Structural determination of ‘cord factor’ from aCorynebacterium diphtheriae strain by a combination of mass spectral ionization methods: Field desorption cesium cationization and electron impact mass spectrometry studies

Puzo, Germain; Tissié, G.; Lacave, Charlotte; Aurelle, Hélène; Promé, Jean‐Claude

doi:10.1002/bms.1200051212

Cited by 13 publications

(7 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…On gas-liquid chromatography, the long-chain ketones isolated from B3 and B2 behaved similarly and palmitone was the main component. The same behaviour was observed for corynomycolic acids isolated from B2 and B1 ('true' cord factor) [4] by gas-liquid chromatography either under pyrolytic conditions of the methyl esters (a mixture of saturated and unsaturated aldehyde and esters containing 16, 14 and 12 carbon atoms on the aliphatic chain was identified) or as the 3-0-trimethylsilyl-methyl ester derivatives. This latter analysis showed four main constituents : the major one possessed the same retention time as the synthetic methyl 3-trimethylsilyloxy-2-tetradecyl-octadecanoate.…”

Section: Structural Study Of B2supporting

confidence: 67%

“…The former series lost a trimethylsilanol molecule to give more intense peaks at rnje = 765, 737, 709, 681 and 653, corresponding to the different homologues. As it was previously shown [4], the homologues series (oxonium -MesSiOH)' did not interfere with another series resulting from an H transfer during the splitting of the glycosidic bond [lo]. Therefore, this series could be used for the determination of the unsaturation degree of the homologues.…”

Section: Structural Study Of B3mentioning

confidence: 85%

“…The compound from the upper spot (RF 0.9, called B1) was identified as the 'true' cord factor i.e. 6,6'-bis(corynomycoloyl)-a-~-trehalose, and its structure has been described in a separate paper [4]. The compounds giving the middle spot (RF0.7) and the lower spot (RF 0.5), called respectively B2 and B3, both gave a-D-trehalose as the only detectable watersoluble component after alkaline hydrolysis (identified by gas-liquid chromatography of the Me3Si ether and by hydrolysis into D-glucose).…”

Section: Separation Isolation and Characterization Of (3-ouo-atyl) -mentioning

confidence: 99%

“…The fragmentation pathways of the two glucosyl moieties of the bis acyl-trehalose appeared to be independent, and the presence of oxonium ions bearing either a 3-0x0-ester group or a corynomycolate ester indicated the acylation of the two glucosyl moieties. Thus, the electron impact spectrum of B2 behaved as the superimposition of two spectra, one from a (3-oxoacyl)-glucosyl compound, such as B3, and the other from a corynomycoloyl-glucosyl species such as B1 [4].…”

Section: Structural Study Of B2mentioning

confidence: 99%

“…Me3Si, trimethylsilyl. differing by the chain length and the unsaturation degree, was presented elsewhere [4]. …”

mentioning

confidence: 99%

See 4 more Smart Citations

Occurrence of 3‐Oxo‐acyl Groups in the 6,6′‐Diesters of α‐d‐Trehalose

Puzo¹,

Tissié²,

Aurelle³

et al. 1979

European Journal of Biochemistry

View full text Add to dashboard Cite

The 'cord factor' from Corynebacterium diphtheriae was fractionated by thin-layer chromatography of the trimethylsilyl derivatives into three main components: the true 'cord factor' B1 which has been described previously and two compounds, B2 and B3, containing 3-0x0-acyl esters. These products were analyzed by mass spectrometry (field desorption/cationization and electron impact of the trimethylsilyl derivatives) and by chemical degradation.Compound B3 is a 6,6'-bis(3-oxo-acyl)-cc-~-trehalose. The 3-0x0-acyl groups are mainly saturated species, branched at C-2, containing 24, 26, 28, 30 or 32 carbon atoms. The main component of B3 is the 6,6'-bis(3-oxo-2-tetradecyl-octadecanoyl)-cc-~-trehalose.Compound B2 is a 6,6'-mixed ester of cc-D-trehalose with the preceding 3-0x0 acids and with corynomycolic acids. These latter contain an even number of carbon atoms from 24 to 32 and are a mixture of saturated, mono-unsaturated and di-unsaturated species. The main compound of B2 is 6-(3-oxo-2-tetradecyl-octadecanoyl)-6'-(3-hydroxy-2-tetradecyl-octadecanoyl)-cc-~-trehalose.The metabolic implications of this structural study, together with results already published, suggest that condensation of the common fatty acids by a Claisen-like process via the 0x0 ester pathway may not be the only way for the synthesis of corynomycolic acids.In previous papers, it was shown that both whole cells and cell-free extracts of Corynehacteriunz diphtheriae catalyzed the transformation of palmitic acid into C32 3-0x0 esters [1,2] by a Claisen-like condensation reaction. By pulse-labelling experiments the first condensation product has been detected [l], and further isolated and identified as 6-0-(3-0x0-2-tetradecyl-octadecanoy1)-cc-D-trehalose [3]. Some other compounds containing 3-0x0-ester groups were also detected: after labelling of whole cells by means of [l -'4C]palmitic acid, radioactive palmitone was isolated from products of alkaline hydrolysis of lipid fractions. For instance, the 'cord factor' fraction, which was an apparently homogenous fraction by chromatography, gave a small quantity of palmitone after hydrolysis, together with a large amount of corynomycolic acid. This work describes an exhaustive fractionation of 'cord factor' preparation and the isolation of new 3-0x0-ester-containing compounds. A structural study of the 'true' cord factor isolated from this fraction, namely a mixture of 6,6'-bis corynomycoloyl)-sc-D-trehaloses (compound B1 in Fig. 1) Ahhreviotion. Me3Si, trimethylsilyl. differing by the chain length and the unsaturation degree, was presented elsewhere [4]. MATERIALS AND METHODS Physical MethodsThe infrared spectra were obtained with a Perkin Elmer 177 instrument.The mass spectra were measured on a Varian MAT 31 1A mass spectrometer equipped with a combined electron impact/field ionization/field desorption ion source. For field desorption measurements, the cationization method with caesium iodide [5] was used, by the two-step procedure : a carbon-activated tungstenwire emitter was first dipped into a 0.5 ...

