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Reclassification of Micrococcus agilis (Ali-Cohen 1889) to the genus Arthrobacter as Arthrobacter agilis comb. nov. and emendation of the genus Arthrobacter.

Phylogenetic evidence derived from a 16S ribosomal DNA analysis indicated that the type strain of Micrococcus agilis, DSM 20550 (= ATCC 966 = CCM 2390), is less closely related to the type species of the genus Micrococcus, Micrococcus luteus, than to the type species of the genus Arthrobacter, Arthrobacter globiformis, and related Arthrobacter species. The phylogenetic position of M. agilis is supported by the presence of peptidoglycan variation A3 alpha and by the presence of MK-9(H2) as the major isoprenolog, a characteristic also found in strains of A. globiformis, Arthrobacter crystallopoietes, Arthrobacter atrocyaneus, Arthrobacter citreus, Arthrobacter aurescens, Arthrobacter ilicis, Arthrobacter ureafaciens, Arthrobacter oxydans, Arthrobacter histidinolovorans, and Arthrobacter nicotinovorans. The last six species and M. agilis are characterized by the presence of threonine in the interpeptide bridge of the peptidoglycan. Threonine has not been found in the peptidoglycans of other Arthrobacter species or in members of the genus Micrococcus. Despite the fact that a morphological life cycle is not known, these data support the proposal that M. agilis should be transferred to the genus Arthrobacter as Arthrobacter agilis comb. nov.

Arthrobacter↗

Isolation of Arthrobacter spp. from clinical specimens and description of Arthrobacter cumminsii sp. nov. and Arthrobacter woluwensis sp. nov.

Arthrobacter spp. are very widely distributed in the environment (e.g., soil) but have not been described as causing disease in humans. Over a 6-year period, two reference laboratories isolated or received 11 strains which were eventually identified as belonging to the genus Arthrobacter. These strains had been initially identified as Centers for Disease Control and Prevention coryneform group B-1 and B-3 bacteria (whitishgrayish colonies of 2 mm or greater in diameter after 24 h of incubation, respiratory metabolism, absent or weak acid production from sugars, and hydrolysis of gelatin). However, chemotaxonomic investigations revealed lysine as the diamino acid of the cell wall and the presence of branched cellular fatty acids (with anteiso-pentadecanoic acid predominating) which was compatible with an assignment of the 11 isolates to the genus Arthrobacter only. Peptidoglycan and 16S rRNA gene sequence analyses demonstrated that three of the strains studied were representatives of a new Arthrobacter species for which the name Arthrobacter cumminsii sp. nov. is proposed and that one other strain represented a second new Arthrobacter species for which the name Arthrobacter woluwensis sp. nov. is proposed. This report is the first on the isolation of Arthrobacter spp. from clinical specimens.

Arthrobacter↗

Identification of Arthrobacter oxydans, Arthrobacter luteolus sp. nov., and Arthrobacter albus sp. nov., isolated from human clinical specimens.

Five Arthrobacter isolates from clinical specimens were studied by phenotypic, chemotaxonomic, and genetic characterization. Two strains had characteristics consistent with those of Arthrobacter oxydans. One strain was related to A. citreus; however, DNA-DNA hybridization and phenotypic characteristics indicated that this strain belongs to a new species, for which the name Arthrobacter luteolus sp. nov. is proposed. Two strains were closely related to A. cumminsii by 16S rRNA gene sequencing, but DNA-DNA hybridization, peptidoglycan type, and some phenotypic features indicated that they should be assigned to a new species, for which the name Arthrobacter albus sp. nov. is proposed. The type strain of A. luteolus is CF25 (DSM 13067). The type strain of A. albus is CF43 (DSM 13068).

Arthrobacter↗

Arthrobacter bergerei sp. nov. and Arthrobacter arilaitensis sp. nov., novel coryneform species isolated from the surfaces of cheeses.

Fourteen isolates of two different bacterial species isolated from the surface of smear-ripened cheeses were found to exhibit many characteristics of the genus Arthrobacter. The isolates were aerobic, Gram-positive, catalase-positive, non-spore-forming and non-motile. The cell-wall peptidoglycan contained lysine, alanine and glutamic acid. rrs sequence analysis indicated that the new isolates Re117T and Ca106T are closely related to the Arthrobacter nicotianae group and showed highest sequence similarity (>98 %) to Arthrobacter nicotianae and Arthrobacter protophormiae. However, DNA-DNA hybridization studies indicated that the strains represented two novel genomic species within the genus Arthrobacter and did not belong to A. nicotianae or A. protophormiae (<43 % DNA-DNA relatedness). On the basis of the phylogenetic and phenotypic distinctiveness of the new isolates, these bacteria should be classified as two novel Arthrobacter species, for which the names Arthrobacter bergerei sp. nov. and Arthrobacter arilaitensis sp. nov. are proposed. Type strains have been deposited in culture collections as Arthrobacter bergerei Ca106T (=CIP 108036T=DSM 16367T) and Arthrobacter arilaitensis Re117T (=CIP 108037T=DSM 16368T).

