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Electron spin resonance study of copper biosorption by bacteria.

The biosorption of copper by bacteria was studied by using ESR spectroscopy. Among bacteria tested, Arthrobacter nicotianae has the most excellent ability to sorb copper. The biosorption of copper by Arthrobacter cells was so rapid, affected by the solution pH, and obeys the Langmuir adsorption equilibrium. The electron spin resonance (ESR) spectra of Cu(II) in bacterial cells are axial type, having a major absorption to higher field at gperpendicular and lesser absorption to lower field at gparallel with four lines. The ESR parameters showed that Cu(II) in the cells has the tetragonally distorted octahedral structure with nitrogen and oxygen as ligand atoms, which suggests that most of copper in bacterial cells combined with amino acid residues in the cell surface proteins. The variation of spectral patterns among bacteria could explain as the change of ligand circumstances caused by the pH of the cell surface.

Absorption↗

Dimethylphthalate hydrolysis by specific microbial esterase.

TWO BACTERIAL STRAINS: Arthrobacter sp. and Sphingomonas paucimobilis were isolated from soil by enrichment cultures using dimethylphthalate (DMP) or monomethylphthalate (MMP) as sole carbon source, respectively. DMP was rapidly transformed by an Arthrobacter sp. culture with formation of MMP and phthalic acid (PA) which is further degraded. This strain was unable to hydrolyse MMP. A mechanism of degradation of DMP was proposed with two ways: DMP-->PA and DMP-->MMP. The S. paucimobilis strain hydrolyses only MMP and a coculture of the two strains allowed a complete degradation of DMP.

Arthrobacter↗

Quantification of bacterial lead resistance via activity assays.

The level of microbial resistance to heavy metals is an important issue for the microbial ecology of heavy metal-contaminated habitats. However, assays based upon growth in nutrient media will overestimate the resistance level due to metal ion interactions with inorganic and organic components. The analysis of Pb-resistant bacteria isolated from soils containing up to 38 mmol total Pb x kg(-1) indicated that PYT80B medium which did not contain inorganic salts, contained low amounts of organic matter, and was buffered with a molecule that did not interact with metal ions (2-N-morpholinoethanesulfonic acid (MES)) provided the lowest estimates of lead resistance. However, better results were obtained by assaying metabolic activity (aerobic respiration) of resting cells suspended in 10 mM MES. By this criterion, 50% inhibition of Arthrobacter JS7 was found at 37 microM Pb(NO3)2. The effects of Pb+2 concentrations upon respiration of resting cells and growth rate in PYT80B medium were similar. The activity assay also showed that metal resistance was induced to higher levels when Arthrobacter JS7 was grown in the presence of Pb.

Arthrobacter↗

Optimization of protease activity of alkaliphilic bacteria isolated from an alkaline lake in India.

Alkaliphilic bacteria belonging to the genera Bacillus, Staphylococcus, Micrococcus, Pseudomonas and Arthrobacter isolated from sediment samples of the alkaline Lonar lake situated in the Buldhana District of Maharashtra State, India, were studied for the production of protease activity. Among the 54 isolates obtained, Arthrobacter ramosus and Bacillus alcalophilus exhibited high protease activity using soyacake as a sole source of carbon and nitrogen. Protease activity was optimum at 1% initial substrate concentration, at 30 degrees C and under shake culture condition for both organisms. The enzyme was thermostable (65 degrees C), stable at pH 12 and also active in the presence of commercial detergent. This enzyme removed blood stains from cotton fabric indicating its potential use in detergent formulations.

Arthrobacter↗

Metabolism of nitrate esters by a consortium of two bacteria.

The products of condensation of organic alcohols and nitric acid are nitrate esters with the general structure C-O-NO2. These products are widely employed as vasodilators and explosives, and are true xenobiotic compounds, as they do not occur in nature. We have isolated and characterized a consortium of two microorganisms, Arthrobacter ilicis and Agrobacterium radiobacter, that mineralized recalcitrant ethylene glycol dinitrate. The Arthrobacter strain was the actual degrading microorganism, although the second microbe facilitated mineralization. The biodegradation of ethylene glycol dinitrate by A. ilicis involved the progressive elimination of the nitro groups from the organic molecule to generate ethylene glycol, which was then mineralized. Waters polluted with ethylene glycol dinitrate have been shown amenable to biological treatment in a pilot plant with wastewaters generated during the synthesis of the chemical in a factory.

