along an altitudinal gradient. Biodiversity of arbuscular mycorrhizal fungi (AMF) and associated bacteria was analyzed considering altitude and grasses photosynthetic pathways (metabolic type C₃, C₄). Cultivation-dependent approaches were applied to obtain further information about the phylogeny of the dominating cultivable aerobic–heterotrophic bacteria communities present in rhizospheric soil samples. In average, the bacterial count ranged between 1.30 × 10² and 8.66 × 10⁴ CFU g⁻¹ of dry weight of soil. Individual bacterial colonies of aerobic heterotrophic bacteria grown on R2A medium were morphologically grouped and identified as typical soil bacteria belonging to the genera Bacillus, Pseudomonas, and Arthrobacter. Ten AMF taxa were found: Acaulospora sp., A. laevis, A. spinosa, Gigaspora sp., Gi. ramisporophora, Glomus sp., Gl. aggregatum, Gl. ambisporum, Gl. sinuosum, and Scutellospora biornata. AMF diversity decreased with altitude.     

Investigation of the synthesis of xylose/glucose isomerases in sixArthrobacter sp. strains

The substrate specificity of isomerases produced by six strains ofArthrobacter sp. was studied. The role of utilizable carbon sources in controlling enzyme biosynthesis was established. All of the strains studied were found to produce xylose isomerases efficiently, converting D-xylose into D-xylulose and D-glucose into D-fructose. All but A.ureafaciens B-6 strains showed low activity toward D-ribose,Arthrobacter sp. B-5 was slightly active toward L-arabinose, andA. ureafaciens B-6 andArthrobacter sp. B-2239, toward L-rhamnose. InArthrobacter sp. B-5, the synthesis of xylose/glucose isomerase was constitutive (i.e., it was not suppressed by readily metabolizable carbon sources. The synthesis of xylose/glucose isomerase induced by D-xylose inArthrobacter sp. strains B-2239, B-2240, B-2241, and B-2242 and by D-xylose and xylitol inA. ureafaciens B-6 was suppressed by readily metabolizable carbon sources in a concentration-dependent manner. The data obtained suggest that D-xylose and/or its metabolites are involved in the regulation of xylose/glucose isomerase synthesis in theArthrobacter sp. strains B-5, B-2239, B-2240, and B-2241.     

Thermophilic methanogenesis from pectin by a mixed defined bacterial culture

Thermophilic degradation of pectin was studied in batch cultures at 55°C by different associations of anaerobic bacteria, includingClostridium thermocellum, Methanobacterium sp., andMethanosarcina sp.Clostridium thermocellum alone produced large amounts of methanol along with some isopropanol and H₂. The inoculation ofMethanobacterium sp. in the culture did not affect the metabolism ofC. thermocellum; this demonstrates the absence of interspecies hydrogen transfer. In the presence of the methylotrophicMethanosarcina sp., methanol was reduced to methane without effect on pectin hydrolysis; a small amount of the H₂ produced was also used to reduce methanol.     

Microbial biodiversity and in situ bioremediation of endosulfan contaminated soil

Molecular characterization based on 16s rDNA gene sequence analysis of bacterial colonies isolated from endosulfan contaminated soil showed the presence of Ochrobacterum sp, Burkholderia sp, Pseudomonas alcaligenes, Pseudomonas sp and Arthrobacter sp which degraded 57–90% of α-endosulfan and 74–94% of β-endosulfan after 7days. Whole cells of Pseudomonas sp and Pseudomonas alcaligenes showed 94 and 89% uptake of α-isomer and 86 and 89% of β-endosulfan respectively in 120 min. In Pseudomonas sp, endosulfan sulfate was the major metabolite detected during the degradation of α-isomer, with minor amount of endosulfan diol while in Pseudomonas alcaligenes endosulfan diol was the only product during α-endosulfan degradation. Whole cells of Pseudomonas sp also utilized 83% of endosulfan sulfate in 120 min. In situ applications of the defined consortium consisting of Pseudomonas alcaligenes and Pseudomonas sp (1:1) in plots contaminated with endosulfan showed that 80% of α-endosulfan and 65% of β-endosulfan was degraded after 12 weeks of incubation. Endosulfan sulfate formed during endosulfan degradation was subsequently degraded to unknown metabolites. ERIC-PCR analysis indicated 80% survival of introduced population of Pseudomonas alcaligenes and Pseudomonas sp in treated plots.     

Trophic interactions in soils as they affect energy and nutrient dynamics. II. Physiological responses of selected rhizosphere bacteria

Comparative microbial functions in the plant root zone were studied by evaluating rhizosphere-derivedPseudomonas andArthrobacter growth in chemostat culture and responses to root-exudate-related nutrients after varied starvation periods. These organisms were chosen to represent zymogenous and autochthonous microbes, respectively. In chemostat culture, thePseudomonas isolate showed increased energy charge and decreased populations with higher growth rates, whereas theArthrobacter had lower energy charge and cell population values which did not change appreciably with growth rate. The responses of these two types of organisms also differed with starvation. ThePseudomonas lost its ability to respire efficiently in the presence of several known root exudate components, whereas theArthrobacter isolate, in comparison, maintained a lower but more consistent ability to utilize these nutrients with increased starvation. TheArthrobacter also showed increased utilization of several substrates after starvation, suggesting its potential ability to function under restricted nutrient availability conditions. These results suggest thatPseudomonas-type organisms in the rhizosphere may best function in periods of more intense exudate release, whereas organisms of theArthrobacter- type may be more efficient at nutrient utilization during periods of lesser nutrient flux. Based on these data the rhizosphere-derivedPseudomonas isolate was considered to be an appropriate bacterium to use in more complex rhizosphere microcosm experiments where nutrient flux dynamics would be emphasized.