响应面法优化工业化独立发酵凝固型发酵乳的发酵工艺

黄娟; 黄燕燕; 彭小霞

doi:10.13386/j.issn1002-0306.2021120031

响应面法优化工业化独立发酵凝固型发酵乳的发酵工艺

doi: 10.13386/j.issn1002-0306.2021120031

黄娟^{1, 2, ,},
黄燕燕³,
彭小霞²

1.
华南理工大学食品科学与工程学院，广东广州 510640
2.
广东燕塘乳业股份有限公司，广东广州 511356
3.
佛山科学技术学院食品科学与工程学院，广东佛山 528225

详细信息

作者简介:
黄娟（1991−），女，博士，助理工程师，研究方向：微生物利用与乳品加工，E-mail：1185537582@qq.com

通讯作者:
黄娟（1991−），女，博士，助理工程师，研究方向：微生物利用与乳品加工，E-mail：1185537582@qq.com

中图分类号: TS201.1
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- 文章访问数: 19
- HTML全文浏览量: 4
- PDF下载量: 8
- 被引次数: 0
出版历程
- 收稿日期: 2021-12-06
- 网络出版日期: 2022-06-19
- 刊出日期: 2022-08-03

Optimization of the Fermentation Process of Industrialized Independent Fermentation Coagulated Fermented Milk by Response Surface Methodology

1.
School of Food Science and Engineering, South China University of Technology, Guangzhou 510640, China
2.
Guangdong Yantang Dairy Co., Ltd., Guangzhou 511356, China
3.
School of Food Science and Engineering, Foshan University, Foshan 528225, China

摘要

摘要: 工业化独立发酵凝固型发酵乳受发酵温度波动、发酵时长及发酵后剪切破坏等因素的影响，易出现成品乳清析出量过多和凝固状态不佳等现象，本研究通过单因素实验初步探究了发酵温度波动、发酵温度、发酵时长、发酵菌种配比及模拟工业化灌装后破坏对产品质构和风味的影响，在单因素实验的基础上，设计响应面法Box-Behnken组合试验，对发酵温度、发酵时长、发酵菌种配比工艺参数进行了优化。单因素实验结果显示：灌装后发酵2 h内剪切破坏影响不大，但超出时间后将严重影响产品质构和酸化速度，灌装后发酵温度波动将显著影响产品的酸化速度和不良率，这提示工业化发酵过程中应当尽量缩短灌装后段液态乳在常温环境中的暂存时间，由于机器故障导致的灌装延迟超过2 h的液态乳应直接发酵为成品再进行二次利用，不适宜再经管道剪切后进行灌装。响应面模型最优解的结果表明，在实际发酵温度42 ℃，发酵时长5.6 h，菌种配比1:1最佳工艺条件条件下制备的工业化独立发酵凝固型发酵乳的质构完好率为100%，终止发酵的酸度为68.56°T，冷库后放置12 h的酸度为70.2°T，发酵乳的综合感官评分为8.24分，与预测值接近，证明模型有效。流变学特性曲线结果显示该发酵乳的抗剪切能力较弱，200 s⁻¹高剪切后的酸奶黏度恢复率仅为25.55%，符合脆软嫩滑形式的凝固型发酵乳的感官特点。响应面法可有效拟合该凝固型发酵乳的发酵温度、发酵时长、发酵菌种配比之间的关系，为工业化生产中不可避免的菌种配比称量误差、发酵温度堆热传热波动、人为取样时机导致的发酵时长误差等情况给予了一定的理论指导，方便在实际操作中，快速提出相应的对策。
- 凝固型发酵乳 /
- 感官评价 /
- 工业化 /
- 独立发酵 /
- 剪切破坏 /
- 温度波动
Abstract: Industrialized independent fermentation coagulated fermented milk is prone to the phenomenon of excessive whey precipitation and poor coagulation state of the finished product affected by factors such as fermentation temperature fluctuation, fermentation time and shear damage after fermentation. The effects of fermentation temperature fluctuation, fermentation temperature, fermentation time, fermentation strain ratio, and simulated industrial filling damage on product texture and flavor were studied. On the basis of single-factor experiments, a response surface methodology Box-Behnken combination experiment was designed. The process parameters of fermentation temperature, fermentation time and fermentation strain ratio were optimized. The single-factor experiment results showed that the shear damage had little effect within 2 hours of fermentation after filling, but it could seriously affect the product texture and acidification speed when the time exceeded, suggesting that in the industrial fermentation process, the temporary storage time of the liquid milk in the room temperature environment should be shortened as much as possible, and the liquid milk waiting for filling delayed over 2 hours due to machine failure should be directly fermented into finished products for secondary use. It wasn’t suitable for filling after cutting through the pipeline. The results of the optimal solution of the response surface model showed that the texture integrity rate of the industrialized independent fermented coagulated fermented milk prepared under the optimal technological conditions of the actual fermentation temperature of 42 ℃, the fermentation time of 5.6 h, and the bacterial species ratio of 1:1 was 100%, the acidity of terminating fermentation was 68.56°T, and the acidity of 12 h after cold storage was 70.2°T. The comprehensive sensory score of fermented milk was 8.24, which was close to the predicted value, proving the validity of the model. The results of the rheological characteristic curve showed that the fermented milk had weak shear resistance, and the yogurt viscosity recovery rate after 200 s⁻¹ high shear was only 25.55%, which was in line with the sensory characteristics of the coagulated fermented milk in the form of crisp, soft and smooth. The response surface methodology can effectively fit the relationship among the fermentation temperature, fermentation time and fermentation strain ratio of the coagulated fermented milk. It provides certain theoretical guidance for the unavoidable error of strain proportioning and weighing, the fluctuation of fermentation temperature and heat transfer, and the error of fermentation time caused by artificial sampling opportunity in industrial production, which is convenient for putting forward corresponding countermeasures quickly in practical operation.
- coagulated fermented milk /
- sensory evaluation /
- industrialization /
- independent fermentation /
- shear failure /
- temperature fluctuation

