黑豆粕饮料发酵工艺的优化及其抗氧化活性评价

陈惠敏; 吴晓平; 汪少芸

doi:10.13386/j.issn1002-0306.2022040291

黑豆粕饮料发酵工艺的优化及其抗氧化活性评价

doi: 10.13386/j.issn1002-0306.2022040291

福州大学生物科学与工程学院，福建福州 350108

基金项目: 宁夏回族自治区重点研发计划（2021BEF02013）；中央引导地方科技发展项目（2020L3004）。

详细信息

作者简介:
陈惠敏（1993−），女，博士研究生，研究方向：生物活性多肽与功能食品，E-mail：358245371@qq.com

通讯作者:
汪少芸（1970−），女，博士，教授，研究方向：食品生物化学，E-mail：shywang@fzu.edu.cn

中图分类号: TS278
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出版历程
- 收稿日期: 2022-04-25
- 网络出版日期: 2023-01-12
- 刊出日期: 2023-03-01

Optimization of Fermentation Technology and Evaluation of Antioxidant Activity of Black Soybean Meal Beverage

College of Food Science and Engineering, Fuzhou University, Fuzhou 350108, China

摘要

摘要: 以黑豆粕为原料，通过乳酸菌发酵制备具有抗氧化活性的乳酸饮料。实验采用单因素和响应面分析法优化乳酸菌发酵工艺，以DPPH•清除率为指标，考察料液比、接种量、发酵温度、发酵时间对发酵过程的影响；再通过DPPH•清除实验、•OH清除实验、ABTS⁺•清除实验、脂质过氧化抑制实验来评价发酵产物的抗氧化功效。结果表明：最佳发酵菌株为嗜热链球菌，最佳的发酵工艺为料液比1:5 g/mL、接种量5%、温度37.6 ℃、时间36 h。在最佳发酵工艺下，发酵液的DPPH•清除率为71.56%。黑豆粕发酵产物的DPPH、OH、ABTS自由基的半抑制浓度分别为2.43、1.30和0.37 mg/mL，且有效抑制脂质过氧化反应，说明发酵产物具有较好的体外抗氧化活性。饮料的感官评价可达90.21，悬浮稳定性为96.8%。本研究将为开发以黑豆粕为原料的功能性食品提供一个新思路，也为实现农副产品的进一步开发提供参考。
- 黑豆粕 /
- 发酵 /
- 抗氧化 /
- 嗜热链球菌 /
- 功能性乳酸饮料
Abstract: Black soybean meal was used as raw material to prepare lactic acid beverage with antioxidant activity through lactic acid bacteria fermentation. Single factor and response surface methodology were used to optimize the fermentation process of lactic acid bacteria. The effects of the solid-liquid ratio, inoculation quantity, fermentation temperature, and fermentation time on the fermentation process were investigated with DPPH radical scavenging ratio as an index. The DPPH radical scavenging experiment, OH radical scavenging experiment, ABTS radical scavenging experiment, and lipid peroxidation inhibition experiment were tested to assess the antioxidant effect of fermented products. The results showed that Streptococcus thermophilus was the best fermentation strain, and the optimum fermentation processes were solid-liquid ratio 1:5 g/mL, inoculation quantity 5%, fermentation temperature 37.6 ℃, and fermentation time 36 h, respectively. Under the optimal fermentation conditions, the DPPH radical scavenging ratio was 71.56%. The semi-inhibitory concentrations of DPPH, OH and ABTS free radicals in the fermentation products of black soybean meal were 2.43, 1.30 and 0.37 mg/mL, respectively, and the lipid peroxidation was effectively inhibited, indicating that the fermentation products had good antioxidant activity in vitro. The sensory evaluation and suspension stability of beverage were 90.21 and 96.8%, respectively. This study provides a new idea for the development of functional foods with black soybean meal as raw material, and also provides a reference for the further development of agricultural by-products.
- black soybean meal /
- fermentation /
- antioxidant activity /
- Streptococcus thermophilus /
- functional lactic acid beverage

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图 1 菌种生长曲线

Figure 1. Growth curve of strains

注：（a）嗜热链球菌；（b）发酵乳杆菌；（c）德氏乳杆菌。

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图 2 料液比对黑豆粕发酵的影响

Figure 2. Effect of solid-liquid ratio on fermentation of black soybean meal

注：不同字母表示差异显著（P<0.05），图3~图5同。

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图 3 接种量对黑豆粕发酵的影响

Figure 3. Effect of inoculation quantity on fermentation of black soybean meal

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图 4 发酵温度对黑豆粕发酵的影响

Figure 4. Effect of fermentation temperature on fermentation of black soybean meal

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图 5 发酵时间对黑豆粕发酵的影响

Figure 5. Effect of fermentation time on fermentation of black soybean meal

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图 6 料液比和接种量交互作用对发酵液DPPH•清除率影响的响应面图

Figure 6. Response surface plot showing the effect of solid-liquid ratio and inoculation quantity of the scavenging ratio of DPPH radical of fermentation forth

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图 7 料液比和发酵温度交互作用对发酵液DPPH•清除率影响的响应面图

Figure 7. Response surface plot showing the effect of solid-liquid ratio and fermentation temperature of the scavenging ratio of DPPH radical of fermentation forth

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图 8 接种量和发酵温度交互作用对发酵液DPPH•清除率影响的响应面图

