Determination of average degree of polymerisation and distribution of oligosaccharides in a partially acid-hydrolysed homopolysaccharide:: A comparison of four experimental methods applied to mannuronan

被引:33
作者
Campa, C
Oust, A
Skjåk-Bræk, G
Paulsen, BS
Paoletti, S
Christensen, BE [1 ]
Ballance, S
机构
[1] Norwegian Univ Sci & Technol, NOBIPOL, Dept Biotechnol, NO-7491 Trondheim, Norway
[2] Univ Trieste, Dept Biochem Biophys & Macromol Chem, I-34127 Trieste, Italy
[3] Univ Oslo, Sch Pharm, Dept Pharmacognosy, NO-0316 Oslo, Norway
关键词
degree of polymerisation; alginate; oligosaccharides; mannuronan;
D O I
10.1016/j.chroma.2003.11.045
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The average degree of polymerisation (DP) and distribution of oligosaccharides in partially acid hydrolysed mannuronans were quantitatively evaluated by H-1 NMR, electrospray ionisation mass spectrometry (ESI-MS), micellar electrokinetic capillary chromatography with UV detection (MEKC-UV), and high-pressure anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD). Our investigation shows that H-1 NMR, MEKC-UV and, in particular, HPAEC-PAD can be used as quantitative tools to aid the investigation of polysaccharide structure, function and synthesis. For the latter two techniques, especially, this represents a significant new development as it enables calculation of the quantity of individual oligomers of nominal DP by direct analysis of a defined oligomer mixture. Appropriate statistical averages of number and weight distributions were also calculated and found to fit very well to predicted Kuhn distributions that assume random depolymerisation. (C) 2003 Elsevier B.V. All rights reserved.
引用
收藏
页码:271 / 281
页数:11
相关论文
共 30 条
[1]   HIGH-PERFORMANCE ANION-EXCHANGE CHROMATOGRAPHY WITH PULSED AMPEROMETRIC DETECTION OF LINEAR AND BRANCHED GLUCOSE OLIGOSACCHARIDES [J].
AMMERAAL, RN ;
DELGADO, GA ;
TENBARGE, FL ;
FRIEDMAN, RB .
CARBOHYDRATE RESEARCH, 1991, 215 (01) :179-192
[2]  
Baker D. R., 1995, CAPILLARY ELECTROPHO
[3]   PREPARATIVE CHROMATOGRAPHY OF OLIGOGALACTURONIC ACIDS [J].
DONER, LW ;
IRWIN, PL ;
KURANTZ, MJ .
JOURNAL OF CHROMATOGRAPHY, 1988, 449 (01) :229-239
[4]  
Ertesvåg H, 1999, METH BIOTEC, V10, P71
[5]   Time-resolved 1H and 13 CNMR spectroscopy for detailed analyses of the Azotobacter vinelandii mannuronan C-5 epimerase reaction [J].
Hartmann, M ;
Duun, AS ;
Markussen, S ;
Grasdalen, A ;
Valla, S ;
Skjåk-Bræk, G .
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, 2002, 1570 (02) :104-112
[6]   INFLUENCE OF BORATE COMPLEXATION ON THE ELECTROPHORETIC BEHAVIOR OF CARBOHYDRATES IN CAPILLARY ELECTROPHORESIS [J].
HOFFSTETTERKUHN, S ;
PAULUS, A ;
GASSMANN, E ;
WIDMER, HM .
ANALYTICAL CHEMISTRY, 1991, 63 (15) :1541-1547
[7]   ANALYSIS OF OLIGOGALACTURONIC ACIDS WITH 50 OR FEWER RESIDUES BY HIGH-PERFORMANCE ANION-EXCHANGE CHROMATOGRAPHY AND PULSED AMPEROMETRIC DETECTION [J].
HOTCHKISS, AT ;
HICKS, KB .
ANALYTICAL BIOCHEMISTRY, 1990, 184 (02) :200-206
[8]   ANALYSIS OF PECTATE LYASE-GENERATED OLIGOGALACTURONIC ACIDS BY HIGH-PERFORMANCE ANION-EXCHANGE CHROMATOGRAPHY WITH PULSED AMPEROMETRIC DETECTION [J].
HOTCHKISS, AT ;
HICKS, KB .
CARBOHYDRATE RESEARCH, 1993, 247 :1-7
[9]   Isolation of oligogalacturonic acids up to DP 20 by preparative high-performance anion-exchange chromatography and pulsed amperometric detection [J].
Hotchkiss, AT ;
Lecrinier, SL ;
Hicks, KB .
CARBOHYDRATE RESEARCH, 2001, 334 (02) :135-140
[10]   Quantitative analysis of amylopectin unit chains by means of high-performance anion-exchange chromatography with pulsed amperometric detection [J].
Koch, K ;
Andersson, R ;
Åman, P .
JOURNAL OF CHROMATOGRAPHY A, 1998, 800 (02) :199-206