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PubMed2001

Complete denitration of nitroglycerin by bacteria isolated from a washwater soakaway.

Four axenic bacterial species capable of biodegrading nitroglycerin (glycerol trinitrate [GTN]) were isolated from soil samples taken from a washwater soakaway at a disused GTN manufacturing plant. The isolates were identified by 16S rRNA gene sequence homology as Pseudomonas putida, an Arthrobacter species, a Klebsiella species, and a Rhodococcus species. Each of the isolates utilized GTN as its sole nitrogen source and removed nitro groups sequentially from GTN to produce glycerol dinitrates and mononitrates (GMN), with the exception of the Arthrobacter strain, which achieved removal of only the first nitro group within the time course of the experiment. The Klebsiella strain exhibited a distinct preference for removal of the central nitro group from GTN, while the other five strains exhibited no such regioselectivity. All strains which removed a second nitro group from glycerol 1,2-dinitrate showed regiospecific removal of the end nitro group, thereby producing glycerol 2-mononitrate. Most significant was the finding that the Rhodococcus species was capable of removing the final nitro group from GMN and thus achieved complete biodegradation of GTN. Such complete denitration of GTN has previously been shown only in mixed bacterial populations and in cultures of Penicillium corylophilum Dierckx supplemented with an additional carbon and nitrogen source. Hence, to the best of our knowledge, this is the first report of a microorganism that can achieve complete denitration of GTN.

Arthrobacter↗

PubMed2001

Dual-bioaugmentation strategy to enhance remediation of cocontaminated soil.

Although metals are thought to inhibit the ability of microorganisms to degrade organic pollutants, several microbial mechanisms of resistance to metal are known to exist. This study examined the potential of cadmium-resistant microorganisms to reduce soluble cadmium levels to enhance degradation of 2,4-dichlorophenoxyacetic acid (2,4-D) under conditions of cocontamination. Four cadmium-resistant soil microorganisms were examined in this study. Resistant up to a cadmium concentration of 275 microg ml(-1), these isolates represented the common soil genera Arthrobacter, Bacillus, and Pseudomonas. Isolates Pseudomonas sp. strain H1 and Bacillus sp. strain H9 had a plasmid-dependent intracellular mechanism of cadmium detoxification, reducing soluble cadmium levels by 36%. Isolates Arthrobacter strain D9 and Pseudomonas strain I1a both produced an extracellular polymer layer that bound and reduced soluble cadmium levels by 22 and 11%, respectively. Although none of the cadmium-resistant isolates could degrade 2,4-D, results of dual-bioaugmentation studies conducted with both pure culture and laboratory soil microcosms showed that each of four cadmium-resistant isolates supported the degradation of 500-microg ml(-1) 2,4-D by the cadmium-sensitive 2,4-D degrader Ralstonia eutropha JMP134. Degradation occurred in the presence of up to 24 microg of cadmium ml(-1) in pure culture and up to 60 microg of cadmium g(-1) in amended soil microcosms. In a pilot field study conducted with 5-gallon soil bioreactors, the dual-bioaugmentation strategy was again evaluated. Here, the cadmium-resistant isolate Pseudomonas strain H1 enhanced degradation of 2,4-D in reactors inoculated with R. eutropha JMP134 in the presence of 60 microg of cadmium g(-1). Overall, dual bioaugmentation appears to be a viable approach in the remediation of cocontaminated soils.

2,4-Dichlorophenoxyacetic Acid↗

PubMed2003

mRNA differential display in a microbial enrichment culture: simultaneous identification of three cyclohexanone monooxygenases from three species.

mRNA differential display has been used to identify cyclohexanone oxidation genes in a mixed microbial community derived from a wastewater bioreactor. Thirteen DNA fragments randomly amplified from the total RNA of an enrichment subculture exposed to cyclohexanone corresponded to genes predicted to be involved in the degradation of cyclohexanone. Nine of these DNA fragments are part of genes encoding three distinct Baeyer-Villiger cyclohexanone monooxygenases from three different bacterial species present in the enrichment culture. In Arthrobacter sp. strain BP2 and Rhodococcus sp. strain Phi2, the monooxygenase is part of a gene cluster that includes all the genes required for the degradation of cyclohexanone, while in Rhodococcus sp. strain Phi1 the genes surrounding the monooxygenase are not predicted to be involved in this degradation pathway but rather seem to belong to a biosynthetic pathway. Furthermore, in the case of Arthrobacter strain BP2, three other genes flanking the monooxygenase were identified by differential display, demonstrating that the repeated sampling of bacterial operons shown earlier for a pure culture (D. M. Walters, R. Russ, H. Knackmuss, and P. E. Rouvière, Gene 273:305-315, 2001) is also possible for microbial communities. The activity of the three cyclohexanone monooxygenases was confirmed and characterized following their expression in Escherichia coli.

