Cell-Wall Polysaccharides in Coryneform Bacteria

Takeuchi, Mariko; Yokota, Akira

doi:10.2323/jgam.35.233

Cited by 36 publications

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“…Cell walls were hydrolyzed with 2 N HCl at 100°C for 2 h, dried in vacuo, and then analyzed by the method of Mikami and Ishida (10) by using an HPLC apparatus (model LC-5A; Shimadzu Co., Ltd.) equipped with a Shim-pack ISA 07/S2504 column (250 by 4 mm) and a Shimadzu model RE-530 spectrofluorometer (20). Glycolyl analysis.…”

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confidence: 99%

Proposal of Two New Species in the Genus Microbacterium: Microbacterium dextranolyticum sp. nov. and Microbacterium aurum sp. nov.

Yokota

Takeuchi

Weiss

1993

International Journal of Systematic Bacteriology

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The taxonomic positions of Flavobacterium sp. strain IF0 14592= (= M-73T) (T = type strain), a dextran-a-l,2-debranching enzyme producer, and Microbacterium sp. strain IF0 15204T (= H-5T), an isolate obtained from corn steep liquor, were investigated; on the basis of the results of chemotaxonomic and phenetic studies and DNA-DNA similarity data, we propose that these bacteria should be classified in the genus Microbacterium as Microbacterium dextranolyticum sp. nov. and Microbacterium aurum sp. nov., respectively.The type strain of M. dextranolyticum is strain IF0 14592, and the type strain of M. aurum is strain IF0 15204.The genus Microbacterium was proposed by Orla-Jensen (15), and its description was emended by Collins et al. (1). Four species have been described previously: Microbacterium lacticum, Microbacterium imperiale, Microbacterium laevaniformans, and Microbacterium arborescens (1 , 5, 6).During a taxonomic study of Flavobacterium strains in the Institute for Fermentation at Osaka (IFO) culture collection, we found that Flavobacterium sp. strain I F 0 14592T (= M-73=) (T = type strain), a dextran-cx-1,2-debranchin enisolate obtained from corn steep liquor, belong to the genus Microbacterium (21). To determine the taxonomic positions of these organisms, we examined their physiological and chemotaxonomic characteristics and compared these characteristics with those of previously described species of the genus Microbacterium .In this paper we describe characterization of these two strains and propose that they are representatives of two new species of the genus Microbacterium on the basis of the results of chemotaxonomic, phenetic, and DNA-DNA hybridization studies.zyme producer (8, 13), and strain IF0 15204= (=H-5 ?g ), an MATERIALS AND METHODSBacterial strains and culture conditions. The bacterial strains which we studied are listed in Table 1. In addition to the type strains, reference strains of M. lacticum and M. luevunifonnans were included. All strains were cultured at 28°C with aerobic shaking in a peptone-yeast extract medium supplemented with brain heart infusion (PY-BHI medium), which contained 1% peptone, 0.2% yeast extract, 0.2% Bacto brain heart infusion (Difco Laboratories), 0.2% NaCl, and 0.2% D-glucose (pH 7.2). Cells were harvested by centrifugation, washed with water, and then lyophilized.Morphological and phenotypic characteristics. Cell morphology was determined by using cells grown on PY-BHI medium. Motility was determined with a light microscope by the hanging drop method. Unless otherwise indicated, all tests were carried out at 28°C. Catalase activity was determined by bubble formation in a 3% hydrogen peroxide solution. Oxidase activity was determined by the oxidation * Corresponding author.of 1% tetramethyl-p-phenylenediamine on filter paper. Acid production from carbohydrates was studied in a medium containing 0.3% peptone, 0.25% NaCl, 0.003% bromcresol purple, and 0.5% carbohydrate (pH 7.2). Assimilation of organic acids was studied in a medium containing 0.5% organic acid (s...

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confidence: 99%

Proposal of Two New Species in the Genus Microbacterium: Microbacterium dextranolyticum sp. nov. and Microbacterium aurum sp. nov.

