粘细菌TA菌株、植物内生真菌AF001、EYR61菌株次级代谢产物的初步研究
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摘要
本论文对粘细菌TA菌株、植物内生真菌AF001和EYR61菌株的发酵代谢产物进行了分离纯化,运用核磁共振、质谱的方法进行结构鉴定,对部分化合物进行X-Ray单晶衍射分析、红外光谱、紫外可见光谱和旋光值的测定,并通过抗菌、抗肿瘤实验对部分化合物进行活性检测。
对TA菌株次级代谢产物的分离纯化,共得到5个化合物TA7、TA9、TA10、TA11和ITA12。通过对~1H-NMR的分析,鉴定出这5个化合物都为聚醚类化合物,推测其为液体发酵过程中添加的消泡剂。
AF001菌株为番荔枝植物内生真菌,通过形态学和分子生物学方法鉴定为球座菌(Guignardia mangiferae)。从其固体平板发酵产物中分离得到2个脂肪酸类化合物LAF15和LAF16,1个甾醇类化合物LAF18,以及5个全新结构的杂合萜类化合物LAF1、LAF4、LAF11、LAF12和LAF13。对这5个杂合萜进行了抗菌、抗肿瘤实验以及红外光谱、紫外可见光谱、旋光值的测定;并通过X-Ray单晶衍射分析确定化合物LAF1的相对立体构型。抗菌活性实验结果表明,化合物LAF4、LAF11和LAF13对白假丝酵母菌有微弱的抑制活性,在50μg/mL浓度下抑制率分别为12.9%、33.7%和21.5%;细胞毒性测定结果表明,在浓度为20μg/mL时,5个化合物对人肝癌HepG2细胞表现出微弱的生长抑制活性,抑制率分别为1 1.35%、8.88%、6.71%、4.34%和5.03%。
EYR61菌株为马尾松植物内生真菌,通过形态学和分子生物学方法鉴定其可能为嗜热子囊菌(Thermoascus crustaceus)。从固体平板发酵产物中分离鉴定了两个真菌聚酮类化合物EYR1和EYR15,分别鉴定为10-去甲基藜芦酸甲酯(methyl 10-demethylveratrate)和降红没药酯(Norbisabolide),其中EYR1也是新化合物。另外正在对化合物EYR3、EYR7、EYR8和EYR13的结构进行解析。
本论文的实验结果表明,植物内生真菌、特别是药用植物的内生真菌,能够产生众多结构新颖的次级代谢产物,其中不乏具有生物活性的物质,因而植物内生真菌的代谢产物是开发新药的重要资源。
In this study,the secondary metabolites of strain TA of myxobacteria, endophytic fungi strains AF001 and EYR61 were studied.Their secondary metabolites were separated and purified,structure of compounds were elucidated by extensive spectroscopic data analyses.In the same time,some compounds were elucidated by X-ray crystallographic,Infrared spectrum, UV-visible spectrum and Optical rotations analysis,and antimicrobe, anticancer activities were also be studied.
The secondary metabolites of the strain TA were studied and five compounds TA7,TA9,TA10,TA11 and TA12 were obtained.They were all polyethers elucidated by ~1H-NMR analysis.We consider that they are foam suppressor which added during liquid fermentation.
The strain AF001 is an endophytic fungi of sweetsop,it was identified by morphological and molecule biological methods.The result indicated that the strain AF001 belonged to Guignardia mangiferae.The secondary,metabolites of its solid fermentation were studied,we got LAF15 and LAF16 two fatty acids and one sterols compound LAF18,moreover we found five new compounds LAF1,LAF4,LAF11,LAF12 and LAF13,these new compounds were elucidated by Infrared spectrum,UV-visible spectrum,Optical rotations analysis,and their antimicrobe,anticancer activities were also be studied,and we also did X-ray crystallographic analysis of compound LAF1.The result indicated that LAF4,LAF11 and LAF13 exhibited weak antimicrobe activities against Candida albicans at 50μg/mL,their suppression ratio were 12.9%, 33.7%and 21.5%respectively,five new compounds all exhibited weak anticancer activities against human liver cancer cell HepG2 at 20μg/mL,their suppression ratio were 11.35%,8.88%,6.71%,4.34%and 5.03%respectively.
The strain EYR61 is an endophytic fungi of redpine,it was identified by morphological and molecule biological methods.The result indicated that the strain EYR61 belonged to Thermoascus crustaceus.Two compounds EYR1 and EYR15 were identified from secondary metabolites of EYR61 solid fermentation.EYR1 was a new compound,and the structure of EYR3,EYR7, EYR8 and EYR13 are being analyzed now.