Arthrobacter↗

Biomineralization of an organophosphorus pesticide, Monocrotophos, by soil bacteria.

AIMS: To study biomineralization of Monocrotophos (MCP) and identify the metabolites formed during biodegradation. METHODS AND RESULTS: Two cultures, namely Arthrobacter atrocyaneus MCM B-425 and Bacillus megaterium MCM B-423, were isolated by enrichment and adaptation culture technique from soil exposed to MCP. The isolates were able to degrade MCP to the extent of 93% and 83%, respectively, from synthetic medium containing MCP at the concentration of 1000 mg x l(-1), within 8 d, under shake culture condition at 30 degrees C. The cultures degraded MCP to carbon dioxide, ammonia and phosphates through formation of one unknown compound--Metabolite I, valeric or acetic acid and methylamine, as intermediate metabolites. The enzymes phosphatase and esterase, reported to be involved in biodegradation of organophosphorus compounds, were detected in both the organisms. CONCLUSIONS: Arthrobacter atrocyaneus MCM B-425 and B. megaterium MCM B-423 isolated from soil exposed to MCP were able to mineralize MCP to carbon dioxide, ammonia and phosphates. SIGNIFICANCE AND IMPACT OF THE STUDY: Pathway for biodegradation of MCP in plants and animals has been reported. A microbial metabolic pathway of degradation involving phosphatase and esterase enzymes has been proposed. The microbial cultures could be used for bioremediation of wastewater or soil contaminated with Monocrotophos.

Ammonia↗

Switching substrate preference of thermophilic xylose isomerase from D-xylose to D-glucose by redesigning the substrate binding pocket.

The substrate specificity of thermophilic xylose isomerase from Clostridium thermosulfurogenes was examined by using predictions from the known crystal structure of the Arthrobacter enzyme and site-directed mutagenesis of the thermophile xylA gene. The orientation of glucose as a substrate in the active site of the thermophilic enzyme was modeled to position the C-6 end of hexose toward His-101 in the substrate-binding pocket. The locations of Met-87, Thr-89, Val-134, and Glu-180, which contact the C-6-OH group of the substrate in the sorbitol-bound xylose isomerase from Arthrobacter [Collyer, C.A., Henrick, K. & Blow, D. M. (1990) J. Mol. Biol. 212, 211-235], are equivalent to those of Trp-139, Thr-141, Val-186, and Glu-232 in the thermophilic enzyme. Replacement of Trp-139 with Phe reduced the Km and enhanced the kcat of the mutant thermophilic enzyme toward glucose, whereas this substitution reversed the effect toward xylose. Replacement of Val-186 with Thr also enhanced the catalytic efficiency of the enzyme toward glucose. Double mutants with replacements Trp-139----Phe/Val-186----Thr and Trp-139----Phe/Val-186----Ser had a higher catalytic efficiency (kcat/Km) for glucose than the wild-type enzyme of 5- and 2-fold, respectively. They also exhibited 1.5- and 3-fold higher catalytic efficiency for D-glucose than for D-xylose, respectively. These results provide evidence that alteration in substrate specificity of factitious thermophilic xylose isomerases can be achieved by designing reduced steric constraints and enhanced hydrogen-bonding capacity for glucose in the substrate-binding pocket of the active site.

Aldose-Ketose Isomerases↗

Biotreatment of hydrogen sulfide- and ammonia-containing waste gases by fluidized bed bioreactor.