Arthrobacter↗

Biochemical and phylogenetic analyses of psychrophilic isolates belonging to the Arthrobacter subgroup and description of Arthrobacter psychrolactophilus, sp. nov.

During our work on psychrophilic microorganisms we obtained a large collection of new isolates. In order to identify six of these, we examined their growth properties, cell wall compositions, and their 16S rRNA gene sequences. The results showed that all of the isolates are gram-positive, aerobic, contain lysine in their cell walls, and belong to the high mol% G+C Arthrobacter subgroup. Phylogenetic analysis of the 16S rRNA genes grouped five isolates obtained from a small geographical region into a monophyletic clade. Isolate B7 had a 16S rRNA sequence that was 94.3% similar to that of Arthrobacter polychromogenes and 94.4% similar to that of Arthrobacter oxydans. Primary characteristics that distinguish isolate B7 from the Arthrobacter type strain (Arthrobacter globiformis) and A. polychromogenes include lack of growth at 37 degrees C, growth at 0-5 degrees C, the ability to use lactose as a sole carbon source, and the absence of blue pigments. Because of these differences, isolate B7 was chosen as a type strain representing a new Arthrobacter species, Arthrobacter psychrolactophilus. The sixth isolate, LV7, differed from the other five because it did not have the rod/ coccus morphological cycle and was most closely related to Arthrobacter agilis.

Arthrobacter↗

Enzymes of dimethylsulfone metabolism and the phylogenetic characterization of the facultative methylotrophs Arthrobacter sulfonivorans sp. nov., Arthrobacter methylotrophus sp. nov., and Hyphomicrobium sulfonivorans sp. nov.

Novel methylotrophic Arthrobacter and Hyphomicrobium species are described. Constitutive membrane-associated dimethylsulfone- and dimethylsulfoxide-reductases were found in Arthrobacter methylotrophus strain TGA and Hyphomicrobium sulfonivorans strain S1. Enzyme activities increased during growth with dimethylsulfone or dimethylsulfoxide, respectively, and different ratios of activity with different growth substrates indicated that they are separate enzymes. SDS-PAGE showed some membrane-associated polypeptides to be enhanced during growth with dimethylsulfone (54 kDa in H. sulfonivorans, 21-24 kDa, 54 kDa and 80 kDa in A. methylotrophus). Western blotting with anti-dimethylsulfoxide-reductase antibody showed cross-reaction with 54- and 21-kDa polypeptides in A. methylotrophus. All strains contained rhodanese and sulfur oxygenase after growth with dimethylsulfone. Sulfite was oxidized in the Arthrobacter species by APS reductase and sulfite dehydrogenase. H. sulfonivorans oxidized sulfite with APS reductase, which is unusual for an alpha-proteobacterium. The Arthrobacter species were distinguished from each other and from other Arthrobacter and Micrococcus species by 16S rRNA gene sequence analysis. The menaquinone and fatty acid profiles of the Arthrobacter species were similar. Their peptidoglycan structures were L-Lys- L-Ser- L-Thr- L-Ala for A. sulfonivorans and L-Lys- L-Ala(2-4) for A. methylotrophus. H. sulfonivorans exhibited gross morphology typical for Hyphomicrobium, but possessed helically twisted prosthecae. 16S rRNA gene sequence analysis showed it to be distinct from all the other Hyphomicrobium, Filomicrobium and Pedomicrobium species sequenced to date. Formal descriptions of the new species are given.

Arthrobacter↗

IS1473, a putative insertion sequence identified in the plasmid pAO1 from Arthrobacter nicotinovorans: isolation, characterization, and distribution among Arthrobacter species.

A putative insertion sequence (IS1473) has been cloned and sequenced. The 1087-bp element was found between the moaA and the ndhA genes in the upstream region of the nicotine dehydrogenase (ndh) operon in the 160-kb pAO1 plasmid of Arthrobacter nicotinovorans. It is flanked by an imperfect repeat of 33 bp and carries two overlapping open reading frames which, by programmed -1 translational frameshifting, may produce a transposase of 36.735 Da with a pI = 10. 18. The deduced protein is similar to the transposases IS481 and IS1002 from Bordetella and IS476 from Xanthomonas campestris, all members of the IS3 family. The putative insertion element was found as a single copy in the pAO1 plasmid and absent on the chromosome of the A. nicotinovorans genome. Similar sequences were detected by hybridization on total DNA from Arthrobacter globiformis, Arthrobacter ramosus, and Arthrobacter ureafaciens.

