虫草素的抑菌活性及机理研究

doi:10.13560/j.cnki.biotech.bull.1985.2020-1169

摘要/Abstract

摘要：

虫草素为蛹虫草等食药保健品的主要活性成分。为进一步评价分析虫草素的抑菌活性及机理,通过最低抑菌浓度及抑菌圈实验测定、扫描电镜观察虫草素对6种常见细菌的抑菌谱及效果,测定供试菌的生长曲线、胞内紫外吸收物质泄露和菌体形态变化,分析虫草素抑制菌体生长及破坏细胞结构机理。结果表明,在溶解度范围内虫草素对3种G⁺菌有明显抑菌效果,而对3种G^-菌无明显抑菌效果。生长测定及电镜观察发现经虫草素处理后供试G⁺菌菌体形态严重受损,胞内紫外吸收物质外泄,细菌生长进程受阻;而部分供试G^-菌菌体表面有轻微破损并伴随少量内容物泄露,但细菌生长进程不受影响。虫草素具有一定的抑菌选择性,更易破坏G⁺菌的壁膜系统起到抑菌作用,对G^-菌整体生长进程无明显影响。

关键词: 虫草素, 最低抑菌浓度, 抑菌圈, 生长曲线, 菌体形态, 抑菌机理

Abstract:

Cordycepin is the main active component of Cordyceps militaris and other food and health products,the purpose of the study is to further evaluate and analyze the main antibacterial activity and mechanism of cordycepin. The bacteriostatic spectrum and effect of cordycepin on six bacteria were determined by minimum inhibitory concentration,bacteriostatic zone test and electron microscopic observation,and the growth curve,intracellular ultraviolet absorption substances leakage and morphological changes of the tested bacteria were further determined to analyze the mechanism of cordycepin inhibiting cell growth and destroying cell structure. The results showed that cordycepin had obvious bacteriostatic effect on three G⁺bacteria in the range of solubility,but had no obvious bacteriostatic effect on three G^-bacteria. Growth assay and electron microscopic observation showed that the morphology of G⁺ bacteria was seriously damaged after cordycepin treatment,the intracellular ultraviolet absorption substances were leaked out,and the bacterial growth process was blocked. While the surface of some tested G^-bacteria was slightly damaged and accompanied by a small amount of content leaked,but the bacterial growth process was not affected. Cordycepin presents certain selectivity in bacteriostasis,and it is easier to destroy the wall membrane system of the tested G⁺ bacteria and play a bacteriostatic role,but has no significant effect on the overall growth process of the tested G^-bacteria.

Key words: cordycepin, minimum inhibitory concentration, inhibitory zone, growth curve, bacterial morphology, antibacterial mechanism

高苏, 马婕馨, 刘警鞠, 赵国柱. 虫草素的抑菌活性及机理研究[J]. 生物技术通报, 2021, 37(4): 137-144.

GAO Su, MA Jie-xin, LIU Jing-ju, ZHAO Guo-zhu. Study on Antibacterial Activity and Mechanism of Cordycepin[J]. Biotechnology Bulletin, 2021, 37(4): 137-144.

图/表 7

表1 虫草素MIC测定结果

Table1 Determination of cordycepin MIC

供试菌株 Tested strains	虫草素浓度 Concentration of cordycepin/（mg·mL^-1）
供试菌株 Tested strains	0	0.625	1.25	2.5	5	10
B. subtilis BS-1	+++	++	+	-	-	-
B. thuringiensis BT-1	+++	+++	++	+	-	-
S. aureus SA-1	+++	+++	++	+	-	-
E. coli EC-1	+++	+++	+++	+++	+++	+
S. enteritidis SE-1	+++	+++	+++	+++	+++	+
P. aeruginosa PA-1	+++	+++	+++	+++	+++	++

图1 虫草素对6种供试菌抑菌圈 A为对照孔,B、C、D分别为2.5 mg/mL、5 mg/mL、10 mg/mL虫草素组

Fig.1 DIZ of cordycepin against six tested strains A is the control well. B, C and D are 2.5 mg/mL, 5 mg/mL and 10 mg/mL. respectively cordycepin group

表2 虫草素DIZ测定结果

Table 2 Determination of cordycepin DIZ

供试菌株 Tested strains	DIZ/mm
供试菌株 Tested strains	2.5 mg/mL 虫草素cordycepin	5 mg/mL 虫草素 cordycepin	10 mg/mL 虫草素 cordycepin
B. subtilis BS-1	3.6 ± 0.17^d	5.2 ± 0.16^b	8.75 ± 0.21^a
B. thuringiensis BT-1	0	3.52 ± 0.23^d	5.25 ± 0.15^b
S. aureus SA-1	0	2.97 ± 0.20^e	4.15 ± 0.31^c
E. coli EC-1	0	0	0
S. enterica SE-1	0	0	0
P. aeruginosa PA-1	0	0	0

图2 虫草素对6种供试菌生长曲线的影响

Fig.2 Effects of cordycepin on the growth curves of six tested strains

图3 虫草素对6种供试菌胞内紫外吸收物质泄露的影响

Fig.3 Effects of cordycepin on the leakage of intracellular ultraviolet absorbents of six tested strains

图4 场发射扫描电镜下的供试G+菌形态大写字母表示各实验组,小写字母表示各对照组;A,a：B. subtilis BS-1;B,b：B. thuringiensis BT-1;C,c：S. aureus SA-1;放大倍数：8000×;比例尺5 μm

Fig.4 Morphology of tested G+ bacteria under field emission scanning electron microscope Capital letters refer to each experimental group, and lowercase letters refer to each control group. A, a: B.Subtilis BS-1. B, b: B. thuringiensis BT-1. C, c: S.aureus SA-1. Magnification: 8000×. Scale: 5 μm

图5 场发射扫描电镜下的供试G-菌形态大写字母表示各实验组,小写字母表示各对照组;A,a：E. coli EC-1;B,b：S. enteritidis SE-1;C,c：P. aeruginosa PA-1;放大倍数：8 000×;比例尺5 μm

Fig.5 Morphology of tested G- bacteria under field emission scanning electron microscope Capital letters refer to each experimental group, and lowercase letters refer to each control group. A, a: E. coli EC-1. B, B: S. enteritidis SE - 1. C, c: P.aeruginosa PA-1.Magnification: 8000×. Scale: 5 μm

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