野皂荚多糖氧化降解产物的体外代谢过程研究

doi:10.3969/j.issn.0253-2417.2020.03.013

摘要/Abstract

摘要：

采用不同体积分数（0.3%、0.6%、1.5%、3.0%和4.5%）H₂O₂溶液对野皂荚半乳甘露聚糖（GMG）进行氧化降解预处理，得到的产物（GMG-0.3、GMG-0.6、GMG-1.5，GMG-3.0和GMG-4.5）用于模拟体外远端盲肠代谢实验，探究SD大鼠肠道菌群对GMG的体外代谢情况。结果表明：经过不同体积分数H₂O₂预处理后，GMG相对分子质量差异明显，由未处理（GMG）的1 039 400降至4.5% H₂O₂处理（GMG-4.5）的32 060；红外光谱分析发现H₂O₂预处理并没有改变GMG的结构。测定了代谢0、6、12、24和48 h时代谢产物的pH值、总糖消耗率、还原糖以及短链脂肪酸（SCFAs）和支链脂肪酸（BCFAs）浓度，综合评价了肠道微生物菌群在48 h内对半乳甘露聚糖的利用情况，结果发现：预处理GMG实验组代谢情况显著优于空白对照组和未经过预处理GMG实验组，GMG-0.6代谢情况最佳，代谢产生乙酸浓度较高为25.36 mmoL/L，总SCFAs浓度为45.05 mmol/L，总糖消耗率最大为（52.86±3.11）%，还原糖质量浓度先上升后下降至（0.002±0.003）g/L。研究结果表明氧化降解野皂荚半乳甘露聚糖可作为降低血糖、调控胰岛素分泌、改善肠道健康的一种功能性食品添加剂。

关键词: 野皂荚半乳甘露聚糖, 过氧化氢, 体外代谢, 相对分子质量, 短链脂肪酸

Abstract:

The 0.3%, 0.6%, 1.5%, 3.0% and 4.5% H₂O₂ solutions were used to pretreat the Gleditsia microphylla galactomannan (GMG). The obtained products (GMG-0.3, GMG-0.6, GMG-1.5, GMG-3.0 and GMG-4.5) were used to simulate the metabolism of distal cecum and investigate the in vitro metabolism of GMG by intestinal microflora of SD rats. The results showed that after pretreatment with different mass fractions of H₂O₂, the relative molecular weights of GMG were significantly different. The molecular weight decreased from 1 039 400 to 32 060 after treatment with 4.5% H₂O₂; and the infrared spectroscopy analysis found that the pretreatment with H₂O₂ did not change the structure of GMG. By analyzing the pH value, total sugar consumption rate, reducing sugar, and short-chain fatty acid (SCFAs) and branched-chain fatty acid (BCFAs) concentrations in the metabolism broth during 48 hours, the utilization of galactomannan by the intestinal microflora was calculated. The results showed that the metabolism of the pretreated GMG experimental groups was significantly better than that of the blank control group and the non-pretreated GMG experimental group, and the metabolism of GMG-0.6 experimental group was the best. The higher concentration of acetic acid produced by metabolism was 25.36 mmoL/L, the total SCFAs concentration was 45.05 mmol/L, the consumption rate of total sugar was (52.86±3.11)%, and the mass concentration of reducing sugar first increased and then decreased to (0.002±0.003) g/L. The experimental results showed that oxidative degradation of GMG could be used as a functional food additive to decrease blood glucose, regulate insulin secretion and improve intestinal health.

Key words: Gleditsia microphylla galactomannan (GMG), hydrogen peroxide, in vitro metabolism, molecular weight, short-chain fatty acid

中图分类号:

TQ35

王鑫纯,徐伟,蒋建新,朱莉伟. 野皂荚多糖氧化降解产物的体外代谢过程研究[J]. 林产化学与工业, 2020, 40(3): 99-107.

Xinchun WANG,Wei XU,Jianxin JIANG,Liwei ZHU. In vitro Metabolism Process of Oxidative Degradation Products from Gleditsia microphylla Galactomannan[J]. Chemistry and Industry of Forest Products, 2020, 40(3): 99-107.

图/表 6

图1

图2

表1

GMG代谢不同时间代谢产物中SCFAs及BCFA含量变化1)"

