Microdetermination of Hexosamines

Tsuji, Akio; Kinoshita, Toshio; Hoshino, Masanori

doi:10.1248/cpb.17.217

Cited by 62 publications

(40 citation statements)

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“…Fractions (5 ml) were collected and assayed for total hexose (0) and total hexosamine ( 0 ) . Pooled fractions are indicated by bars by the method of Tsuji et al [24] with glucosamine as a standard after N-deacylation of samples by treatment in 2 M HCl at 100°C for 2 h. Phosphorus was determined by the method of Lowry et al [25], and 3-deoxyoctulosonic acid by the thiobarbituric acid method [26, 271. Descending paper chromatography was carried out on Toyo no.…”

Section: Other Methods and Materialsmentioning

confidence: 99%

Characterization of a polysaccharide component of lipopolysaccharide from Pseudomonas aeruginosa IID 1008 (ATCC 27584) as D‐rhamnan

Yokota

Kaya

Sawada

et al. 1987

European Journal of Biochemistry

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Structural studies were carried out on a rhamnose‐rich polysaccharide isolated from the O‐polysaccharide fraction of lipopolysaccharide in Pseudomonas aeruginosa IID 1008 (ATCC 27584) after destruction of the major O‐specific chain by alkaline treatment. The isolated polysaccharide contained rhamnose, 3‐O‐methyl‐6‐deoxyhexose, glucose, xylose, alanine, galactosamine and phosphorus in a molar ratio of 67:6.9:4.3:2.1:1.1:1.0:4.1. Data from analysis involving Smith degradation, methylation, 1H‐NMR spectroscopy and optical rotation measurement showed that the polysaccharide was built up of three moieties, a rhamnan chain composed of about 70 D‐rhamnose residues, the core chain and an oligosaccharide chain comprising 3‐O‐methyl‐6‐deoxyhexose, xylose, rhamnose and probably glucose. The repeating unit of the rhamnan chain was indicated to have the following structure: →3)D‐Rha(α1 → 3)D‐Rha(α1 → 2)D‐Rha(α1 →. This structure is identical with that proposed previously for the repeating unit of the side chain of lipopolysaccharide from plant pathogenic bacteria Pseudomonas syringae pv. morsprunorum C28 [Smith, A. R. W., Zamze, S. E., Munro, S. M., Carter, K. J. and Hignett, R. C. (1985) Eur. J. Biochem. 149, 73–78].

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Section: Other Methods and Materialsmentioning

confidence: 99%

Characterization of a polysaccharide component of lipopolysaccharide from Pseudomonas aeruginosa IID 1008 (ATCC 27584) as D‐rhamnan

Yokota

Kaya

Sawada

et al. 1987

European Journal of Biochemistry

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“…Analyses were carried out by standard methods as follows: phosphorus (Chen, Toribara & Warner, 1956); carbohydrate (Dubois et a/., 1956); reducing sugars (Somogyi, 1952); 2-keto-3-deoxyoctonic acid (KDO) (Ellwood, 1970); heptose (Wright & Rebers, 1972); amino compounds (Moore & Stein, 1954); amino sugars (Tsuji, Kinoshita & Hoshino, 1969); uronic acids (Bitter & Muir, 1962) after hydrolysis in 0.5 M-H,SO, at 100 "C for 13 h; N-acetylamino sugars (Reissig, Strominer & Leloir, 1955), after hydrolysis in 1 M-HCl at 100 "C for 5 min. Amino compounds were tentatively identified on a Jeol amino acid autoanalyser.…”

Section: Extraction Of Lipopolysaccharide (Lps)mentioning

confidence: 99%

Cell Envelope Components of Strains Belonging to the Genus Thermus

Pask-Hughes¹,

Williams

1978

Journal of General Microbiology

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Envelopes of Thermus strains had a multilayered Gram-negative appearance. Peptidoglycan contained significant amounts of glycine, and ornithine was the dibasic amino acid. Lipopolysaccharide (LPS) did not contain heptose and 2-keto-3-deoxyoctonate was not positively identified. The polysaccharide from LPS contained glucose, glucosamine, galactose, galactosamine and, in two strains, mannose. The fatty acids identified in lipopolysaccharide were C14, C16 and CIS, both saturated and unsaturated.

