碱提甘蔗皮多糖提取工艺、初步结构及其对α-葡萄糖苷酶的抑制作用

doi:10.3864/j.issn.0578-1752.2021.12.015

Abstract

Abstract:

【Objective】 The objectives of this research were to establish the optimized technology, further explore the structural characteristics and evaluate α-glucosidase inhibitory effect of polysaccharide with alkaline-extracted from sugarcane peel.【Method】Single factor experiment and response surface methodology (RSM) were used to optimize the extraction process of alkali-extracted sugarcane peel polysaccharide (SPAP). The method of phenol sulfuric acid was used to determine the content of polysaccharide. After deproteinization by Sevag method and depigmentation by AB-8, the structure of SPAP was characterized. The molecular weight distribution, monosaccharide compositions and characteristic functional groups of SPAP were detected by HPLC, GC-MS and FT-IR, respectively. Finally, the method of 4-nitrophenol-2-β-D-glucopyranoside (PNPG) was used to determine the α-glucosidase inhibitory effect of polysaccharide extract. 【Result】 The optimum extraction conditions were as follows: extraction temperature 37℃, sodium hydroxide concentration 5%, ratio of material to solvent 1﹕46 (g·mL^-1), and extraction times 4 times. Under these conditions, the yield of SPAP reached 10.84%. After removing protein and pigment, the content of SPAP reached 86.54%. Furthermore, SPAP was mainly composed of arabinose, xylose, glucose and galactose, with molecular weight of 3.03×10³ kD. It might be a pyran hetero-polysaccharide with α or β configuration. In addition, SPAP showed good inhibitory effect on α-glucosidase. The inhibition rate of SPAP reached 78.31%.【Conclusion】 The process optimization of alkali-extracted of polysaccharides from sugarcane peel could effectively utilize raw materials and improve the yield and efficiency. The structural characteristics of SPAP were preliminarily described by HPLC, FT-IR and GC-MS. At the same time, SPAP showed good inhibitory effect on α-glucosidase and potential hypoglycemic activity. These results provided a theoretical basis for further study on the structure-activity relationship of SPAP.

Key words: polysaccharide with alkaline-extracted from sugarcane peel, response surface analysis, structural characterization, α-glucosidase inhibition

WANG XuanXuan,LIU ChunYu,XIE BeiYu,ZHANG ShuShu,WANG DanYang,ZHU ZhenYuan. Extraction Technology, Preliminary Structure and α-glucosidase Inhibition of Polysaccharide with Alkaline-Extracted from Sugarcane Peel[J].Scientia Agricultura Sinica, 2021, 54(12): 2653-2665.

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References 39

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因素 Factor	水平Level
因素 Factor	-1	0	1
提取温度 Extraction temperature A (℃)	30	35	40
氢氧化钠浓度 NaOH concentration B (%)	5	6	7
料液比 Ratio of material to solvent C (g·mL^-1)	1﹕45	1﹕50	1﹕55
提取次数 D (次)	3	4	5

方差来源 Source of variance	平方和 Sum of square	自由度 Degree of freedom	均方 Mean square	F值 F value	Pr>F	显著性 Significance
模型 Model	1.39	14	0.099	9.50	< 0.0001	**
A	0.10	1	0.10	9.85	0.0073	*
B	0.052	1	0.052	4.99	0.0423	*
C	0.022	1	0.022	2.08	0.1713
D	0.069	1	0.069	6.62	0.0221	*
AB	5.625E-003	1	5.625E-003	0.54	0.4747
AC	2.500E-005	1	2.500E-005	2.398E-003	0.9616
AD	0.027	1	0.027	2.61	0.1284
BC	0.16	1	0.16	14.97	0.0017	*
BD	0.35	1	0.35	33.96	< 0.0001	**
CD	6.250E-004	1	6.250E-004	0.060	0.8101
A²	0.081	1	0.081	7.77	0.0145	*
B²	0.14	1	0.14	13.86	0.0023	*
C²	0.077	1	0.077	7.43	0.0164	*
D²	0.51	1	0.51	49.39	< 0.0001	**
残差 Residual	0.15	14	0.010
失拟项 Lack of fit	0.14	10	0.014	5.15	0.0641	不显著 No significance
纯误差 Pure error	0.011	4	2.630E-003
总和 Sum	1.53	28

Extraction Technology, Preliminary Structure and α-glucosidase Inhibition of Polysaccharide with Alkaline-Extracted from Sugarcane Peel

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	样品 Sample (μL)	阿卡波糖 Acarbose (μL)	α-葡萄糖苷酶 α-glucosidase (μL)	PBS (μL)	PNPG (μL)
样品组 Sample group	40		40		20
样品背景组 Sample background group	40			40	20
阳性对照组 Positive control group		40	40		20
阳性背景组 Positive background group		40		40	20
空白组 Blank group			40	40	40

RUN	A	B	C	D	Y (%)
1	0	0	0	0	10.86
2	0	0	0	0	10.86
3	0	1	1	0	10.72
4	-1	1	0	0	10.43
5	0	0	0	0	10.95
6	-1	-1	0	0	10.62
7	0	0	0	0	10.85
8	-1	0	1	0	10.69
9	0	0	1	1	10.54
10	0	1	-1	0	10.29
11	0	-1	1	0	10.51
12	0	0	-1	-1	10.34
13	0	-1	0	-1	10.09
14	-1	0	-1	0	10.58
15	-1	0	0	1	10.23
16	1	0	0	1	10.75
17	0	1	0	1	10.30
18	0	1	0	-1	10.59
19	-1	0	0	-1	10.34
20	0	1	0	1	10.99
21	1	1	0	0	10.57
22	0	-1	-1	0	10.87
23	1	0	0	-1	10.53
24	0	0	-1	1	10.41
25	1	0	-1	0	10.71
26	0	0	0	0	10.81
27	1	0	1	0	10.83
28	0	0	1	-1	10.42
29	1	-1	0	0	10.61

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