Arthrobacter↗

PubMed1999

Characterization of the bacterial community of a zinc-polluted soil.

The bacterial community of a zinc-contaminated soil (Maatheide soil in Lommel, Belgium) was studied using cultivation as well as cultivation-independent techniques. Colony-forming units (CFU) were determined by plating on media with or without metals. Dominant isolates were characterized by fatty acid methyl ester analysis (FAME analysis) and PCR fingerprinting using repetitive extragenic palindromic sequences as primers. DNA was directly extracted from soil samples and used as a template for the PCR amplification of the 16S rDNA (8-1511) or a 16S rDNA fragment (968-1401). Clones resulting from cloning the 16S rDNA from soil DNA were sequenced. Temperature gradient gel electrophoresis (TGGE analysis) was performed for 16S rDNA fragments (968-1401) amplified from the dominant isolates, the clones, and the total soil DNA extracted according to two protocols differing in strength of lysis. Total CFU ranged from 10(4) to 10(5)/g soil. The majority of the isolates were identified by FAME analysis as Arthrobacter spp. (18 out of 23). None of the isolates were identified as a Ralstonia eutropha like strain (formerly Alcaligenes eutrophus). Metalloresistant Rastomia eutropha like strains were previously shown to be dominant in the analyzed biotope. Most of the isolates were zinc tolerant but only seven could be considered zinc resistant. Sequences of the 16S rDNA clones obtained from total soil DNA were affiliated with genes of different bacteria such as alpha-proteobacteria, beta-proteobacteria, and the Cytophaga-Flexibacter-Bacteroides group. None of the sequenced clones aligned with the Ralstonia eutropha 16S rRNA gene. TGGE analysis of the 16S rDNA fragments (968-1401) amplified from the dominant strains, the clones, and the total soil DNA showed that isolates and clones represented only a part of the bands present in the TGGE pattern from total DNA. The 968-1401 fragment amplified from all Arthrobacter strains had a similar electrophoretic mobility. This band was seen as a major band in the pattern of DNA extracted from soil using a harsh cell lysis, whereas it did not appear, or appeared only as a weak band, in patterns obtained from soil DNA extracted using gentle lysis. The previously reported predominance of a Ralstonia eutropha like strain in this soil was no longer observed. This may suggest a population replacement by less resistant bacteria, concomitant with a progressive decrease of the zinc toxicity in the Maatheide soil.

Arthrobacter↗

PubMedDate not supplied

Marine biosurfactants, I. Screening for biosurfactants among crude oil degrading marine microorganisms from the North Sea.

Three bacterial strains of marine origin were isolated during a screening for biosurfactants among n-alkane degrading microorganisms. One strain-identified as Alcaligenes sp. MM1-produced a novel glucose lipid. In the case of Arthrobacter sp. EK 1 the well-known trehalose tetraester was found as major component. From another pure culture classified as Arthrobacter sp. SI 1, extracellular emulsifying agents with properties indicating high molecular weight substances were detected. Furthermore trehalose corynomycolates were found at up to 2 g/l. The isolated biosurfactants showed good interfacial and emulsifying properties.

Alcaligenes↗

PubMed1985

Sulindac oxidation/reduction by microbial cultures; microbial models of mammalian metabolism.

The oxidation and reduction of the sulphoxide moiety of the anti-inflammatory agent sulindac was investigated to explore microbial systems exhibiting parallels of known mammalian metabolism. Of 24 cultures initially screened, four catalysed the expected reactions in analytical studies. Arthrobacter species (ATCC 19140) and Sporobolomyces pararoseus (ATCC 11386) produced sulindac sulphide, Aspergillus alliaceus (NRRL 315) produced sulindac sulphone, and Nocardia corallina (ATCC 19070) produced both the sulphide and sulphone. Preparative-scale production and full structural elucidation of metabolites was accomplished for sulindac sulphide with Arthrobacter species, and sulindac sulphone with A. alliaceus and N. corallina. N. corallina also exhibited an aeration-dependent, reversible reduction of sulindac to the sulphide, and further oxidation to the sulphone. This organism thus parallels the composite of major phase-I redox transformations of this drug observed in mammals.

