超声辅助酶改性小球藻蛋白工艺优化及其乳化特性的分析

杨清馨; 刘少伟; 韩莉君

doi:10.13386/j.issn1002-0306.2022080075

超声辅助酶改性小球藻蛋白工艺优化及其乳化特性的分析

doi: 10.13386/j.issn1002-0306.2022080075

华东理工大学食品科学与工程系生物反应器工程国家重点实验室，上海 200237

详细信息

作者简介:
杨清馨（1998−），女，硕士研究生，研究方向：食品蛋白加工、食品生物技术等，E-mail：dorayqx@163.com

通讯作者:
刘少伟（1972−），男，博士，教授，研究方向：食品加工、谷物、乳制品食品开发等，E-mail：swliu@ecust.edu

中图分类号: TS201.2
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出版历程
- 收稿日期: 2022-08-09
- 刊出日期: 2023-05-15

Optimization of Ultrasound-Assisted Enzyme Modification Process of Chlorella Protein and Analysis of Its Emulsification Properties

The State Key Laboratory of Bioreactor Engineering, Department of Food Science and Engineering, East China University of Science and Technology, Shanghai 200237, China

摘要

摘要: 本文以小球藻蛋白为研究对象，探究了超声辅助酶改性条件优化及改性后乳化特性的变化。选择超声时间、超声功率、酶添加量、酶反应时间为考察因素，使用层次分析法对乳化活性、乳化稳定性和乳析指数分别赋予权重，并以综合加权得分为响应值，采用四因素三水平的响应面法探究超声辅助酶改性小球藻蛋白乳化特性的最佳改性条件。结果表明，最佳改性工艺条件为：超声时间31 min、超声功率209 W、酶添加量3.1%、酶反应时间3 h。在此优化条件下，乳化性能的最佳评分可以达到98.99，与预测值接近。同时，改性小球藻蛋白的溶解性和起泡性与原蛋白相比分别提高了68.30%和49.44%，但持水性和持油性的改善效果并不明显。综上，该研究为小球藻蛋白在食品乳化体系中的应用提供了一定的理论基础。
- 小球藻蛋白 /
- 乳化特性 /
- 超声改性 /
- 酶改性 /
- 响应面 /
- 层次分析法
Abstract: The optimized conditions of ultrasound-assisted enzyme modification and the effects of modification on the emulsification properties of Chlorella protein were investigated in this paper. The ultrasonic time, ultrasonic power, enzyme addition and enzyme reaction time were selected as the investigating factors. The analytic hierarchy process (AHP) was used to weight the emulsification activity, emulsification stability and creaming index, and the comprehensive weighted score was used as the response value. A four-factor, three-level response surface method was used to investigate the optimal modification conditions for the emulsification properties of ultrasound-assisted enzyme-modified Chlorella protein. The results showed that the optimal modification conditions were as follows: Ultrasonic time 31 min, ultrasonic power 209 W, enzyme addition 3.1% and enzyme reaction time 3 h. Under the optimized conditions, the best score of emulsification performance was 98.99, which was close to the predicted value. Compared with the original protein, the solubility and foaming ability of modified Chlorella protein increased by 68.30% and 49.44%, respectively. However, the modification of water-holding and oil-holding properties showed no significant effect. In summary, this study could provide certain theoretical basis for the application of Chlorella protein in food emulsification systems.
- Chlorella protein /
- emulsifying characteristics /
- ultrasonic modification /
- enzymatic modification /
- response surface methodology /
- analytical hierarchy process

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图 1 超声时间对小球藻蛋白乳化性能及其综合得分的影响

Figure 1. Effects of ultrasonic time on the emulsifying property and comprehensive score of Chlorella protein

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图 2 超声功率对小球藻蛋白乳化性能及其综合得分的影响

Figure 2. Effects of ultrasonic power on the emulsifying property and comprehensive score of Chlorella protein

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图 3 酶添加量对小球藻蛋白乳化性能及其综合得分的影响

Figure 3. Effects of enzyme addition on the emulsifying property and comprehensive score of Chlorella protein

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图 4 酶反应时间对小球藻蛋白乳化性能及其综合得分的影响

Figure 4. Effects of enzyme reaction time on the emulsifying property and comprehensive score of Chlorella protein

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图 5 各因素交互作用对小球藻蛋白乳化特性影响的响应面图

Figure 5. Response surface diagram of the interaction of various factors on the emulsifying characteristics of Chlorella protein

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表 1 响应面试验因素水平表

Table 1. Factors and levels for response surface test

水平	因素
水平	A超声时间（min）	B超声功率（W）	C酶浓度（%）	D酶反应时间（h）
−1	20	150	2	2.5
0	30	200	3	3.0
1	40	250	4	3.5

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表 2 目标树层次评分标准^[23]

Table 2. Evaluation standard of target tree at different levels^[23]

