纳豆芽孢杆菌发酵菜用大豆中总黄酮的提取及其降血脂效果评价

李秀凉 倪庆圆 杨宸 宋永 韩晓云 孙庆申

李秀凉,倪庆圆,杨宸,等. 纳豆芽孢杆菌发酵菜用大豆中总黄酮的提取及其降血脂效果评价[J]. 食品工业科技,2021,42(7):1−9. doi:  10.13386/j.issn1002-0306.2020070003
引用本文: 李秀凉,倪庆圆,杨宸,等. 纳豆芽孢杆菌发酵菜用大豆中总黄酮的提取及其降血脂效果评价[J]. 食品工业科技,2021,42(7):1−9. doi:  10.13386/j.issn1002-0306.2020070003
LI Xiuliang, NI Qingyuan, YANG Chen, et al. Extraction of Total Flavonoids in Vegetable Soybean Fermented by Bacillus natto and Evaluation of Its Effect on Lowering Blood Lipid[J]. Science and Technology of Food Industry, 2021, 42(7): 1−9. (in Chinese with English abstract). doi:  10.13386/j.issn1002-0306.2020070003
Citation: LI Xiuliang, NI Qingyuan, YANG Chen, et al. Extraction of Total Flavonoids in Vegetable Soybean Fermented by Bacillus natto and Evaluation of Its Effect on Lowering Blood Lipid[J]. Science and Technology of Food Industry, 2021, 42(7): 1−9. (in Chinese with English abstract). doi:  10.13386/j.issn1002-0306.2020070003

纳豆芽孢杆菌发酵菜用大豆中总黄酮的提取及其降血脂效果评价

doi: 10.13386/j.issn1002-0306.2020070003
基金项目: 黑龙江省自然科学基金面上项目(C2006049);哈尔滨市应用技术研究与开发项目(青年后备人才 A类RC2017QN020010)资助
详细信息
    通讯作者:

    孙庆申(1977−),男,博士,教授,研究方向:食品与药品活性物质的挖掘,E-mail:sunqingshen@hlju.edu.cn

  • 中图分类号: TS201.2

Extraction of Total Flavonoids in Vegetable Soybean Fermented by Bacillus natto and Evaluation of Its Effect on Lowering Blood Lipid

  • 摘要: 本文以菜用大豆等外品为原料,通过纳豆枯草芽孢杆菌发酵来提高其中总黄酮的含量,为提高菜用大豆产品的附加值提供理论支持。实验利用纳豆芽孢杆菌对菜用大豆等外品进行发酵,以发酵产品中总黄酮含量为评价指标确定纳豆发酵的最优条件,通过D101型大孔树脂对提取的总黄酮进行纯化,采用芦丁比色法测定黄酮含量。体内实验分为肥胖预防实验和缓解实验:以C57BJ/6L小鼠为模型,预防实验在饲喂高脂饲料的同时,灌胃总黄酮提取物溶液(50 mg/kg·d);缓解实验是利用高脂饲料建立起肥胖小鼠模型以后,灌胃低、中、高剂量的总黄酮提取物溶液(分别为20、50和100 mg/kg·d)。通过小鼠体重、血脂以及促炎因子等指标评价总黄酮溶液的降脂效果。结果表明,菜用大豆发酵生产黄酮的最佳条件为:枯草芽孢杆菌接菌量为1.15 × 1011 CFU/100 g菜用大豆,发酵温度37 ℃,发酵时间24 h,后熟时间12 h,此时纳豆总黄酮得率为4.30 mg/g菜用大豆;测得该粗提物中黄酮含量为22.1%;预防实验组给予黄酮处理后,小鼠体重和Lee's指数与正常对照组相比均无显著差异。缓解实验中高、中、低剂量黄酮提取物处理组小鼠血清中甘油三酯(TG)含量分别为313.1 ± 10.12、310.39 ± 31.76和310.1 ± 10.20 nmol/L,与正常对照组(330.8 ± 34.36 nmol/L)无显著差异(P>0.05);黄酮各组灌胃干预后,小鼠血清总胆固醇(TC)、白介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)含量均较高脂对照组显著下降(P <0.05)。结论:利用纳豆芽孢杆菌发酵菜用大豆,提高了其中的总黄酮含量,该黄酮提取物具有预防和缓解高脂饮食小鼠血脂升高的效果。
  • 图  1  发酵时间(A)、发酵温度(B)、接菌量(C)和后熟时间(D)对纳豆中总黄酮含量的影响

    注:图中标记小写字母不同者表示差异显著(p<0.05)。

    Figure  1.  Effect of fermentation time (A), fermentation temperature(B), inoculation dose(C) and ripening time(D) on the content of total flavonoids in natto

