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Novel Polyoxyethylene-Containing Glycolipids Are Synthesized in Corynebacterium matruchotii and Mycobacterium smegmatis Cultured in the Presence of Tween 80.

Wang C, Mahrous EA, Lee RE, Vestling MM, Takayama K - J Lipids (2010)

Bottom Line: Correspondingly the trehalose dicorynomycolate content was reduced.MALDI mass spectrometry, MS-MS, (1)H-NMR, and (13)C-NMR were used to characterize the series-2 glycolipids.Series-2A glycolipid is trehalose 6-C(36:2)-corynomycolate-6'-polyoxyethylenate and series-2B glycolipid is trehalose 6-C(36:2)-corynomycolate-6'-furan ring-containing polyoxyethylenate.

View Article: PubMed Central - PubMed

Affiliation: Mycobacteriology Research Laboratory, William S. Middleton Memorial Veterans Hospital, Madison, WI 53705, USA.

ABSTRACT
The addition of polyoxyethylene sorbitan monooleate (Tween 80) to a culture of mycobacteria greatly influences cell permeability and sensitivity to antibiotics but very little is known regarding the underlying mechanism. Here we show that Corynebacterium matruchotii (surrogate of mycobacteria) converts Tween 80 to a structural series of polyoxyethylenic acids which are then used to form novel series-2A and series-2B glycolipids. Minor series-3 glycolipids were also synthesized. The polyoxyethylenic acids replaced corynomycolic acids in the cell wall. Correspondingly the trehalose dicorynomycolate content was reduced. MALDI mass spectrometry, MS-MS, (1)H-NMR, and (13)C-NMR were used to characterize the series-2 glycolipids. Series-2A glycolipid is trehalose 6-C(36:2)-corynomycolate-6'-polyoxyethylenate and series-2B glycolipid is trehalose 6-C(36:2)-corynomycolate-6'-furan ring-containing polyoxyethylenate. Mycobacterium smegmatis grown in the presence of Tween 80 also synthesizes series-2 type glycolipids. The synthesis of these novel glycolipids in corynebacteria and mycobacteria should result in gross changes in the cell wall permeability and drug sensitivity.

No MeSH data available.


Related in: MedlinePlus

Predicted products of degradation of Tween 80 by C. matruchotii.  Structure A is polyoxyethylenic acid derived from the furan ring of Tween 80 (http://www.sigma-aldrich.com/) and structure B is the furan ring-associated polyoxyethylenic acid.  Structures C and D are further degradation products of B.  The terminal hydroxyl group is thought to be oxidized to the carboxyl group and all of these products are used by the bacteria to synthesize series-2A and -2B glycolipids.  Alternative to structure B, the carboxylic acid group could be on the other polyoxyethylenate unit.  The proposed structures of series-2A and -2B glycolipids are shown in upper right and lower right panels, respectively.
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fig3: Predicted products of degradation of Tween 80 by C. matruchotii. Structure A is polyoxyethylenic acid derived from the furan ring of Tween 80 (http://www.sigma-aldrich.com/) and structure B is the furan ring-associated polyoxyethylenic acid. Structures C and D are further degradation products of B. The terminal hydroxyl group is thought to be oxidized to the carboxyl group and all of these products are used by the bacteria to synthesize series-2A and -2B glycolipids. Alternative to structure B, the carboxylic acid group could be on the other polyoxyethylenate unit. The proposed structures of series-2A and -2B glycolipids are shown in upper right and lower right panels, respectively.

Mentions: We concluded that the series-2 glycolipid contains trehalose, C36:2-corynomycolic acid, and two structural series of polyoxyethylenic acids (which are the variables). We then attempted to isolate the polyoxyethylenic acids from series-2 glycolipid by mild base hydrolysis with TBAH. We were unsuccessful when we used the much stronger KOH reagent. Purified series-2 glycolipid was hydrolyzed in 15% TBAH at 100°C, the fatty acids were methylated and analyzed by MALDI mass spectrometry (Figure 2). The spectrum showed two series of molecular ion peaks occurring at intervals of 44 amu. They were MNa+ at m/z (A) 317, 361, 449, and 537; (B) 611, 655 and 699 as remnants of degradation. We could not associate the structures of these molecular ions with the polyoxyethylenic acids as shown in Figure 3. The molecular ion at m/z 585 represents methyl C36:2-corynomycolate. This is additional evidence that series-2 glycolipid contains polyoxyethylenate, replacing one corynomycolate in the structure of TDCM.


