松针精油的协同抑菌效应及机制

doi:10.3969/j.issn.1000-2006.201902016

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南京林业大学学报（自然科学版） ›› 2020, Vol. 44 ›› Issue (2): 98-104.doi: 10.3969/j.issn.1000-2006.201902016

松针精油的协同抑菌效应及机制

刘文丽¹(), 包怡红¹^,²^,^*()

1.东北林业大学林学院,黑龙江哈尔滨 150040
2.黑龙江省森林食品资源利用重点实验室,黑龙江哈尔滨 150040

收稿日期:2019-02-20 修回日期:2019-12-28 出版日期:2020-03-30 发布日期:2020-04-01
通讯作者: 包怡红
基金资助:
黑龙江省自然科学基金资助项目(ZD2019C002);国家重点研发计划(2017YFC1601901)

Synergistic antimicrobial effect and mechanism of pine needle essential oil

LIU Wenli¹(), BAO Yihong¹^,²^,^*()

1. College of Forestry,Northeast Forestry University,Harbin 150040,China
2. Key Laboratory of Forest Food Resources Utilization of Heilongjiang Province,Harbin 150040,China

Received:2019-02-20 Revised:2019-12-28 Online:2020-03-30 Published:2020-04-01
Contact: BAO Yihong

摘要/Abstract

摘要：

【目的】研究3种松针精油对供试菌的协同抑菌效应和机制,利用松针精油天然抑菌物质抑制微生物的生长特性,为将松针精油应用于食品、化妆品等领域提供理论依据。【方法】通过微量二倍稀释法测定黑皮油松松针精油(PTEO)、樟子松松针精油(PSEO)、红松松针精油(PKEO)对大肠杆菌(Escherichia coli)、金黄色葡萄球菌(Staphylococcus aureus)、枯草芽孢杆菌(Bacillus subtilis)的抑菌效果,通过棋盘稀释法来测定其协同效果及最佳复配比,以最佳精油复配比研究其对3种供试菌的抑菌机理。【结果】3种精油单独作用时均表现出对3种供试菌较好的抑菌效果,部分复配后效果得到明显提高,针对不同微生物复配最佳抑菌配方为:E.coli用抑菌精油为PTEO和PSEO,浓度均为0.31 μL/mL; S. aureus用抑菌精油为PKEO和PTEO,浓度均为0.16 μL/mL; B. subtilis用抑菌精油为PKEO和PSEO,浓度分别为0.16、0.08 μL/mL。通过复配精油对3种供试菌的抑菌机理研究发现复配精油能够破坏菌体的正常形态,破坏细胞膜的渗透性,导致核酸等大分子物质的泄露,并且对细胞菌体蛋白的合成和积累有干扰作用。【结论】3种精油复配后表现出更好的抑菌作用,精油之间的协同作用可以减少到抑菌效果时精油的用量。

关键词: 松针精油, 协同作用, 抑菌机理, 黑皮油松, 樟子松, 红松

Abstract:

【Objective】Synergistic bacteriostatic effects of pine needle essential oil from three pine species on bacteria and the underlying mechanism were studied; moreover, a theoretical basis was provided for the use of pine needle essential oil as a natural bacteriostatic substance to inhibit the growth of microorganisms and its application in food and cosmetics.【Method】The antibacterial effects of pine needle oil from Pinus tabulaeformis var. mukdensis(PTEO), P. sylvestris var. mongolica Litv. (PSEO), and P. koraiensis Sieb. et Zucc. (PKEO) on Escherichia coli, Staphylococcus aureus, and Bacillus subtilis were determined by using a micro-dilution method. The synergistic effects and the optimal compounding ratio were determined by using a checkerboard dilution method. The antibacterial mechanism against Escherichia Coli, Staphylococcus aureus, and Bacillus subtilis was studied using the optimal essential oil ratio. 【Result】The results revealed that the three essential oils showed good antibacterial effects against the three tested bacteria. After compounding, the effect was evidently improved, and the composition of and bacteriostatic effect on different microorganisms also differed. The highest antibacterial effects after compounding were obtained with the following ratios: E. coli (PTEO and PSEO, MIC combination 0.31 μL/mL each), S. aureus (PKEO and PTEO, MIC combination 0.16 μL/mL each), andB. subtilis (PKEO and PSEO, MIC combination: 0.16 and 0.08 μL/mL, respectively). The investigation of the antibacterial mechanism of the compounded oils on the three tested bacteria indicated that compounded essential oils can disrupt the normal morphology of bacterial cells and permeability of cell membranes, leading to the leakage of macromolecules such as nucleic acids, destruction of the permeability of the cell membrane proteins; moreover, they interfere with the synthesis and accumulation of cell proteins. 【Conclusion】The combination of three pine needle essential oils has a good antibacterial effect, and the synergistic effect of the essential oils can reduce the amount of essential oils required for the antibacterial effect. The study provides a preliminary understanding of the antibacterial mechanism of the pine needle essential oil.

Key words: pine needle essential oil, synergistic effect, bacteriostatic mechanism, Pinus tabulaeformis var. mukdensis, P. sylvestris var. mongolica Litv., P. koraiensis Sieb. et Zucc.

中图分类号:

TS201.3

刘文丽,包怡红. 松针精油的协同抑菌效应及机制[J]. 南京林业大学学报（自然科学版）, 2020, 44(2): 98-104.

LIU Wenli, BAO Yihong. Synergistic antimicrobial effect and mechanism of pine needle essential oil[J].Journal of Nanjing Forestry University (Natural Science Edition), 2020, 44(2): 98-104.DOI: 10.3969/j.issn.1000-2006.201902016.

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菌种 strain	MIC/(μL·mL^-1)
菌种 strain	PKEO	PSEO	PTEO
大肠杆菌 E.coli	2.50	0.63	2.50
金黄色葡萄球菌 S.aureus	1.25	0.31	0.63
枯草芽孢杆菌 B.subtilis	1.25	0.16	0.31

菌种 strain	处理 treatment	PTEO-PSEO		PKEO-PSEO		PKEO-PTEO
菌种 strain	处理 treatment	PTEO	PSEO	PKEO	PSEO	PKEO	PTEO
大肠杆菌 E. coli	MIC_单独	2.50	0.63	2.50	0.63	2.50	2.5
	MIC_混合	0.31	0.31	2.5	0.08	1.25	1.25
	FIC	0.13	0.50	1.0	0.13	0.50	0.50
	FICI	0.63		1.13		1
金黄色葡萄球菌 S. aureus	MIC_单独	0.63	0.31	1.25	0.31	1.25	0.63
	MIC_混合	0.31	0.08	0.16	0.16	0.16	0.16
	FIC	0.50	0.25	0.13	0.50	0.13	0.25
	FICI	0.75		0.63		0.38
枯草芽孢杆菌 B. subtilis	MIC_单独	0.31	0.16	1.25	0.16	1.25	0.31
	MIC_混合	0.04	0.16	0.16	0.08	0.63	0.08
	FIC	0.13	1.00	0.13	0.50	0.50	0.25
	FICI	1.13		0.63		0.75

菌种	0	MIC_联合	2MIC_联合
大肠杆菌 E.coli	0.031±0.017 a	0.23±0.005 b	0.390±0.013 c
金黄色葡萄球菌 S.aureus	0.019±0.003 a	0.206±0.021 b	0.374±0.007 c
枯草芽孢杆菌 B.subtilis	0.037±0.004 a	0.189±0.004 b	0.34±0.006 c

松针精油的协同抑菌效应及机制

Synergistic antimicrobial effect and mechanism of pine needle essential oil

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