An efficient procedure for studying pectin structure which combines limited depolymerization and 13C NMR

被引:53
作者
Catoire, L
Goldberg, R
Pierron, M
Morvan, C
du Penhoat, CH
机构
[1] Ecole Normale Super, Dept Chim, CNRS, URA 1679, F-75231 Paris 05, France
[2] Inst Jacques Monod, Lab Enzymol Milieu Struct, F-75251 Paris, France
[3] Univ Rouen, Fac Sci, SCUEOR CNRS URA 203, F-76821 Mt St Aignan, France
来源
EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS | 1998年 / 27卷 / 02期
关键词
pectins; C-13; NMR; relaxation data; thermally-induced depolymerization; pectinmethylesterase;
D O I
10.1007/s002490050118
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
A protocol for partial thermally-induced depolymerization of differently methoxylated pectin samples is described. The resulting macromolecules have been fully characterized with various complementary techniques, such as size exclusion chromatography (SEC), potentiometry, viscometry and C-13 NMR. Optimum conditions afford samples at 50-80% yield with weight-average molecular weights in the 4 to 20 kDa range. The major fraction of these polysaccharides adopts the random-coil conformation and such samples are suitable for C-13 NMR structural studies at room temperature. The methoxyl distributions of two apple pectin samples with a degree of esterification (DE) between 54 and 74% and a citrus pectin (DE, 72%) were shown to be random in nature, whereas that of a lightly methoxylated apple pectin (DE 39%) was partially blockwise. The carbon relaxation parameters of the depolymerized pectins attain asymptotic values for M-w > 4 kDa. The M-w values estimated from intrinsic viscosity data with the Mark-Houwink relationship reported for native pectins are in good agreement with those obtained by either end-group analysis (NMR) or SEC. Thus, all the physicochemical data indicate that the secondary structure of the isolated chains of depolymerized pectin is closely related to that of the parent polymers. Finally, pectinmethylesterase activity towards the depolymerized pectins was similar to that of the untreated samples.
引用
收藏
页码:127 / 136
页数:10
相关论文
共 28 条
[1]   SEQUENTIAL STRUCTURE BY H-1-NMR AS A DIRECT ASSAY FOR PECTINESTERASE ACTIVITY [J].
ANDERSEN, AK ;
LARSEN, B ;
GRASDALEN, H .
CARBOHYDRATE RESEARCH, 1995, 273 (01) :93-98
[2]   GEL-PERMEATION CHROMATOGRAPHY AND THE MARK-HOUWINK RELATION FOR PECTINS WITH DIFFERENT DEGREES OF ESTERIFICATION [J].
ANGER, H ;
BERTH, G .
CARBOHYDRATE POLYMERS, 1986, 6 (03) :193-202
[3]  
ANGER H, 1985, NAHRUNG, V29, P397, DOI 10.1002/food.19850290425
[4]  
Aspinall GO, 1982, POLYSACCHARIDES, P36
[5]   STRUCTURE OF CITRUS PECTINS AND VISCOMETRIC STUDY OF THEIR SOLUTION PROPERTIES [J].
AXELOS, MAV ;
THIBAULT, JF ;
LEFEBVRE, J .
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, 1989, 11 (03) :186-191
[6]   TRENDS IN MOLECULAR-MOTION FOR A SERIES OF GLUCOSE OLIGOMERS AND THE CORRESPONDING POLYMER PULLULAN AS MEASURED BY C-13 NMR RELAXATION [J].
BENESI, AJ ;
BRANT, DA .
MACROMOLECULES, 1985, 18 (06) :1109-1116
[7]   IMMOBILIZED AND FREE APOPLASTIC PECTINMETHYLESTERASES IN MUNG BEAN HYPOCOTYL [J].
BORDENAVE, M ;
GOLDBERG, R .
PLANT PHYSIOLOGY, 1994, 106 (03) :1151-1156
[8]   An NMR study of the dynamic single-stranded conformation of sodium pectate [J].
Catoire, L ;
Derouet, C ;
Redon, AM ;
Goldberg, R ;
duPenhoat, CH .
CARBOHYDRATE RESEARCH, 1997, 300 (01) :19-29
[9]   ON THE INTRINSIC-VISCOSITY OF ANIONIC AND NONIONIC RODLIKE POLYSACCHARIDE SOLUTIONS [J].
CHUN, MS ;
PARK, OO .
MACROMOLECULAR CHEMISTRY AND PHYSICS, 1994, 195 (02) :701-711
[10]  
Cros S, 1996, BIOPOLYMERS, V39, P339, DOI 10.1002/(SICI)1097-0282(199609)39:3<339::AID-BIP6>3.0.CO