Optimization of Enzymatic Hydrolysis Conditions for the Production of Crisp Plum Juice by Response Surface Methodology
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摘要: 目的：研究酶解提取酥李果汁的最佳工艺条件，为李深加工利用提供理论参考。方法：以酥李出汁率为指标，在单因素实验基础上采用响应面试验优化，对单一果胶酶、单一纤维素酶、复合酶（果胶酶和纤维素酶）提取酥李果汁的工艺条件分别进行优化。结果：不同加酶方式中对酥李出汁率的影响因素顺序均为酶解温度>加酶量>酶解pH>酶解时间；果胶酶酶解提取酥李果汁的最佳工艺条件为：加酶量0.45 g/L、酶解温度38 ℃、酶解pH3.8、酶解时间72 min，出汁率提高27.13%；维素酶酶解提取酥李果汁的最佳工艺条件为：加酶量0.55 g/L、酶解温度41 ℃、酶解pH4.2、酶解时间105 min，出汁率提高20.18%；复合酶酶解提取酥李果汁的最佳工艺条件为：果胶酶添加量0.45 g/L、纤维素酶添加量0.55 g/L、酶解温度41 ℃、酶解pH4.0、酶解时间87 min，出汁率提高31.79%。三种加酶方式中，回归模型均能较好地反应相应酶制备酥李果浆的出汁率，所得工艺合理可靠。结论：在酶法提取酥李果汁过程中，果胶酶和纤维素酶的不同添加方式均能有效提高酥李出汁率，其中采用复合酶提取酥李果汁效果最佳。本研究成果为贵州李产品开发提供了一定的技术参考。Abstract: Objective: To investigate the optimum enzymolysis extraction process conditions of plum fruit, and to provide theoretical bases fordeep processing andutilizationof itsresources. Method: The juice yield was used as an indicator. On the basis of single-factor experiments, response surface methodology (RSM) was employed to optimize the enzymatic hydrolysis pretreatment conditions for extracting fruit juice from crisp plum with methods of adding enzymes, including pectinases, cellulase and complex enzyme consisted of pectinases and cellulase. Result: The results suggested that the extraction parameters in the order of significance were enzymolysis temperature, enzyme concentration, enzymolysis pH, and enzymolysis time by response surface analysis. The optimum conditions of enzymolysis with pectinases were pectinases concentration 0.45 g/L, enzymolysis temperature 38 ℃, enzymolysis pH3.8 and enzymolysis time 72 min, in which the average juice yield of crisp plum was 78.63%. The optimum conditions of enzymolysis with cellulase were cellulase concentration 0.55 g/L, enzymolysis temperature 41 ℃, enzymolysis pH4.2 and enzymolysis time 105 min, in which the average juice yield of crisp plum was 74.33%. The optimum conditions of enzymolysis with complex enzyme were pectinases concentration 0.45 g/L, cellulase concentration 0.55 g/L, enzymolysis temperature 41 ℃, enzymolysis pH4.0 and enzymolysis time 87 min, in which the average juice yield of crisp plum was 81.51%. Conclusion: In the extract process of crisp plum juice, juice yields all increased when extracting juice, by adding pectinases, cellulase or complex enzyme consisted of pectinases and cellulose on the corresponding optimal conditions of enzymatic hydrolysis. The study provides technical references for the product development of Guizhou plum fruit.