Amino Acid Sequence↗

Reclassification of two strains of Arthrobacter oxydans and proposal of Arthrobacter nicotinovorans sp. nov.

Arthrobacter oxydans DSM 419 and DSM 420 have chemical and microbiological properties that are consistent with assignment to the genus Arthrobacter. Both organisms have the lysine-alanine-threonine-alanine peptidoglycan type. DNA-DNA pairing studies indicated that A. oxydans DSM 419 should be reclassified as Arthrobacter ureafaciens and that A. oxydans DSM 420T forms the nucleus of a distinct genomic species. We propose that A. oxydans DSM 420 should be reclassified as Arthrobacter nicotinovorans sp. nov. The type strain is strain DSM 420.

Amino Acid Sequence↗

Arthrobacter gangotriensis sp. nov. and Arthrobacter kerguelensis sp. nov. from Antarctica.

Two coryneform bacteria were isolated from a penguin rookery soil sample collected in Antarctica, near the Indian station Dakshin Gangotri (strain Lz1y(T)), and from sea water from Kerguelen island, Antarctica (strain KGN15(T)). They have morphological and chemotaxonomic properties (peptidoglycan A4alpha type; major menaquinones MK-8, MK-9 and MK-10; predominant fatty acids anteiso-C(15 : 0) and anteiso-C(17 : 0)) that are characteristic of members of the genus Arthrobacter. The isolates shared 97.8 % 16S rRNA gene sequence similarity to each other and were most closely related to Arthrobacter sulfureus (about 98.5 % sequence similarity). DNA-DNA hybridization experiments revealed 50 % relatedness between the isolates, while the levels of DNA-DNA relatedness between strains Lz1y(T) and KGN15 (T) and their phylogenetic relative, A. sulfureus, were respectively 54 and 12 %. Based on the above data and distinct phenotypic differences between the isolates and A. sulfureus, two novel species are proposed, Arthrobacter gangotriensis sp. nov. (type strain Lz1y(T)=DSM 15796(T)=JCM 12166(T)) and Arthrobacter kerguelensis sp. nov. (type strain KGN15(T)=DSM 15797(T)=JCM 12165(T)).

Antarctic Regions↗

Transformation of Arthrobacter and studies on the transcription of the Arthrobacter ermA gene in Streptomyces lividans and Escherichia coli.

We report the development of a plasmid-mediated transformation system for Arthrobacter sp. NRRLB3381, using the Streptomyces cloning vector pIJ702. Our procedure gives a transformation frequency of 10(3)/micrograms of plasmid DNA. In addition we have explored the expression of the Arthrobacter ermA gene in Streptomyces lividans and Escherichia coli, and shown that the ermA promoter is recognized in S. lividans not E. coli. The relationship between Arthrobacter, Streptomyces and E. coli promoters is discussed.

Arthrobacter↗

Plasmids for nicotine-dependent and -independent gene expression in Arthrobacter nicotinovorans and other arthrobacter species.

The first inducible Arthrobacter overexpression system, based on the promoter/operator and the repressor of the 6-D-hydroxynicotine oxidase gene of Arthrobacter nicotinovorans, is described here. Nicotine-dependent overproduction and affinity purification of recombinant proteins are presented. The system will allow the production of complex enzymes and genetic complementation studies in Arthrobacter species.

Arthrobacter↗

Myceloid growth of Arthrobacter globiformis and other Arthrobacter species.

Transitory myceloid growth occurs in certain complex media with Arthrobacter globiformis strain ATCC 8010. This type of growth, however, was not observed in a medium which contained an array of metal ions but did not contain agents able to complex metal ions. Addition of metal-complexing agents to this medium caused an interruption in the life cycle of strain 8010 so that growth occurred only as the myceloid form. It appeared that manganese was the critical metal that was removed by the metal-complexing agents. During growth, the myceloid cells started to fragment, but wall septation was incomplete. A. globiformis strain ATCC 4336 and several other Arthrobacter species and soil isolates, but not Arthrobacter crystallopoietes, responded to metal-complexing agents as did strain 8010. Biotin and vitamin B12 were not involved in this myceloid growth.

Arthrobacter↗

Characteristics of Arthrobacter cumminsii, the most frequently encountered Arthrobacter species in human clinical specimens.

