响应面法优化蛋壳豆渣发酵工艺生产高钙饲料

陈劭舒; 董丽婷; 夏凤腾; 刘玉洁; 罗灿; 李怡成; 王征

doi:10.13386/j.issn1002-0306.2022070321

响应面法优化蛋壳豆渣发酵工艺生产高钙饲料

doi: 10.13386/j.issn1002-0306.2022070321

陈劭舒^1,,
董丽婷²,
夏凤腾¹,
刘玉洁¹,
罗灿¹,
李怡成¹,
王征^1,*, ,

1.
湖南农业大学生物科学技术学院，湖南长沙 410128
2.
湖南畅想农业科技有限公司，湖南张家界 427000

基金项目: 湖南省长沙市自然科学基金（kq2202218）。

详细信息

作者简介:
陈劭舒（1998−），女，硕士研究生，研究方向：微生物资源研究及利用，E-mail：c986316054@163.com

通讯作者:
王征（1967−），女，博士，教授，研究方向：植物天然产物与动物肠道健康及无抗替代，E-mail：wz8918@126.com

中图分类号: TQ920
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- 文章访问数: 22
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出版历程
- 收稿日期: 2022-08-02
- 刊出日期: 2023-05-01

Optimized the Fermentation Process of Eggshell and Soybean Dregs to Produce High Calcium Feed by Response Surface Methodology

1.
College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha 410128, China
2.
Hunan Imagination Agricultural Science and Technology, Zhangjiajie 427000, China

摘要

摘要: 大量废弃的蛋壳和湿豆渣是良好的钙和氮素资源，为探索一条解决两类农业废弃物合理利用的新途径，本实验选用枯草芽孢杆菌、植物乳杆菌和粪肠球菌固体生料发酵鸡蛋壳、豆渣混合物。以乳酸钙含量为指标，利用单因素实验研究了豆渣蛋壳比例、固水比、葡萄糖添加量、接种量、混合菌种比例、发酵温度和发酵时间对发酵产乳酸钙的影响，并利用响应面试验优化发酵产乳酸钙的工艺条件。实验结果表明：在豆渣蛋壳比例90:10、枯草芽孢杆菌:植物乳杆菌:粪肠球菌为1:1:1（体积比）、固水比1:2.5、葡萄糖添加量15.6%、接菌量14.7%、发酵温度37 ℃、发酵6 d的条件下，蛋壳豆渣混合物中的乳酸钙含量达到13.58%。与发酵前相比，发酵后豆渣饲料中的游离氨基酸、乳酸钙含量显著提高（P<0.05），pH、粗脂肪、可溶性蛋白显著降低（P<0.05）。综上所述，利用益生菌发酵蛋壳与豆渣，可以制备一种高钙饲料。
- 豆渣 /
- 鸡蛋壳 /
- 固体发酵 /
- 乳酸钙 /
- 高钙饲料
Abstract: Discarded eggshells and wet soybean dregs are rich sources of calcium and nitrogen. This study aimed to establish a novel optimized method for the utilization of agricultural wastes. Eggshells and soybean dregs were the primary raw materials used in this study. Bacillus subtilis, Lactobacillus plantarum, and Enterococcus faecalis were the probiotic strains selected for mixed fermentation. Harman’s single-factor test based on the assessment of calcium lactate content in the fermented forage was used to screen the following process parameters: Ratio of soybean dregs to eggshells, solid to water ratio, addition of glucose, inoculation amount, ratio of probiotic strains, fermentation temperature and duration. Optimum fermentation conditions were determined using response surface methodology. The results revealed that the conditions required for optimum fermentation were as follows: Soybean dregs to eggshells ratio of 90:10, solid to water ratio of 1:2.5, addition of 15.6% glucose, inoculation amount of 14.7%, Bacillus subtilis, Lactobacillus plantarum and Enterococcus faecalis in the ratio of 1:1:1 (v/v), and fermentation duration of 6 days at 37 °C. The amount of calcium lactate in the fermented forage was 13.58%. Free amino acid and calcium lactate concentrations in the fermented forage were significantly higher (P<0.05) in comparison to their concentrations in the raw materials. A significant decrease in pH and concentrations of crude fat and soluble protein in the fermented forage was also observed (P<0.05). In conclusion, this study established a novel optimized method for the production of high calcium feed from eggshells and soybean dregs using probiotic fermentation.
- soybean dregs /
- eggshells /
- solid fermentation /
- calcium lactate /
- high calcium feed