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Key words:
- crisp plum juice /
- juice yield /
- pectinase /
- cellulase /
- response surface methodology
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图 2 不同酶解温度对酥李出汁率的影响
Figure 2. Effects of different enzyme hydrolysis temperature on juice yield of crisp plum
图 3 不同单酶酶解pH对酥李出汁率的影响
Figure 3. Effects of different enzyme hydrolysis pH on juice yield of crisp plum
图 4 不同酶解时间对酥李出汁率的影响
Figure 4. Effect of different enzymolysistime on juice yield of crisp plum
表 1 响应面分析法优化果胶酶、纤维素酶酶解工艺
Table 1. Optimization of pectinase and cellulose extraction by response surface methodolog
酶种类 水平 因素 A加酶量（g/L） B酶解温度（℃） C酶解pH D酶解时间（min） 果胶酶 −1 0.25 32 3.6 60 0 0.35 35 3.9 75 1 0.45 38 4.2 90 纤维素酶 −1 0.35 35 3.9 75 0 0.45 38 4.2 90 1 0.55 41 4.5 105 
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表 2 响应面分析法优化复合酶酶解工艺
Table 2. Optimization of complex enzyme extraction by response surface methodology
水平 因素 A果胶酶添加量（g/L） B纤维素酶添加量（g/L） C酶解温度（℃） D酶解pH E酶解时间（min） −1 0.25 0.35 35 3.6 50 0 0.35 0.45 38 3.8 70 1 0.45 0.55 41 4.0 90
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表 3 果胶酶制备酥李果汁工艺优化响应面试验结果
Table 3. Results of response surface method for pectinase extraction of crisp plum juice
实验号 A B C D 出汁率（%） 1 −1 0 0 −1 69.54 2 0 1 0 1 70.73 3 0 0 1 −1 73.54 4 1 −1 0 0 69.81 5 0 0 −1 −1 76.43 6 0 −1 1 0 69.32 7 0 1 −1 0 69.41 8 0 0 −1 1 69.41 9 0 0 0 0 68.49 10 0 −1 0 1 77.32 11 0 1 1 0 69.98 12 0 0 1 1 69.98 13 0 −1 −1 0 68.15 14 1 0 −1 0 73.66 15 0 0 0 0 70.62 16 1 1 0 0 68.15 17 0 1 0 −1 71.21 18 0 0 0 0 71.21 19 −1 1 0 0 78.72 20 0 0 0 0 77.32 21 −1 −1 0 0 72.21 22 −1 0 −1 0 74.67 23 0 −1 0 −1 72.36 24 1 0 0 −1 67.34 25 −1 0 1 0 68.99 26 0 0 0 0 68.99 27 −1 0 0 −1 73.65 28 0 1 0 1 68.66 29 0 0 1 −1 72.98
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表 4 出汁率回归模型的方差分析
Table 4. Variance analysis of multiple regression model of juice yield
方差来源 平方和 自由度 均方差 F值 P值 显著性 模型 253.84 14 18.13 19.42 <0.0001 ** A 27.76 1 27.76 29.73 <0.0001 ** B 188.73 1 188.73 202.14 <0.0001 ** C 1.07 1 1.07 1.14 0.3029 D 4.800E-003 1 4.800E-003 5.141E-003 0.9439 AB 0.98 1 0.98 1.05 0.3229 AC 5.625E-003 1 5.625E-003 6.025E-003 0.9392 AD 6.400E-003 1 6.400E-003 6.855E-003 0.9352 BC 4.00 1 4.00 4.28 0.0574 BD 6.250E-004 1 6.250E-004 6.694E-004 0.9797 * CD 0.13 1 0.13 0.14 0.7113 A2 0.44 1 0.44 0.48 0.5019 B2 10.73 1 10.73 11.49 0.0044 ** C2 14.16 1 14.16 15.17 0.0016 ** D2 0.26 1 0.26 0.28 0.6059 残差 13.07 14 0.93 失拟项 7.55 10 0.76 0.55 0.7999 纯误差 5.52 4 1.38 总和 266.92 28 R2 0.9510 R2Adj 0.9021 注：“*”表示对结果影响显著（P<0.05），“**”表示对结果影响极显著（P<0.01）；表6、表8同。
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表 5 纤维素酶制备酥李果汁工艺优化响应面试验结果
Table 5. Results of response surface method for cellulose extraction of crisp plum juice
实验号 A B C D 出汁率（%） 1 −1 0 0 1 65.32 2 −1 0 0 −1 65.59 3 −1 1 0 0 70.45 4 −1 −1 0 0 65.42 5 −1 0 −1 0 67.77 6 −1 0 1 0 64.44 7 0 1 0 1 73.21 8 0 0 1 −1 65.1 9 0 0 −1 −1 67.19 10 0 −1 1 0 64.27 11 0 1 −1 0 73.1 12 0 0 −1 1 65.76 13 0 0 0 0 65.76 14 0 −1 0 1 64.93 15 0 1 1 0 69.44 16 0 0 1 1 66.4 17 0 −1 −1 0 65.15 18 0 0 0 0 66.99 19 0 1 0 −1 73.1 20 0 0 0 0 67.99 21 0 0 0 0 68.14 22 0 −1 0 −1 66.77 23 0 0 0 0 68.76 24 1 0 1 0 66.51 25 1 0 0 1 69.32 26 1 −1 0 0 66.19 27 1 0 −1 0 66.99 28 1 1 0 0 74.5 29 1 0 0 −1 69.43