In conclusion,endophytic fungi,especially from offcinal,can produce numerous new and unique stucture secondary metabolites,and many of them have biological activity,so the secondary metabolites of endophytic fungi are an important source of new drug discovery.
对TA菌株次级代谢产物的分离纯化,共得到5个化合物TA7、TA9、TA10、TA11和ITA12。通过对~1H-NMR的分析,鉴定出这5个化合物都为聚醚类化合物,推测其为液体发酵过程中添加的消泡剂。
AF001菌株为番荔枝植物内生真菌,通过形态学和分子生物学方法鉴定为球座菌(Guignardia mangiferae)。从其固体平板发酵产物中分离得到2个脂肪酸类化合物LAF15和LAF16,1个甾醇类化合物LAF18,以及5个全新结构的杂合萜类化合物LAF1、LAF4、LAF11、LAF12和LAF13。对这5个杂合萜进行了抗菌、抗肿瘤实验以及红外光谱、紫外可见光谱、旋光值的测定;并通过X-Ray单晶衍射分析确定化合物LAF1的相对立体构型。抗菌活性实验结果表明,化合物LAF4、LAF11和LAF13对白假丝酵母菌有微弱的抑制活性,在50μg/mL浓度下抑制率分别为12.9%、33.7%和21.5%;细胞毒性测定结果表明,在浓度为20μg/mL时,5个化合物对人肝癌HepG2细胞表现出微弱的生长抑制活性,抑制率分别为1 1.35%、8.88%、6.71%、4.34%和5.03%。
EYR61菌株为马尾松植物内生真菌,通过形态学和分子生物学方法鉴定其可能为嗜热子囊菌(Thermoascus crustaceus)。从固体平板发酵产物中分离鉴定了两个真菌聚酮类化合物EYR1和EYR15,分别鉴定为10-去甲基藜芦酸甲酯(methyl 10-demethylveratrate)和降红没药酯(Norbisabolide),其中EYR1也是新化合物。另外正在对化合物EYR3、EYR7、EYR8和EYR13的结构进行解析。
本论文的实验结果表明,植物内生真菌、特别是药用植物的内生真菌,能够产生众多结构新颖的次级代谢产物,其中不乏具有生物活性的物质,因而植物内生真菌的代谢产物是开发新药的重要资源。
In this study,the secondary metabolites of strain TA of myxobacteria, endophytic fungi strains AF001 and EYR61 were studied.Their secondary metabolites were separated and purified,structure of compounds were elucidated by extensive spectroscopic data analyses.In the same time,some compounds were elucidated by X-ray crystallographic,Infrared spectrum, UV-visible spectrum and Optical rotations analysis,and antimicrobe, anticancer activities were also be studied.
The secondary metabolites of the strain TA were studied and five compounds TA7,TA9,TA10,TA11 and TA12 were obtained.They were all polyethers elucidated by ~1H-NMR analysis.We consider that they are foam suppressor which added during liquid fermentation.
The strain AF001 is an endophytic fungi of sweetsop,it was identified by morphological and molecule biological methods.The result indicated that the strain AF001 belonged to Guignardia mangiferae.The secondary,metabolites of its solid fermentation were studied,we got LAF15 and LAF16 two fatty acids and one sterols compound LAF18,moreover we found five new compounds LAF1,LAF4,LAF11,LAF12 and LAF13,these new compounds were elucidated by Infrared spectrum,UV-visible spectrum,Optical rotations analysis,and their antimicrobe,anticancer activities were also be studied,and we also did X-ray crystallographic analysis of compound LAF1.The result indicated that LAF4,LAF11 and LAF13 exhibited weak antimicrobe activities against Candida albicans at 50μg/mL,their suppression ratio were 12.9%, 33.7%and 21.5%respectively,five new compounds all exhibited weak anticancer activities against human liver cancer cell HepG2 at 20μg/mL,their suppression ratio were 11.35%,8.88%,6.71%,4.34%and 5.03%respectively.
The strain EYR61 is an endophytic fungi of redpine,it was identified by morphological and molecule biological methods.The result indicated that the strain EYR61 belonged to Thermoascus crustaceus.Two compounds EYR1 and EYR15 were identified from secondary metabolites of EYR61 solid fermentation.EYR1 was a new compound,and the structure of EYR3,EYR7, EYR8 and EYR13 are being analyzed now.
In conclusion,endophytic fungi,especially from offcinal,can produce numerous new and unique stucture secondary metabolites,and many of them have biological activity,so the secondary metabolites of endophytic fungi are an important source of new drug discovery.
引文
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