Yokota

Takeuchi

Weiss

1993

International Journal of Systematic Bacteriology

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“…The cell wall structures of the cystites differed from those of vegetative cells, and cystites were not produced by the addition of tetracycline at logarithmic and stationary phases. (Takeuchi and Yokota, 1989). Therefore the components of polysaccharides in cell walls would not correlate with cystite formation, and the production of polysaccharides of the cell walls of cystites would have an effect more distinctly on cystite formation and the KOH reaction than the production of peptidoglycans would.…”

Section: Discussionmentioning

confidence: 99%

“…Polysaccharides in crude cell walls were prepared by the sonication-SDS method (Takeuchi and Yokota, 1989). Neutral sugars in polysaccharides were determined by HPLC (Shimadzu LC-10A series, Shimadzu Corp., Kyoto, Japan) with an Asahipak NH2P-50 4E column (4.6ϫ250 mm, Showa Denko K.K., Tokyo, Japan) and an RID-10A detector (Shimadzu Corp.).…”

Section: Introductionmentioning

confidence: 99%

“…The eluent was 2 mM CuSO 4 , and the column was kept at 35°C. The type of peptidoglycan of Arthrobacter species was referred to the papers of Schleifer and Kandler (1972), Claus andBerkeley (1986), andFunke et al (1996).Polysaccharides in crude cell walls were prepared by the sonication-SDS method (Takeuchi and Yokota, 1989). Neutral sugars in polysaccharides were determined by HPLC (Shimadzu LC-10A series, Shimadzu Corp., Kyoto, Japan) with an Asahipak NH2P-50 4E column (4.6ϫ250 mm, Showa Denko K.K., Tokyo, Japan) and an RID-10A detector (Shimadzu Corp.).…”

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The formation and structures of cystites of Arthrobacter ureafaciens NRIC 0157T induced by antibiotics.

Tanaka

Uchimura

Komagata

2001

J. Gen. Appl. Microbiol.

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The cystite formation of Arthrobacter ureafaciens NRIC 0157 T was induced by some antibiotics, and the addition of tetracycline at a lag phase was effective for cystite formation in YPM liquid medium. Cystites differed from vegetative cells in the cell wall structure, protein content, and water content. Furthermore, the characteristics and structures of cystites induced by tetracycline were similar to those of cystites produced by nutritional imbalance in CT medium. Consequently, various triggers would induce cystite formation. It is interesting that cystite formation was found in part of the Arthrobacter strains and seemed to correlate with the type of peptidoglycan.Key Words--Arthrobacter; Arthrobacter ureafaciens; cell wall; cystites J. Gen. Appl. Microbiol., 47, 85-97 (2001) * Address reprint requests to: Dr. Naoto Tanaka, Laboratory of General and Applied Microbiology, Department of Applied Biology and Chemistry, Faculty of Applied Bioscience, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156-8502, Japan. E-mail: QWP07736@nifty.ne.jp T : Indicates the type strain of a species. Full PaperThis paper deals with the effect of antibiotics on the cystite formation of A. ureafaciens NRIC 0157T and the structures of cell walls and the intracellular structures of cystites and vegetative cells. A correlation between the imbalance growth and the cystite formation of A. ureafaciens NRIC 0157 T is discussed. Materials and MethodsBacterial strains. The following strains were used in this study: (Tanaka et al., 2000), and was composed of 1.0% beef extract (Difco Laboratories), 1.0% peptone (Mikuni Kagaku Sangyo Ltd.), 0.5% NaCl, and 1.5% agar. The pH was adjusted to 7.0 with NaOH.Cultivation. Strains were aerobically precultured in YPM liquid medium with 30 ml L-form test tubes or 500 ml Erlenmeyer flasks at 28 or 37°C. Cells at a late logarithmic phase were collected by centrifugation at 1,600ϫg for 10 min at 20°C and washed twice with saline. The washed cells were suspended in saline, and the suspension was inoculated with 1% (v/v) into fresh YPM liquid medium, CT liquid medium, or other liquid media with a pipette. The liquid media were aerobically incubated at 28 or 37°C for 1 or 2 days up to full growth.Determination of bacterial growth. Bacterial growth in liquid media was determined by absorbance at 660 nm with a single-beam spectrophotometer (NO-VASPEC II, Amersham Pharmacia Biotech, Tokyo, Japan).Investigation of cell shapes. Cell shapes were investigated with a phase-contrast microscope (BX-50, oil immersion lens, Olympus, Tokyo, Japan). The change of cell shapes was monitored at various intervals during the growth.Determination of the Gram reaction by the KOH reaction. An incubated liquid medium was centrifuged at 1,600ϫg for 10 min, and precipitates were used to detect the KOH reaction (Powers, 1995).Cell counts. A total cell number was determined with a Thoma-Zeiss counting chamber, and a viable cell number was determined by the number of colonies appearing on nutrient agar plates...

