Trennung einfacher zucker auf cellulose-schichten

Schweiger, Anton

doi:10.1016/s0021-9673(00)80803-1

Cited by 84 publications

(12 citation statements)

References 5 publications

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“…Chromatograms were developed for 16 h in ethyl acetate-acetic acid-water (3:1:1, vol/vol/vol), and the carbohydrates were visualized by spraying the chromatogram with p-anisidine and phthalic acid, which were dissolved in absolute ethanol, and heated to 100°C for 5 min (20). Preparation of cross-absorbed antiserum.…”

Section: Methodsmentioning

confidence: 99%

Characterization of an outer membrane mannanase from Bacteroides ovatus

Gherardini¹,

Salyers²

1987

J Bacteriol

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Bacteroides ovatus utilizes guar gum, a high-molecular-weight branched galactomannanan, as a sole source of carbohydrate. No extracellular activity was detectable. Approximately 30% of the total cell-associated mannanase activity partitioned with cell membranes. When inner and outer membranes of B. ovatus were separated on sucrose gradients, the mannanase activity was associated mainly with fractions containing outer membranes. Enzyme activity was solubilized by 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) or by Triton X-100 at a detergent-to-protein ratio of 1:1. The enzyme was stable for only 4 h at 37C and for 50 to 60 h at 4°C. Analysis of the products of the CHAPS-solubilized mannanase on Bio-Gel A-5M and Bio-Gel P-10 gel filtration columns indicated that the enzyme breaks guar gum into high-molecular-weight fragments. The CHAPS-solubilized mannanase was partially purified by chromatography on a FPLC Mono Q column. The partially purified mannanase preparation contained three major polypeptides (Mr 94,500, 61,000, and 43,000) and several minor ones. High mannanase activity was seen only when B. ovatus was grown on guar gum. Cross-absorbed antiserum detected two other guar gum-associated outer membrane proteins: a CHAPS-extractable 49,000-dalton polypeptide and a 120,000-dalton polypeptide that was not solubilized by CHAPS. Neither of these polypeptides was detectable in the partially purified mannanase preparation. These results indicate that there are at least two guar gum-associated outer membrane polypeptides other than the mannanase.

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Section: Methodsmentioning

confidence: 99%

Characterization of an outer membrane mannanase from Bacteroides ovatus

Gherardini¹,

Salyers²

1987

J Bacteriol

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“…Phosphorus was estimated by the procedure of Bartlett (1), which was adapted for use with the Technicon Autoanalyzer. A ninhydrin spray was used for the detection of amino nitrogen, and the presence of sugars on chromatograms was determined by the method of Schweiger (24) and by use of an anilinediphenylamine reagent (Sigma Chemical Co.). The presence of fatty acids on chromatograms was visualized by dipping the chromatogram into rhodamine 6G, followed by long-wave length ultraviolet examination.…”

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confidence: 99%

Occurrence of Phosphonosphingolipids in Bdellovibrio bacteriovorus Strain UKi2

1973

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The major phospholipids of two strains of Bdellovibrio bacteriovorus were characterized. Both strain UKil, which is obligately saprophytic, and strain UKi2, which is facultatively parasitic, contained phosphatidylethanolamine and phosphatidylglycerol as their major glycerophosphatides. A branched, 15-carbon fatty acid is the major component of these alkali-labile lipids. Absent from UKil but present in UKi2 were three alkali-stable lipids (compounds 8, 9, and 11) which appear to be phosphosphingolipids. After acid hydrolysis, both compound 8 and 9 yield the identical phosphorus-containing substance that is water soluble, dipolar ionic, and ninhydrin positive. This substance appears to contain a C-P bond since P1 could not be released from this substance by treatment with alkaline phosphatase or by very harsh mineral acid treatment. Based on chromatographic comparisons, this phosphonate appears to be a novel lipid constituent. Upon degradation, compound 8 yields 1 mol of dihydroxy long-chain base and compound 9 yields 1 mol of a trihydroxy long-chain base. These bases appear to have a 17-carbon, possibly branched, structure based on gas-liquid chromatography retention times. Degradation of both sphingolipids yields a mixture of hydroxy fatty acids, the major component being a branched, 15-carbon hydroxy acid. 1971).MATERIALS AND METHODS Growth of bacteria. B. bacteriovorus strains UKil and UKi2 (4) were grown on 1% (wt/vol) peptone-0.3% (wt/vol) yeast extract on a rotary shaker at 30 C for 15 h. In some experiments the medium was supplemented with "2P-orthophosphate (carrier-free, New England Nuclear Corp., or ICN Corp.).Extraction of lipid. The phospholipids were extracted according to a modification of the method of Lester and Steiner (17). '2P-lieled cells were harvested from 25 ml of medium by centrifugation and suspended in 1.0 ml of water. A 5-ml amount of (CH2CH2)20-CH,CH2OH (3:1, vol/vol) was added, on July 16, 2020 by guest

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“…The solvent system was n-butanol, pyridine and water (6: 4: 3, by volume). The carbohydrates were detected with anisidine-phthalate (APT) reagent 12 ) for reducing sugar, urea-phosphoric acid (UPA) reagent for ketose,13) and alkaline triphenyltetrazolium chloride (TTC) reagent 14 ) reacting specifically' with a-keto alcohol groups (-CO-CHOH-) of sugars.…”

Section: Methodsmentioning

confidence: 99%