甜玉米芯多糖铬配合物的制备、结构表征及体外活性的研究

修伟业; 王鑫; 李雨蒙; 遇世友; 黎晨晨; 罗钰; 周卓; 马永强

doi:10.13386/j.issn1002-0306.2022080338

甜玉米芯多糖铬配合物的制备、结构表征及体外活性的研究

doi: 10.13386/j.issn1002-0306.2022080338

黑龙江省谷物食品与谷物资源重点实验室，哈尔滨商业大学食品工程学院，黑龙江哈尔滨 150028

基金项目: 黑龙江省应用技术研究与开发计划（GA20B301）；哈尔滨商业大学“青年创新人才”支持计划（2019CX31）

详细信息

作者简介:
修伟业（1996−），男，博士研究生，研究方向：农产品加工及精深利用，E-mail：xiuweiye0451@163.com

通讯作者:
王鑫（1984−），女，博士，高级工程师，研究方向：农产品精深加工及利用，E-mail：wangxinfood@163.com

中图分类号: TS201.1
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出版历程
- 收稿日期: 2022-08-31
- 网络出版日期: 2023-05-21
- 刊出日期: 2023-07-01

Preparation, Structural Characterization and in Vitro Activity of Chromium Complexes of Sweet Corn Cob Polysaccharide

Key Laboratory of Cereal Food and Cereal Resources in Heilongjiang Province, School of Food Engineering, Harbin University of Commerce, Harbin 150028, China

摘要

摘要: 为提升甜玉米芯多糖生物学活性，制备一种甜玉米芯多糖铬配合物（SCP80-Cr），以单因素实验为基础，以响应面试验优化制备工艺，并进行表征及探究体外活性。结果表明，SCP80-Cr制备最佳工艺条件为pH10.1、反应温度74 ℃、多糖浓度3 mg/mL，反应时间60 min。SCP80-Cr中Cr元素重量百分比为21.08%，元素百分比为6.67%。对SCP80-Cr进行表征发现，SCP80-Cr几乎不含有蛋白、核酸、色素类物质，而Cr是以Cr-O及Cr-OH的形式与SCP80缔合，并具有较强的分子团聚特性。单糖组成测定说明SCP80-Cr单糖组成摩尔百分比为甘露糖:葡萄糖:半乳糖:木糖:阿拉伯糖=0.58:89.50:4.65:1.33:3.58。体外降糖实验结果表明，SCP80-Cr对α-淀粉酶及α-葡萄糖苷酶抑制率的IC₅₀分别为4.70±0.38 mg/mL和3.99±0.26 mg/mL；体外抗氧化实验结果表明，SCP80-Cr清除DPPH自由基及羟基自由基IC₅₀分别为4.80±0.34 mg/mL和4.99±0.28 mg/mL，说明SCP80-Cr相比于SCP80具有更好的体外降糖及抗氧化活性。研究结果为甜玉米芯多糖在功能性食品中的研究提供一定参考价值。
- 甜玉米芯多糖铬配合物 /
- 表征 /
- 体外降糖 /
- 体外抗氧化
Abstract: To enhance the biological activity of sweet corn cob polysaccharide, a sweet corn cob polysaccharide chromium complex (SCP80-Cr) was prepared, characterized and its in vitro activity was also explored. The results showed that the optimal process conditions for the preparation were pH10.1, reaction temperature 74 ℃, polysaccharide concentration of 3 mg/mL, and the reaction time of 60 min. The weight percentage of Cr(III) in SCP80-Cr was 21.08% and the elemental percentage was 6.67%. The characterization revealed that SCP80-Cr contained almost no protein, nucleic acid or pigment, while Cr was conjugated with SCP80 in the form of Cr-O and Cr-OH and had strong molecular agglomeration properties. The molar percentage of the monosaccharide composition of SCP80-Cr was mannose: glucose: galactose: xylose: arabinose = 0.58: 89.50: 4.65: 1.33: 3.58. The results of in vitro hypoglycemia experiments showed that the IC₅₀ of SCP80-Cr on α-amylase and α-glucosidase inhibitions were 4.70±0.38 mg/mL and 3.99±0.26 mg/mL, respectively. Moreover, the results of in vitro antioxidant experiments showed that SCP80-Cr scavenged DPPH radicals and hydroxyl radicals with IC₅₀ of 4.80±0.34 mg/mL and 4.99±0.28 mg/mL, respectively, indicating that SCP80-Cr had better in vitro hypoglycemic and antioxidant activities compared with SCP80. The results of the study provided some references for the study of sweet corn cob polysaccharides in functional foods.
- sweet corn cob polysaccharide-chromium complex /
- characterization /
- in vitro hypoglycemia activity /
- in vitro antioxidant activity

