响应曲面法优化漳州芽孢杆菌液态发酵桑叶粉营养品质的研究

doi:10.11924/j.issn.1000-6850.casb2021-0205

中国农学通报 ›› 2022, Vol. 38 ›› Issue (16): 145-154.doi: 10.11924/j.issn.1000-6850.casb2021-0205

所属专题：生物技术；植物保护

响应曲面法优化漳州芽孢杆菌液态发酵桑叶粉营养品质的研究

姜文强¹(), 缪凌鸿¹^,², 高亮³, 朱悦洁¹, 林艳², 罗晨皓¹, 钱琳洁¹, 陈士友¹, 张维娜³, 施大林³, 刘波¹^,², 沈怀舜¹^,², 戈贤平¹^,²()

¹南京农业大学无锡渔业学院,江苏无锡 214081
²中国水产科学研究院淡水渔业研究中心/农业农村部淡水渔业和种质资源利用重点实验室,江苏无锡 214081
³江苏省苏微微生物研究有限公司,江苏无锡 214063

收稿日期:2021-02-05 修回日期:2021-09-28 出版日期:2022-06-05 发布日期:2022-06-09
通讯作者: 戈贤平
作者简介:姜文强,男,1996年出生,山东烟台人,博士研究生,从事水产动物营养与饲料研究。通信地址：214081 江苏无锡薛家里69号南京农业大学无锡渔业学院,E-mail： 2463540256@qq.com。
基金资助:
国家现代农业产业建设基金“国家大宗淡水鱼产业技术体系建设项目”(CARS-45-10);中国水产科学研究院基本科研业务费资助“青虾用发酵磷虾粉功能性饲料添加剂的研制”(2020XT1203);中国水产科学研究院创新团队项目(2020TD59)

Study on Nutrients and Quality Improvement of Liquid Fermented Mulberry Leaf Meal by Bacillus zhangzhouensis Using Response Surface Methodology

JIANG Wenqiang¹(), MIAO Linghong¹^,², GAO Liang³, ZHU Yuejie¹, LIN Yan², LUO Chenhao¹, QIAN Linjie¹, CHEN Shiyou¹, ZHANG Weina³, SHI Dalin³, LIU Bo¹^,², SHEN Huaishun¹^,², GE Xianping¹^,²()

¹Wuxi Fisheries College, Nanjing Agricultural University, Wuxi, Jiangsu 214081
²Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture and Rural Affairs/ Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi, Jiangsu 214081
³Jiangsu Limited Company of Suwei Microbiology, Wuxi, Jiangsu 214063

Received:2021-02-05 Revised:2021-09-28 Online:2022-06-05 Published:2022-06-09
Contact: GE Xianping

摘要/Abstract

摘要：

研究旨在探讨响应面法优化漳州芽孢杆菌液态发酵桑叶粉的工艺及发酵之后产物的营养品质。以桑叶粉为原料、漳州芽孢杆菌为发酵菌种,以粗蛋白含量为响应值,通过响应面优化桑叶粉的液态发酵工艺。结果表明,漳州芽孢杆菌液态发酵桑叶粉的最优条件为料水比5.8%、装液量11.7%、发酵时间61 h、发酵温度34℃。在此条件下,漳州芽孢杆菌发酵桑叶粉所得产物中,分子量在1000 Da以下的肽百分比为97.4%。与原桑叶粉相比,漳州芽孢杆菌发酵桑叶粉所得产物的粗蛋白含量为17.456%,比原桑叶粉粗蛋白含量(14.029%)显著提高24.4%,小肽含量显著增加21.61%,必需氨基酸及总氨基酸含量显著升高,单宁含量显著降低34.71%,抗氧化活性显著增强。总体而言,漳州芽孢杆菌在优化的液态发酵条件下所制得的发酵桑叶粉,营养成分和品质、抗氧化活性显著提高,单宁含量降低,适口性提高。

关键词: 响应曲面法, 桑叶粉, 漳州芽孢杆菌, 营养物质, 品质

Abstract:

This study aims to optimize the fermentation process and the nutritional quality of fermented mulberry leaf meal by Bacillus zhangzhouensis through response surface methodology (RSM). In this study, mulberry leaf meal and Bacillus zhangzhouensis were selected as the raw material and microbial strain, and the crude protein content was the response value for liquid fermentation. The liquid fermentation process of mulberry leaf meal was optimized through response surface methodology (RSM) analysis. The results showed that the optimum condition for liquid fermentation of mulberry leaf meal by Bacillus zhangzhouensis was as follows: the material water ratio of 5.8%, the liquid volume of 11.7%, the duration time of 61 h, and the temperature of 34℃. After being fermented under the above condition, the percentage of peptides with molecular weight below 1000 Da was 97.4%. Compared to the mulberry leaf meal, the content of crude protein and small peptides in fermented mulberry leaf meal significantly increased by 24.4% and 21.61%, respectively. In addition, the content of essential amino acids and total amino acids significantly increased, and the tannin content decreased by 34.71%. The antioxidant activity of fermented mulberry leaf meal was significantly higher than that of unfermented one. In general, the nutritional quality, the antioxidant activities and the palatability were improved, while the content of tannin decreased in fermented mulberry leaf meal after liquid fermentation using Bacillus zhangzhouensis under the optimized condition.

