杏鲍菇528深层发酵生物合成麦角硫因的培养基优化

汤葆莎; 吴俐; 翁敏劼; 赖谱富; 李怡彬

doi:10.19303/j.issn.1008-0384.2021.06.016

杏鲍菇528深层发酵生物合成麦角硫因的培养基优化

doi: 10.19303/j.issn.1008-0384.2021.06.016

汤葆莎^{1, 2,},
吴俐^{1, 2},
翁敏劼^{1, 2},
赖谱富^{1, 2},
李怡彬^{1, 2, ,}

1.
福建省农业科学院农业工程技术研究所，福建　福州　350003
2.
福建省农产品（食品）加工重点实验室，福建　福州　350003

基金项目: 国家重点研发计划项目（2018YFD0400200）；福建省科技计划公益类专项（2019R1032-11）；福建省农业科学院对外合作项目（DEC201821204）；福建省农业科学院创新团队建设项目（STIT2017-3-5、STIT2017-1-10）

详细信息

作者简介:
汤葆莎（1971−），女，硕士，高级农艺师，研究方向：农产品加工技术及食用菌栽培研究（E-mail：tbsty@126.com）

通讯作者:
李怡彬（1981−），男，硕士，研究员，研究方向：食品科学与工程（E-mail：lyb9951@163.com）

中图分类号: S 646.1+4
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出版历程
- 收稿日期: 2020-05-20
- 修回日期: 2021-06-23
- 刊出日期: 2021-06-28

Optimization of medium for biosynthesis of ergothioneine by submerged fermentation of Pleurotus eryngii 528

TANG Baosha^{1, 2
,},
WU Li^{1, 2},
WENG Minjie^{1, 2},
LAI Pufu^{1, 2},
LI Yibin^{1, 2
, ,}

1.
Institute of Agri-engineering Technology, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian　350003, China
2.
Fujian Key Laboratory of Agricultural Product (Food) Processing, Fuzhou, Fujian　350003, China

摘要

摘要: 目的为提高杏鲍菇528发酵物的麦角硫因含量，筛选杏鲍菇528培养的营养配方，以期为开发高含量的麦角硫因杏鲍菇深层液体发酵物提供理论基础。方法以固体培养菌丝生长速度和菌丝形态探讨杏鲍菇528菌丝生长的适宜碳氮源；采用单因素试验，以发酵物干重、沉淀物情况和初始pH值为指标，研究杏鲍菇528液体深层发酵菌丝生长的营养配方，探究不同氨基酸对深层液体培养发酵物麦角硫因含量的影响；最后通过L₉（3⁴）正交试验分析提高杏鲍菇528发酵物麦角硫因含量的深层发酵培养基配方。结果杏鲍菇528固体培养菌丝生长的适宜碳、氮源分别为玉米糁粉和蛋白胨；在杏鲍菇528液体发酵培养中，玉米糁粉2.0%和蛋白胨2.0%组合碳氮源培养基液体培养的发酵物干重值达33.30±0.81 g·L⁻¹，显著高于其他浓度比例的碳氮源组合，且沉淀物情况和初始pH更适宜杏鲍菇528液体培养；天冬氨酸、半胱氨酸、组氨酸、精氨酸、谷氨酸这5种氨基酸均可明显提高杏鲍菇528发酵物麦角硫因产率，其中组氨酸诱导培养的杏鲍菇液体发酵物中麦角硫因含量最高，是对照组的3.73倍，且明显高于其他氨基酸。结论杏鲍菇528深层发酵生物合成麦角硫因的最佳培养基配方为：玉米糁粉2.0%、蛋白胨1.5%、组氨酸0.15%、麦麸细粉1.0%、KH₂PO₄ 0.1%、MgSO₄ 0.1%、谷氨酸0.1%和维生素B₆ 0.1%，在此条件下，杏鲍菇发酵物麦角硫因含量达20.50±1.80 mg·L⁻¹。
- 杏鲍菇528 /
- 液体发酵 /
- 麦角硫因 /
- 组氨酸
Abstract: Objective In order to increase the ergothioneine content of Pleurotus eryngii 528 fermented products, the nutritional formula of Pleurotus eryngii 528 was screened to provide a theoretical basis for the development of high content of ergothioneine in submerged fermentation of Pleurotus eryngii . Method The suitable carbon and nitrogen sources for Pleurotus eryngii 528 mycelium growth were discussed based on the solid culture mycelium growth rate and mycelium morphology. A single factor test was conducted to study the nutritional formula of mycelium growth in Pleurotus eryngii 528 submerged fermentation, with the indexes of the dry weight of the fermented product, the sedimentation and the initial pH value of the medium. The effects of different amino acids on ergothioneine content in submerged fermentation broth were investigated. Finally, L₉(3⁴) orthogonal test was used to analyze the formula of submerged fermentation medium for increasing ergothioneine content in Pleurotus eryngii 528. Result Corn grits and peptone were the suitable carbon and nitrogen sources for the growth of Pleurotus eryngii 528. In the liquid fermentation of Pleurotus eryngii 528, the dry weight of the fermentation product in the liquid culture medium of corn grits powder 2.0% and peptone 2.0% reached 33.30±0.81 g·L⁻¹ , which was significantly higher than that of other carbon and nitrogen source combinations, and the sediment condition and initial pH value were more suitable for Pleurotus eryngii 528 liquid culture. Aspartic acid, cysteine, histidine, arginine and glutamic acid improved significantly the ergothioneine yield of Pleurotus eryngii 528 fermented product. The content of ergothioneine in the liquid fermentation of Pleurotus eryngii induced by histidine was the highest, which was 3.73 times of the control group, and was significantly higher than other amino acids. Conclusion The optimum medium formula of ergothioneine biosynthesis by submerged fermentation of Pleurotus eryngii 528 was as follows: corn grits 2.0%, peptone 1.5%, histidine 0.15%, wheat bran powder 1.0%, KH₂PO₄0.1%, MgSO₄0.1%, glutamic acid 0.1% and vitamin B6 0.1%. Under these conditions, the content of ergothioneine in Pleurotus eryngii fermentation reached 20.50±1.80 mg·L⁻¹.
- Pleurotus eryngii 528 /
- liquid fermentation /
- ergothioneine /
- histidine