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图 1 发酵温度和发酵时长对发酵酸度、pH的影响

Figure 1. The influence of fermentation temperature and fermentation time on fermentation acidity and pH

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图 2 菌种比例对产品酸化速度的影响

Figure 2. The influence of strain ratio on the acidification speed of products

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图 3 各因素间交互作用对凝固型发酵乳感官评分影响的响应面及等高线

Figure 3. Response surface and contour lines of the interaction of various factors on the sensory score of coagulated fermented milk

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图 4 最优条件下凝固型发酵乳的酸化曲线

Figure 4. Acidification curve of coagulated fermented milk under optimal condition

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图 5 最优条件下凝固型发酵乳的流变学三段式剪切扫描曲线

Figure 5. Rheological three-section shear scan curve of coagulated fermented milk under optimal condition

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表 1 工业化独立发酵凝固型发酵乳的温度波动实验设计

Table 1. Experimental design of temperature fluctuation of industrialized independent fermentation coagulated fermented milk

组别	10 ℃	25 ℃	30 ℃	42 ℃
1	1 h	0	0	6 h
2	1 h	0	1 h	6 h
3	1 h	0	1 h	30→42 ℃（6 h）^*
4	0	0	1 h	6 h
5	0	1 h	0	6 h
6	0	0	0	6 h
注：每个组别均是从左至右进行的温度波动模拟实验，*表示该温度波动过程为培养箱温度由30 ℃改设置为42 ℃的缓慢升温变化过程，非直接将产品放入已升温至42 ℃的环境中。

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表 2 工业化独立发酵凝固型发酵乳发酵工艺的BBD因素水平设计

Table 2. BBD factor level design of industrialized independent fermentation coagulated fermented milk fermentation process

水平编码值	因素
水平编码值	A发酵温度（℃）	B发酵时长（h）	C菌种比例（DCU/DCU）
−1	38	4.0	1:2
0	40	5.0	1:1
1	42	6.0	3:2
注：C菌种比例是指复配菌种型号1和型号2菌种的比例，DCU为丹尼斯克公司的菌活力标识单位。

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表 3 凝固型发酵乳产品的感官评分标准

Table 3. Sensory scoring standard for coagulated fermented milk products

评价项目	计分权重w_i	单一评价项目得分x_i
评价项目	计分权重w_i	1~4分	5~7分	8~10分
镜面光滑度	10%	不能接受	一般接受	非常接受
香气（自然浓郁程度）	5%	平淡	一般	浓郁
冷藏后搅拌破坏后的质构	15%	碎块较多	一般丝滑，存在些许碎块	非常丝滑
味道（按喜好程度打分）	20%	酸、涩等不良风味	一般接受	酸甜香感整体怡人
发酵过程状态（是否有明显分层或乳清析出等情况）	10%	严重析水，产品离壁	轻度不佳	无任何异常
冷藏后熟化状态（是否有明显分层或乳清析出等情况）	25%	严重析水，产品离壁	轻度不佳	无任何异常
粘稠度等触感（按喜好程度打分）	15%	糊口等不良感官	基本满意	非常满意

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表 4 不同条件下制备的工业化独立发酵凝固型发酵乳的感官评分及不同摇晃次数发酵7 h的产品酸度

Table 4. Sensory scores of industrialized independent fermentation coagulated fermented milk prepared under different conditions and acidity of products fermented for 7 hours with different shaking times

