Thin-layer chromatography of carbohydrates on silica gel impregnated with sodium acetate, monosodium phosphate and disodium phosphate

The cell wall compositions of two strains of "true" nocardiae, Nocardia asteroides R 399 and N. caviae IM 1381, and two strains of "so-called nocardiae," N . piracicabensis and N. mediterranei, were determined. Two different methods were used for preparing the cell walls. In the one, the bacteria were sonically treated, and the cell walls were obtained by differential centrifugation; in the other, the bacteria were delipidated before sonic treatment. The cell walls of true nocardiae contain nocardic acids, identified by comparison with nocardic acids isolated from the whole cell. The peptidoglycan structures of the "true" and "so-called" nocardiae studied showed some important differences. I n the true nocardiae, the glycan strand is constituted of P-N-acetylglucosaminyl ( 1 -+ 4) N-glycolylmuramic acid, and muramic acid is N-acetylated in the strains of "so-called nocardiae," as it is in the majority of bacteria. In addition, the peptide monomers of true nocardiae are diamidated on both the a-carboxyl group of wglutamic acid and the (L) carboxyl group of meso-diaminopimelic acid, whereas the peptide monomers of the "socalled nocardiae" have only one amide substituent on the carboxyl group of mesodiaminopimelic acid. I t is difficult to differentiate without ambiguity members of the genus Nocardia from the mass of the other actinomycetes which contain major amounts of meso-diaminopimelic acid. Our results suggest that true nocardiae may be those actinomycetes whose cell walls contain, in addition to arabinose and galactose, N-glycolylmuramic acid in the glycan part of the peptidoglycan and diamidated peptides in the peptide part of the peptidoglycan. In addition, true nocardiae contain nocardic acids.The differentiation between nocardiae and related organisms is often quite difficult. Morphologically the genus Nocardia contains filamentous, branching bacteria (19). The validity of this characteristic as a differential criterion has been criticized in past years, and biochemical or chemical tests have been proposed to establish a more precise classification (20). Since the initial work of Cummins (7, 8), cell wall composition has proven to be most useful in classifying organisms belonging to the order Actinomycetales. For example, LL-diaminopimelic acid (Dpm) is found in the cell walls of streptomycetes and rneso-Dpm in the cell walls of nocardiae (2).Moreover, some sugars, such as arabinose and galactose, are found in the celi walls of nocardiae but not in those of streptomycetes. Recently it has been proposed that the specific lipids present in whole cells of nocardiae be used to differentiate nocardiae from streptomycetes (27) and from mycobacteria (17,21). Since the first isolation of nocardic acids from Nocardia asteroides (25) and the determination of their structure (3, 4), nocardic acids with related structures have been found in all the species of Nocardia so far examined. Nocardic acids are a-branched, P-hydroxylated acids of the mycolic type; they have recently been found in the cell walls of Nocardia ...

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“…The sugars were detected by spraying with an ethanol-sulfuric acid solution of naphthoresorcinol (18).…”

Section: Methodsmentioning

confidence: 99%

Cell Walls of Nocardiae and Related Actinomycetes: Identification of the Genus Nocardia by Cell Wall Analysis

Karahjoli

Gateau

et al. 1972

International Journal of Systematic Bacteriology

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“…After assay, the PANIG-GA samples were washed and stored until new test. The starch hydrolysis products obtained after 15 and 30 min of reaction were analyzed by thin layer chromatography (TLC), as described by Lato et al [12].…”

Section: Properties Of Immobilized Glucoamylasementioning

confidence: 99%

Immobilization of Aspergillus niger glucoamylase onto a polyaniline polymer

Silva

Asquieri

Fernandes

2005

Process Biochemistry

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“…After the incubation samples were concentrated by speed vacuum and spotted onto a thin-layer chromatography plates (TLC). The chromatogram was developed in a solution of n-butanolacetic acidether -H 2 0 (9:6:3:1) and sugars were visualized by spraying the plate with a solution consisting of 2% aniline, 2% diphenylalanine, and 15 % phosphoric acid dissolved in acetone, followed by incubation for 1 h at 60°C (Lato et al 1969). …”

Section: -mentioning

confidence: 99%

Purification and characterization of an extracellular chitinase from the entomopathogen Metarhizium anisopliae

Pinto¹,

Barreto²,

Vainstein³

et al. 1997

Can. J. Microbiol.

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Chitinases are produced by Metarhizium anisopliae when it is grown in the presence of chitin. A chitinase from the culture filtrate of Metarhizium anisopliae was successively purified by precipitation with ammonium sulphate, followed by anion-exchange chromatography on DEAE-Sephacel. The purified enzyme, which has a molecular mass of approximately 30 kDa by sodium dodecyl sulphatepolyacrylamide gel electrophoresis, catalyses the hydrolysis of p-nitrophenyl 0-N-diacetylchitobiose with an apparent K,,, of 0.537 mmol and V,',,, of 4.86 nmol . mL-I . min-I. The optimum pH and temperature were 4.5 -5.0 and 40 -45 "C, respectively. RCsumC : Le Metarhizium anisopliae produit des chitinases lorsqu'il est cultivC en prksence de chitine. Une chitinase prCsente dans un filtrat de culture de Metarhizium anisopliae a Ct C purifiCe par prkcipitation au sulfate d'ammonium et chromatographie Cchangeuse d'anions sur DEAE-Sephacel. L'enzyme purifiCe dont la masse molCculaire dCterminCe par Clectrophor&se sur gel de polyacrylamide avec dodCcylsulfate de sodium Ctait d'environ 30 kDa catalysait l'hydrolyse de la p-nitrophCny1 0-N-diacCtylchitobiose selon un K,,, de 0.537 mmol et une V, , de 4.86 nmol . mL-I . min-'. Le pH optimal oscillait entre 4.5 et 5.0 et la tempCrature optimale entre 40 et 45OC.

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Thin-layer chromatography of carbohydrates on silica gel impregnated with sodium acetate, monosodium phosphate and disodium phosphate

Cited by 55 publications

References 3 publications

Cell Walls of Nocardiae and Related Actinomycetes: Identification of the Genus Nocardia by Cell Wall Analysis

Cell Walls of Nocardiae and Related Actinomycetes: Identification of the Genus Nocardia by Cell Wall Analysis

Immobilization of Aspergillus niger glucoamylase onto a polyaniline polymer

Purification and characterization of an extracellular chitinase from the entomopathogen Metarhizium anisopliae

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