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图 1 不同碳、氮源培养基对杏鲍菇528菌丝生长的影响（第9 d，左为碳源，右为氮源）

Figure 1. Effects of different carbon and nitrogen source culture media on mycelial growth of Pleurotus eryngii 528 (At 9^th day, left as carbon sources, right as nitrogen sources)

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图 2 不同碳、氮源培养基杏鲍菇528菌丝生长速率变化（A为碳源，B为氮源）

Figure 2. Variation of mycelial growth rate of Pleurotus eryngii 528 with different carbon and nitrogen sources (A is carbon sources, B is nitrogen sources)

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图 3 不同氨基酸对发酵物麦角硫因含量的影响

Figure 3. Effects of different amino acids on the content of ergothioneine in fermentation product

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表 1 正交试验设计因素及水平

Table 1. Factors and levels of orthogonal experimental design

水平 Level	因素 Factor%
水平 Level	玉米糁粉 Corngrits A	蛋白胨 Peptone B	组氨酸 Histidine C
1	1.5	1.5	0.10
2	2.0	2.0	0.15
3	2.5	2.5	0.20

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表 2 不同比例玉米糁粉和蛋白胨对杏鲍菇528深层发酵培养液的沉淀物情况、初始pH及发酵物干重影响

Table 2. Effects of different proportions of corn grits and peptone on the sediment condition, initial pH in culture medium and dry weight of fermentation products of Pleurotus eryngii 528 submerged fermentation

配方 Formula	玉米糁粉 Corn grits/%	蛋白胨 Peptone/%	麦麸细粉 Wheat bran powder/%	谷氨酸 Glutamic acid/%	维生素B₆ Vitamin B₆/%	KH₂PO₄/%	MgSO₄/%	沉淀物情况 Sediment condition	pH	发酵物干重 Dry weight of fermentation/ (g·L⁻¹)
A	1.0	1.0	1.0	0.1	0.1	0.1	0.1	+	4.96	24.60±0.17 D
B	1.0	2.0	1.0	0.1	0.1	0.1	0.1	+	5.10	31.20 ±0.99 C
C	1.0	3.0	1.0	0.1	0.1	0.1	0.1	+	5.11	31.50 ±1.19 C
D	2.0	1.0	1.0	0.1	0.1	0.1	0.1	++	4.96	24.70±0.11 D
E	2.0	2.0	1.0	0.1	0.1	0.1	0.1	++	5.10	33.30±0.81 A
F	2.0	3.0	1.0	0.1	0.1	0.1	0.1	++	5.11	32.90±1.10 B
G	3.0	1.0	1.0	0.1	0.1	0.1	0.1	+++++	4.95	24.90±0.25 D
H	3.0	2.0	1.0	0.1	0.1	0.1	0.1	+++++	5.11	33.10±0.70 AB
I	3.0	3.0	1.0	0.1	0.1	0.1	0.1	+++++	5.12	32.80±0.64 B
注：字母不同表示差异显著（P＜0.05）；+表示沉淀程度：+为沉淀少，++为沉淀较多，+++++沉淀多。 Note: Different letters indicate significant differences (P＜0.05); + indicates the degree of precipitation, + means less precipitation, ++ means more precipitation, +++++means the most precipitation.

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表 3 L₉（3⁴）正交试验结果

Table 3. The results of L₉(3⁴)orthogonal test

序列号 Serial number	因素 Factor/%				麦角硫因含量 Ergothioneine content/ (mg·L ⁻¹)
序列号 Serial number	玉米糁粉 Corn grits A	蛋白胨 Peptone B	组氨酸 Histidine C	空列 Null column D	麦角硫因含量 Ergothioneine content/ (mg·L ⁻¹)
1	1.5	1.5	0.10	1	10.39
2	1.5	2.0	0.15	2	13.60
3	1.5	2.5	0.20	3	16.41
4	2.0	1.5	0.15	3	19.48
5	2.0	2.0	0.20	1	18.49
6	2.0	2.5	0.10	2	11.91
7	2.5	1.5	0.20	2	19.41
8	2.5	2.0	0.10	3	9.98
9	2.5	2.5	0.15	1	16.92
$ {{\bar K}_1}$	13.47	16.43	10.76	15.27
$ {{\bar K}_2}$	16.63	14.03	16.67	14.97
$ {{\bar K}_3}$	15.44	15.08	18.11	15.29
R	3.16	2.40	7.35	0.32

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表 4 正交试验方差分析

Table 4. Variance analysis of orthogonal experiment

变异来源 Source of variation	偏差平方和 Deviation sum of squares	自由度 Degrees of freedom	F值 F value	显著性 Significance
A	15.29	2	1.90
B	8.67	2	1.08
C	90.95	2	11.30	*
空列 Null column	0.19	2	0.02
误差 Error	24.16	6

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