响应面法优化鸡肉蛋白酶解工艺

于亚辉; 陈沁雯; 李晓婷; 方婷

doi:10.19303/j.issn.1008-0384.2020.01.012

响应面法优化鸡肉蛋白酶解工艺

doi: 10.19303/j.issn.1008-0384.2020.01.012

福建农林大学食品科学学院，福建　福州　350002

基金项目: 国家自然科学基金项目(31401597、31601393)

详细信息

作者简介:
于亚辉（1993−），男，硕士，研究方向：食品风味化学（E-mail：2951588827@qq.com）

通讯作者:
方婷（1981−），女，博士，教授，研究方向：食品加工理论与应用（fangting930@163.com）

中图分类号: TS 254.1
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出版历程
- 收稿日期: 2019-09-22
- 修回日期: 2019-10-25
- 刊出日期: 2020-01-01

Optimized Proteolysis of Chicken Protein for Food Flavoring

College of Food Science, Fujian Agriculture and Forestry University, Fuzhou, Fujian　350002, China

摘要

摘要: 目的采用双酶分步酶解方法优化鸡肉蛋白酶解工艺，为制备品质较佳的鸡肉香精提供技术参考。方法以动物蛋白酶与复合蛋白酶为目标酶，游离氨基酸、多肽含量为评价指标，在单因素试验的基础上，采用Plackett-Burman设计筛选出重要影响因素。接着采用最陡爬坡试验逼近最大响应区域，确定响应面优化试验中心点，最后选取游离氨基酸、多肽含量以及热反应产物风味评分3个指标为评价依据，采用Box-Behnken响应面优化法，探讨鸡肉蛋白双酶分步酶解工艺及其对制备鸡肉香精风味的影响，确定重要参数的最佳水平。结果总加酶量、总时长、固液比为主要影响因素，通过响应面及岭脊分析确定参数值分别为0.82%、7.26 h、1 ∶ 4.40，预测综合得分为89.41，实际得分88.97，模型可靠。结论通过多指标响应面法优化了鸡肉蛋白酶解条件，提高酶解效果，并制备出风味较佳的鸡肉香精。
- 双酶 /
- 分步酶解工艺 /
- Plackett-Burman设计 /
- 响应面优化
Abstract: Objective The stepwise enzymatic hydrolysis was applied to hydrolyze chicken protein for the preparation of a food flavoring ingredient with improved quality. Method An animal and a compound proteolytic enzyme were used to digest the chicken protein. Free amino acid and peptide contents in the hydrolyzed material were employed as the criteria to initially identify the critical process evaluation parameters based on a single factor Plackett-Burman design. A steepest ascent test was conducted to approximate the maximum response area and arrive at the center point in a response surface experiment. Then, according to the selected indicators for the Box-Behnken experiment, optimized process conditions were determined. Result The selected indicators for the Box-Behnken optimization included the free amino acid and polypeptide contents as well as the flavor scores on the thermal reaction products of the hydrolyzed compounds. The optimal processing conditions were found to include the addition of 0.82% enzymes on the chicken protein substrate, a solid-liquid ratio of 1 ∶ 4.40, and a reaction duration of 7.26 h to achieve an actual composite evaluation score of 88.97 in comparison to the predicted 89.41. The method was deemed reliable. Conclusion The multi-indicator response surface method effectively optimized the enzymatic proteolysis process of chicken protein materials in producing a high quality flavoring ingredient for food applications.
- enzymatic proteolysis /
- stepwise hydrolysis /
- Plackett-Burman design /
- response surface optimization

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图 1 单因素试验结果

Figure 1. Single factor test results

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图 2 总加酶量、总时长和固液比交互作用对游离氨基酸浓度及多肽浓度影响的响应面和等高线

Figure 2. Response surface and contour lines on interactions among enzyme applied/process duration/solid-liquid ratio for process and free amino acid/peptide concentrations of hydrolysate

