混菌固态发酵菜籽粕工艺优化

doi:10.3864/j.issn.0578-1752.2019.24.014

Abstract

Abstract:

【Objective】 The aim of this study was to optimize the solid state fermentation conditions for rapeseed meal with mixed strains and to improve the feeding value of rapeseed meal. It would provide a reference for the application of fermented rapeseed meal in animal husbandry in China.【Method】The rapeseed meal used in this study was conventional rapeseed meal. Three strains of Lactobacillus acidophilus, Bacillus subtilis and Saccharomyces cerevisiae were used, and the glucosinolates degradation rate (X%), total acid increase rate (Y%), polypeptide increase rate (Z%) were used as evaluation indexes. Method M was used as the comprehensive evaluation index (M=0.7*X+0.15*Y+0.15*Z). In the experiment one, the effects of different strains addition levels and the best mixed ratio of the three strains in fermented rapeseed meal were investigated through L₉ (3 ⁴) three-factors and three-levels orthogonal array design methods. Based on the result of experiment one, an L₁₆ (4 ⁵) four-factors and four-levels of orthogonal array design experiment was designed to explore the optimal conditions of mixed strains solid state fermentation of rapeseed meal, and the fermentation temperature, feed water ratio, time, and inoculation quantity were selected as experimental factors. 【Result】The results showed as follow: (1) L₉ (3 ⁴) orthogonal array design showed that the optimal mixed strains ratio of fermented rapeseed meal were Lactobacillus acidophilus : Bacillus subtilis : Saccharomyces cerevisiae = 1:3:2. Under these condition, the contents of glucosinolates was degraded by 23.5%, the total acid and polypeptide increase rate were 179.2% and 375.0%, respectively. (2) Based on the result of experiment one, the range analysis of experiment two showed that the optimal conditions of mixed strains solid state fermentation of rapeseed meal were as follows: fermentation temperature was 33 °C, feed water ratio was 1:1, time was 84h and inoculation quantity was 6%. The pilot test of this conditions showed that the contents of glucosinolates was degraded by 48.8%, the total acid increase rate was 499.7%, the polypeptide increase rate was 148.4%, and the total change rate M was 131.4%. The results were in line with our expectation. The lowest total change rate M was noted when the temperature was 31 °C, and the total change rate M in temperature 33 °C was higher than 35 °C and 37 °C, but the difference between groups was not significant (P ＞ 0.05). The higher M was also noted when feed water ratio was 1:1.0 and fermentation time was 84h. No significant differences in M were observed between conditions in different levels of inoculation quantity. (3) The fermentation of rapeseed meal by mixed strains also increased the content of CP (37.05% vs. 40.90%) and decreased the concentration of crude fiber (17.47% vs. 16.72%). The amino acid composition of rapeseed meal and fermented rapeseed meal showed that fermentation increased the content of several amino acids in rapeseed meal, especially Asp, Thr, Ser, Glu, Pro, Ala, and Lys. The greatest change caused by fermentation was for glucosinolates, which decreased from 36.08 μmol·g ^-1 to 18.48 μmol·g ^-1. The fermentation of rapeseed meal increased the content of polypeptides (from 0.84% to 2.09%) and increased the content of total acid (from 1.01% to 6.05%) compared to unfermented rapeseed meal. The concentration of crude fat was similar in rapeseed meal and FRSM (4.31% and 4.39%).【Conclusion】The optimal fermentation conditions could effectively degrade the glucosinolates and increase the content of polypeptide and total acid in rapeseed meal, so the nutritional value of rapeseed meal was significantly improved.

Key words: rapeseed meal, mixed strains, solid state fermentation, orthogonal design

ZhengKe WU,GuoHua LIU,Yang LI,AiJuan ZHENG,WenHuan CHANG,ZhiMin CHEN,HuiYi CAI. Optimization of Solid State Fermentation for Rapeseed Meal with Mixed Strains[J].Scientia Agricultura Sinica, 2019, 52(24): 4603-4612.

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水平 Level	因素Factors
水平 Level	嗜酸乳杆菌Lactobacillus acidophilus	枯草芽孢杆菌 Bacillus subtilis	酿酒酵母 Saccharomyces cerevisiae
1	5%	5%	5%
2	10%	10%	10%
3	15%	15%	15%

水平 Level	因子Factors			结果Result
水平 Level	嗜酸乳杆菌 Lactobacillus acidophilus	枯草芽孢杆菌 Bacillus subtilis	酿酒酵母 Saccharomyces cerevisiae	硫甙降解率X Degradation rate of GS (%)	总酸增加率Y Increase rate of total acid (%)	多肽增加率Z Increase rate of polypeptide (%)	总变化率M Total rate of change (%)
1	3	3	3	24.19	287.13	197.62	89.65
2	1	3	2	23.45	179.21	375.00	99.55
3	3	1	2	23.22	204.95	176.19	73.43
4	2	1	3	22.89	140.59	238.10	72.83
5	2	3	1	21.33	107.92	330.95	80.76
6	2	2	2	19.39	184.16	328.57	90.48
7	1	1	1	20.45	147.52	282.14	78.76
8	1	2	3	21.04	90.10	342.86	79.67
9	3	2	1	22.80	206.93	171.43	72.71
标准误SEM				1.83	18.32	21.37	2.25
P值 P value				0.68	0.03	0.04	0.06
K1	85.99	75.01	77.41
K2	81.36	80.96	87.82
K3	78.59	89.98	80.71
优水平Optimal levels	A1	K3	N2
极差(RJ)	7.40	14.97	10.41
排序 Rank	枯草芽孢杆菌（Bacillus subtilis）＞酿酒酵母（Saccharomyces cerevisiae）＞嗜酸乳杆菌（Lactobacillus acidophilus）

