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【目的】为发现天然的防污损物质,从分离自海绵Haliclona sp.的细菌Pseudomonas putida中寻找具有抗硅藻附着活性的化合物。【方法】综合菌落生长形态、扫描电镜及16S rDNA序列分析,鉴定细菌种属;采用活性(抗硅藻附着)-化学(薄层色谱、二极管阵列高效液相和核磁共振氢谱)导向法对其中的活性组分进行分离纯化,波谱分析确定结构;采用抗硅藻附着活性模型对单体化合物进行活性复筛。【结果】该细菌鉴定为Pseudomonas putida;从其发酵液中分离得到6个环二肽,分别鉴定为环(亮氨酸-脯氨酸)(1)、环(亮氨酸-丙氨酸)(2)、环(苯丙氨酸-丙氨酸)(3)、环(缬氨酸-酪氨酸)(4)、环(丙氨酸-酪氨酸)(5)、环(丙氨酸-色氨酸)(6);化合物3和6在浓度为50μg/mL时具有明显的抑制硅藻附着活性(抑制率分别为50%和85%)。【结论】海洋细菌Pseudomonas putida中具有抗硅藻附着的活性化合物为环(苯丙氨酸-丙氨酸)和环(丙氨酸-色氨酸)。
[Purpose] To find out natural anti-fouling substances, a bacterium Pseudomonas putida isolated from the sponge Haliclona sp. Was searched for compounds having anti-diatom-attachment activity. 【Method】 The bacterial species were identified by the growth pattern of colonies, scanning electron microscopy and 16S rDNA sequence analysis. The active (anti-diatom) -chemical (TLC, diode array and 1H-NMR) Among them, the active components were isolated and purified, and their structures were determined by spectral analysis. The active compounds of the resveratrol were screened by anti-diathesis activity model. 【Result】 The strain was identified as Pseudomonas putida. Six cyclic dipeptides were isolated from the fermentation broth and identified as cyclic (leucine-proline) (1), cyclic (leucine-alanine) (3), cyclo (valine-tyrosine) (4), cyclo (alanine-tyrosine) (5), cyclo Tryptophan) (6). Compounds 3 and 6 showed significant inhibitory activity against diatoms (50% and 85% inhibition, respectively) at a concentration of 50 μg / mL. 【Conclusion】 The active compounds against marine bacteria Pseudomonas putida that are resistant to diatoms are cyclic (phenylalanine-alanine) and cyclic (alanine-tryptophan).