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从采集自南极FAO 48.1海域海水样品中分离纯化到一株产吡咯喹啉醌(PQQ)海洋细菌,对其进行了细菌分类学鉴定、摇瓶培养发酵、发酵产物提取及HPLC定量分析,并对纯化的PQQ进行了针对一氧化氮(NO)抑制活性及羟自由基(.OH)清除抗氧化活性评价。结果表明,该菌株与副球菌属(Paracoccus sp.)内菌株的16S rRNA基因序列同源性较高。综合多个分类学指标的分析结果,将其鉴定为属于副球菌属,命名为Paracoccus sp.PQ-08。生物活性评价结果表明,PQQ可显著抑制人脐静脉内皮(ECV-304)模式细胞内诱导型一氧化氮合成酶(iNOS)在mRNA转录水平及蛋白水平的表达量,明显降低胞内的NO释放量。当其作用浓度为100μM时,对iNOS蛋白表达的最高抑制率为67.9%(P<0.05)。此外,实验还证实PQQ具有较强的清除.OH的抗氧化活性。
A bacterial strain of PQQ was isolated and purified from seawater samples collected from the 48.1 sea area of Antarctic. The bacteria were identified by taxonomy, shake flask culture, fermentation product extraction and HPLC quantitative analysis. Purified PQQ was evaluated against nitric oxide (NO) inhibitory activity and hydroxyl radical (.OH) scavenging antioxidant activity. The results showed that the 16S rRNA gene of Paracoccus sp. Was highly homologous. Based on the analysis of multiple taxonomic indicators, it was identified as Paracoccus sp., Named Paracoccus sp. PQ-08. The results of bioassay showed that PQQ could significantly inhibit the expression of inducible nitric oxide synthase (iNOS) in human umbilical vein endothelial cells (ECV-304) at mRNA level and protein level, and significantly reduce intracellular NO release the amount. The highest inhibition of iNOS protein expression was 67.9% (P <0.05) at a concentration of 100 μM. In addition, the experiment also confirmed that PQQ has strong anti-oxidative activity of OH.