show abstract

Section: Structural Study Of B2supporting

confidence: 67%

Section: Structural Study Of B3mentioning

confidence: 85%

Section: Separation Isolation and Characterization Of (3-ouo-atyl) -mentioning

confidence: 99%

Section: Structural Study Of B2mentioning

confidence: 99%

“…Me3Si, trimethylsilyl. differing by the chain length and the unsaturation degree, was presented elsewhere [4]. …”

mentioning

confidence: 99%

See 3 more Smart Citations

Occurrence of 3‐Oxo‐acyl Groups in the 6,6′‐Diesters of α‐d‐Trehalose

Puzo¹,

Tissié²,

Aurelle³

et al. 1979

European Journal of Biochemistry

View full text Add to dashboard Cite

show abstract

Field desorption mass spectral fragmentation of monosaccharide isomers

Deutsch

1980

Org. Mass Spectrom.

View full text Add to dashboard Cite

Field desorption ionization of selected monosaccharide isomers by potassium ion attachment in high fields is described. The ionization is obtained with a tungsten wire activated with carbon needles and covered with KH,PO, salt. In addition to the potassium cationized molecular ions, reproducible fragmentation was obtained at 10 mA and 20 mA typical of the various monosaccharides. For instance, D-glucose tends to lose a water molecule, whereas D-fructose loses primarily methanol under similar conditions. Methanol elimination is suggested to be characteristic of furanoidal ketoses and water elimination for pyranoidal aldoses, whereas glycol elimination occurs for furanoidal aldoses.Field ionization (FI) and field desorption mass spectrometry (FDMS) of some monosaccharides such as D-glucose, D-fructose and D-ribose have been reported by several authors since 1973.'*2 The occurrence of distinct fragmentation patterns in the FD mass spectra of monosaccharides could provide a reliable tool for the characterization of these compounds. However, the FD mass spectra of the sugars studied showed mainly quasimolecular ions with very little fragmentation.An advance towards the characterization of carbohydrates by FDMS was made by ionization of carbohydrates by alkali ion attachment on the surface of salt layers, which had been deposited on emitter^.^-^ This technique increased the sensitivity of the FDMS method and also gives some fragmentation patterns. Of special interest is the fact that carbohydrates can produce cationized species with cations from the emitters previously treated with alkali solutions.6This study focuses on the ability of carbohydrate isomers to give potassium cationized molecular ions with emitters pretreated with a saturated solution of KH,PO,. With this treatment, the carbohydrates gave reproducible structure dependent fragmentation at different wire currents, as well as the usual quasimolecular ions.A comparison of the F D mass spectra of D-glucose obtained with a treated emitter at various wire currents (Fig. l) shows striking differences. In addition to the protonated molecular ion at m/z 181 a potassium cationized ion shifted by 38 u appears with high abundance at m/z 219. This peak is accompanied by the 4'K isotope peak. An increase in the wire current to 15 mA changes the spectrum, giving rise to an additional peak at m/z 163 due to elimination of water. At 2 0 m A three more peaks appear at mlz 61, 91 and 119. This fragmentation is readily reproducible.The FD mass spectrum of D-fructose under the same conditions is shown in Fig. 2. The major fragmentation at 1 0 m A leads to a peak at m/z 149 corresponding to elimination of a neutral fragment with the composition of methanol (CH,O). The peak at m/z 163 found in the FD mass spectrum of Dglucose, corresponding to water elimination, is absent or of lower abundance in the spectrum, and is replaced by the m/z 149 peak. An increase in the wire current gives more fragmentation which qualitatively resembles the fragmentation of D-glucose. Similar beh...

show abstract

Fatty Acids and Complex Lipids

Asselineau

1984

Gas Chromatography Mass Spectrometry Applications in Microbiology

View full text Add to dashboard Cite

Structural determination of ‘cord factor’ from aCorynebacterium diphtheriae strain by a combination of mass spectral ionization methods: Field desorption cesium cationization and electron impact mass spectrometry studies

Cited by 13 publications

References 17 publications

Occurrence of 3‐Oxo‐acyl Groups in the 6,6′‐Diesters of α‐d‐Trehalose

Occurrence of 3‐Oxo‐acyl Groups in the 6,6′‐Diesters of α‐d‐Trehalose

Field desorption mass spectral fragmentation of monosaccharide isomers

Fatty Acids and Complex Lipids

Contact Info

Product

Resources

About