阿舒假囊酵母合成的精油及其抗真菌活性与机理

doi:10.7503/cjcu20180477

高等学校化学学报 ›› 2019, Vol. 40 ›› Issue (2): 272.doi: 10.7503/cjcu20180477

阿舒假囊酵母合成的精油及其抗真菌活性与机理

何峰^1,²(), 白金海¹, 陈淑贤¹, 谭小蓓¹

1. 经济林木种质改良与资源综合利用湖北省重点实验室(黄冈师范学院), 黄冈 438000
2. 大别山特色资源开发湖北省协同创新中心, 黄冈 438000;

收稿日期:2018-07-04 出版日期:2019-02-10 发布日期:2018-11-15
作者简介:
联系人简介: 何峰, 男, 博士, 副教授, 主要从事微生物、植物资源挖掘与开发方面的研究. E-mail: hfeng@hust.edu.cn
基金资助:
湖北省高等学校优秀中青年科技创新团队项目(批准号: T201820)资助

Antifungal Activity and Mechanism of an Essential Oil from Eremothecium ashbyii^†

HE Feng^1,^2,^*(), BAI Jinhai¹, CHEN Shuxian¹, TAN Xiaobei¹

1. Hubei Key Laboratory of Economic Forest Germplasm Improvement and Resources Comprehensive Utilization, Huanggang Normal University, Huanggang 438000, China
2. Hubei Collaborative Innovation Center for the Characteristic Resources Exploitation of Dabie Mountains, Huanggang 438000, China

Received:2018-07-04 Online:2019-02-10 Published:2018-11-15
Contact: HE Feng E-mail:hfeng@hust.edu.cn
Supported by:
† Supported by the Outstanding Young and Middle-aged Scientific and Technological Innovation Team of Colleges and Universities in Hubei Province, China(No.T201820)

摘要/Abstract

摘要：

利用发酵法对阿舒假囊酵母进行发酵, 用萃取法和水蒸气蒸馏法从发酵液中提取精油; 通过气相色谱-质谱(GC-MS)法确定其成分; 分别利用抑菌圈和二倍稀释法实验法测定了精油的抗真菌能力、最小抑菌浓度和最小杀菌浓度; 利用荧光标记法、精油处理酵母后细胞内容物检测法及透射电子显微镜研究了精油的抗菌机理. 结果表明, 阿舒假囊酵母发酵法得到的精油产率为0.54 g/L. 用GC-MS法鉴定了22种化学成分, 占精油总量的98.82%. 精油主要由单萜、倍半萜、芳香醇和倍半萜烃类物质组成. 精油对白色念珠菌、光滑念珠菌、热带念珠菌、季也蒙念珠菌、新型隐球菌和酿酒酵母具有良好的抗菌活性, 其最小抑菌浓度分别为31.25, 31.25, 62.5, 31.25, 15.625和15.625 μg/mL; 最小杀菌浓度分别为62.5, 62.5, 125.0, 62.5, 31.25和31.25 μg/mL. 结果表明, 精油的抗菌机理可能是破坏真菌的细胞壁、细胞膜, 使胞内物质外泄, 最终导致真菌死亡. 该研究结果为发酵法合成精油及其应用提供了新思路.

关键词: 阿舒假囊酵母, 精油, 抗真菌活性, 抗菌机理

Abstract:

The essential oil was obtained from the broth of Eremothecium ashbyii H4565 by solvent extraction and steam distillation. The chemical components of essential oil were investigated by gas chromatography-mass(GC-MS) spectrometry. Antifungal activities were studied by inhibition zone method. Two fold dilution method was used to determine the minimum inhibitory concentration(MIC) and minimum bactericidal concentration(MBC). Cytoplasmic membrane permeability, protein and potassium ion leakage, and transmission electron microscope were used to determine antifungal mechanism. The results showed that the yield of essential oil was 0.54 g/L. Twenty two compounds, about 98.82% constituents of the total, were identified by GC-MS analysis. The essential oil was composed mainly of monoterpenes, sesquiterpenes, aromatic alcohol, and sesquiterpene hydrocarbons. The essential oil exhibited strong antibacterial activities against C. albicans, C. guilliermondii, C. tropicalis, C. glabrata, C. neoformans and S. cerevisiae. The MIC of the essential oil against these microorganisms were 31.25, 31.25, 62.5, 31.25, 15.625 and 15.625 μg/mL, respectively. The MBC of the essential oil against these microorganisms were 62.5, 62.5, 125.0, 62.5, 31.25 and 31.25 μg/mL, respectively. Based on analyses of cytoplasmic membrane permeability, protein and potassium ion leakage and transmission electron microscope, the possible targets of the essential oil were cell wall and cytoplasmic membrane. The results showed a new light on development and application of the essential oil from microorganism.

Key words: Eremothecium ashbyii, Essential oil, Antifungal activity, Antibacterial mechanism

中图分类号:

O621
Q599

TrendMD:

何峰, 白金海, 陈淑贤, 谭小蓓. 阿舒假囊酵母合成的精油及其抗真菌活性与机理. 高等学校化学学报, 2019, 40(2): 272.

HE Feng,BAI Jinhai,CHEN Shuxian,TAN Xiaobei. Antifungal Activity and Mechanism of an Essential Oil from Eremothecium ashbyii^†. Chem. J. Chinese Universities, 2019, 40(2): 272.