Figure 8. Response surface plot showing the effect of inoculation quantity and fermentation temperature of the scavenging ratio of DPPH radical of fermentation forth

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图 9 不同浓度的发酵产物对DPPH•清除率的影响

Figure 9. Effect of concentration of fermented products on scavenging ratio of DPPH radical

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图 10 不同浓度的发酵产物对•OH清除率的影响

Figure 10. Effect of concentration of fermented products on scavenging ratio of OH radical

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图 11 不同浓度的发酵产物对ABTS⁺•清除率的影响

Figure 11. Effect of concentration of fermented products on scavenging ratio of ABTS radical

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图 12 不同浓度的发酵产物在亚油酸体系中的抗氧化作用

Figure 12. Antioxidant effect of different concentration of fermented products in linoleic acid system

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表 1 响应面试验因素水平表

Table 1. Factors and levels of response surface experiment

因素	水平
因素	−1	0	1
A 料液比（g/mL）	1:10	1:5	3:10
B 接种量（%）	4	5	6
C 发酵温度（℃）	34	37	40
D 发酵时间（h）	24	30	36

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表 2 饮料的感官评价方法

Table 2. Sensory evaluation methods for beverages

评价项目	评价指标	得分（分）
色泽形态	浅黄色，无分层	21~30
	色泽稍深或稍浅，放置一段时间出现分层	11~20
	色泽过深或过浅，分层现象明显	0~10
香味	豆子香味浓郁，清爽的酸味	21~30
	豆子香味较淡，酸味较重	11~20
	酸味过重	0~10
滋味	入口有豆沙的绵密感，酸甜可口	31~40
	滋味偏甜或偏酸，但有原材料的滋味	16~30
	滋味过甜或过酸，原材料滋味被掩盖	0~15

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表 3 不同菌种对黑豆粕发酵情况

Table 3. Fermentation of black soybean meal by different strains

菌种	多肽浓度（μg/mL）	水解度（%）	DPPH自由基清除率（%）
嗜热链球菌	944.18±4.36^a	13.13±0.46^a	64.24±2.62^c
发酵乳杆菌	1508.01±3.98^c	47.03±0.63^c	45.95±2.10^a
德氏乳杆菌	1124.33±2.11^b	26.25±3.24^b	57.35±0.50^b
注：同列不同小写字母表示差异显著（P<0.05）。

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表 4 响应面试验设计与结果

Table 4. Experimental design and results for response surface analysis

试验号	A 料液比（g/mL）	B 接种量（%）	C 发酵温度（℃）	D 发酵时间（h）	DPPH自由基清除率（%）
1	1:10	4	37	30	42.29
2	3:10	4	37	30	56.97
3	1:10	6	37	30	52.49
4	3:10	6	37	30	45.77
5	1:5	5	34	24	58.47
6	1:5	5	40	24	60.19
7	1:5	5	34	36	57.08
8	1:5	5	40	36	62.50
9	1:10	5	37	24	55.97
10	3:10	5	37	24	60.70
11	1:10	5	37	36	54.98
12	3:10	5	37	36	62.69
13	1:5	4	34	30	55.68
14	1:5	6	34	30	46.17
15	1:5	4	40	30	45.37
16	1:5	6	40	30	58.56
17	1:10	5	34	30	52.67
18	3:10	5	34	30	37.59
19	1:10	5	40	30	33.80
20	3:10	5	40	30	59.72
21	1:5	4	37	24	62.19
22	1:5	6	37	24	60.70
23	1:5	4	37	36	61.69
24	1:5	6	37	36	62.69
25	1:5	5	37	30	69.91
26	1:5	5	37	30	70.60
27	1:5	5	37	30	71.30
28	1:5	5	37	30	70.37
29	1:5	5	37	30	69.91

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表 5 发酵工艺模型回归方程方差分析

Table 5. ANOVA of regression equation for fermentation process

变异来源	平方和	自由度	均方和	F值	P值	显著性水平
模型	2633.89	14	188.13	165.67	< 0.0001	***
A 料液比	81.33	1	81.33	71.62	< 0.0001	***
B 接种量	0.40	1	0.40	0.35	0.5625
C 发酵温度	12.98	1	12.98	11.43	0.0045	***
D 发酵时间	0.97	1	0.97	0.85	0.3713
AB	114.49	1	114.49	100.82	< 0.0001	***
AC	420.25	1	420.25	370.07	< 0.0001	***
AD	2.22	1	2.22	1.95	0.1838
BC	128.82	1	128.82	113.44	< 0.0001	***
BD	1.55	1	1.55	1.36	0.2622
CD	3.42	1	3.42	3.01	0.1045
A²	1047.76	1	1047.76	922.65	< 0.0001	***
B²	451.51	1	451.51	397.60	< 0.0001	***
C²	812.13	1	812.13	715.15	< 0.0001	***
D²	0.65	1	0.65	0.57	0.4615
残差	15.90	14	1.14
失拟	14.57	10	1.46	4.38	0.0836	不显著
净误差	1.33	4	0.33
注：*代表P<0.01，代表P<0.05。

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表 6 饮料成分分析与评价

Table 6. Component analysis and evaluation of beverage

组分	蛋白质（%）	粗脂肪（%）	总糖（%）	乳酸（mg/mL）	悬浮稳定性（%）	感官评分（分）
含量	8.69±0.19	6.06±0.85	21.2±1.23	0.36±0.07	96.8±1.2	90.21±2.2

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