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图 1 单因素实验结果

Figure 1. Results of single-factor test

注：图中不同小写字母表示组间具有显著性差异（P<0.05）。

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图 2 两因素交互作用对铁离子还原力影响的等高线与曲面图

Figure 2. Contour and surface diagram of the influence of the interaction of two factors on the reduction force of iron ions

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3 扫描电镜图像及EDS元素映射图像

3. SEM image and corresponding EDS elemental mapping image

注：A~C：SCP80 SEM图像（A：×10 k，B：×20 k，C：×40 k）；D~F：SCP80-Cr SEM图像（A：×10 k，B：×20 k，C：×40 k）；G：SCP80-Cr EDS图像；H：SCP80-Cr 元素mapping分布图像。

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图 4 SCP80-Cr紫外（A）及红外（B）光谱

Figure 4. Ultraviolet spectrum (A) and Infrared spectrum (B) of SCP80-Cr

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图 5 SCP80-Cr X射线光电子能谱图

Figure 5. XPS of SCP80-Cr

注：A~C：SCP80 X射线光电子能谱（A：全谱图；B：C 1s；C：O 1s）；D~G：SCP80-Cr X射线光电子能谱（D：全谱图；E：C 1s；F：O 1s；G：Cr 2p）。

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图 6 SCP80-Cr的热稳定性分析

Figure 6. Thermal stability analysis of SCP80-Cr

注：A：TG；B：DTG。

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图 7 SCP80-Cr的2D（A）及3D（B）AFM图像

Figure 7. AFM image of SCP80-Cr (A：2D；B：3D)

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图 8 SCP80-Cr单糖组成分析

Figure 8. Monosaccharide composition of SCP80-Cr

注：A：14种单糖标准品液相色谱曲线；B：SCP80-Cr单糖组成液相色谱曲线。其中1：L-古洛糖醛酸；2：D-甘露糖醛酸；3：D-甘露糖；4：D-核糖；5：L-鼠李糖；6：D-氨基葡萄糖；7：D-葡萄糖醛酸；8：D-半乳糖醛酸；9：D-葡萄糖；10：D-氨基半乳糖；11：D-半乳糖；12：D-木糖；13：L-阿拉伯糖；14：L-岩藻糖。

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图 9 SCP80-Cr体外活性分析

Figure 9. Analysis of the in vitro activity of SCP80-Cr

注：A：α-淀粉酶抑制率；B：α-葡萄糖苷酶抑制率；C：DPPH自由基清除率；D：羟基自由基清除率。

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表 1 星点设计因素与水平

Table 1. Factors and levels used for the central composite design

水平	因素
水平	A pH	B温度（℃）	C多糖浓度（mg/mL）
−1.682	9.00	70.00	2.00
−1	9.41	72.03	2.41
0	10.00	75.00	3.00
1	10.59	77.97	3.59
1.682	11.00	80.00	4.00

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表 2 液相梯度洗脱条件

Table 2. HPLC gradient elution conditions

时间（min）	流动相（%）
时间（min）	A	B
0	86	14
9	83	17
28	78	22
29	50	50
31	50	50

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表 3 星点设计-响应面法试验设计及结果

Table 3. Experimental design and results of central design response surface method (CCD)

试验号	因素			Y 铁离子还原力（A）
试验号	A pH	B 温度	C 多糖浓度	Y 铁离子还原力（A）
1	0	0	−1.682	0.301
2	−1	−1	1	0.173
3	1	−1	−1	0.189
4	−1	−1	−1	0.306
5	1	−1	1	0.404
6	0	−1.682	0	0.417
7	0	0	0	0.539
8	0	1	1	0.310
9	0	0	0	0.513
10	0	1.682	0	0.291
11	0	0	1.682	0.276
12	0	0	0	0.396
13	0	0	0	0.491
14	−1	1	−1	0.136
15	1.682	0	0	0.252
16	−1.682	0	0	0.107
17	0	0	0	0.497
18	−1	1	1	0.101
19	1	1	−1	0.231
20	0	0	0	0.521