Gas mixtures of H2S and NH3 are the focus of this study of research concerning gases generated from animal husbandry and treatments of anaerobic wastewater lagoons. A heterotrophic microflora (a mixture of Pseudomonas putida for H2S and Arthrobacter oxydans for NH3) was immobilized with Ca-alginate and packed into a fluidized bed reactor to simultaneously decompose H2S and NH3. This bioreactor was continuously supplied with H2S and NH3 separately or together at various ratios. The removal efficiency, removal rate, and metabolic product of the bioreactor were studied. The results showed that the efficiency remained above 95% when the inlet H2S concentration was below 30 ppm at 36 L/hr. Furthermore, the apparent maximum removal and the apparent half-saturation constant were 7.0 x 10(-8) g-S/cell/day and 76.2 ppm, respectively, in this study. The element sulfur as a main product prevented acidification of the biofilter, which maintained the stability of the operation. As for NH3, the greater than 90% removal rate was achieved as long as the inlet concentration was controlled below 100 ppm at a flow rate of 27 L/hr. In the NH3 inlet, the apparent maximum removal and the apparent half-saturation constant were 1.88 x 10(-6) g-N/cell/day and 30.5 ppm, respectively. Kinetic analysis showed that 60 ppm of NH3 significantly suppressed the H2S removal by Pseudomonas putida, but H2S in the range of 5-60 ppm did not affect NH3 removal by Arthrobacter oxydans. Results from bioaerosol analysis in the bioreactor suggest that the co-immobilized cell technique applied for gas removal creates less environmental impact.

Ammonia↗

The structure of resting bacterial populations in soil and subsoil permafrost.

The structure of individual cells in microbial populations in situ of the Arctic and Antarctic permafrost was studied by scanning and transmission electron microscopy methods and compared with that of cyst-like resting forms generated under special conditions by the non-spore-forming bacteria Arthrobacter and Micrococcus isolated from the permafrost. Electron microscopy examination of microorganisms in situ revealed several types of bacterial cells having no signs of damage, including "dwarf" curved forms similar to nanoforms. Intact bacterial cells in situ and frozen cultures of the permafrost isolates differed from vegetative cells by thickened cell walls, the altered structure of cytoplasm, and the compact nucleoid, and were similar in these features to cyst-like resting forms of non-spore-forming "permafrost" bacterial strains of Arthrobacter and Micrococcus spp. Cyst-like cells, being resistant to adverse external factors, are regarded as being responsible for survival of the non-spore-formers under prolonged exposure to subzero temperatures and can be a target to search for living microorganisms in natural environments both on the Earth and on extraterrestrial bodies.

Arthrobacter↗

Action of chondroitinases. I. The mode of action of two chondroitinase-AC preparations of different origin.

The modes of action of two chondroitinases-AC [EC 4.2.2.5] from Arthrobacter aurescens and Flavobacterium heparinum were examined. By comparison of the increase of viscosity and by analyses of digests using paper chromatography and gel filtration, it was shown that the Arthrobacter enzyme (A-Chase) degrades the substrate by a stepwise attack, while the Flavobacterium enzyme (F-Chase) exhibits a more random attack, though the first attack of both enzymes is of endo-type. Further study was carried out of the initial rate and final extent of the enzymic degradation of various mucopolysaccharides. The order of the initial rates at which the native mucopolysaccharides are degraded was similar for both enzymes. For A-Chase, the initial rate and final extent of degradation of modified chondroitin sulfate C or chondroitin methyl ester, however, were low compared with those of F-Chase. These results also suggested that A-Chase degrades the substrate by stepwise attack and F-Chase by random attack.

Arthrobacter↗

Cold-active citrate synthase: mutagenesis of active-site residues.

