产紫丁香苷内生真菌CJ7的鉴定及发酵条件研究

doi:10.11924/j.issn.1000-6850.casb2021-0888

摘要/Abstract

摘要：

为高效利用刺五加内生真菌资源，推动紫丁香苷的生产，采用HPLC法，以紫丁香苷为对照，分析内生真菌CJ7的次生代谢产物;采用琼脂糖扩散法，对内生真菌CJ7进行抑菌活性分析，并采用单因素实验及正交实验法对内生真菌CJ7发酵条件进行优选;采用形态学观察及ITS序列分析法对内生真菌CJ7进行菌种鉴定。结果表明，在内生真菌CJ7发酵液中含有紫丁香苷。抑菌活性实验结果表明，内生真菌CJ7对10种致病菌均有较好的抑制作用，尤其对金黄色葡萄球菌抑菌效果较为明显。经优化，CJ7的最佳发酵工艺为培养基中葡萄糖质量浓度2.5%，牛肉膏质量浓度2.0%，初始pH 7.0，装液量40%，接种量2%，培养8天，摇床转数120 r/min，发酵温度28℃。经鉴定，内生真菌CJ7为产烟曲霉(Aspergillus fumigatus)。上述研究说明，内生真菌CJ7为紫丁香苷产生菌。研究结果可为采用微生物发酵法生产紫丁香苷提供参考。

关键词: 紫丁香苷, 内生真菌, 抑菌活性, 发酵条件, 鉴定

Abstract:

The paper aims to efficiently utilize endophytic fungi of Acanthopanax senticosus for the production of syringin. The fermentation broth of endophytic fugus CJ7 was analyzed by HPLC with syringin as reference. The antimicrobial activities of CJ7 were examined by agar diffusion. Meanwhile, the fermentation condition of CJ7 was screened by single factor combined with orthogonal experiments, and the endophytic fungus CJ7 was identified by morphological observation and ITS sequence analysis. The results showed that syringin was detected in the fermentation broth of CJ7, and good inhibitory effects of CJ7 on 10 common pathogens were recognized, especially on Staphylococcus aureus. After being optimized, the best culture condition was as follows: 2.5% glucose and 2.0% beef extract fix in PDA culture with initial pH value of 7.0, medium liquid volume of 40% and inoculation amount of 2%. Besides, the fermentation period was 8 d, and the table rotational speed was set at 120r/min. The strain of CJ7 was identified as Aspergillus fumigatus. As a conclusion, the endophytic fungus CJ7 is a syringin-producing one, and the study can lay a foundation for the production of syringin by microbial fermentation.

Key words: syringin, endophytic fungus, antimicrobial activity, fermentation condition, identification

中图分类号:

R931

张赫, 尤梦瑶, 万璐, 闫佳佳, 刘松梅, 郑春英. 产紫丁香苷内生真菌CJ7的鉴定及发酵条件研究[J]. 中国农学通报, 2022, 38(25): 143-150.

ZHANG He, YOU Mengyao, WAN Lu, YAN Jiajia, LIU Songmei, ZHENG Chunying. Identification and Fermentation Conditions of Syringin-producing Endophytic Fungus CJ7[J]. Chinese Agricultural Science Bulletin, 2022, 38(25): 143-150.

图/表 15

参考文献 29

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水平	因素
水平	A.氮源浓度/%	B.碳源浓度/%	C.接种量(v/v)/%	D.发酵时间/d
1	1.5	1.5	1	6
2	2.0	2.0	2	8
3	2.5	2.5	3	10

水平	因素
水平	A.氮源浓度/%	B.碳源浓度/%	C.接种量(v/v)/%	D.发酵时间/d
1	1.5	1.5	1	6
2	2.0	2.0	2	8
3	2.5	2.5	3	10

样品	大肠杆菌	枯草芽孢杆菌	金黄色葡萄球菌	短小芽孢杆菌	单增李斯特菌	鲍曼不动杆菌	肺炎克雷伯菌	铜绿假单孢菌	屎肠球菌	白假丝酵母
CJ7	15.02±0.32	14.79±0.80	18.05±0.17	14.93±0.13	14.02±0.14	18.55±0.09	15.91±0.18	16.53±0.28	17.10±0.24	12.57±0.19
甲醇	-	-	-	-	-	-	-	-	-	-
链霉素	26.71±0.31	27.14±0.30	28.52±0.07	19.13±0.21	24.20±0.35	28.07±0.05	26.50±0.01	28.72±0.30	25.00±0.08	-
制霉素	-	-	-	-	-	-	-	-	-	19.90±0.05

样品	大肠杆菌	枯草芽孢杆菌	金黄色葡萄球菌	短小芽孢杆菌	单增李斯特菌	鲍曼不动杆菌	肺炎克雷伯菌	铜绿假单孢菌	屎肠球菌	白假丝酵母
CJ7	15.02±0.32	14.79±0.80	18.05±0.17	14.93±0.13	14.02±0.14	18.55±0.09	15.91±0.18	16.53±0.28	17.10±0.24	12.57±0.19
甲醇	-	-	-	-	-	-	-	-	-	-
链霉素	26.71±0.31	27.14±0.30	28.52±0.07	19.13±0.21	24.20±0.35	28.07±0.05	26.50±0.01	28.72±0.30	25.00±0.08	-
制霉素	-	-	-	-	-	-	-	-	-	19.90±0.05

组别	因素				抑菌圈直径/mm
组别	A	B	C	D	抑菌圈直径/mm
1	1	1	1	1	25.83±0.30
2	1	2	2	2	28.33±0.30
3	1	3	3	3	29.33±0.30
4	2	1	2	3	30.50±0.00
5	2	2	3	1	29.17±0.70
6	2	3	1	2	30.67±0.70
7	3	1	3	2	23.50±0.00
8	3	2	1	3	21.67±0.40
9	3	3	2	1	26.67±0.20
K1	83.49	81.83	80.17	81.67	ΣY=249.67 CT= 6926.12
K2	94.34	79.17	87.50	84.50
K3	71.84	88.67	82.00	83.50
R	7.50	3.17	2.45	0.95
S	84.41	16.02	9.71	1.38