产物 products	时间/h time	不同多糖液的代谢产物浓度metabolites concentration of different polysaccharides solution/(mmol·L^-1)
产物 products	时间/h time	空白 control	GMG	GMG-0.3	GMG-0.6	GMG-1.5	GMG-3.0	GMG-4.5
乙酸 acetic acid	0	0^a	0^a	0^a	0^a	0^a	0^a	0^a
	6	0^a	4.92±0.18^b	0^a	0^a	0^a	0^a	0^a
	12	0^a	10.50±0.05^c	2.31±0.07^b	3.61±0.06^b	3.42±0.04^b	3.84±0.02^b	4.01±0.10^b
	24	2.71±0.12^b	12.17±0.24^c	17.27±0.18^c	17.06±0.09^c	17.35±0.17^c	16.69±0.15^c	12.26±0.16^c
	48	2.78±0.05^b	15.89±0.20^c	21.72±0.09^c	25.36±0.20^c	21.07±0.01^c	21.36±0.02^c	15.21±0.16^c
丙酸 propionic acid	0	1.93±0.01^a	1.82±0.01^a	1.89±0.01^a	1.91±0.02^a	1.92±0.01^a	1.91±0.02^a	1.80±0.02^a
	6	1.71±0.05^b	0.88±0.09^b	2.16±0.00^b	2.13±0.05^b	2.06±0.13^b	1.96±0.02^b	2.12±0.05^b
	12	1.92±0.01^b	1.08±0.01^b	2.38±0.03^b	2.31±0.04^b	2.32±0.00^b	2.93±0.03^c	2.32±0.01^b
	24	1.61±0.03^c	2.53±0.02^c	6.34±0.03^c	5.91±0.05^c	6.90±0.09^c	5.94±0.11^d	5.16±0.13^c
	48	1.11±0.02^d	4.05±0.04^d	10.25±0.01^c	10.07±0.15^d	9.73±0.03^c	9.51±0.03^e	6.82±0.10^c
正丁酸 n-butyric acid	0	0^a	0^a	0^a	0^a	0^a	0^a	0^a
	6	0^a	0^a	5.91±0.01^b	6.85±0.09^b	6.88±0.14^b	6.17±0.05^b	7.80±0.01^b
	12	0^a	0.96±0.00^b	6.53±0.04^c	6.49±0.02^b	6.74±0.03^b	6.03±0.03^b	7.59±0.05^b
	24	0^a	2.96±0.00^c	6.83±0.03^c	6.07±0.02^c	6.49±0.05^c	6.13±0.01^b	7.24±0.06^c
	48	0.56±0.01^b	3.96±0.01^c	8.57±0.09^d	9.62±0.16^d	11.09±0.09^d	9.43±0.05^c	7.85±0.02^d
总SCFAs total SCFAs	0	1.93±0.02^a	0^a	1.89±0.02^a	1.91±0.03^a	1.92±0.03^a	1.91±0.03^a	1.80±0.03^a
	6	1.71±0.07^b	5.79±0.33^b	8.07±0.01^b	8.98±0.04^b	8.94±0.01^b	8.14±0.09^b	9.92±0.08^b
	12	1.92±0.01^b	12.54±0.07^c	11.21±0.09^b	12.41±0.01^b	12.48±0.10^b	12.79±0.05^b	13.92±0.23^b
	24	4.33±0.21^c	14.16±0.23^c	30.44±0.16^c	29.05±0.24^c	30.74±0.17^c	28.75±0.08^c	24.67±0.04^c
	48	4.45±0.03^c	13.84±0.25^c	40.54±0.23^c	45.05±0.31^c	41.89±0.06^c	40.29±0.16^c	29.88±0.06^c
BCFAs (异丁酸 isobutyric acid)	0	0^a	0^a	0^a	0^a	0^a	0^a	0^a
	6	0^a	1.56±0.02^b	1.34±0.02^b	1.36±0.03^b	1.34±0.04^b	1.32±0.01^b	1.37±0.03^b
	12	0^a	1.52±0.01^b	1.49±0.01^b	1.47±0.01^b	1.42±0.02^b	1.48±0.02^b	1.34±0.01^b
	24	0^a	1.58±0.03^b	0.57±0.02^c	0.58±0.03^c	0.54±0.01^c	0.58±0.03^c	0.84±0.00^c
	48	0^a	1.76±0.02^c	0^c	0^c	0^c	0^c	1.38±0.02^d

表1

表2

表3

图3

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6	6.99±1.36^a	5.92±0.95^a	6.63±0.99^a	6.19±0.64^a	6.70±2.36^a	6.82±1.08^a
12	8.06±1.65^b	7.84±0.53^b	12.16±0.75^b	7.89±0.75^b	9.92±0.96^b	9.83±0.18^b
24	23.90±1.13^c	33.04±0.06^c	38.05±2.13^c	35.58±0.23^c	32.53±0.19^c	34.80±0.03^c
48	37.27±0.08^c	46.87±0.10^c	52.86±3.11^d	45.08±0.57^c	47.72±0.41^c	48.85±1.32^c

时间/h time	GMG	GMG-0.3	GMG-0.6	GMG-1.5	GMG-3.0	GMG-4.5
6	0.014±0.001^a	0.015±0.003^a	0.015±0.003^a	0.014±0.006^a	0.014±0.007^a	0.014±0.002^a
12	0.007±0.007^b	0.022±0.010^b	0.026±0.000^b	0.023±0.025^b	0.026±0.003^b	0.021±0.002^b
24	0.003±0.001^c	0.002±0.002^c	0.003±0.001^c	0.003±0.001^c	0.001±0.001^c	0.007±0.004^c
48	0.002±0.000^c	0.001±0.003^c	0.002±0.003^c	0.002±0.003^c	0.000±0.000^c	0.005±0.000^c

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