Arthrobacter↗

PubMedDate not supplied

[Enzymes of intermediary metabolism in coryneform bacteria].

Enzymes of the intermediate metabolism were studied in ten strains of Corynebacterium-like organisms belonging to the genera Arthrobacter, Brevibacterium, Corynebacterium and Nocardia. All of these were found to contain enzymes of the glycolytic pathway, and nine strains among ten had dehydrogenases of the pentose phosphate shunt. The activity of enzymes of the citric acid cycle was low: alpha-ketoglutarate dehydrogenase was not found in Arthrobacter, Corynebacterium, Brevibacterium linens and Nocardia minima. Eight strains possessed the activity of the key enzyme of the gamma-aminobutyrate shunt, i.e. gamma-aminobutyrate aminotransferase. The activity of enzymes of the glyoxylate shunt was found in nine strains, and their level was rather high even during growth on glucose. Therefore, it is possible to study the taxonomic structure of this group of microorganisms by analyzing the composition and the level of enzymes involved in the intermediate metabolism. The competence of the Brevibacterium genus is corroborated by the typical species Brevibact. linens, as well as the reality of saprophytic representatives of the Corynebacterium genus, and a special taxonomic position of the group Brevibact. ammoniagenes--Brevibact. stationis.

Actinomycetales↗

PubMed2000

[Two different fermentation techniques of steriod 1,4-dehydrogenation and 11 alpha-hydroxylation].

Two kinds of micro-organism, Arthrobacter sp. AX86(1,4-dehdrogenator) and Absidia sp. A28(11 alpha-hydroxylator) were used in this experiment. Two different fermentation techniques were performed to accomplish the multiple conversional reactions for producing 16 beta-methyl-11 alpha,17 alpha,21-trihydroxy-1,4-pregnadiene-3,20-dione(III) from 16 beta-methyl-3 beta,17 alpha,21-trihydroxy-5 alpha-pregnane-20-one-21-acetate(I): 1) To produce product(III) by means of a two-step fermentation method which were independently performed first by Arthrobacter and next by Absiaia, and 2) the product was obtained by a sequential fermentation system of aforesaid two micro-organisms in a single fermentor without isolation of the intermediates from the mixture. Our results showed that in both fermentation systems high yield of product was obtained. However, according to the technical simplicity, shorter duration of fermentation cycle and efficient yield of product, the second method is better than the first one.

Absidia↗

PubMedDate not supplied

Differentiation of genera of the coryneform bacteria.

A scheme of the generic structure of the group of coryneform bacteria, including the genera Arthrobacter, Brevibacterium, Cellulomonas, Corynebacterium, and Rhodococcus, is suggested. Morphological, chemotaxonomic (presence and stereochemical form of diaminopimelic acid, lipid A, and L-arabinose), and physiological features were used as diagnostic criteria. The position of Microbacterum and Mycococcus and of coryneforms with a nocardial wall but giving a positive Hugh-Leifson anaerobic test, and also of Arthrobacter pascens, which may be identical with Mycoplana sp., remains uncertain.

Actinomycetales↗

PubMed1992

[Biotransformation of zearalenone].

The conversion of Zearalenone by some strains of microorganisms was investigated. When the selective strains of Rhodotorula sp., Arthrobacter sp., Saccharomyces sp., and Candida sp. were incubated by shaking or standing at 28 degrees C for 72 h with an alcoholic solution of Zearalenone as the substrate at a concentration of 2-10 mg/ml ethanol (50-100 micrograms/ml medium), it was readily converted to give Zearalenols, consisting either mainly of the alpha-isomer (e.g. 96% in case of Rhodotorula sp. and 84% in case of Arthrobacter sp. as determined by HPLC) or beta-isomer (e.g. 91% and 92% in Saccharomyces sp. and Candida sp., respectively). The structure of the product was confirmed by 13C-NMR, MS and HPLC.

Arthrobacter↗

PubMedDate not supplied

[The microbial transformation of phenanthrene and anthracene].

The transformation of phenanthrene and anthracene by Rhodococcus rhodnii 135, Pseudomonas fluorescens 26K, and Arthrobacter sp. K3 is studied. Twenty-one intermediates of phenanthrene and anthracene transformation are identified by HPLC, mass spectrometry, and NMR spectroscopy. P. fluorescens 26K and Arthrobacter sp. K3 are found to produce a wide range of intermediates, whereas R. rhodnii 135 oxidizes phenanthrene, resulting in the formation of a sole product, 3-hydroxyphenanthrene. Putative transformation pathways of phenanthrene and anthracene are proposed for the three bacterial strains studied. These strains can be used to obtain valuable compounds (such as hydroxylated polycyclic aromatic hydrocarbons) that are difficult to produce by chemical synthesis.

