Response Surface Optimization on Enzymatic Extraction Process of Turmeric Starch
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摘要:
目的 优化姜黄淀粉酶法提取工艺,提高姜黄淀粉提取率。 方法 以新鲜姜黄为原材料,姜黄淀粉提取率为指标,在几种蛋白酶中筛选最佳酶,采用单因素试验进一步确定提取酶解时间、酶解温度、酶解pH和中性蛋白酶添加量等4个因素影响姜黄淀粉提取率的优化范围。在单因素试验基础上,采用Box-Behnken方法,进行4因素3水平响应面优化设计,共进行29组处理,每组处理3次重复,考察酶解时间、酶解温度、酶解pH和中性蛋白酶添加量等4个因素对姜黄淀粉提取率的影响,从而确定姜黄淀粉酶法提取的最佳工艺条件。 结果 姜黄淀粉提取最佳工艺参数为:酶解pH6.83、酶解温度51.45 ℃、酶解时间6.20 h、中性蛋白酶添加量0.13%,姜黄淀粉提取率的理论值为62.00%。考虑实际操作的简便,确定姜黄淀粉提取的最佳工艺参数为:酶解时间6 h,酶解温度52 ℃,酶解pH6.8,中性蛋白酶添加量0.13%,在此条件下实际验证值为60.42%,拟合得到的模型与实际吻合良好。 结论 通过响应面法优化了姜黄淀粉的酶法提取工艺条件,提高了姜黄淀粉提取量,为姜黄淀粉的工业化生产提供了理论依据。 Abstract:Objective Optimization on the enzymatic extraction process of starch from turmeric was improved. Method Using fresh turmeric as raw material, the best enzyme was screened among several proteases according to the starch extraction rate. A single-factor test was conducted to define the range of the time, digestion temperature, pH and the amount of neutral protease for extraction. Based on the single-factor test results, a 4-factor, 3-level Box-Behnken response surface optimization experiment was designed on 29 treatments with 3 replicates each to determine the optimal processing conditions. Results The optimized process applied 0.13% added neutral protease in a solution at pH 6.83 held at 51.45 °C for 6.20 h to achieve 62.00% turmeric starch extraction. For practical application the conditions were modified to be pH 6.8, 52°C, and 6 h to obtain a turmeric extraction rate of 60.42%. The experimental results fitted well with the prediction model. The fitted model was in good agreement with the actual one. Conclusion The enzymatic extraction of turmeric starch was optimized by response surface methodology in preparation for industrialized production. -
Key words:
- Turmeric /
- starch /
- extraction /
- response surface methodology /
- neutral protease
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表 1 蛋白酶最适条件
Table 1. Optimum conditions for protease digestion
蛋白酶种类
Protease types最适pH值
Optimum
pH最适温度
Optimum
temperature/
°C蛋白酶添加量
Protease
addition/
(U·mL−1)中性蛋白酶
Neutral protease6.8 60 300 碱性蛋白酶
Alkaline protease11.0 50 300 木瓜蛋白酶
Papain5.7 55 300 胰蛋白酶
Trypsin8.1 37 300 胃蛋白酶
Pepsin3.0 37 300 空白(CK)
Blank(CK)6.8 25 — 表 2 响应面试验的因素和水平
Table 2. Factors and levels of response surface experiments
水平
Level因素
FactorsA:酶解pH
Enzymatic pHB:酶解时间
Enzymatic
digestion
time /hC:酶解温度
Enzymatic
digestion
temperature/°CD:中性蛋白酶添加量
Neutral
protease
addition/%−1 6.4 4 40 0.100 0 6.8 6 50 0.125 1 7.2 8 60 0.150 表 3 响应面试验设计与结果
Table 3. Response surface test design and results
序号
Serial numberA B C D 提取率
Extraction rate/%1 6.4 4 50 0.125 52.14 2 7.2 6 50 0.15 52.67 3 6.8 6 40 0.15 48.39 4 6.8 6 40 0.1 47.65 5 6.8 6 60 0.15 56.44 6 6.8 8 60 0.125 53.62 7 6.8 6 50 0.125 62.18 8 6.8 4 50 0.15 51.44 9 6.8 4 40 0.125 51.34 10 6.8 6 60 0.1 48.66 11 7.2 6 60 0.125 55.23 12 6.8 8 40 0.125 54.69 13 6.8 6 50 0.125 63.24 14 6.4 6 60 0.125 50.74 15 7.2 6 40 0.125 52.46 16 7.2 6 50 0.1 50.69 17 6.8 6 50 0.125 61.57 18 6.8 4 60 0.125 52.07 19 6.4 6 50 0.1 46.78 20 6.4 6 50 0.15 53.48 21 6.8 4 50 0.1 46.64 22 6.8 6 50 0.125 60.24 23 7.2 8 50 0.125 53.3 24 6.4 8 50 0.125 53.19 25 6.4 6 40 0.125 50.09 26 6.8 6 50 0.125 60.89 27 6.8 8 50 0.15 54.18 28 7.2 4 50 0.125 52.94 29 6.8 8 50 0.1 47.84 表 4 回归模型的方差分析
Table 4. Analysis of variance for regression models
方差来源
Source of variance平方和
Sum of squares自由度
Freedom均方
Mean SquareF值
F valueP值
P-value显著性
Significance模型 Model 570.86 14 40.78 21.41 <0.0001 ** A 9.85 1 9.85 5.17 0.0393 * B 8.76 1 8.76 4.60 0.0501 C 12.28 1 12.28 6.45 0.0236 * D 66.93 1 66.93 35.14 <0.0001 ** AB 0.12 1 0.12 0.062 0.8062 AC 1.12 1 1.12 0.59 0.4552 AD 5.57 1 5.57 2.92 0.1093 BC 0.81 1 0.81 0.43 0.5249 BD 0.59 1 0.59 0.31 0.5857 CD 12.39 1 12.39 6.50 0.0231 * A2 124.22 1 124.22 65.21 <0.0001 ** B2 126.50 1 126.50 66.41 <0.0001 ** C2 141.31 1 141.31 74.18 <0.0001 ** D2 294.00 1 294.00 154.34 <0.0001 ** 残差 Residual 26.67 14 1.90 失拟项 Lack of fit 21.29 10 2.13 1.58 0.3487 纯误差 Pure error 5.38 4 1.34 总和 Cor total 597.53 28 相关系数 R2 0.9554 校正相关系数 R2 0.9107 **P<0.01,表示极显著,*P<0.05,表示显著,P>0.05,表示不显著。
** indicates extremely significant at P<0.01, * indicates significant at P<0.05, and no sign indicates not significant at P>0.05. -
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