Sulfated Arabinose from Codium fragile: Preparation and Extensive Structural Characterization†

doi:10.7503/cjcu20180583

Abstract

Abstract:

The crude polysaccharide was obtained from Codium fragile by water extracted and alcohol precipitated, and further 6 fractions purified by Q-Sepharose fast flow column were named CFP1—CFP6, respectively. CFP6 is a higher purity arabinose of them. The absolute molecular mass of the main fraction CFP6 was determined by high performance gel permeation chromatography and laser scattering instrument. The monosaccharide composition was analyzed by reversed phase high-performance liquid chromatography(RP-HPLC) labeled with 1-phenyl-3-methyl-5-pyrazolone(PMP). The results showed that CFP6 is a homogeneous polysaccharide component with a molecular weight of 79290 and consists of arabinose(Ara) and galactose(Gal) with a molar ratio of 14.8:1.0. Subsequently, the structural features and oligosaccharide sequence composition were identified by multi-dimensional nuclear magnetic resonance spectroscopy and liquid chromatography-mass spectrometry, further to elucidate the fine structure of the complex polysaccharide. The results revealed that the backbone of CFP6 is β-(1→3)-linked arabinan and sulfate group is located at the C4 or C2 positions. CFP6 is a higher purity arabinose and its extensive structure information can enrich the marine carbohydrate data and provides a methodological reference for the study of complex polysaccharide structures.

Key words: Codium fragile, Polysaccharide, Structural analysis, Oligosaccharide sequence analysis, MS/MS spectrometry

CLC Number:

O629

TrendMD:

JIN Xin,JI Guoli,ZHAO Xiaoliang,LI Guoyun,YU Guangli. Sulfated Arabinose from Codium fragile: Preparation and Extensive Structural Characterization^†[J]. Chem. J. Chinese Universities, 2019, 40(4): 733.

Figures/Tables 5

Fig.1 NMR spectra of polysaccharide CFP6 obtained from C. fragile in D2O at 25 ℃ (A) 1H-1H COSY spectrum; (B) 1H-13C HMQC spectrum; (C) 1H-13C HMBC spectrum.

Table 1 Chemical shifts(δ) for the resonances of glycosyl residues of CFP6 in 1H and 13C NMR spectra

Residue	δ
Residue	H1/C1	H2/C2	H3/C3	H4/C4	H5/C5	H6/C6
Ara(A)	5.15/95.05	3.77/69.09	4.01/68.88	3.61/70.63	4.08/63.66	——
Gal(B)	4.84/91.97	4.24/70.63	4.13/69.15	3.96/71.14	3.75/69.00	4.14/61.53
Ara2S(C)	5.36/96.53	4.71/73.27	4.29/70.14	4.08/68.88	3.91/69.09	——
Ara4S(D)	5.36/96.53	4.59/72.79	4.25/70.44	4.85/74.85	3.79/72.33	——

Fig.2 HILIC-FTMS profiling of CFP6 oligosaccharides generated by free radical depolymerization (A)Total ion chromatography of CFP6 oligosaccharides; (B) semiquantitative analysis of CFP6 oligosaccharides. A: Ara, S: sulfation, the following number represents the number of the previous residues.

Fig.3 ESI-MS/MS spectra of CFP6 oligosaccharides and reduced oligosaccharides A2S1(A), A2S2(B), A3S1(C), A3S2(D), A4S2(E) and A5S2(F)(free acid form of oligosaccharides)

Fig.4 ESI-MS/MS spectra of CFP6 oligosaccharides and reduced oligosaccharides A2S1(A), A2S2(B), A3S1(C), A3S2(D), A4S2(E) and A5S2(F)(alditol form of reduced oligosaccharides)

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