A comparison of the crystal structure of the dimeric enzyme citrate synthase from the psychrophilic Arthrobacter strain DS2-3R with that of the structurally homologous enzyme from the hyperthermophilic Pyrococcus furiosus reveals a significant difference in the accessibility of their active sites to substrates. In this work, we investigated the possible role in cold activity of the greater accessibility of the Arthrobacter citrate synthase. By site-directed mutagenesis, we replaced two alanine residues at the entrance to the active site with an arginine and glutamate residue, respectively, as found in the equivalent positions of the Pyrococcus enzyme Also, we introduced a loop into the active site of the psychrophilic citrate synthase, again mimicking the situation in the hyperthermophilic enzyme. Analysis of the thermoactivity and thermostability of the mutant enzymes reveals that cold activity is not significantly compromised by the mutations, but rather the affinity for one of the substrates, acetyl-CoA, is dramatically increased. Moreover, one mutant (Loop insertion/K313L/A361R) has an increased thermostability but a reduced temperature optimum for catalytic activity. This unexpected relationship between stability and activity is discussed with respect to the nature of the dependence of catalytic activity on temperature.

Acetyl Coenzyme A↗

Facultative wood-digesting bacteria from the hind-gut of the termite Reticulitermes hesperus.

Among the facultative bacteria capable of growth on mesquite wood which were isolated from the asceptically dissected hind-gut of the termite Reticulitermes hesperus were two strains of Bacillus cereus, one strain each of Arthrobacter, Alcaligenes and Serratia, and a very small Gram-negative fermentative rod. The B. cereus strains, the Serratia marcescens strain and the Arthrobacter sp. grew well on a mineral salts alpha-cellusose agar. One of the Bacillus cereus strains and Serratia marcescens hydrolysed gels of carboxymethylcellulose. All isolates grew well with mesquite wood as the carbon source. The Serratia marcescens isolate produced prodigiosin but differed from a typed strain both in size and in some physiological characteristics.

Alcaligenes↗

Coryneform bacteria in semen of chronic prostatitis patients.

Our aim was to compare the presence and species composition of coryneform bacteria in chronic prostatitis patients and controls. Semen of 50 men with inflammatory prostatitis and 59 controls (without pelvic pain/discomfort complaints and leukocytospermia) was investigated. First-catch urine was additionally investigated in 36 men (30 with and 6 without prostatitis). Coryneform bacteria were found in semen of 76% men with inflammatory prostatitis and 83% controls. More than half of the isolates were identified as Corynebacterium seminale. Prostatitis patients with severe leukocytospermia (>1 million white blood cells per ml) harboured significantly more Corynebacterium group G (33% vs. 2%, p = 0.0003) and Arthrobacter sp. (17% vs. 2%, p = 0.03) in comparison with controls. Nine species of coryneforms with high concentration (>or=10,000 CFU per ml) were found in prostatitis patients as against only four species in controls. Half of the men harboured corynebacteria in semen as well as in urine, 22% of men in semen only, and 3% in urine only. The total concentration of coryneforms was greater in semen than in urine (median 5000 vs. 100 CFU per ml, p = 0.053). We suggest that although coryneforms are generally considered as saprophytes, they are not uniform and some species (Corynebacterium group G and Arthrobacter sp.) may be associated with inflammatory prostatitis.

Adult↗

Attachment of different soil bacteria to arbuscular mycorrhizal fungal extraradical hyphae is determined by hyphal vitality and fungal species.

Attachment of certain bacteria to living arbuscular mycorrhizal fungal extraradical hyphae may be an important prerequisite for interactions between these microorganisms, with implications for nutrient supply and plant health. The attachment of five different strains of gfp-tagged soil bacteria (Paenibacillus brasilensis PB177 (pnf8), Bacillus cereus VA1 (pnf8), Pseudomonas fluorescens SBW25 :: gfp/lux, Arthrobacter chlorophenolicus A6G, and Paenibacillus peoriae BD62 (pnf8)) to vital and nonvital extraradical hyphae of the arbuscular mycorrhizal fungi Glomus sp. MUCL 43205 and Glomus intraradices MUCL 43194 was examined. Arthrobacter chlorophenolicus did not attach to hyphae, whereas the other bacterial strains did to a varying degree. Only P. brasilensis showed greater attachment to vital hyphae than nonvital hyphae of both Glomus species tested. Pseudomonas fluorescens showed a higher attachment to vital compared with nonvital Glomus sp. MUCL 43205 hyphae, whereas this relationship was opposite for attachment to G. intraradices. Both B. cereus and P. peoriae showed higher attachment to nonvital hyphae. This study provides novel evidence that under laboratory conditions soil bacteria differ in their ability to colonize vital and nonvital hyphae and that this can also be influenced by the arbuscular mycorrhizal fungal species involved. The significance of bacterial attachment to mycorrhizal fungal extraradical hyphae is discussed.