编号 Number	因子Factors				结果Result (%)
编号 Number	温度 Temperature (℃)	料水比 Feed water ratio	发酵时间 Time (h)	接种量 Inoculation quantity (%)	硫甙降解率X Degradation rate of GS	总酸增加率Y Increase rate of total acid	多肽增加率Z Increase rate of polypeptide	总变化率M Total rate of change
1	1	1	1	1	31.15	220.64	184.85	82.63
2	1	2	2	2	31.55	318.98	148.96	92.28
3	1	3	3	3	36.16	363.23	117.67	97.45
4	1	4	4	4	41.82	214.42	122.50	79.81
5	2	1	2	3	29.75	483.75	148.16	115.61
6	2	2	1	4	53.58	508.91	134.70	134.04
7	2	3	4	1	53.92	471.88	114.88	125.76
8	2	4	3	2	56.49	541.77	79.19	132.69
9	3	1	3	4	48.21	451.77	154.93	124.76
10	3	2	4	3	48.77	488.55	131.40	127.13
11	3	3	1	2	46.37	468.02	118.30	120.40
12	3	4	2	1	47.26	452.96	56.60	109.51
13	4	1	4	2	41.69	432.11	120.58	112.08
14	4	2	3	1	42.06	529.86	148.41	131.19
15	4	3	2	4	43.23	494.08	114.10	121.49
16	4	4	1	3	41.66	447.40	62.62	105.66
标准误SEM					1.15	19.94	5.67	6.35
P值 P value					0.01	0.04	0.07	0.06
K1	88.04	108.77	110.68	112.27
K2	130.78	121.16	109.72	118.11
K3	120.45	116.27	125.27	111.46
K4	117.60	110.67	111.20	115.02
优水平 Optimal levels	2	2	3	2
极差 RJ	42.73	12.39	15.55	6.65
排序 Rank	温度 Temperature＞时间 Time＞料水比 Feed water ratio＞接种量 Inoculation quantity

水平 Level	结果Result
水平 Level	温度 Temperature(℃)	料水比 Feed water ratio	发酵时间 Time(h)	接种量 Inoculation quantity（%）
1	88.04 a	108.77	110.68	112.27
2	130.78b	121.16	109.72	118.11
3	120.45b	116.27	125.27	111.46
4	117.60 b	110.67	111.20	115.02
标准误SEM	4.35	4.35	4.35	4.35
P值 P value	0.02	0.67	0.79	0.99

水平 Level	结果Result
水平 Level	温度 Temperature (℃)	料水比 Feed water ratio	发酵时间 Time (h)	接种量 Inoculation quantity (%)
1	35.17a	37.70	43.19	43.59
2	48.43b	43.99	37.94	44.02
3	47.65b	44.92	45.73	39.08
4	42.16ab	46.80	46.55	46.71
标准误SEM	2.04	2.04	2.04	2.04
P值 P value	0.06	0.46	0.47	0.66

Optimization of Solid State Fermentation for Rapeseed Meal with Mixed Strains

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项目 Item	菜籽粕 Rapeseed meal	发酵菜籽粕 Fermented rapeseed meal	标准误 SEM
粗蛋白CP	38.36	40.67	1.09
总能GE (kcal·kg^-1)	4923.51	5063.45	36.56
硫甙 μmol·g^-1	36.08	18.48	0.22
粗纤维 Crude fiber	17.47	16.72	1.23
粗脂肪 Crude fat	4.31	4.39	0.15
多肽 Polypeptides	0.84	2.09	0.09
总酸Total acids	1.01	6.05	0.10
天冬氨酸Asp (%)	2.99	3.14	0.12
苏氨酸Thr (%)	1.83	1.99	0.08
丝氨酸Ser (%)	1.64	1.82	0.11
谷氨酸Glu (%)	7.06	7.77	0.23
脯氨酸 Pro (%)	2.69	3.26	0.26
甘氨酸 Gly (%)	2.20	2.38	0.13
丙氨酸 Ala (%)	1.92	2.21	0.15
缬氨酸 Val (%)	2.22	2.48	0.27
异亮氨酸 Ile (%)	1.81	1.92	0.06
亮氨酸 Leu (%)	3.12	3.33	0.11
酪氨酸 Tyr (%)	1.17	1.22	0.09
苯丙氨酸Phe (%)	1.68	1.80	0.10
组氨酸 His (%)	1.19	1.27	0.05
赖氨酸 Lys (%)	2.70	2.92	0.09
精氨酸 Arg (%)	2.46	2.54	0.11
胱氨酸 Cys (%)	1.07	1.13	0.03
蛋氨酸 Met (%)	0.93	0.98	0.04

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