Novel Polyoxyethylene-Containing Glycolipids Are Synthesized in Corynebacterium matruchotii and Mycobacterium smegmatis Cultured in the Presence of Tween 80.

Wang C, Mahrous EA, Lee RE, Vestling MM, Takayama K - J Lipids (2010)

Predicted products of degradation of Tween 80 by C. matruchotii.  Structure A is polyoxyethylenic acid derived from the furan ring of Tween 80 (http://www.sigma-aldrich.com/) and structure B is the furan ring-associated polyoxyethylenic acid.  Structures C and D are further degradation products of B.  The terminal hydroxyl group is thought to be oxidized to the carboxyl group and all of these products are used by the bacteria to synthesize series-2A and -2B glycolipids.  Alternative to structure B, the carboxylic acid group could be on the other polyoxyethylenate unit.  The proposed structures of series-2A and -2B glycolipids are shown in upper right and lower right panels, respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3066834&req=5

fig3: Predicted products of degradation of Tween 80 by C. matruchotii. Structure A is polyoxyethylenic acid derived from the furan ring of Tween 80 (http://www.sigma-aldrich.com/) and structure B is the furan ring-associated polyoxyethylenic acid. Structures C and D are further degradation products of B. The terminal hydroxyl group is thought to be oxidized to the carboxyl group and all of these products are used by the bacteria to synthesize series-2A and -2B glycolipids. Alternative to structure B, the carboxylic acid group could be on the other polyoxyethylenate unit. The proposed structures of series-2A and -2B glycolipids are shown in upper right and lower right panels, respectively.
Mentions: We concluded that the series-2 glycolipid contains trehalose, C36:2-corynomycolic acid, and two structural series of polyoxyethylenic acids (which are the variables). We then attempted to isolate the polyoxyethylenic acids from series-2 glycolipid by mild base hydrolysis with TBAH. We were unsuccessful when we used the much stronger KOH reagent. Purified series-2 glycolipid was hydrolyzed in 15% TBAH at 100°C, the fatty acids were methylated and analyzed by MALDI mass spectrometry (Figure 2). The spectrum showed two series of molecular ion peaks occurring at intervals of 44 amu. They were MNa+ at m/z (A) 317, 361, 449, and 537; (B) 611, 655 and 699 as remnants of degradation. We could not associate the structures of these molecular ions with the polyoxyethylenic acids as shown in Figure 3. The molecular ion at m/z 585 represents methyl C36:2-corynomycolate. This is additional evidence that series-2 glycolipid contains polyoxyethylenate, replacing one corynomycolate in the structure of TDCM.

Bottom Line: Correspondingly the trehalose dicorynomycolate content was reduced.MALDI mass spectrometry, MS-MS, (1)H-NMR, and (13)C-NMR were used to characterize the series-2 glycolipids.Series-2A glycolipid is trehalose 6-C(36:2)-corynomycolate-6'-polyoxyethylenate and series-2B glycolipid is trehalose 6-C(36:2)-corynomycolate-6'-furan ring-containing polyoxyethylenate.

View Article: PubMed Central - PubMed

Affiliation: Mycobacteriology Research Laboratory, William S. Middleton Memorial Veterans Hospital, Madison, WI 53705, USA.

ABSTRACT
The addition of polyoxyethylene sorbitan monooleate (Tween 80) to a culture of mycobacteria greatly influences cell permeability and sensitivity to antibiotics but very little is known regarding the underlying mechanism. Here we show that Corynebacterium matruchotii (surrogate of mycobacteria) converts Tween 80 to a structural series of polyoxyethylenic acids which are then used to form novel series-2A and series-2B glycolipids. Minor series-3 glycolipids were also synthesized. The polyoxyethylenic acids replaced corynomycolic acids in the cell wall. Correspondingly the trehalose dicorynomycolate content was reduced. MALDI mass spectrometry, MS-MS, (1)H-NMR, and (13)C-NMR were used to characterize the series-2 glycolipids. Series-2A glycolipid is trehalose 6-C(36:2)-corynomycolate-6'-polyoxyethylenate and series-2B glycolipid is trehalose 6-C(36:2)-corynomycolate-6'-furan ring-containing polyoxyethylenate. Mycobacterium smegmatis grown in the presence of Tween 80 also synthesizes series-2 type glycolipids. The synthesis of these novel glycolipids in corynebacteria and mycobacteria should result in gross changes in the cell wall permeability and drug sensitivity.

No MeSH data available.


Related in: MedlinePlus