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“…In order to clarify whether the sugar compositions of coryneform bacteria are characteristic of the species or not, it was necessary to compare those of many strains belonging to the same species. In the previous study, we analyzed the sugar composition of the polysaccharides obtained from cell walls (12), but in this study, we compared the sugar composition of the cell walls, because it was confirmed that sugar compositions of cell-wall polysaccharides are basically the same as those of cell walls (11).…”

Section: Takeuchimentioning

confidence: 99%

Evaluation of cell-wall sugar composition as a taxonomic marker of some coryneform bacteria.

Takeuchi

Yokota

1993

J. Gen. Appl. Microbiol.

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Cell-wall amino acid composition has proven to be a useful taxonomic criterion at the generic level for classifying Gram-positive bacteria (2,10), but the taxonomic significance of cell-wall sugars remains to be resolved. Nevertheless, in Bergey's Manual of Systematic Bacteriology (4), cell-wall sugar composition based on the results of Keddie and Cure (6) was described. Coryneform bacteria were considered to contain no sugars characteristic of species or genera, except for the genera Corynebacterium, Mycobacterium, Gordona, Tsukamurella and Rhodococcus with arabinogalactan as the cell-wall polysaccharide (5,11).In the previous paper (12), we reported on the taxonomic values of the polysaccharides and teichoic acids from the cell walls of coryneform bacteria, which were characterized in relation to their taxonomy. We found that the kind of cell-wall polymer, a neutral polysaccharide, teichoic acid or arabinogalactanmycolic acid complex, was homogeneous within each genus, and hence concluded that cell-wall polymers could be used as a valuable chemotaxonomic marker for classifying coryneform bacteria at generic level (12). However, comparison of sugar compositions of cell-wall polymers among genera revealed that there were no sugar profiles characteristic of the genus.Recently, we analyzed the chemical structures of cell-wall polysaccharides from four species of the genus Micro bacterium, Microbacterium lacticum, Micro bacterium laevaniformans, Microbacterium imperiale and Microbacterium arborescens, to determine whether or not there were structural similarities in the peptidoglycanlinked polysaccharides among these bacteria. We have found that the chemical structures of the species of the genus Microbacterium were diverse, but the structure of the strains within a species may be basically similar, and therefore, the heteroge-* Address reprint requests to: Ms .

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Cell-Wall Polysaccharides in Coryneform Bacteria

Cited by 36 publications

References 24 publications

Proposal of Two New Species in the Genus Microbacterium: Microbacterium dextranolyticum sp. nov. and Microbacterium aurum sp. nov.

Proposal of Two New Species in the Genus Microbacterium: Microbacterium dextranolyticum sp. nov. and Microbacterium aurum sp. nov.

The formation and structures of cystites of Arthrobacter ureafaciens NRIC 0157T induced by antibiotics.

Evaluation of cell-wall sugar composition as a taxonomic marker of some coryneform bacteria.

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