Key words: response surface methodology, mulberry leaf meal, Bacillus zhangzhouensis, nutrients, quality

中图分类号:

S888.2

姜文强, 缪凌鸿, 高亮, 朱悦洁, 林艳, 罗晨皓, 钱琳洁, 陈士友, 张维娜, 施大林, 刘波, 沈怀舜, 戈贤平. 响应曲面法优化漳州芽孢杆菌液态发酵桑叶粉营养品质的研究[J]. 中国农学通报, 2022, 38(16): 145-154.

JIANG Wenqiang, MIAO Linghong, GAO Liang, ZHU Yuejie, LIN Yan, LUO Chenhao, QIAN Linjie, CHEN Shiyou, ZHANG Weina, SHI Dalin, LIU Bo, SHEN Huaishun, GE Xianping. Study on Nutrients and Quality Improvement of Liquid Fermented Mulberry Leaf Meal by Bacillus zhangzhouensis Using Response Surface Methodology[J]. Chinese Agricultural Science Bulletin, 2022, 38(16): 145-154.

图/表 9

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来源	平方和	自由度	均方	F值	P值	显著性	来源	平方和	自由度	均方	F值	P值	显著性
模型	34.23	14	2.45	3.36	0.0152	^*	CD	0.61	1	0.61	0.84	0.3752
A	3.18	1	3.18	4.38	0.0551		A²	7.66	1	7.66	10.53	0.0059	^**
B	2.12	1	2.12	2.91	0.1102		B²	4.3	1	4.30	5.90	0.0291	^*
C	0.39	1	0.39	0.54	0.4737		C²	13.05	1	13.05	17.94	0.0008	^**
D	0.91	1	0.91	1.26	0.2811		D²	0.56	1	0.56	0.76	0.3968
AB	2.2	1	2.20	3.03	0.1039		残差	10.18	14	0.73
AC	2.28	1	2.28	3.13	0.0985		失拟项	8.12	10	0.81	1.57	0.352
AD	0.36	1	0.36	0.50	0.4911		纯误差	2.07	4	0.52
BC	3.13	1	3.13	4.31	0.0569		总和	44.42	28
BD	0.11	1	0.11	0.15	0.7079

来源	平方和	自由度	均方	F值	P值	显著性	来源	平方和	自由度	均方	F值	P值	显著性
模型	34.23	14	2.45	3.36	0.0152	^*	CD	0.61	1	0.61	0.84	0.3752
A	3.18	1	3.18	4.38	0.0551		A²	7.66	1	7.66	10.53	0.0059	^**
B	2.12	1	2.12	2.91	0.1102		B²	4.3	1	4.30	5.90	0.0291	^*
C	0.39	1	0.39	0.54	0.4737		C²	13.05	1	13.05	17.94	0.0008	^**
D	0.91	1	0.91	1.26	0.2811		D²	0.56	1	0.56	0.76	0.3968
AB	2.2	1	2.20	3.03	0.1039		残差	10.18	14	0.73
AC	2.28	1	2.28	3.13	0.0985		失拟项	8.12	10	0.81	1.57	0.352
AD	0.36	1	0.36	0.50	0.4911		纯误差	2.07	4	0.52
BC	3.13	1	3.13	4.31	0.0569		总和	44.42	28
BD	0.11	1	0.11	0.15	0.7079

指标	桑叶粉	发酵桑叶粉	显著性	指标	桑叶粉	发酵桑叶粉	显著性
粗蛋白/%	14.03±0.02	17.46±0.06	^*	赖氨酸/%	0.48±0.01	0.50±0.02
粗脂肪/%	8.16±0.56	20.18±0.12	^**	精氨酸/%	0.61±0.01	0.62±0.03
灰分/%	10.73±0.26	11.87±0.71		非必需氨基酸/%	5.63±0.08	6.02±0.13
总能/(MJ/kg)	17.26±0.01	18.33±0.06		天门冬氨酸/%	1.24±0.01	1.25±0.03
必需氨基酸/%	4.91±0.01	5.24±0.09	^*	谷氨酸/%	1.56±0.01	1.62±0.04
苏氨酸/%	0.59±0.01	0.62±0.01	^*	丝氨酸/%	0.56±0.01	0.59±0.01	^*
缬氨酸/%	0.70±0.01	0.75±0.02		甘氨酸/%	0.69±0.00	0.74±0.02	^*
蛋氨酸/%	0.10±0.04	0.15±0.03		丙氨酸/%	0.73±0.01	0.81±0.01	^**
异亮氨酸/%	0.56±0.01	0.60±0.02		酪氨酸/%	0.38±0.02	0.43±0.01
亮氨酸/%	0.95±0.04	1.02±0.06		胱氨酸/%	0.03±0.00	0.05±0.01
苯丙氨酸/%	0.67±0.01	0.74±0.01	^**	脯氨酸/%	0.43±0.08	0.54±0.06
组氨酸/%	0.24±0.01	0.25±0.01		总氨基酸/%	10.55±0.08	11.26±0.22	^*