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表 1 鸡肉风味基料评价标准

Table 1. Evaluation standards on chicken flavor

评价项目 Evaluation project	满分标准 Full score standard	评价分数范围 Evaluation score range
香味 Fragrance	香气浓郁、饱满、圆润与炖煮鸡肉香气较为一致 The aroma is rich, full, round and the chicken aroma is more consistent	0～30
色泽 Color	红褐色、色泽均匀无明显异色 Reddish brown, uniform color, no obvious color	0～30
滋味 Taste	口感柔和具有鸡肉滋味、无苦味等不适感 The palate is soft and has a taste of chicken and no bitterness.	0～20
异味 Odor	无异味、焦糊味 No odor, burnt smell	0～20

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表 2 单因素试验设计

Table 2. Single factor test design

因素 Factor	处理组 Group
因素 Factor	1	2	3	4	5
总加酶量 Total enzyme amount /%	0.2	0.4	0.6	0.8	1.0
加酶比 Enzyme ratio	1∶1	1∶2	2∶1	1∶3	3∶1
固液比 Solid-liquid ratio	1∶1	1∶2	1∶3	1∶4	1∶5
总时长 Total length/h	4	5	6	7	8
时长比 Duration ratio	1∶1	1∶2	2∶1	2∶3	3∶2
NaCl添加量 NaCl addition amount/%	0.4	0.6	0.8	1.0	1.2

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表 3 Plackett-Burman设计试验参数和水平

Table 3. Factors and levels of PIackett-Burman design

编号 Code	变量 Variable	水平（−1） Level （−1）	水平（1） Level (1)
A	总加酶量 Total enzyme amount /%	0.4	0.8
B	加酶比 Enzyme ratio	1∶1	3∶1
C	总时长 Total length/h	5	7
D	时长比 Duration ratio	3∶2	2∶1
E	固液比 Solid-liquid ratio	1∶2	1∶4
F	NaCl添加量 NaCl addition amount/%	0.6	1.0

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表 4 Plackett-Burman试验设计及结果

Table 4. Plackett-Burman design and experimental results

试验号 Test number	A	B	C	D	E	F	氨基酸含量 Amino acid content /(mg·g⁻¹)	多肽含量 Peptide content/ (mg·hg⁻¹)	综合评分 Comprehensive score (Y)
1	1	1	−1	1	1	1	337.73	278.67	103.46
2	1	−1	−1	−1	1	−1	320.65	266.78	98.65
3	−1	−1	1	−1	1	1	257.71	225.08	81.33
4	−1	1	1	−1	1	1	265.43	235.96	84.56
5	−1	−1	−1	1	−1	1	222.03	168.44	65.19
6	1	−1	1	1	1	−1	342.45	280.19	104.45
7	−1	1	1	1	−1	−1	234.04	185.24	70.19
8	−1	1	−1	1	1	−1	247.69	215.66	78.04
9	1	1	1	−1	−1	−1	287.58	248.37	90.22
10	1	1	−1	−1	−1	1	263.58	221.64	81.54
11	1	−1	1	1	−1	1	300.36	245.71	91.6
12	−1	−1	−1	−1	−1	−1	207.75	156.49	60.78

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表 5 Plackett-Burman试验显著性因子选择模型方差分析

Table 5. ANOVA of Plackett-Burman test on selected factorial model

变异来源 Source of variation	自由度 Degree of freedom	效应 Effect	F值 F value	P值 P value	显著性 Significance
总模型 Total model	6	84.17	95.15	＜0.000 1	**
A	1	10.82	361.09	＜0.000 1	**
B	1	0.50	0.77	0.419 3
C	1	2.89	25.78	0.003 8	**
D	1	1.32	5.38	0.068 1
E	1	7.58	177.28	＜0.000 1	**
F	1	0.45	0.61	0.469 0
注：：P＜0.05显著；：P＜0.01极显著。 Note: : P＜0.05 significant; **: P＜0.01 extremely significant.