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图 1 豆渣和蛋壳比例对乳酸钙含量的影响

注：不同小写字母表示组间差异显著（P<0.05）；图2~图7同。

Figure 1. Effects of ratio of soybean dregs and eggshells on the content of calcium lactate

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图 2 固水比对乳酸钙含量的影响

Figure 2. Effect of solid-water ratio on the content of calcium lactate

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图 3 葡萄糖添加量对乳酸钙含量的影响

Figure 3. Effects of glucose addition on the content of calcium lactate

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图 4 菌种比例对乳酸钙含量的影响

Figure 4. Effects of different mixed starter cultures on the content of calcium lactate

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图 5 接菌量对乳酸钙含量的影响

Figure 5. Effects of inoculation amount on the content of calcium lactate

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图 6 发酵温度对乳酸钙含量的影响

Figure 6. Effects of fermentation temperature on the content of calcium lactate

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图 7 发酵时间对乳酸钙含量的影响

Figure 7. Effects of fermentation time on the content of calcium lactate

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图 8 各因素交互作用对乳酸钙含量影响的响应面曲图

Figure 8. The response surface diagram of interaction among various factors on the content of calcium lactate

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表 1 响应面试验因素与水平

Table 1. Factors and levels of response surface methodology

因素	水平
因素	−1	0	1
A 葡萄糖添加量（%）	8	12	16
B 接菌量（%）	10	15	20
C 温度（℃）	34	37	40

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表 2 响应面分析方案与试验结果

Table 2. Program and experimental data of response surface methodology

试验号	A 葡萄糖添加量	B 接菌量	C 温度	乳酸钙含量（%）
1	−1	−1	0	8.36
2	1	−1	0	13.36
3	−1	1	0	8.73
4	1	1	0	12.00
5	−1	0	−1	8.36
6	1	0	−1	13.09
7	−1	0	1	8.91
8	1	0	1	12.55
9	0	−1	−1	10.82
10	0	1	−1	11.59
11	0	−1	1	11.36
12	0	1	1	10.82
13	0	0	0	12.36
14	0	0	0	12.18
15	0	0	0	12.36
16	0	0	0	12.55
17	0	0	0	12.64

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表 3 回归模型方差分析

Table 3. Variance analysis of regression model

方差来源	平方和	Df	均方值	F值	P值	显著性
模型	45.70	9	5.08	110.59	<0.0001	**
A 葡萄糖添加量	34.60	1	34.60	753.53	<0.0001	**
B 接菌量	0.0747	1	0.0747	1.63	0.2430
C 温度	0.0065	1	0.0065	0.1406	0.7188
AB	0.7460	1	0.7460	16.25	0.0050	**
AC	0.2976	1	0.2976	6.48	0.0383	*
BC	0.4345	1	0.4345	9.46	0.0179	*
A²	5.21	1	5.21	113.50	<0.0001	**
B²	2.02	1	2.02	43.92	0.0003	**
C²	1.41	1	1.41	30.68	0.0009	**
残差	0.3214	7	0.0459
失拟项	0.1958	3	0.0653	2.08	0.2459
纯误差	0.1256	4	0.0314
总离差	46.02	16
R²=0.993	R²_Adj=0.984
注：“*”表示影响极显著，P<0.01；“”表示影响显著，P<0.05。

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表 4 发酵前后蛋壳豆渣混合物营养成分的变化

Table 4. Comparison of nutrients between fermented material and original material

营养成分	发酵前	发酵后
粗灰分（%）	13.38±0.81^a	14.43±0.18^a
粗蛋白（%）	14.19±0.98^a	14.42±0.46^a
粗纤维（%）	16.67±1.38^a	16.77±0.89^a
粗脂肪（%）	3.00±0.18^a	2.40±0.13^b
可溶性蛋白（%）	4.57±0.10^a	2.68±0.02^b
pH	6.76±0.05^a	5.61±0.04^b
游离氨基酸（%）	2.81±0.05^b	7.87±0.14^a
乳酸钙含量（%）	0.97±0.10^b	13.58±0.10^a
注：同行不同字母代表差异显著（P<0.05）。

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