The biosynthesis of alginate by a mucoid strain of Pseudomonas aeruginosa, isolated from a cystic-fibrosis patient, was monitored by using 13C-n.m.r. spectroscopy of bacterial cultures incubated with 1-13C- or 2-13C-enriched fructose. When 1-13C- or 2-13C-enriched fructose was used as the precursor of alginate, enrichment with 13C in the constituent uronic acid monomers of the polysaccharide could only be detected in C-1 or C-2 respectively, indicating that alginate is synthesized in Ps. aeruginosa directly from fructose, with the hexose molecule being retained intact; this rules out the involvement of C3 intermediates, which occurs when glucose is the alginate precursor. The absence of detectable poly-L-gluluronate block sequences from the alginate of Ps. aeruginosa was confirmed, and it was shown that there is no modification of the arrangement of the constituent uronic acids between polymerization to form alginate and the appearance of the mature alginate in the extracellular medium. The 13C-n.m.r. data also provided independent evidence for acetylation on D-mannuronate residues and for the ratio of D-mannuronate to L-guluronate residues in newly synthesized alginate, which had previously been determined only for material secreted from bacteria into the extracellular medium.
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May 1990
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Research Article|
May 01 1990
The use of 13C-n.m.r. spectroscopy to monitor alginate biosynthesis in mucoid Pseudomonas aeruginosa
A Narbad;
A Narbad
*Department of Biochemistry, University of Wales College of Cardiff, P.O. Box 903, Cardiff CFI IST, U.K.
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M J E Hewlins;
M J E Hewlins
†School of Chemistry and Applied Chemistry, University of Wales College of Cardiff, P.O. Box 912, Cardiff CFI 3TB, Wales, U.K.
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P Gacesa;
P Gacesa
*Department of Biochemistry, University of Wales College of Cardiff, P.O. Box 903, Cardiff CFI IST, U.K.
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N J Russell
N J Russell
*Department of Biochemistry, University of Wales College of Cardiff, P.O. Box 903, Cardiff CFI IST, U.K.
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Publisher: Portland Press Ltd
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© 1990 London: The Biochemical Society
1990
Biochem J (1990) 267 (3): 579–584.
Citation
A Narbad, M J E Hewlins, P Gacesa, N J Russell; The use of 13C-n.m.r. spectroscopy to monitor alginate biosynthesis in mucoid Pseudomonas aeruginosa. Biochem J 1 May 1990; 267 (3): 579–584. doi: https://doi.org/10.1042/bj2670579
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