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表 6 最陡爬坡试验设计及结果

Table 6. Design and results of steepest ascent experiment

试验号 Test number	A/%	C/h	E	氨基酸含量 Amino acid content /(mg·g⁻¹)	多肽含量 Peptide content/ (mg·hg⁻¹)	综合评 Comprehensive score (Y)
1	0.2	4	1∶1	129.48	87.31	34.49
2	0.4	5	1∶2	223.59	168.63	62.74
3	0.6	6	1∶3	326.27	261.65	94.33
4	0.8	7	1∶4	360.43	301.25	106.38
5	1	8	1∶5	351.96	286.77	102.56

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表 7 Box-Behnken Design试验因素与水平

Table 7. Levels and factors of Box-Behnken experiment

编号 Code	因素 Factor	水平 Level
编号 Code	因素 Factor	−1	0	1
A	总加酶量 Total enzyme amount /%	0.6	0.8	1
B	总时长 Total length / h	6	7	8
C	固液比 Solid-liquid ratio	1∶3	1∶4	1∶5

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表 8 Box-Behnken Design试验设计及结果

Table 8. Box-Behnken design and experimental results

序号 Serial number	A	B	C	氨基酸含量 Amino acid content/(mg·g⁻¹)	多肽含量 Peptide content/(mg·hg⁻¹)	感官评分 Sensory score (P)	综合评分 Comprehensive score (Y)
1	−1	0	−1	321.79	264.82	46.33	67.46
2	0	0	0	351.96	281.77	83.15	89.37
3	0	1	1	358.28	286.36	69.53	83.38
4	−1	0	1	320.54	260.88	70.65	79.21
5	−1	1	0	333.88	265.63	56.38	73.39
6	0	0	0	352.55	280.92	78.18	86.86
7	0	0	0	354.02	281.96	79.65	87.78
8	0	−1	1	340.34	270.52	68.47	80.29
9	0	0	0	353.88	283.03	80.25	88.16
10	−1	−1	0	350.08	275.15	44.87	69.55
11	0	0	0	351.64	283.77	81.37	88.62
12	1	−1	0	355.25	288.32	43.77	70.45
13	1	0	1	356.28	290.63	61.28	79.47
14	0	−1	−1	344.65	265.58	50.33	71.11
15	1	1	0	365.99	292.06	62.43	80.84
16	0	1	−1	355.29	282.05	64.67	80.39
17	1	0	−1	345.37	270.93	55.05	73.96

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表 9 响应面回归模型方差分析

Table 9. ANOVA on response surface regression model

方差来源 Source of variance	平方和 Sum of square	自由度 Degree of freedom	均方 Mean square	F值 F value	P值 P value	显著性 Significance
总模型 Total model	853.19	9	94.80	88.54	＜0.000 1	**
A	28.55	1	28.55	26.66	0.001 3	**
B	88.38	1	88.38	82.54	＜0.000 1	**
C	108.26	1	108.26	101.11	＜0.000 1	**
AB	10.71	1	10.71	10.00	0.015 9	*
AC	9.73	1	9.73	9.09	0.019 5	*
BC	9.57	1	9.57	8.93	0.020 3	*
A²	355.03	1	355.03	331.58	＜0.000 1	**
B²	123.55	1	123.55	115.39	＜0.000 1	**
C²	65.67	1	65.67	61.34	0.000 1	**
残差 Residual	7.49	7	1.07
失拟项 Missing item	3.97	3	1.32	1.50	0.342 8
绝对误差 Absolute error	3.53	4	0.88
总和 Sum	860.68	16
相关系数 Correlation coefficient	0.991 3
校正决定系数 Correction decision coefficient	0.980 1
变异系数 Coefficient of variation	1.30
信噪比 Signal to noise ratio	25.528＞4
注：：P＜0.05显著；：P＜0.01极显著。 Note: : P＜0.05 significant; **: P＜0.01 extremely significant.

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