野生和人工培养红贝俄氏孔菌化学成分及抑菌活性测定

【目的】考察不同培养基对红贝俄氏孔菌乙酸乙酯提取物化学成分及抑菌活性的影响,为红贝俄氏孔菌的人工培养提供理论依据。【方法】分别用葡萄糖酵母膏蛋白胨培养基(GYP)和综合马铃薯培养基(CPDA)对红贝俄氏孔菌进行人工培养,采用乙酸乙酯提取菌丝体和发酵液化学成分,通过气相色谱—质谱联用技术(GC-MS)对比分析野生和人工培养红贝俄氏孔菌乙酸乙酯提取物的化学成分,通过微量稀释培养法评价野生菌和人工培养红贝俄氏孔菌乙酸乙酯提取物对金黄色葡萄球菌和大肠杆菌的抑菌活性。【结果】GC-MS分析结果表明,野生菌和培养菌丝体乙酸乙酯提取物的主要成分为棕榈酸甲酯、棕榈酸和油酸等,其中,从野生菌检测出13种成分,含量较高的主要成分是亚油酸甲酯(18.522%)、棕榈酸甲酯(15.976%)和棕榈酸(11.164%);从GYP培养菌丝体共检出13种成分,含量较高的主要成分是棕榈酸(28.677%)、油酸(15.108%)和棕榈酸甲酯(8.983%);从CPDA培养菌丝体共检出14种成分,含量较高的主要成分是棕榈酸甲酯(22.575%)、棕榈酸(17.643%)和硬脂酸甲酯(4.787%);有7种相同成分共同存在于3种提取物中,有3种成分仅在野生菌中检测到,有7种成分仅在培养菌丝体中发现。从GYP培养液中共检出9种成分,含量较高的主要成分是十八甲基环壬硅氧烷(4.663%)、1-十九碳烯(2.423%)和(E)-3-十八碳烯(1.785%);从CPDA培养液中共检出14种成分,含量较高的主要成分是棕榈酸(12.913%)、(Z,E)-2,13-十八碳二烯-1-醇(5.985%)和棕榈酸甲酯(5.591%)。体外抑菌试验结果表明,GYP培养液提取物对金黄色葡萄球菌的抑菌活性最强,其最低抑菌浓度(MIC)为0.80 mg/m L,其余提取物对金黄色葡萄球菌和大肠杆菌的MIC均为1.60 mg/m L。【结论】可采用GYP和CPDA作为培养基对野生红贝俄氏孔菌进行规模培养,以解决野生红贝俄氏孔菌资源不足的问题。

Chemical components and antibacterial activity of wild and artificially cultured Earliella scabrosa (Pers.) Gilb.& Ryvarden

[Objective] The effects of different media on the chemical components and antibacterial activity of ethyl acetate extract from Earliella scabrosa (Pers.) Gilb.& Ryvarden were investigated to provide theoretical basis for the artificial culture of E.scabrosa.[Method] E.scabrosa was artificially cultured in two different media,glucose-yeast-peptone medium (GYP) and comprehensive potato-dextrose-agar medium (CPDA) respectively.Ethyl acetate was used to extract the chemical components of mycelia and fermentation broths.The chemical components of ethyl acetate extracts from wild and artificially cultured E.scabrosa were comparatively analyzed by gas chromatography-mass spectrometry (GC-MS).The antibacterial activity of these extracts against Staphylococcus aureus and Escherichia coli were tested by microdilution method.[Result] GC-MS analysis showed that the main chemical components of the ethyl acetate extracts from wild and artificially cultured mycelia were methyl hexadecanoate,hexadecanoic acid,oleic acid,etc.Thirteen components were identified from the wild mycelia,and among them,the main components with high content were methyl linoleate (18.522％),methyl hexadecanoate (15.976％) and hexadecanoic acid (11.164％).Thirteen components were identified from the mycelia in GYP medium,and among them,the main components with high content were hexadecanoic acid (28.677％),oleic acid (15.108％),and methyl hexadecanoate (8.983％).Fourteen components were identified from the mycelia in CPDA medium,and among them,the main components with high content were methyl hexadecanoate (22.575％),hexadecanoic acid(17.643％),and methyl octadecanoate(4.787％).There were seven identical components existing in three extracts,and three components were only found in wild mycelia,and seven components were only detected in cultured mycelia.Nine components were identified in GYP culture medium,and among them,the main components with high content were octadecamethylcyclononasiloxane(4.663％),1-nonadecene(2.423％) and (E)-3-octadecene (1.785％).Fourteen components were identified in CPDA culture medium,and among them,the main components with high content were hexadecanoic acid (12.913 ％),(Z,E)-2,13-octadecadien-1-ol (5.985％),and methyl hexadecanoate (5.591％).Antibacterial test in vitro showed that the ethyl actate extracts of the GYP culture medium had the strongest antibacterial activity against S.aureus with minimum inhibitory concentration(MIC) value of 0.80 mg/mL,while the MIC of other extracts against S.aureus and E.coli were all 1.60 mg/mL.[Conclusion]Wild E.scabrosa can be cultured in scale in GYP and CPDA media to solve the problem of wild fungus resource shortage.