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中國精品科技期刊2020

酪蛋白源降膽固醇肽的酶解制備工藝優化

梁小慧 王孝治 趙佳園 金庭飛 黎旭 羅夢帆 譚東虎 劉明真 羅海波 郭宇星

梁小慧,王孝治,趙佳園,等. 酪蛋白源降膽固醇肽的酶解制備工藝優化[J]. 食品工業科技,2022,43(19):280?287. doi:  10.13386/j.issn1002-0306.2022010165
引用本文: 梁小慧,王孝治,趙佳園,等. 酪蛋白源降膽固醇肽的酶解制備工藝優化[J]. 食品工業科技,2022,43(19):280?287. doi:  10.13386/j.issn1002-0306.2022010165
LIANG Xiaohui, WANG Xiaozhi, ZHAO Jiayuan, et al. Optimization of Preparation of Casein-derived Cholesterol-lowering Peptide by Enzymatic Hydrolysis[J]. Science and Technology of Food Industry, 2022, 43(19): 280?287. (in Chinese with English abstract). doi:  10.13386/j.issn1002-0306.2022010165
Citation: LIANG Xiaohui, WANG Xiaozhi, ZHAO Jiayuan, et al. Optimization of Preparation of Casein-derived Cholesterol-lowering Peptide by Enzymatic Hydrolysis[J]. Science and Technology of Food Industry, 2022, 43(19): 280?287. (in Chinese with English abstract). doi:  10.13386/j.issn1002-0306.2022010165

酪蛋白源降膽固醇肽的酶解制備工藝優化

doi: 10.13386/j.issn1002-0306.2022010165
基金項目: 江蘇省農業科技自主創新資金項目(CX(20)3050);國家自然科學基金面上項目(31972093);江蘇省林業科技創新與推廣項目(LYKJ[2022]12)。
詳細信息
    作者簡介:

    梁小慧(1996?),女,碩士研究生,研究方向:食品功能因子,E-mail:1318237522@qq.com

    通訊作者:

    郭宇星(1981?),女,博士,教授,研究方向:食品功能因子,E-mail:guoyuxing1981@163.com

  • 中圖分類號: TS201.2

Optimization of Preparation of Casein-derived Cholesterol-lowering Peptide by Enzymatic Hydrolysis

  • 摘要: 以酪蛋白為原料,采用中性蛋白酶、堿性蛋白酶以及胰蛋白酶對酪蛋白進行水解,確定制備降膽固醇肽的最佳蛋白酶;通過單因素實驗和響應面試驗,研究水解pH、水解溫度、酶與底物比、底物濃度和水解時間對酪蛋白水解度和膽固醇膠束溶解度抑制率的影響,確定最佳水解條件;而后通過超濾和凝膠過濾層析確定降膽固醇肽的初步分離工藝。結果表明:制備酪蛋白源降膽固醇肽的最佳水解工具酶是中性蛋白酶,其最佳酶解條件為反應溫度51.3 ℃,酶與底物濃度比6.47%,pH6.34,底物濃度5 g/100 mL,反應時間3.5 h,膽固醇抑制率為58.25%±0.59%;Sephadex G-10分離酪蛋白降膽固醇肽條件為上樣濃度80 mg/mL,上樣體積2.5 mL,洗脫速度3.5 mL/min;經酶解、超濾及層析后制備的酪蛋白源降膽固醇肽峰1和峰2樣品在100 μg/mL的膽固醇溶解度抑制率為24.2%±0.24%和4.3%±0.16%。經酶解制備分離后,獲得具有抑制降固醇膠束溶解活性的降膽固醇肽,為降膽固醇肽的開發提供理論研究基礎。
  • 圖  1  不同工具酶酶解酪蛋白水解度和膽固醇膠束抑制率比較

    Figure  1.  Comparison of enzymatic hydrolysis degree of and cholesterol micellar inhibition rate with different enzymes

    注:不同字母表示同一指標差異顯著,P<0.05;圖2~圖6同。

    圖  2  pH對制備降膽固醇活性肽的影響

    Figure  2.  Effect of pH on preparation cholesterol-lowering peptide

    圖  3  溫度對制備降膽固醇活性肽的影響

    Figure  3.  Effect of temperature on preparation cholesterol-lowering peptide

    圖  4  酶比對制備降膽固醇活性肽的影響

    Figure  4.  Effect of enzyme rate on preparation cholesterol-lowering peptide

    圖  5  底物濃度對制備降膽固醇活性肽的影響

    Figure  5.  Effect of concentration of substrate on preparation cholesterol-lowering peptide

    圖  6  反應時間對制備降膽固醇活性肽的影響

    Figure  6.  Effect of reaction time on preparation cholesterol-lowering peptide

    圖  7  交互變量對膽固醇抑制率的相應曲面和等高線圖

    Figure  7.  Response surface plots and coutour plots for effect of interactive variables on cholesterol inhibition rate

    圖  8  Sephadex G-10分離酪蛋白肽條件的優化

    Figure  8.  Optimization of separation conditions of casein peptide by Sephadex G-10

    注:A-上樣濃度;B-上樣體積;C-洗脫速度。

    表  1  響應面因素水平編碼表

    Table  1.   Factors and levels of response surface text

    因素代碼編碼
    ?101
    溫度(℃)X1455055
    E/S(% (m/v))X2567
    pHX366.57
    下載: 導出CSV

    表  2  響應面試驗設計與結果表

    Table  2.   Design and result of Box-Behnken experient

    實驗號不同水平變量的編碼值Y(膽固醇抑制率%)
    溫度(X1E/S(X2pH(X3
    110?151.71
    201145.05
    30?1152.81
    40?1?152.67
    500058.27
    6?1?1047.79
    7?10?144.92
    800059.24
    900060.93
    1001?156.48
    1100057.32
    1210147.46
    13?10141.18
    14?11048.91
    1511055.73
    161?1053.19
    1700058.60
    下載: 導出CSV

    表  3  膽固醇抑制率的方差分析表

    Table  3.   ANOVA for cholesterol inhibition rate

    變異來源平方和均方FP
    總模型523.3958.1525.590.0002*
    X179.9579.9535.180.0006*
    X20.0110.0110.00460.9477
    X346.4646.4620.450.0027*
    X1X20.50.50.220.652
    X1X30.0650.0650.0290.8705
    X2X333.4733.4714.730.0064*
    X12175.22175.2277.11<0.0001**
    X224.354.351.910.2092
    X32156.85156.8569.03<0.0001**
    殘差15.912.27
    失擬項8.692.91.610.3214
    純誤差7.221.8
    合計539.3
    S=1.51R2=0.9705R2Adj=0.9326
    注:**差異極顯著(P<0.0001);*差異顯著(P<0.05)。
    下載: 導出CSV
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  • 收稿日期:  2022-01-20
  • 網絡出版日期:  2022-08-18
  • 刊出日期:  2022-09-23

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