土壤中拮抗菌株铜绿假单胞菌HBD-12次级代谢产物的分离鉴定及其体外抗菌抗肿瘤活性检测

Isolation, structural identification of secondary metabolites from Pseudomonas aeruginosa HBD-12 with antibacterial and antitumor activities

摘要:

为从土壤中筛到具有抗菌和抗肿瘤等生物活性的菌株，以孕烯醇酮作为唯一碳源进行筛菌，经分子生物学鉴定及菌株发酵液抑菌活性测定，发现一株铜绿假单胞菌HBD-12对大肠杆菌、苏云金芽孢杆菌、指状青霉、意大利青霉具有较好抑菌效果。运用柱层析法分离纯化该菌株发酵液成分，采用波谱法解析所得单体化合物结构，并使用HTRF激酶检测试剂盒测定其抗肿瘤活性。结果显示：分离得到的单体化合物1-羟基-9,10-二氮杂菲和3-羟基-9,10-二氢二氮杂菲均具有显著的抗肿瘤活性。在浓度为20 μg/mL时，1-羟基-9,10-二氮杂菲和3-羟基-9,10-二氢二氮杂菲对Aurora激酶A的抑制率分别为78.39%±2.29%和60.34%±8.35%。由此可见该菌株的次级代谢产物对新型抗菌、抗肿瘤药物的研发具有很好的利用价值。

Abstract:

To screen strains with antibacterial and antitumor activity, pregnenolone was used as the sole carbon source for screening bacteria from soil. Based on bacteriostatic activity assay, Pseudomonas aeruginosa HBD-12 was found to be effectively inhibiting the growth of Escherichia coli, Bacillus thuringiensis, Penicillium digitatum and Penicillium italicum, and its fermentation broth was separated and purified using column chromatography. Then, structure of the obtained monomeric compounds was analyzed by spectrum analysis, and their antitumor activity was measured using HTRF kinase detection kit. The isolated monomeric compounds 1-hydroxy-9,10-phenanthroline and 3-hydroxy-9,10-dihydrophenanthroline had significant antitumor activity. At 20 μg/mL, 1-hydroxy-9,10-phenanthroline and 3-hydroxy-9,10-dihydrophenanthroline inhibited 78.39±2.29% and 60.34±8.35% Aurora kinase A, respectively. Therefore, the secondary metabolites of Pseudomonas aeruginosa HBD-12 have the potential to develop antibacterial and antitumor drugs.