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表 4 回归方程的方差分析结果

Table 4. Analysis of variance of regression model

差异来源	平方和	自由度	均方	F值	P	显著性
模型	0.37	9	0.041	16.65	<0.0001	**
A	0.032	1	0.032	13.14	0.0046	**
B	0.019	1	0.019	7.68	0.0198	*
C	5.161×10⁻⁴	1	5.161×10⁻³	0.21	0.6555
AB	4.513×10⁻³	1	4.513×10⁻³	1.85	0.2038
AC	0.027	1	0.027	10.93	0.0079	**
BC	1.805×10⁻⁴	1	1.805×10⁻⁴	0.074	0.7912
A²	0.20	1	0.20	80.83	<0.0001	**
B²	0.044	1	0.044	18.06	0.0017	**
C²	0.089	1	0.089	36.35	0.0001	**
残差	0.024	10	2.441×10⁻³
失拟项	0.012	5	2.336×10⁻³	0.92	0.5365	不显著
纯误差	0.013	5	2.546×10⁻³
总和	0.39	19
注：代表影响显著（P<0.05）；*代表影响极显著（P<0.01）。

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参考文献(30)

[1]	BI Z, ZHAO Y, HU J, et al. A novel polysaccharide from Lonicerae japonicae caulis: Characterization and effects on the function of fibroblast-like synoviocytes[J]. Carbohydrate Polymers,2022,292:119674. doi: 10.1016/j.carbpol.2022.119674
[2]	WANG S, LI G, ZHANG X, et al. Structural characterization and antioxidant activity of Polygonatum sibiricum polysaccharides[J]. Carbohydrate Polymers,2022,291:119524. doi: 10.1016/j.carbpol.2022.119524
[3]	SONG J, WU Y, MA X, et al. Structural characterization and α-glycosidase inhibitory activity of a novel polysaccharide fraction from Aconitum coreanum[J]. Carbohydrate Polymers,2020,230(C):115586.
[4]	LIU W, HU C, LIU Y, et al. Preparation, characterization, and α-glycosidase inhibition activity of a carboxymethylated polysaccharide from the residue of Sarcandra glabra (Thunb.) Nakai[J]. International Journal of Biological Macromolecules, 2017, 99: 454−464.
[5]	LU K, CHEN S, LIN Y, et al. An antidiabetic nutraceutical combination of red yeast rice (Monascus purpureus), bitter gourd (Momordica charantia), and chromium alleviates dedifferentiation of pancreatic β cells in db/db mice[J]. Food Science & Nutrition, 2020, 8(12): 6718-6726.
[6]	YE H, SHEN Z, CUI J, et al. Hypoglycemic activity and mechanism of the sulfated rhamnose polysaccharides chromium(III) complex in type 2 diabetic mice[J]. Bioorganic Chemistry, 2019, 88: 102942.
[7]	WANG X, XIU W, HAN Y, et al. Structural characterization of a novel polysaccharide from sweet corncob that inhibits glycosylase in STZ-induced diabetic rats: Structural characterization of a novel polysaccharide[J]. Glycoconjugate Journal,2022,39(3):413−427.
[8]	WANG X, WANG Z, ZHANG K, et al. Effects of sweet corncob polysaccharide on pancreatic protein expression in type 2 diabetic rats[J]. Journal of Functional Foods, 2022, 88.
[9]	MA Y, WANG X, GAO S. Hypoglycemic activity of polysaccharides from sweet corncob on streptozotocin-induced diabetic rats.[J]. Journal of Food Science,2017,82(1):208−213. doi: 10.1111/1750-3841.13554
[10]	WANG X, XIU W, HAN Y. et al. Structural characterization of a non-starch polysaccharide from sweet corn cobs[J]. Journal of Food Processing and Preservation, 2022, 46(9).
[11]	秦月. 虎眼万年青多糖及其铬复合物降血糖作用的研究[D]. 哈尔滨: 黑龙江中医药大学, 2018: 30−32 QIN Y. Hypoglycemic effect of Ornithogalum caudatum polysaccharide and its chromium complex[D]. Harbin: Heilongjiang University of Chinese Medicine, 2018: 30−32.
[12]	公衍玲, 郭遥遥, 刘洋. 樱桃核乙醇提取物体外抗氧化及降糖降脂研究[J]. 青岛科技大学学报(自然科学版),2017,38(6):14−17. [GONG Y L, GUO Y Y, LIU Y. Study on in vitro antioxidant, hypoglycemic and lipid-lowering activities of cherry kernel ethanol extracts[J]. Journal of Qingdao University of Science and Technology(Natural Science Edition),2017,38(6):14−17. GONG Y, GUO Y, LIU Y. Study on in vitro antioxidant, hypoglycemic and lipid-lowering activities of cherry kernel ethanol extracts[J]. Journal of Qingdao University of Science and Technology(Natural Science Edition), 2017, 38(6): 14-17.