Anthracenes↗

PubMed2003

[The product identification study on the isolating process of strains produced trehalose].

In the isolating process of stains whose endocellular enzymes can produce trehalose on starch or maltooligosaccharides, we discovered the components of enzymatic reactant were complicated and it was difficult to purify them each other, however, we have to know quickly whether there was tehalose in the enzymatic reactant above. In order to make it clearly, thinner layer chromatography, high performance liquid chromatography-electrospray ionization mass spectrometry and 13C-nuclear magnetic resonance were adopted. The unknown oligosaccharide produced by Arthrobacter nicotinovorus D-97 was identified rapidly and this unknown oligosaccharide needn't be purified from the enzymatic reactant solely. The production on starch or maltooligosaccharides by endocellular enzymes of Arthrobacter nicotinovorus D-97 is trehalose. This research method in our paper can be applied in the isolating process of production trehalose and other functional glucobioses strains.

Arthrobacter↗

PubMed1989

Sialidase from different sources compared for electrophoretically separating serum alkaline phosphatase fractions from liver and bone.

We compared sialidase (neuraminidase; EC 3.2.1.18) from Vibrio cholerae, Clostridium perfringens, and Arthrobacter ureafaciens, seeking to improve the electrophoretic separation of the liver and bone isoenzymes of alkaline phosphatase (EC 3.1.3.1) on cellulose acetate membranes. Resolution is decisively determined by the type and activity of sialidase used in the preincubation of serum sample. Sialidase from Arthrobacter ureafaciens is not suited for this method. For optimal separation of the two isoenzymes we recommend the use of sialidase from Vibrio cholerae, determination of its activity with a standard procedure such as described here (mucin or sialyl lactose as substrates), and a final concentration of sialidase activity of 2.0 or 2.9 U/L (measured with mucin or sialyl lactose) in the incubation mixture.

Alkaline Phosphatase↗

PubMed1986

Enzymes of vitamin B6 degradation. Purification and properties of 4- and 5-pyridoxolactonases.

4-Pyridoxolactone and 5-pyridoxolactone, formed by dehydrogenation of pyridoxal or isopyridoxal during the bacterial degradation of vitamin B6 by Pseudomonas MA-1 and Arthrobacter Cr-7, respectively, are hydrolyzed to the corresponding acids by distinct inducible lactonases which were purified to homogeneity. 4-Pyridoxolactonase from Pseudomonas MA-1 has an Mr of 54,000 and contains two probably identical subunits of Mr = 28,600. It has a pH optimum of 7.0, a Km of 5.9 microM, and a Vmax at 25 degrees C of 35.2 mumol X min-1 X mg-1. 5-Pyridoxolactonase from Arthrobacter Cr-7 has an Mr of 65,200 and also contains two probably identical subunits of Mr = 32,800. It has a pH optimum of 7.1-7.7, a Km of 300 microM, and a Vmax at 25 degrees C of 21.5 mumol-1 X min-1 X mg-1. The two lactonases require no added cofactors or metal ions; their activities are inhibited by sulfhydryl reagents but are not affected by metal-chelating reagents. Although the two lactonases are entirely specific for their respective substrates, 4-pyridoxolactone is a competitive inhibitor (KI = 52 microM) for 5-pyridoxolactonase, and 5-pyridoxolactone is a competitive inhibitor (KI = 48 microM) for 4-pyridoxolactonase.

Arthrobacter↗

PubMed1974

Microbial Decomposition of alpha-Picoline.

An organism, which degrades alpha-picoline but also utilizes 2-ethylpyridine or piperidine as alternative growth substrates, has been isolated from soil and characterized as arthrobacter sp. alpha-picoline-grown cells oxidize 2-ethylpyridine and vice versa. Other pyridine derivatives tested are neither utilized as growth substrates nor oxidized by the organism. alpha-Picolinate and 2-hydroxy-6-methylpyridine are not metabolized, indicating that degradation is neither initiated by methyl oxidation nor by hydroxylation in the 6-position of pyridine ring. Succinate semi-aldehyde and pyruvate accumulate when alpha-picoline oxidation by resting cell suspensions is blocked by semicarbazide. The Arthrobacter grown on alpha-picoline rapidly oxidizes succinate semi aldehyde...