相对重要性	定义
1	同等重要
3	稍微重要
5	明显重要
7	强烈重要
9	绝对重要
2,4,6,8	以上相邻指标的中间值
倒数	若i与j的重要性之比为A_ij，则j与i的重要性之比为1/A_ij

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表 3 3种指标的比较矩阵表

Table 3. Decision matrix of paried comparison on three indexes

指标	乳化活性	乳化稳定性	乳析指数	特征向量	权重值（%）	最大特征值	CI值	CR值
乳化活性	1	1	5/3	1.154	38.462	3.001	0.0005	0.0009
乳化稳定性	1	1	5/3	1.154	38.462
乳析指数	3/5	3/5	1	0.692	23.077
注：CI=（最大特征根−n）/（n−1）；CR=CI/RI。

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表 4 响应面设计方案与结果

Table 4. The design and results of response surface

实验编号	A	B	C	D	EAI（m²/g）	ESI（min）	CI（%）	综合得分
1	−1	−1	0	0	25.77	109.22	25.55	75.13
2	1	−1	0	0	28.21	115.23	23.17	82.42
3	−1	1	0	0	29.54	111.74	22.73	83.29
4	1	1	0	0	30.13	115.49	20.97	87.20
5	0	0	−1	−1	24.88	112.78	24.72	76.30
6	0	0	1	−1	28.83	114.92	22.64	83.63
7	0	0	−1	1	26.06	112.69	22.42	80.32
8	0	0	1	1	28.86	114.17	20.28	85.42
9	−1	0	0	−1	26.33	116.78	22.49	81.76
10	1	0	0	−1	27.91	114.96	22.08	83.36
11	−1	0	0	1	28.64	111.49	22.02	83.14
12	1	0	0	1	30.19	113.96	19.81	88.22
13	0	−1	−1	0	24.72	110.34	25.52	74.72
14	0	1	−1	0	25.44	112.24	23.13	78.67
15	0	−1	1	0	27.90	113.02	23.98	80.44
16	0	1	1	0	30.69	116.74	20.13	89.19
17	−1	0	−1	0	25.55	112.98	25.76	75.79
18	1	0	−1	0	28.43	114.77	23.77	81.78
19	−1	0	1	0	29.16	115.34	22.15	84.70
20	1	0	1	0	27.02	117.01	20.13	85.44
21	0	−1	0	−1	26.83	109.77	25.65	76.28
22	0	1	0	−1	29.73	111.84	22.43	83.88
23	0	−1	0	1	24.12	110.77	19.97	80.70
24	0	1	0	1	29.07	112.67	19.85	86.60
25	0	0	0	0	36.79	121.82	19.18	98.28
26	0	0	0	0	35.62	126.22	19.56	97.93
27	0	0	0	0	36.15	125.71	19.03	98.97
28	0	0	0	0	35.08	125.13	18.87	97.88
29	0	0	0	0	35.27	124.98	19.14	97.70

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表 5 响应面方差分析

Table 5. Variance analysis of response surface

方差来源	平方和	自由度	均方	F	P	显著性
模型	1472.82	14	105.13	158.33	<0.0001	**
A-超声时间	50.47	1	50.47	76.01	<0.0001	**
B-超声功率	127.66	1	127.66	192.26	<0.0001	**
C-酶添加量	141.73	1	141.73	213.44	<0.0001	**
D-酶反应时间	30.69	1	30.69	46.22	<0.0001	**
AB	2.86	1	2.86	4.30	0.0570
AC	6.89	1	6.89	10.38	0.0062	*
AD	3.03	1	3.03	4.56	0.0509
BC	5.76	1	5.76	8.67	0.0106	*
BD	0.72	1	0.72	1.09	0.3146
CD	1.24	1	1.24	1.87	0.1928
A²	323.59	1	323.59	487.33	<0.0001	**
B²	499.67	1	499.67	752.51	<0.0001	**
C²	530.30	1	530.30	798.64	<0.0001	**
D²	354.30	1	354.30	533.58	<0.0001	**
残差	9.30	14	0.67
失拟项	8.28	10	0.83	3.27	0.1356
纯误差	1.01	4	0.25
总误差	1481.12	28
R²=0.9937 R²_Adj=0.9874 CV（%）=0.9609
Adequate Precision：39.7870
注：*为P<0.0001，差异极显著；为P<0.05，差异显著。

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表 6 超声辅助酶改性对小球藻蛋白功能特性的影响

Table 6. Effect of ultrasound assisted enzymatic modification on the functional properties of Chlorella protein

样品	溶解性（%）	持水性（g·g⁻¹）	持油性（g·g⁻¹）	起泡性（%）	泡沫稳定性（%）
小球藻蛋白	32.75±1.83	1.51±0.03	3.11±0.03	45.83±1.45	133.33±3.98
改性小球藻蛋白	55.12±2.47	1.74±0.01	3.42±0.04	68.49±3.01	120.97±2.66
注：以上经t检验均在P<0.05水平上差异显著。

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