[13]	陈玥彤, 张闪闪, 李文意, 等. 黑木耳多糖的磷酸化修饰、结构表征及体外降糖活性[J]. 食品科学,2022,43(8):29−35. [CHEN Y T, ZHANG S S, LI W Y, et al. Structural characterization and hypoglycemic effect in vitro of phosphorylated Auricularia auriculata polysaccharide[J]. Food Science,2022,43(8):29−35. CHEN Y, ZHANG S, LI W, et al. Structural characterization and hypoglycemic effect in vitro of phosphorylated Auricularia auriculata polysaccharide[J] Food Science, 2022, 43(8): 29-35.
[14]	李春英, 夏依拉·艾尼, 毛建卫, 等. 栀子黄对α-葡萄糖苷酶抑制活性的研究[J]. 新宝登录入口(中国)有限公司,2015,36(24):112−114, 119. [LI C Y, XIAYILA A, MAO J W, et al. Study on the inhibitory effect of gardenia yellow on α-glucosidase activity[J]. Science and Technology of Food Industry,2015,36(24):112−114, 119. LI C, XIAYILA A, MAO, J, et al. Study on the inhibitory effect of gardenia yellow on α-glucosidase activity[J] Science and Technology of Food Industry, 2015, 36(24): 112-114, 119.
[15]	ZUO M, LIU X, LIU D, et al. Extraction, Characterization and antioxidant activity in vitro of proteins from Semen Allii Fistulosi[J]. Molecules,2018,23(12):3235. doi: 10.3390/molecules23123235
[16]	ZHANG Y, CHEN Z, HUANG Z, et al. A comparative study on the structures of Grifola Frondosa polysaccharides obtained by different decolourization methods and their in vitro antioxidant activities[J]. Food & Function,2019,10(10):.6720−6731.
[17]	LI L, XU J, CAO Y, et al. Preparation of Ganoderma lucidum polysaccharide-chromium (III) complex and its hypoglycemic and hypolipidemic activities in high-fat and high-fructose diet-induced prediabetic mice[J]. International Journal of Biological Macromolecules,2019,140(C):782−793.
[18]	QIU J, ZHANG H, WANG Z, et al. Response surface methodology for the synthesis of an Auricularia auricula-judae polysaccharides CDDP complex[J]. International Journal of Biological Macromolecules,2016,93:333−343. doi: 10.1016/j.ijbiomac.2016.06.066
[19]	GUO W, SHI F, LI L, et al. Preparation of a novel Grifola frondosa polysaccharide-chromium (III) complex and its hypoglycemic and hypolipidemic activities in high fat diet and streptozotocin-induced diabetic mice[J]. International Journal of Biological Macromolecules,2019,131:81−88. doi: 10.1016/j.ijbiomac.2019.03.042
[20]	张雯, 张禧庆, 董淑君, 等. 南瓜皮多糖铬的制备与体外消化的分析[J]. 新宝登录入口(中国)有限公司,2022,43(17):223−229. [ZHANG W, ZHANG X Q, DONG S J, et al. Preparation and in vitro digestibility of pumpkin peel polysaccharide chromium(III) complex[J]. Science and Technology of Food Industry,2022,43(17):223−229. doi: 10.13386/j.issn1002-0306.2021110329 ZHANG W, ZHANG X, DONG S, et al. Preparation and in vitro digestibility of pumpkin peel polysaccharide chromium(III) complex[J]. Science and Technology of Food Industry, 2022, 43(17): 223-229. doi: 10.13386/j.issn1002-0306.2021110329
[21]	邰克东, 赵苏茂, 杨紫恒, 等. 高压均质对脂质体囊泡特性和稳定性的影响[J]. 食品科学,2019,40(17):169−177. [TAI K D, ZHAO S M, YANG Z H. et al. Effect of high-pressure homogenization on vesicle characteristics and stability of liposomes[J]. Food Science,2019,40(17):169−177. doi: 10.7506/spkx1002-6630-20180916-158 TAI K, ZHAO S, YANG Z. et al. Effect of high-pressure homogenization on vesicle characteristics and stability of liposomes[J]. Food Science, 2019, 40(17): 169-177. doi: 10.7506/spkx1002-6630-20180916-158