评价项目	计分权重	不同发酵温度（℃）			42 ℃发酵条件不同摇晃次数					42 ℃不同菌种比例			42 ℃不同发酵时间（h）
评价项目	计分权重	43	40	37	0	1	2	3	4	1:0	1:1	0:1	4.5	5.5	6.5
镜面光滑度	10%	6	8	7	9	7^*	6^*	4^**	4^**	4	9	6	9	9	9
香气（自然浓郁程度）	5%	8	8	6	8	8	7	5^**	5^**	4	8	7	6	8	8
冷藏后搅拌破坏后的质构	15%	5	7	8	9	8	6^*	5^**	4^**	4	9	6	7	9	9
味道（按喜好程度打分）	20%	5	8	7	8	8	7	6^*	6^*	4	8	7	7	8	7
发酵过程状态（是否有明显分层或乳清析出等情况）	10%	5	7	7	8	7	6^*	5^**	4^**	4	8	7	8	8	8
冷藏后熟化状态（是否有明显分层或乳清析出等情况）	25%	7	8	7	9	8	7^*	6^**	5^**	4	9	7	7	9	9
粘稠度等触感（按喜好程度打分）	15%	6	7	6	8	7	8	6^*	6^*	7	8	5	9	8	7
综合得分	100%	5.90	7.60	6.95	8.50	7.65^*	6.80^*	5.50^**	5.00^**	4.45	8.50	6.45	7.55	8.50	8.15
发酵7 h的酸度（°T）	−	−	−	−	68.02	65.61^*	65.60^*	64.85^**	63.09^**	−	−	−	−	−	−
注：表中“−”表示不关注其他发酵酸奶样品7 h的酸度且未检测；“”和“*”标识分别表示42 ℃发酵条件摇晃次数≥1的组别分别与不摇晃的组别进行比较的差异显著性水平P<0.05和P<0.01。

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表 5 发酵温度波动对产品不良率及酸化速度的影响

Table 5. The influence of fermentation temperature fluctuation on product defect rate and acidification speed

组别	终点酸度（°T）	终点pH	过夜发酵酸度（°T）	过夜发酵pH	产品不良率（%）
1	59.58±0.53^*	4.38±0.02	80.34±0.65	4.00±0.01	57.14±7.14^*
2	62.35±0.42	4.34±0.01	79.58±0.61^*	4.03±0.01	66.67±8.33^**
3	61.98±0.44^*	4.37±0.01	81.38±0.67	4.02±0.01	71.43±14.29^**
4	63.48±0.26	4.26±0.01	82.75±0.66	3.98±0.02	60.00±6.67^**
5	64.05±0.22	4.30±0.01	83.82±0.71	3.98±0.02	75.00±5.00^**
6	64.96±0.17	4.30±0.01	83.53±0.68	3.98±0.02	38.46±7.69
注：“”和“*”标识分别表示第1~5组（有温度波动）相应数据分别与第6组（无温度波动）进行比较的差异显著性水平P<0.05和P<0.01。

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表 6 基于Box-Behnken设计的不同因素水平组合下的凝固型发酵乳的综合感官得分

Table 6. Comprehensive sensory evaluation of coagulated fermented milk under different factor level combinations based on Box-Behnken design

实验号	A	B	C	综合感官得分
1	0	0	0	8.10
2	−1	−1	0	7.60
3	1	−1	0	7.90
4	−1	0	1	5.80
5	1	0	1	6.10
6	1	0	−1	7.00
7	0	0	0	7.90
8	0	0	0	7.95
9	0	1	1	6.05
10	1	1	0	8.10
11	0	0	0	7.95
12	0	−1	1	5.25
13	0	0	0	7.85
14	0	1	−1	6.70
15	−1	1	0	8.10
16	0	−1	−1	6.30
17	−1	0	−1	6.35

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表 7 基于Box-Behnken设计的响应曲面二次模型的方差分析

Table 7. Analysis of variance for quadratic model of response surface based on Box-Behnken design

来源	平方和	自由度	均方和	F值	P值
模型	14.99	9	1.67	91.91	<0.0001^**
A-发酵温度	0.1953	1	0.1953	10.78	0.0134^*
B-发酵时长	0.4513	1	0.4513	24.90	0.0016^**
C-菌种比例	1.24	1	1.24	68.43	<0.0001^**
AB	0.0225	1	0.0225	1.24	0.3020
AC	0.0306	1	0.0306	1.69	0.2348
BC	0.0400	1	0.0400	2.21	0.1810
A²	0.0475	1	0.0475	2.62	0.1494
B²	0.0725	1	0.0725	4.00	0.0856
C²	12.80	1	12.80	706.36	<0.0001^**
残差	0.1269	7	0.0181
失拟项	0.0919	3	0.0306	3.50	0.1288
净误差	0.0350	4	0.0088
总和	15.12	16
注：差异显著（P<0.05）；*差异极显著（P<0.01）。

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