NMR spectroscopy analysis of oligoguluronates and oligomannuronates prepared by acid or enzymatic hydrolysis of homopolymeric blocks of alginic acid. Application to the determination of the substrate specificity of Haliotis tuberculata alginate lyase.
The 1H and 13C NMR chemicals shifts of the various saturated and unsaturated timers obtained by acid or enzymatic depolymerisation of homopolymeric blocks of alginates are reported. In addition, 13C NMR chemical shifts are assigned for several saturated oligomers of higher polymerisation degrees. Breakdown of alginate and of homopolymeric alginate blocks by Haliotis tuberculata alginate lyase was monitored with 1H NMR spectroscopy and the signals relevant to the identification of the lyase products are pointed out. The enzymes performs beta-elimination on the mannuronic acid residues, independently of their immediately surrounding neighbours. Application of this approach to the analysis of the substrate specificity of alginate lyases is discussed.
Heyraud
A
A
Centre de Recherches sur les Macromolécules Végétales, C.N.R.S. et Université Joseph Fourier, Grenoble, France.
Gey
C
C
Leonard
C
C
Rochas
C
C
Girond
S
S
Kloareg
B
B
D000464:Alginates / Q000737:chemistry*; D000818:Animals; D056890:Eukaryota / Q000737:chemistry; D020723:Glucuronic Acid; D006603:Hexuronic Acids / Q000737:chemistry / Q000302:isolation & purification; D006868:Hydrolysis; D007711:Klebsiella pneumoniae / Q000201:enzymology; D009682:Magnetic Resonance Spectroscopy; D008974:Mollusca / Q000201:enzymology*; D011133:Polysaccharide-Lyases / Q000378:metabolism*; D013379:Substrate Specificity; D014574:Uronic Acids / Q000737:chemistry / Q000302:isolation & purification
D000464:Alginates; D006603:Hexuronic Acids; D014574:Uronic Acids; C007896:guluronic acid; D020723:Glucuronic Acid; C008324:mannuronic acid; D011133:Polysaccharide-Lyases; C016158:poly(beta-D-mannuronate) lyase