图/表 6

参考文献 24

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Compd.	RI^a	RI range^b	CAS number	Content(%)
Limonene	1032	1019—1039	005989-27-5	0.23
Linalool oxide	1077	1061—1090	034995-77-2	0.13
Linalool	1081	1088—1109	000078-70-6	5.30
Phenethyl alcohol	1117	1100—1129	000060-12-8	5.39
Menthone	1153	1125—1174	010458-14-7	0.15
α-Terpineol	1192	1178—1203	010482-56-1	0.42
Citronellol	1234	1215—1237	000106-22-9	34.43
Geraniol	1257	1238—1269	000106-24-1	36.58
Linalyl acetate	1258	1238—1268	000115-95-7	0.19
Geranial	1271	1252—1291	000141-27-5	1.32
Longipinene	1354	1337—1362	005989-08-2	0.47
Eugenol	1359	1345—1375	000097-53-0	0.12
Longicycline	1378	1370—1394	001137-12-8	0.40
β-Elemene	1392	1374—1402	000515-13-9	1.01
Longifolene	1409	1387—1434	000475-20-7	4.22
1-Caryophyllene	1423	1392—1426	000087-44-5	1.77
Thujopsene	1434	1424—1450	000470-40-6	1.11
α-Humulene	1456	1435—1470	006753-98-6	0.32
Germacrene D	1483	1464—1493	023986-74-5	0.34
α-Muurolene	1501	1480—1509	010208-80-7	0.25
Cuparene	1509	1498—1523	016982-00-6	0.11
Nerolidol	1562	1539—1570	007212-44-4	4.56

Compd.	RI^a	RI range^b	CAS number	Content(%)
Limonene	1032	1019—1039	005989-27-5	0.23
Linalool oxide	1077	1061—1090	034995-77-2	0.13
Linalool	1081	1088—1109	000078-70-6	5.30
Phenethyl alcohol	1117	1100—1129	000060-12-8	5.39
Menthone	1153	1125—1174	010458-14-7	0.15
α-Terpineol	1192	1178—1203	010482-56-1	0.42
Citronellol	1234	1215—1237	000106-22-9	34.43
Geraniol	1257	1238—1269	000106-24-1	36.58
Linalyl acetate	1258	1238—1268	000115-95-7	0.19
Geranial	1271	1252—1291	000141-27-5	1.32
Longipinene	1354	1337—1362	005989-08-2	0.47
Eugenol	1359	1345—1375	000097-53-0	0.12
Longicycline	1378	1370—1394	001137-12-8	0.40
β-Elemene	1392	1374—1402	000515-13-9	1.01
Longifolene	1409	1387—1434	000475-20-7	4.22
1-Caryophyllene	1423	1392—1426	000087-44-5	1.77
Thujopsene	1434	1424—1450	000470-40-6	1.11
α-Humulene	1456	1435—1470	006753-98-6	0.32
Germacrene D	1483	1464—1493	023986-74-5	0.34
α-Muurolene	1501	1480—1509	010208-80-7	0.25
Cuparene	1509	1498—1523	016982-00-6	0.11
Nerolidol	1562	1539—1570	007212-44-4	4.56

Microorganism	Diameters of inhibition zone/mm			MIC/ (μg·mL^-1)	MBC/ (μg·mL^-1)
Microorganism	Essential oil	Amphotericin B	n-Hexane	MIC/ (μg·mL^-1)	MBC/ (μg·mL^-1)
C. albicans	28.2±0.3	27.6±0.4	3.1±0.2	31.25	62.5
C. glabrata	28.9±0.5	29.3±0.3	5.2±0.3	31.25	62.5
C. tropicalis	26.1±0.3	26.9±0.2	5.3±0.2	62.5	125.0
C. guilliermondii	27.8±0.2	28.4±0.4	4.2±0.1	31.25	62.5
C. neoformans	31.2±0.2	35.1±0.1	4.4±0.2	15.625	31.25
S. cerevisiae	32.6±0.4	30.4±0.3	4.1±0.3	15.625	31.25

Microorganism	Diameters of inhibition zone/mm			MIC/ (μg·mL^-1)	MBC/ (μg·mL^-1)
Microorganism	Essential oil	Amphotericin B	n-Hexane	MIC/ (μg·mL^-1)	MBC/ (μg·mL^-1)
C. albicans	28.2±0.3	27.6±0.4	3.1±0.2	31.25	62.5
C. glabrata	28.9±0.5	29.3±0.3	5.2±0.3	31.25	62.5
C. tropicalis	26.1±0.3	26.9±0.2	5.3±0.2	62.5	125.0
C. guilliermondii	27.8±0.2	28.4±0.4	4.2±0.1	31.25	62.5
C. neoformans	31.2±0.2	35.1±0.1	4.4±0.2	15.625	31.25
S. cerevisiae	32.6±0.4	30.4±0.3	4.1±0.3	15.625	31.25

阿舒假囊酵母合成的精油及其抗真菌活性与机理

Antifungal Activity and Mechanism of an Essential Oil from Eremothecium ashbyii^†

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阿舒假囊酵母合成的精油及其抗真菌活性与机理

Antifungal Activity and Mechanism of an Essential Oil from Eremothecium ashbyii†

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Antifungal Activity and Mechanism of an Essential Oil from Eremothecium ashbyii^†