葡萄酒泥酵母制备水溶性β-D-葡聚糖工艺优化及其纯化后抗氧化性分析

doi:10.7506/spkx1002-6630-201614005

摘要/Abstract

摘要：

以诱导自溶后的葡萄酒泥酵母细胞壁为试材，采用蛋白酶水解法，通过响应面分析法确定制备水溶性β-D-葡聚糖的最佳工艺条件，将粗品经Sephacryl S-400 HR凝胶柱层析纯化后，对其抗氧化性进行分析。结果表明，酶解制备水溶性β-D-葡聚糖的最佳工艺条件为浸提温度60 ℃、加酶量247 U/mL、pH 7.5、浸提时间1.5 h，得率为80.91%。凝胶柱层析纯化后得到3 种组分，比例依次为组分Ⅰ 68.08%、组分Ⅱ 13.97%、组分Ⅲ 8.79%，其中组分Ⅰ分子质量为100 065.18 D。纯化后的水溶性β-D-葡聚糖清除ABTS＋·、羟自由基、DPPH自由基、O2 － ·和螯合Fe2＋的EC50值分别为2.035、6.352、16.773、5.238、1.061 mg/mL，具有良好的抗氧化活性。

关键词: 葡萄酒泥酵母, 可溶性&beta, -葡聚糖, 提取纯化, 抗氧化

Abstract:

An enzymatic method was used to prepare water-soluble β-D-glucan from the autolyzed cell wall of waste wine
yeast. Single factor method and response surface methodology were applied to optimize the extraction conditions. The watersoluble
β-D-glucan was purified by Sephacryl S-400 HR gel column chromatography to investigate its antioxidant activities.
The optimum conditions for β-D-glucan extraction were determined as follows: temperature, 60 ℃; enzyme dosage, 247 U/mL;
pH, 7.5; and time, 1.5 h. Under these conditions, the yield of water-soluble β-D-glucan was 80.91%. Three components
designated asⅠ, Ⅱ and Ⅲ were eluted from the chromatographic column, the contents of which were 68.08%, 13.97% and
8.79%, respectively. The relative molecular weight of componentⅠ was 100 065.18 D. Antioxidant activity studies showed
that the purified water-soluble β-D-glucan had good antioxidant activities with EC50 values for scavenging, 2’-azinobis-(3-
ethylbenzthiazoline-6-sulphonate (ABTS+·), hydroxyl radical, 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) radical and
superoxide anion radical and chelating Fe2+ of 2.035, 6.352, 16.773, 5.238 and 1.061 mg/mL, respectively.

Key words: wine yeast, water-soluble β-D-glucan, extraction and purification, antioxidant activities

中图分类号:

TS261.9

杨学山，祝霞，李颍，杨婷，马腾臻，韩舜愈. 葡萄酒泥酵母制备水溶性β-D-葡聚糖工艺优化及其纯化后抗氧化性分析[J]. 食品科学, doi: 10.7506/spkx1002-6630-201614005.

YANG Xueshan, ZHU Xia, LI Ying, YANG Ting, MA Tengzhen, HAN Shunyu. Purification and Antioxidant Activities of Water-Soluble β-D-Glucan from Waste Wine Yeast[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201614005.

参考文献

[1] 李莹, 苏婷婷, 王战勇. 葡萄加工副产品的综合利用研究[J]. 中国农

学通报, 2006, 17(4): 106-108.

[2] 郑鹤龄, 张斌, 潘洁, 等. 葡萄酒泥农业应用效果研究[J]. 天津农业

科学, 2006, 21(3): 47-48.

[3] 王淮, 唐治玉, 熊善柏. 废啤酒酵母中β-1,3-D-葡聚糖的提取及成分

分析[J]. 华中农业大学学报, 2006, 24(6): 626-629. DOI:10.13300/

j.cnki.hnlkxb.2005.06.021.

[4] 王静, 戴军, 陈尚卫, 等. 提取酵母细胞壁中β-D-葡聚糖的新方

法[J]. 食品与发酵工业, 2011, 37(1): 189-193. DOI:10.13995/

j.cnki.11-1802/ts.2011.01.043.

[5] LIMBERGER V M, FRANCISCO A D, CHAN A, et al. Barley

β-glucans extraction and partial characterization[J]. Food Chemistry,

2014, 47(4): 84-89.