During a 2-year period, 10 strains of Arthrobacter cumminsii were isolated in or received by a Swiss routine clinical bacteriology laboratory, and 5 further isolates were referred to a Swedish bacteriology reference center over a 5-year period, making A. cumminsii the most frequently encountered Arthrobacter species in these two laboratories. The present report outlines the clinical features of the 15 A. cumminsii strains and presents an extended biochemical characterization of this microorganism. A. cumminsii exhibits a unique cellular fatty acid pattern with the consistent presence of C14:0i and C14:0 fatty acids as well as relatively large amounts of C16:0i and C16:0 fatty acids usually not seen in other Arthrobacter spp. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was found to be a useful tool for confirmation of the identification of A. cumminsii. The MICs of 39 antimicrobial agents were determined, and it was demonstrated that aminoglycosides and quinolones had only weak activities against A. cumminsii strains, in contrast to their activities against most other coryneform bacteria. As a result of the extended characterization of A. cumminsii, an emended description of this species is presented. Due to the lack of A. cumminsii in established identification systems, it is most likely that this species is underdiagnosed in many routine clinical bacteriology laboratories.

Actinomycetales Infections↗

Cloning, nucleotide sequence and expression of a hydantoinase and carbamoylase gene from Arthrobacter aurescens DSM 3745 in Escherichia coli and comparison with the corresponding genes from Arthrobacter aurescens DSM 3747.

The genes encoding hydantoinases (hyuH1) and carbamoylases (hyuC1) from Arthrobacter aurescens DSM 3745 and Arthrobacter aurescens DSM 3747 (hyuH2, hyuC2) were cloned in Escherichia coli and the nucleotide sequences determined. The hydantoinase genes comprised 1,377 base pairs and the carbamoylase genes 1,239 base pairs each. Both hydantoinases, as well as both carbamoylases, showed a high degree of nucleotide and amino acid sequence identity (96-98%). The hyuH and hyuC genes were expressed in E. coli under the control of the rhamnose promoter and the different specific activities obtained in E. coli crude extracts were compared to those produced by the original hosts. For purification the hyuH2 gene was expressed as a maltose-binding protein (MalE) and as an intein-chitin binding domain (CBD) fusion in E. coli. The expression of malE-hyuH2 resulted in the production of more soluble and active protein. With respect to temperature stability, optimal pH and optimal temperature, substrate and stereospecificity, the purified fusion enzyme exhibited properties similar to those of the wild-type enzyme.

ATP-Binding Cassette Transporters↗

Isolation and characterization of Arthrobacter bacteriophages and their application to phage typing of soil arthrobacters.

Seventeen bacteriophages, active against 19 Arthrobacter soil isolates, were isolated from concentrated samples of river water and sewage. Attempts to isolate Arthrobacter bacteriophages from filtrates of broth cultures of the soil isolates or from ultraviolet light-irradiated cultures were unsuccessful. Bacteriophages were not detected in either concentrated or unconcentrated soil extracts. Electron microscopic studies of 11 phages showed morphologies characteristic of Bradley's groups B (exhibited by 9 phages) and C (exhibited by 2 phages). Moles percent guanine plus cytosine, calculated from the deoxyribonucleic acid density of three phages, ranged from 60.2 to 65.3. The phages were characterized by their plague and virion morphology, host range, and serological specificity.

Arthrobacter↗

The application of serological techniques to the taxonomy of Arthrobacter and related organisms.

Antisera were raised against rods of 17 named Arthrobacter and Aureobacterium strains. Antigenic relationships between these strains, other soil bacteria and new Arthrobacter isolates from several soils were studied, using agglutination, immunodiffusion, immunofluorescence and ELISA techniques. Many of the named Arthrobacter species had common antigens, and there were also common antigens amongst named Arthrobacter strains and many fresh Arthrobacter isolates. Agglutination, ELISA and immunofluorescence tests revealed greater antigenic differences between the named strains than did immunodiffusion tests. Serological similarities between the 17 named strains and the fresh Arthrobacter isolates were calculated using SJ coefficients. The occurrence of named strains in serogroups based on immunodiffusion data supported the taxonomic scheme for arthrobacters in Bergey's Manual of Systematic Bacteriology. The distribution of soil isolates in serogroups resembled that in groups based on numerical analysis of diverse characters. This makes it possible to use serological tests to locate particular species in soil samples. Arthrobacter atrocyaneus was serologically distinct from other members of the Ar. globiformis/Ar. citreus group and did not cluster with them phenetically. Serological data suggest that Ar. aurescens, Ar. ureafaciens and Ar. histidinolovorans constitute a single species. Although A. simplex and A. tumescens have been placed in the genus Pimelobacter, they are serologically distinct from one another and more closely resemble Ar. globiformis. Aureobacterium strains were serologically distinct from arthrobacters. Micrococcus roseus showed many cross-reactions with several antisera, supporting the placement of micrococci in the same family as arthrobacters.

Animals↗