[22]	郑成娟, 李珊珊, 陈青. 金樱子果渣中脂肪酸与玉米淀粉包合物的制备及性能[J]. 精细化工,2020,37(12):2482−2489. [ZHENG C J, LI S S, CHEN Q. Preparation and properties of inclusion compound of corn starch and fatty acid in Rasa laevigata pomace[J]. Fine Chemicals,2020,37(12):2482−2489. doi: 10.13550/j.jxhg.20200656 ZHENG C, LI S, CHEN Q. Preparation and properties of inclusion compound of corn starch and fatty acid in Rasa laevigata pomace[J]. Fine Chemicals, 2020, 37(12): 2482-2489. doi: 10.13550/j.jxhg.20200656
[23]	张秀娟, 刘治廷, 杨诗涵, 等. 超声波辅助酶法提取蓝莓果渣花色苷的工艺优化及降解动力学[J]. 精细化工,2022,39(10):2069−2077, 2098. [ZHANG X J, LIU Z T, YANG S H, et al. Condition optimization and degradation kinetics of anthocyanins from blueberry pomace by ultrasonic-assisted enzymatic extraction[J]. Fine Chemicals,2022,39(10):2069−2077, 2098. ZHANG X, LIU Z, YANG S, et al. Condition optimization and degradation kinetics of anthocyanins from blueberry pomace by ultrasonic-assisted enzymatic extraction[J], Fine Chemicals, 2022, 39(10): 2069-2077, 2098.
[24]	ZHANG W, LI L, MA Y, et al. Structural characterization and hypoglycemic activity of a novel pumpkin peel polysaccharide-chromium(III) complex[J]. Foods, 2022, 11(13): 1821-1821.
[25]	全志刚, 王维浩, 赵姝婷, 等. 小米麸皮水溶性膳食纤维-Cr(III)配合物的合成、表征及其体外抗氧化活性[J]. 食品科学,2021,42(21):46−55. [QUAN Z G, WANG W H, ZHAO S T, et al. Synthesis, characterization and antioxidant activity of millet bran soluble dietary fiber-Cr(Ⅲ) complexes[J]. Food Science,2021,42(21):46−55. doi: 10.7506/spkx1002-6630-20200902-023 QUAN Z, WANG W, ZHAO S, et al. Synthesis, characterization and antioxidant activity of millet bran soluble dietary fiber-Cr(Ⅲ) complexes[J]. Food Science, 2021, 42(21): 46-55. doi: 10.7506/spkx1002-6630-20200902-023
[26]	TAO E, CHENG Y, YANG S, et al. Recovering Cr(III) from chromium-containing waste: An in-depth study on mechanism via retaining organic matters[J]. Journal of Environmental Chemical Engineering, 2021, 9(5).
[27]	刘思美, 赵鹏, 张婷婷, 等. 地黄硒多糖的合成、表征及免疫活性分析[J]. 中国中药杂志,2022,47(11):2938−2946. [LIU S M, ZHAO P, ZHANG T T, et al. Synthesis, characterization, and immunological activity of Rehmannia glutinosa seleno-polysaccharides[J]. China Journal of Chinese Materia Medica,2022,47(11):2938−2946. doi: 10.19540/j.cnki.cjcmm.20220224.304 LIU S, ZHAO P, ZHANG T, et al. Synthesis, characterization, and immunological activity of Rehmannia glutinosa seleno-polysaccharides[J]. China Journal of Chinese Materia Medica, 2022, 47(11): 2938-2946. doi: 10.19540/j.cnki.cjcmm.20220224.304
[28]	汤陈鹏, 吕峰, 王蓉琳. 孔石莼多糖锌结构表征与体外降血糖活性[J]. 食品科学,2020,41(7):52−58. [TANG C P, LÜ F, WANG R L. Structural characterization and hypoglycemic activity in vitro of Ulva pertusa polysaccharides-zinc complex[J]. Food Science,2020,41(7):52−58. doi: 10.7506/spkx1002-6630-20190320-263 TANG C, LV F, WANG R. Structural characterization and hypoglycemic activity in vitro of Ulva pertusa polysaccharides-zinc complex[J]. Food Science, 2020, 41(7): 52-58. doi: 10.7506/spkx1002-6630-20190320-263
[29]	WANG C, GAO X, SANTHANAM R, et al. Effects of polysaccharides from Inonotus obliquus and its chromium (III) complex on advanced glycation end-products formation, α-amylase, α-glucosidase activity and H₂O₂ -induced oxidative damage in hepatic L02 cells[J]. Food and Chemical Toxicology, 2018, 116(Pt B): 335−345.
[30]	ZHANG J, CHEN C, FU X. et al. Fructus mori L. polysaccharide-iron chelates formed by self-embedding with iron(iii) as the core exhibit good antioxidant activity[J]. Food & Function,2019,10(6):3150−3160.