[6] LIU X Y, WANG Q, LIU H Z, et al. A new isolation method of

β-D-glucans from spent yeast Saccharomyces cerevisiae[J]. Food

Hydrocolloids, 2008, 22(2): 239-247.

[7] BORCHANIA C, FONTEYNB F, JAMINB G, et al. Enzymatic

process for the fractionation of baker’s yeast cell wall (Saccharomyces

cerevisiae)[J]. Food Chemistry, 2014, 47(1): 108-113.

[8] 董兴叶, 孙楚, 刘瑶, 等. 超声波法对燕麦β-葡聚糖提取及性质

的影响[J]. 食品工业科技, 2014, 35(16): 294-297. DOI:10.13386/

j.issn1002-0306.2014.16.056.

[9] 王英杰, 苏理, 兰文忠, 等. 水溶性β-1,3-葡聚糖制备及应用研究的进

展[J]. 食品与药品, 2014, 23(2): 222-226.

[10] 黄国宏. 酵母β - 1 , 3 -D- 葡聚糖硫酸酯制备工艺的研究[ J ] .

食品工业科技, 2 0 0 9 , 3 0 ( 8 ) : 2 6 1 - 2 6 3 . D O I : 1 0 . 1 3 3 8 6 /

j.issn1002-0306.2009.08.009.

[11] 王庆华, 刘山, 崔炳权, 等. 废弃酵母渣提取β-葡聚糖及其羧甲

基化的研究[J]. 酿酒科技, 2009, 29(8): 120-122. DOI:10.13746/

j.njkj.2009.08.015.

[12] 张学况, 杜丽平, 王超, 等. 酶解法增溶酵母(1→3)-β-D-葡聚糖

的研究[J]. 食品与发酵工业, 2012, 38(7): 48-52. DOI:10.13995/

j.cnki.11-1802/ts.2012.07.012.

[13] 王成龙, 王英珍, 张健, 等. 水溶性酵母葡聚糖的制备及应用研究进

展[J]. 动物医学进展, 2010, 31(12): 128-131.

[14] 余奕珂, 白雪芳, 杜昱光. 可溶性酵母葡聚糖的制备及其活性研究[J].

中国生化药物杂志, 2010, 31(4): 254-257.

[15] 杨婷, 祝霞, 李颍, 等. 葡萄酒泥酵母β-葡聚糖提取工艺条件

优化[J]. 食品工业科技, 2015, 36(18): 286-289. DOI:10.13386/

j.issn1002-0306.2015.18.049.

[16] 许文涛, 张方方, 罗云波, 等. 荞麦水溶性多糖的分离纯化及其分子

质量的测定[J]. 食品科学, 2009, 30(14): 22-24.

[17] 张玉香, 尹卓荣. 甘露糖蛋白的提纯及分子质量测定[J]. 酿酒科技,

2005, 25(2): 72-74. DOI:10.13746/j.njkj.2005.04.027.

[18] 徐韧博. 松塔多糖的分离纯化、结构鉴定及抗氧化活性研究[D]. 哈

尔滨: 哈尔滨工业大学, 2013.

[19] WANG Y, YI L, HU Y. Optimization of polysaccharides extraction

from Trametes robiniophila, and its antioxidant activities[J].

Carbohydrate Polymers, 2014, 111(20): 324-332.

[20] MACHOVA E, BYSTRICKY S. Antioxidant capacities of mannans

and glucans are related to their susceptibility of free radical

degradation[J]. International Journal of Biological Macromolecules,

2013, 61(10): 308-311.

[21] YE C L, HU W L, DAI D H. Extraction of polysaccharides and the

antioxidant activity from the seeds of Plantago asiatica L.[J]. International

Journal of Biological Macromolecules, 2011, 49(6): 466-470.

[22] LEE J, KOO N, MIN D B. Reactive oxygen species, aging, and

antioxidativenutraceuticals[J]. Comprehensive Reviews in Food

Science and Food Safety, 2004, 42(3): 21-33.

[23] 晁正, 冉玉梅, 杨霞, 等.麦麸中低聚木糖的制备及抗氧化活性研究[J].

核农学报, 2014, 21(4): 655-661.

[24] 马国刚, 王建中. 超声波辅助提取青稞β-葡聚糖的工艺条件优

化[J]. 食品科技, 2009, 34(11): 168-174. DOI:10.13684/j.cnki.

spkj.2009.11.061.