Arthrobacter↗

PubMed1984

[H-NMR spectroscopy. Specificity of microbial sialidases against complex substrates].

The specificities of one viral and five bacterial sialidases were investigated by 1H-NMR-spectroscopy with substrates or substrate mixtures containing two sialic acid residues of different linkage types. This technique allows - in contrast to the methods used before - the simultaneous determination of the rates of hydrolysis of both NeuAc linkages in a single experiment. The substrate specificities of the enzymes are discussed on the basis of the relation of the rate constants k/k'. The data obtained are more exact and more informative than those of separate experiments as reported previously. Among the enzymes investigated, i.e. sialidases of fowl plague virus (FPV = VKH), Clostridium perfringens (CP), Vibrio cholerae (VC), Bifidobacterium bifidum var. pennsylvanicum (BBif), Bifidobacterium lactentis (BLac), and Arthrobacter ureafaciens (AU), the activity of the viral sialidase VKH shows the highest, the activities of the Bifidobacterium sialidases the lowest dependence on the nature and on the linkage type of the different substrates. All sialidases preferentially cleave the NeuAc alpha 2-3-Gal linkage with the exception of the enzyme of Arthrobacter ureafaciens (AU) which shows a higher affinity to alpha 2-6 linkages. However, this does not apply to the side-arm-linked NeuAc alpha 2-6 structure in NeuAc alpha 2-3 Gal beta 1-3 (NeuAc alpha 2-6)-GlcNAc beta 1-3Gal beta 1-4Glc (Substrate B). This substrate in generally cleaved very slowly and is hardly affected by the viral enzyme. After the alpha 2-3 linkage, the alpha 2-8 bond in NeuAc alpha 2-8 NeuAc alpha 2-3 Gal beta 1-4Glc(Substrate A) is most susceptible for the sialidases VKH, CP and VC. An elongation of the carbohydrate chain (Substrate D) is accompanied by a reduction of the rate of cleavage for all enzymes. The experiments with alpha 1-acid glycoprotein, fetuin, and with the glycopeptides obtained by proteolytic degradation of the latter, revealed the same specificity towards the alpha 2-3 and the alpha 2-6 linkages as the oligosaccharides. Influenced by the chemical nature and the size of the substrate, NeuAc is released from the native alpha 1-acid glycoprotein more quickly than from the corresponding glycopeptide. All sialidases investigated so far are strictly exo-enzymes as could be demonstrated by the cleavage of NeuAc alpha 2-8 NeuAc alpha 2-3 Gal beta 1-4Glc (Substrate A).

Arthrobacter↗

PubMed1988

Neuraminidase induces capacitation and acrosome reaction in mammalian spermatozoa.

The treatment of epididymal spermatozoa of guinea pig and ejaculated spermatozoa of rabbit with neuraminidase from Arthrobacter ureafaciens induced significant acrosome reaction while the neuraminidase from Cl. perfringens failed to do so. The addition of the neuraminidase inhibitors kept the enzyme induced acrosome reaction to the control level. The zona-free hamster ova test showed that the treatment of spermatozoa with Arthrobacter neuraminidase rendered 82% of the guinea pig and 69% of the rabbit spermatozoa capable of fertilization. Thus, neuraminidase seems to enhance the rate of acrosome reaction by first capacitating spermatozoa in vitro.

Acrosome↗

PubMed1982

Oxygen tolerance of strictly aerobic hydrogen-oxidizing bacteria.

Growth of various bacteria, especially aerobic hydrogen-oxidizing bacteria, in the presence of 2 to 100% (v/v) oxygen in the gas atmosphere was evaluated. The bacterial strains included Alcaligenes eutrophus, A. paradoxus, Aquaspirillum autotrophicum, Arthrobacter spec. strain 11 X, Escherichia coli, Arthrobacter globiformis, Nocardia opaca, N. autotrophica, Paracoccus denitrificans, Pseudomonas facilis, P. putida, and Xanthobacter autotrophicus. Under heterotrophic conditions with fructose or gluconate as substrates neither colony formation on solid medium nor the growth rates in liquid media were drastically impaired by up to 100% oxygen. In contrast, autotrophic growth--with hydrogen, carbon dioxide and up to 80% oxygen in the gas atmosphere--was strongly depressed by high oxygen concentrations. However, only the growth rate, not the viability of the cells, was decreased. Growth retardation was accompanied by a decrease of hydrogenase activity.

Aerobiosis↗