百两金皂苷A的微生物转化及其产物的细胞毒活性研究

Study on biotransformation of ardisiacrispin A and cytotoxic activity of transformed products

目的利用燕麦曲霉Aspergillus avenaceus AA 3.4454对百两金皂苷A进行微生物转化,并对转化产物进行细胞毒活性研究。方法将百两金皂苷A放置燕麦曲霉液体培养基中,28℃、160 r/min条件下共培养3 d后,利用多种柱色谱分离转化产物,采用核磁共振等波谱手段鉴定结构;采用MTT法测定转化产物对肿瘤细胞系的细胞毒活性。结果从百两金皂苷A的燕麦曲霉液体发酵液中分离出3个主要转化产物,其结构分别鉴定为西克拉明皂苷元A-3β-O-{α-D-半乳吡喃糖基-(1→4)-[β-D-木吡喃糖基-(1→2)]-β-D-吡喃葡萄糖基-(1→4)-[β-D-吡喃葡萄糖基-(1→2)]-α-L-阿拉伯吡喃糖苷}(1)、西克拉明皂苷元A-3β-O-{α-D-半乳吡喃糖基-(1→4)-β-D-吡喃葡萄糖基-(1→2)-[β-D-木吡喃糖基-(1→2)-β-D-吡喃葡萄糖基-(1→4)]-α-L-阿拉伯吡喃糖苷}(2)、西克拉明皂苷元A-3β-O-{β-D-木吡喃糖基-(1→2)-β-D-吡喃葡萄糖基-(1→4)-[β-D-吡喃葡萄糖基-(1→2)]-[α-D-半乳吡喃糖基-(1→3)]-α-L-阿拉伯吡喃糖苷}(3)。结论首次利用燕麦曲霉对百两金皂苷A进行微生物转化并分离得到在糖链的不同位置增加了α-D型半乳糖的转化产物,3个转化产物均为新化合物,均具有一定的细胞毒活性,且转化产物1对大细胞肺癌细胞的细胞毒活性略优于底物。

Objective To study the microbial transformation of ardisiacrispin A by Aspergillus avenaceus AA 3.4454 and the cytotoxic activity of the transformed products.Methods Ardisiacrispin A was put into the fluid medium inoculated with the A.avenaceus and cultured in the shaker at 28℃and 160 r/min for 3 d.Three transformed products were separated and purified from the crude extract whose structures were elucidated based on spectrum data.And the cytotoxic activity of the transformed products on tumor cells was determined by MTT assay.Results Three major transformed products were elucidated as cyclamiretin A-3β-O-{α-D-galactopyranosyl-(14)-[β-D-xylopyranosyl-(1→2)]-β-D-glucopyranosyl-(1→4)-[β-D-glucopyranosyl-(1→2)]-α-L-arabinopyranoside}(1),cyclamiretin A-3β-O-{α-D-galactopyranosyl-(1→4)-β-D-glucopyranosyl-(1→2)-[β-D-xylopyranosyl-(1→2)-β-Dglucopyranosyl-(1→4)]-α-L-arabinopyranoside}(2),and cyclamiretin A-3β-O-{β-D-xylopyranosyl-(1→2)-β-D-glucopyranosyl-(1→4)-[β-Dglucopyranosyl-(1→2)]-[α-D-galactopyranosyl-(1→3)]-α-L-arabinopyranoside}(3).Conclusion It is the first time that the three major derivatives could be microbial synthesized from ardisiacrispin A by A.avenaceus AA 3.4454.And the three transformation products are all new compounds which increaseα-D galactose at different positions of the sugar chain.All the transformed products have certain cytotoxic activity,and the cytotoxic activity of the transformed product 1 to NCI-H460 cells is slightly better than the substrate.