A Study of Chemical Structure of Cell Wall Polysaccharides from Two Fibrous Diets and Their Degradation Rule in Digestive Tract of Pigs Using Nuclear Magnetic Resonance Method

YANG Yamin; ZHUANG Su; ZHANG Zhijing; ZHANG Ziheng; LIU Qiang

doi:10.3969/j.issn.1006-267x.2020.07.044

2020 , Vol. 32 >Issue 7: 3343 - 3357

DOI: https://doi.org/10.3969/j.issn.1006-267x.2020.07.044

A Study of Chemical Structure of Cell Wall Polysaccharides from Two Fibrous Diets and Their Degradation Rule in Digestive Tract of Pigs Using Nuclear Magnetic Resonance Method

YANG Yamin ,
ZHUANG Su ,
ZHANG Zhijing ,
ZHANG Ziheng ,
LIU Qiang

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College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China

Received date: 2020-01-15

Online published: 2020-07-15

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Abstract

The objective of this study was to evaluate the feasibility of the application of 2 dimensional heteronuclear single quantum coherence nuclear magnetic resonance (2D HSQC NMR) technique in analyzing the cell wall polysaccharides from trial pig diet (using wheat bran or broccoli leaf and stem meal as single fiber source), terminal chyme of ileum and fecal samples both qualitatively and quantitatively, and to verify its potential in analyzing chemical structure and quantitative analysis and to provide new experimental techniques for cell wall related feed nutrition research. After removing lipid, starch and protein, the cell wall polysaccharide extracts of the diet, ileal digesta and fecal samples were ball milled and dissolved in dimethyl sulfoxide (DMSO)-d₆/pyridine-d₅ (4:1, V/V) to form gel mixture. The HSQC spectra of the gel samples were collected on a 600 MHz nuclear magnetic resonance (NMR) spectrometer equipped with a cryogenically cooled 5-mm TXI gradient probe. The assignment and volume integration of the signals in the anomeric zoon of polysaccharides were performed by using Bruker TopSpin 3.6 with reference to the published data of chemical shift of polysaccharides. The glycosides content and composition of the cell wall polysaccharide extracts were analyzed using gas chromatography (GC) method simultaneously. The results showed that the most of signals in anomeric zoon of spectra could be assigned to corresponding glycoside in polysaccharides referring to the existing references. The arabinfuranoside (Araf) in the cell wall polysaccharide of wheat bran-based diet (WB) were complicated and most of them existed as the form of side chains of polysaccharides. That of broccoli leaf and stem meal-based diet (BR) was relatively simpler and formed arabinan mainly. The relative integral strength of xylopyranoside (Xylp) without side chain ->4)-β-D-Xylp-(1-> in the cell wall polysaccharide of WB was higher significantly than that of BR (P<0.05). The relative integral strengths of galacturonopyranoside (GalpA) and uronic acids glycoside (UA) in the cell wall polysaccharide of BR were significantly higher than those of WB (P<0.05). The relative integral strength of glucopyranosides (Glcp) was higher and mainly ->4)-β-D-Glcp-(1-> in the cell wall polysaccharide of WB and BR, but in BR cell wall polysaccharides, the relative integral strength of ->3)-β-D-Glcp-(1-> was significantly higher than that of WB (P<0.05). The cell wall polysaccharide of WB was mainly degraded in the large intestine of pigs, while that of BR was obviously degraded before entering the terminal ileum. The degrading rates of Araf and Galp in the cell wall polysaccharide of BR was faster than those of WB (P<0.05), but the ->4)-β-D-Xylp-(1-> in the cell wall polysaccharide of WB was more easily degraded in large intestine of pigs than that of WB (P<0.05). The Glcp in the two diets were the same difficult to degrade. It also demonstrated that Araf presented as side chain and the main chain of xylan and galacturonan with side chains were harder to be degraded in pig digestive tract. The relative integral strength of glycosides measured by the NMR method shared the same trend from diets, chyme to fecal samples with their molar percentages determined by GC method. There were significant regressive relationships between above two methods in glycosides with high content (P<0.05). It can be concluded that the 2D HSQC NMR technique illustrated an obvious advantage not only in resolving the detail structures of cell wall polysaccharides but also possess high potential in quantitative analysis. The 2D HSQC NMR method using feed nutrition research still need to be further improved.

Key words： 2D HSQC NMR; cell-wall polysaccharides; pig; wheat bran; broccoli leaf and stem meal

Cite this article

YANG Yamin , ZHUANG Su , ZHANG Zhijing , ZHANG Ziheng , LIU Qiang . A Study of Chemical Structure of Cell Wall Polysaccharides from Two Fibrous Diets and Their Degradation Rule in Digestive Tract of Pigs Using Nuclear Magnetic Resonance Method[J]. Chinese Journal of Animal Nutrition, 2020 , 32(7) : 3343 -3357 . DOI: 10.3969/j.issn.1006-267x.2020.07.044

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