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“…The MBTH assay for the determination of hexosamines requires that the amino sugar have an unsubstituted amino group in the 2-position because the first step in the procedure is a nitrous acid deamination of the amino sugar (12). The greater response of the Pkl HAG would seem to indicate either less N-substitution of the hexosamine residues present or a greater total number of hexosamine residues.…”

Section: Discussionmentioning

confidence: 99%

“…Hexosamine was assayed by the MBTH procedure of Tsuji et al (12,13). Protein was assayed by the method of Lowry et al (14).…”

mentioning

confidence: 99%

Molecular alteration in a Neurospora crassa morphological mutant and its phenocopy.

Springer¹,

Srb²

1978

Proc. Natl. Acad. Sci. U.S.A.

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A procedure using ion exchange chromatography has been developed to detect alterations in a polysaccharide produced by Neurospora crassa. The polysaccharide, isolated from medium that has supported the growth of a culture, is highly responsive to the 3-methyl-2-benzothiazolinone hydrazone assay, indicating a high hexosamine content. The substance elaborated by wild-type N. crassa can be fractionated into two components that appear by rechromatography to be closely related. When isolated from mutants of the peak (pk) locus, the corresponding polysaccharide cannot be resolved into two components. Instead, a single component is consistently found. This variant chromatographic pattern cosegregates with morphological effects of the pk allele after crosses with the wild type. The polysaccharide isolated from a wild-type culture that has been induced by sorbose to phenocopy the hyphal characteristics of pk mutants elutes from the ion exchange column in a manner similar to the corresponding polysaccharide from the pk mutants.Neurospora crassa may undergo two distinct developmental phases. One phase involves the asexual production of conidia that germinate to produce a mycelial mat producing more conidia. The second, the sexual developmental phase, consists of protoperithecia production, fertilization of the protoperithecia, and then maturation of the perithecia, or fruiting bodies. Perithecia maturation includes differentiation of the perithecial wall and internal development of asci containing ascospores. By use of appropriate mutants, loci that regulate development of one phase only (1, 2) as well as loci regulating both phases (3) have been identified. A number of loci of the second type regulate morphology of both asci and vegetative hyphae. Mutations at these various loci offer the potential for dissecting ascus development and providing insight into the relationship of the zygote to the developing ascus. Full utilization of these mutants to study development requires some knowledge of their molecular function. No consistent molecular alteration that can be directly related to alterations in morphology has been previously detected for mutations at any of these loci. The careful examination of a molecular component of Neurospora now reveals a consistent molecular alteration for members of one set of allelic mutants.Reissig and Glasgow (4) reported the isolation of a growthregulating substance both from purified hyphal wall of Neurospora and from medium that had supported a culture. They found no difference between the materials from the two sources. The substance was found to be composed primarily of 2-amino-2-deoxygalactose, with several neutral sugars and amino acids as minor components. A portion of the 2-amino-2-deoxygalactose residues was found to be N-acetylated.Originally referred to as MP (for mucopolysaccharide), the designation was later changed to galactosaminoglycan (5).Mycelial morphology in Neurospora is determined in part by the frequency and location of branching and by growth rate. The galac...

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Microdetermination of Hexosamines

Cited by 62 publications

References 0 publications

Characterization of a polysaccharide component of lipopolysaccharide from Pseudomonas aeruginosa IID 1008 (ATCC 27584) as D‐rhamnan

Characterization of a polysaccharide component of lipopolysaccharide from Pseudomonas aeruginosa IID 1008 (ATCC 27584) as D‐rhamnan

Cell Envelope Components of Strains Belonging to the Genus Thermus

Molecular alteration in a Neurospora crassa morphological mutant and its phenocopy.

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