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表 6 出汁率回归模型的方差分析
Table 6. Variance analysis of multiple regression model of juice yield
方差来源 平方和 自由度 均方差 F值 P值 显著性 模型 212.81 14 15.20 15.88 <0.0001 ** A 16.22 1 16.22 16.95 0.0010 ** B 140.56 1 140.56 146.88 <0.0001 ** C 8.00 1 8.00 8.36 0.0118 * D 0.42 1 0.42 0.44 0.5193 AB 2.69 1 2.69 2.81 0.1158 AC 2.03 1 2.03 2.12 0.1673 AD 6.400E-003 1 6.400E-003 6.688E-003 0.9360 BC 1.93 1 1.93 2.02 0.1772 BD 0.95 1 0.95 0.99 0.3358 CD 1.86 1 1.86 1.95 0.1847 A2 9.041E-003 1 9.041E-003 9.447E-003 0.9239 B2 20.73 1 20.73 21.66 0.0004 ** C2 10.36 1 10.36 10.82 0.0054 ** D2 1.197E-003 1 1.197E-003 1.251E-003 0.9723 残差 13.40 14 0.96 失拟项 7.88 10 0.79 0.57 0.7850 纯误差 5.52 4 1.38 总和 226.21 28 R2 0.9408 R2Adj 0.8815
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表 7 复合酶制备酥李果汁工艺优化响应面试验结果
Table 7. Results of response surface method for complex enzyme extraction of crisp plum juice
实验号 A B C D E 出汁率（%） 1 0 0 1 −1 0 76.49 2 0 0 0 −1 −1 68.8 3 0 0 0 −1 1 68.98 4 0 −1 1 0 0 72.65 5 −1 0 0 0 1 68.31 6 0 0 0 0 0 68.98 7 0 0 −1 0 1 65.92 8 0 1 1 0 0 77.71 9 1 −1 0 0 0 67.65 10 0 0 1 1 0 76.42 11 −1 0 0 1 0 67.75 12 0 0 0 0 0 69.4 13 1 0 1 0 0 76.31 14 0 1 0 0 1 71.43 15 −1 1 0 0 0 71.42 16 −1 −1 0 0 0 67.18 17 −1 0 1 0 0 74.2 18 0 0 −1 −1 0 66.76 19 0 0 1 0 −1 75.09 20 0 −1 0 0 −1 67.18 21 1 0 0 0 1 70.98 22 1 1 0 0 0 72.53 23 1 0 −1 0 0 66.43 24 0 0 0 0 0 69.72 25 0 1 0 0 −1 72.44 26 1 0 0 0 −1 71.35 27 0 0 0 1 −1 70.2 28 −1 0 0 0 −1 68.39 29 0 1 0 1 0 75.09 30 1 0 0 −1 0 70.13 31 0 −1 0 0 1 67.31 32 0 −1 0 −1 0 67.75 33 0 1 0 −1 0 73.66 34 1 0 0 1 0 71.97 35 0 0 −1 1 0 67.09 36 0 0 1 0 1 76.31 37 0 −1 −1 0 0 65.11 38 0 1 −1 0 0 66.76 39 −1 0 −1 0 0 65.92 40 0 −1 0 1 0 67.58 41 0 0 −1 0 −1 66.26 42 0 0 0 0 0 69.98 43 0 0 0 0 0 69.98 44 −1 0 0 −1 0 68.5 45 0 0 0 1 1 71.31 46 0 0 0 0 0 70.75
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表 8 出汁率回归模型的方差分析
Table 8. Variance analysis of multiple regression model of juice yield
方差来源 平方和 自由度 均方差 F值 P值 显著性 模型 485.83 20 24.29 33.04 <0.0001 ** A 15.37 1 15.37 20.90 0.0001 ** B 93.27 1 93.27 126.87 <0.0001 ** C 350.91 1 350.91 477.34 <0.0001 ** D 2.51 1 2.51 3.42 0.0764 E 0.044 1 0.044 0.060 0.8085 AB 0.10 1 0.10 0.14 0.7121 AC 0.64 1 0.64 0.87 0.3597 AD 1.68 1 1.68 2.28 0.1435 AE 0.021 1 0.021 0.029 0.8671 BC 2.91 1 2.91 3.95 0.0578 BD 0.64 1 0.64 0.87 0.3597 BE 0.32 1 0.32 0.44 0.5123 CD 0.040 1 0.040 0.054 0.8175 CE 0.61 1 0.61 0.83 0.3717 DE 0.22 1 0.22 0.29 0.5924 A2 0.59 1 0.59 0.80 0.3789 B2 0.10 1 0.10 0.14 0.7100 C2 10.70 1 10.70 14.56 0.0008 ** D2 2.44 1 2.44 3.31 0.0807 E2 0.21 1 0.21 0.29 0.5941 残差 18.38 25 0.74 失拟项 16.57 20 0.83 2.29 0.1818 纯误差 1.81 5 0.36 总和 504.21 45 R2 0.9635 R2Adj 0.9344
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