食品科学 ›› 2017, Vol. 38 ›› Issue (4): 217-222.doi: 10.7506/spkx1002-6630-201704035

• 工艺技术 • 上一篇    下一篇

黑豆蛋白的分级提取及黑豆花色苷的成分鉴定

江 甜,何 毅,祝振洲,李书艺,何静仁   

  1. 武汉轻工大学食品科学与工程学院,湖北 武汉 430023
  • 出版日期:2017-02-25 发布日期:2017-02-28
  • 基金资助:
    国家自然科学基金面上项目(31371727);国家国际科技合作专项(2014DFG32310); 湖北省科技支撑计划项目(2015BHE015);湖北省自然科学基金项目(2014CFB891)

Fractional Extraction of Protein and Characterization of Anthocyanins from Black Soybean Seeds

JIANG Tian, HE Yi, ZHU Zhenzhou, LI Shuyi, HE Jingren   

  1. College of Food Science and Engineering, Wuhan Polytechnic University, Wuhan 430023, China
  • Online:2017-02-25 Published:2017-02-28

摘要: 以黑豆为原料,添加不同浓度(0、5、10、15、20、25 mmol/L)的不同二价阳离子(Ca2+、Mg2+和Zn2+)分级提取黑豆蛋白,经冷冻干燥后用凯氏定氮法测得蛋白质量浓度,并对所得蛋白级分(7S、11S)的纯度和得率进行分析。结果表明,黑豆提取过程中所加入二价阳离子的类型及浓度显著影响7S和11S球蛋白的得率和纯度。利用综合平衡法,最终选定添加10 mmol/L的Mg2+分级提取黑豆球蛋白,得到7S蛋白纯度和得率分别为(86.29±3.25)%和(2.90±0.14)%,11S蛋白纯度和得率分别为(87.42±3.30)%和(9.11±0.28)%。用乙醇法提取黑豆花色苷,测得黑豆中总花色苷含量为(0.58±0.03)mg/g,总酚含量为(2.22±0.12)mg/g,对黑豆花色苷进行高效液相色谱-串联质谱分析,其组成成分为飞燕草色素-3-O-葡萄糖苷、矢车菊色素-3-O-葡萄糖苷、牵牛花色素-3-O-葡萄糖苷和锦葵花色素-3-O-葡萄糖苷。

关键词: 黑豆, 分级提取, 7S球蛋白, 11S球蛋白, 花色苷

Abstract: Crude protein extract from black soybean seeds was fractionated by addition of different concentrations (0, 5, 10, 15, 20 and 25 mmol/L) of divalent cations (Ca2+, Mg2+ and Zn2+). After the precipitated protein fractions (7S globulin and 11S globulin) were freeze dried, total nitrogen was determined by the Kjeldahl method and the yield and purity of the two fractions were measured. The results revealed that the type and concentration of divalent cations significantly affected the yield and purity of 7S and 11S globulin. Based on comprehensive consideration of yield and purity, 10 mmol/L of Mg2+ was selected to extract black soybean globulin. The purity and yield of the obtained 7S protein were (86.29 ± 3.25)% and (2.90 ± 0.14)%, respectively, while the purity and yield of the obtained 11S protein were (87.42 ± 3.30)% and (9.11 ± 0.28)%, respectively. Anthocyanins from black soybean seeds were extracted with ethanol. It turned out that the content of anthocyanins was (0.58 ± 0.03) mg/g and total phenol was (2.22 ± 0.12) mg/g. By high performance liquid chromatography tandem-mass spectrometry (HPLC-MS/MS) analysis, the isolated anthocyanins were characterized as delphinidin-3-O-glucoside, cyanidin 3-O-glucoside, petunidin 3-O-glucoside and mallow-3-O-glucoside, respectively.

Key words: black soybean, fractional extraction, 7S globulin, 11S globulin, anthocyanin

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