Arthrobacter↗

Reaction mechanism for the conversion of 5-monosubstituted hydantoins to enantiomerically pure L-amino acids.

The specific conversion of D,L-5-monosubstituted hydantoins to optically pure L-amino acids by resting cells of Arthrobacter sp. DSM 7330 has been evaluated. A new nonstereoselective hydantoinase from Arthrobacter sp. DSM 7330 was isolated and characterized. When whole cells were tested, the conversion of D,L-5-methylthioethylhydantoin (D,L-5-MTEH) led to the optically pure intermediate D-carbamoylmethionine (D-CM) and to the optically pure amino acid L-methionine. After purification of the hydantoin hydrolyzing enzyme, the probable reaction mechanism of the conversion of 5-monosubstituted hydantoins to enantiomerically pure L-amino acids could be enlightened.

Amidohydrolases↗

Nitrification in histosols: a potential role for the heterotrophic nitrifier.

Insufficient populations of Nitrosomonas and Nitrobacter were found in a Pahokee muck soil (Lithic medidaprit) to account for the nitrate concentration observed. To determine if heterotrophic nitrifiers could account for some of this discrepancy, a method was developed to measure the levels of heterotrophic nitrifiers in soil. A population of 4.1 X 10(5) Arthrobacter per g of dry fallow soil, capable of producing nitrite and/or nitrate from reduced nitrogenous compounds, was observed. Amendment of the much with 0.5% (wt/wt) sodium acetate and 0.1% (wt/wt) ammonium-nitrogen as ammonium sulfate (final concentrations) not only resulted in the usual increase in autotrophic nitrifiers, but also in a fourfold increase in the heterotrophic nitrifying Arrthrobacter. Amendment of like samples with N-Serve [2-chloro-6(trichloromethyl) pyridinel] prevented the increase in Nitrosomonas, but not that in the heterotrophic nitrifiers. Nitrate production in the presence of the inhibitor was diminished but not prevented. An Arthrobacter sp., isolated from the muck, produced nitrite when inoculated at high densities into sterile soil, unamended or amended with sodium acetate and/or ammomium sulfate. These data suggest that the heterotrophic population may be responsible for some of the nitrate produced in these Histosols.

Acetates↗

Catabolism of 2,4,5-trimethyoxybenzoic acid and 3-methoxycrotonic acid.

4-Methoxygentisic acid was an intermediate formed when Arthrobacter degraded, 2,4,5-trimethoxybenzoic acid. Isolates of Pseudomonas and Arthrobacter from soil grew at the expense of 3-methoxycrotonic acid. Evidence is presented that enzymatic hydration, with elimination of methanol, accounted for replacement of the methoxyl group of 3-methoxycrotonic acid and also of one methoxyl group of 2,4,5-trimethoxybenzoic acid.

Arthrobacter↗

Kinetics of methyl t-butyl ether cometabolism at low concentrations by pure cultures of butane-degrading bacteria.

Butane-oxidizing Arthrobacter (ATCC 27778) bacteria were shown to degrade low concentrations of methyl t-butyl ether (MTBE; range, 100 to 800 microg/liter) with an apparent half-saturation concentration (K(s)) of 2.14 mg/liter and a maximum substrate utilization rate (k(c)) of 0.43 mg/mg of total suspended solids per day. Arthrobacter bacteria demonstrated MTBE degradation activity when grown on butane but not when grown on glucose, butanol, or tryptose phosphate broth. The presence of butane, tert-butyl alcohol, or acetylene had a negative impact on the MTBE degradation rate. Neither Methylosinus trichosporium OB3b nor Streptomyces griseus was able to cometabolize MTBE.

Arthrobacter↗