指标	桑叶粉	发酵桑叶粉	显著性	指标	桑叶粉	发酵桑叶粉	显著性
粗蛋白/%	14.03±0.02	17.46±0.06	^*	赖氨酸/%	0.48±0.01	0.50±0.02
粗脂肪/%	8.16±0.56	20.18±0.12	^**	精氨酸/%	0.61±0.01	0.62±0.03
灰分/%	10.73±0.26	11.87±0.71		非必需氨基酸/%	5.63±0.08	6.02±0.13
总能/(MJ/kg)	17.26±0.01	18.33±0.06		天门冬氨酸/%	1.24±0.01	1.25±0.03
必需氨基酸/%	4.91±0.01	5.24±0.09	^*	谷氨酸/%	1.56±0.01	1.62±0.04
苏氨酸/%	0.59±0.01	0.62±0.01	^*	丝氨酸/%	0.56±0.01	0.59±0.01	^*
缬氨酸/%	0.70±0.01	0.75±0.02		甘氨酸/%	0.69±0.00	0.74±0.02	^*
蛋氨酸/%	0.10±0.04	0.15±0.03		丙氨酸/%	0.73±0.01	0.81±0.01	^**
异亮氨酸/%	0.56±0.01	0.60±0.02		酪氨酸/%	0.38±0.02	0.43±0.01
亮氨酸/%	0.95±0.04	1.02±0.06		胱氨酸/%	0.03±0.00	0.05±0.01
苯丙氨酸/%	0.67±0.01	0.74±0.01	^**	脯氨酸/%	0.43±0.08	0.54±0.06
组氨酸/%	0.24±0.01	0.25±0.01		总氨基酸/%	10.55±0.08	11.26±0.22	^*

指标	桑叶粉	发酵桑叶粉	显著性
小肽/(mg/g)	193.59±2.52	235.46±2.96	^**
单宁/(nmol/g)	1379.14±31.40	900.51±14.41	^**
类黄酮/(mg/g)	11.96±0.53	10.30±0.49	^*
总酚/(mg/g)	13.15±0.13	11.96±0.22	^**
DPPH自由基清除率/%	84.15±1.28	93.26±1.12	^*
羟自由基清除率/%	71.48±0.89	81.84±2.43
总抗/(nmol/g)	253.71±0.37	294.91±0.94	^**
抗超氧阴离子活力/[U/(g·prot)]	10.69±0.13	13.02±0.09	^**
超氧化物歧化酶活力/[U/(mg·prot)]	52.41±0.23	57.97±0.69	^**

响应曲面法优化漳州芽孢杆菌液态发酵桑叶粉营养品质的研究

Study on Nutrients and Quality Improvement of Liquid Fermented Mulberry Leaf Meal by Bacillus zhangzhouensis Using Response Surface Methodology

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水平	因素
水平	A.料水比/%	B.装液量/%	C.发酵时间/h	D.发酵温度/℃
-1	7.5	10	24	30
0	10	20	48	35
1	12.5	30	72	40

编号	A	B	C	D	粗蛋白含量Y	编号	A	B	C	D	粗蛋白含量Y
1	-1	0	0	-1	16.7793	16	0	0	0	0	17.4687
2	0	0	1	-1	16.4067	17	0	-1	-1	0	15.4280
3	0	-1	0	-1	16.5293	18	0	0	0	0	18.1755
4	0	1	0	-1	16.5873	19	0	-1	1	0	15.8513
5	0	0	-1	-1	14.6703	20	1	-1	0	0	14.4777
6	1	0	0	-1	16.5877	21	1	0	-1	0	14.6453
7	-1	0	1	0	17.3123	22	1	1	0	0	14.7737
8	-1	-1	0	0	17.0693	23	1	0	1	0	14.6147
9	-1	1	0	0	14.3980	24	-1	0	0	1	15.9363
10	-1	0	-1	0	14.3234	25	0	0	-1	1	15.1130
11	0	0	0	0	16.4347	26	0	-1	0	1	16.9840
12	0	0	0	0	16.9035	27	0	0	1	1	15.2870
13	0	1	-1	0	16.1343	28	0	1	0	1	16.3897
14	0	1	1	0	13.0180	29	1	0	0	1	14.5383
15	0	0	0	0	17.9333

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