    图  2  缓解实验(A)和预防实验(B)各组小鼠体重变化

    注:图中同一时间内标记小写字母不同者表示差异显著(p<0.05)。

    Figure  2.  Body weight change of mice in remission experiment(A)and prevention experiment(B)

    图  3  缓解实验(A)和预防实验(B)各组小鼠血清甘油三酯变化

    注:图中标记不同小写字母表示差异显著(p<0.05),图4图6同。

    Figure  3.  Changes of triglycerides in mice in remission experiment(A)and prevention experiment(B)

    图  4  缓解实验(A)和预防实验(B)各组小鼠血清总胆固醇变化

    Figure  4.  Changes of total cholesterol in mice in remission experiment(A)and prevention experiment(B)

    图  5  缓解实验(A)和预防实验(B)各组小鼠高密度脂蛋白变化

    Figure  5.  Changes of High density lipoprotein in mice in remission experiment(A)and prevention experiment(B)

    图  6  缓解实验(A、C、E)和预防实验(B、D、F)各组小鼠血清炎性因子含量的变化

    Figure  6.  Changes of proinflammatory factors in mice serum in alleviating experiment(A, C, E)and prevention experiment(B, D, F)

    表  1  纳豆发酵工艺优化正交实验因素与水平

    Table  1.   Factors and levels of orthogonal experiment for optimization of natto fermentation process

    水平A发酵时间(h)B接菌量(CFU /100 g)C后熟时间(h)
    1245.75 × 10106
    2361.15 × 101112
    3481.73 × 101118
    下载: 导出CSV

    表  2  纳豆发酵工艺优化正交实验结果

    Table  2.   Orthogonal experiment result for optimization of natto fermentation process

    实验号ABCY:总黄酮含量(mg/g)
    11113.34 ± 0.17
    21224.30 ± 0.12
    31333.70 ± 0.24
    42123.66 ± 0.27
    52233.79 ± 0.21
    62312.99 ± 0.21
    73133.30 ± 0.26
    83213.23 ± 0.28
    93322.40 ± 0.26
    k13.783.433.19
    k23.483.773.45
    k32.983.033.59
    R0.800.740.41
    下载: 导出CSV

    表  3  正交实验方差分析

    Table  3.   Analysis of orthogonal experimental variance

    因素SS自由度F值Fa
    A0.989323.874F0.05(2,4) = 6.944
    B0.829423.248
    C0.26042
    误差e0.25032
    总变异2.329
    下载: 导出CSV

    表  4  各组小鼠体长及Lee’s指数

    Table  4.   Body length and Lee’s index of mice in different groups

    组别缓解实验组别预防实验
    体长(cm)Lee’s指数体长(cm)Lee’s指数
    正常对照组(A)8.98 ± 0.33a344.94 ± 5.82b正常对照组8.63 ± 0.24a356.58 ± 3.79b
    阳性对照组(B)8.90 ± 0.23a349.73 ± 10.19b阳性对照组8.31 ± 0.39a363.58 ± 11.18b
    模型组(C)8.98 ± 0.36a363.56 ± 18.18a模型组8.57 ± 0.62a382.03 ± 11.15a
    总黄酮高剂量组(D)8.73 ± 0.15a350.20 ± 7.17b总黄酮组8.34 ± 0.38a362.54 ± 6.76b
    总黄酮中剂量组8.81 ± 0.08a351.15 ± 4.96b
    总黄酮低剂量组(F)8.84 ± 0.28a348.65 ± 12.22b
    注:表中同一列标记小写字母不同者表示差异显著(p<0.05),表5同。
    下载: 导出CSV

    表  5  缓解实验及预防实验各组小鼠肝脏及脾脏系数

    Table  5.   Liver and spleen coefficients of mice in alleviating experiment and prevention experiment

    组别缓解试验组别预防实验
    肝脏指数脾脏指数肝脏指数脾脏指数
    正常对照组3.55 ± 0.13a0.24 ± 0.03a正常对照组3.57 ± 0.39a2.57 ± 0.25a
    阳性对照组3.73 ± 0.33a0.27 ± 0.04a阳性对照组3.65 ± 0.20a2.58 ± 0.43a
    模型组3.25 ± 0.37b0.23 ± 0.05b模型组3.38 ± 0.33b2.46 ± 0.42a
    总黄酮高剂量3.53 ± 0.20a0.26 ± 0.05a异黄酮组3.62 ± 0.11a2.63 ± 0.19a
    总黄酮中剂量组3.51 ± 0.63a0.27 ± 0.04a
    总黄酮低剂量组3.58 ± 0.33a0.26 ± 0.05a
    下载: 导出CSV
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  • 收稿日期:  2020-07-02

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