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A novel rhamnolipid-producing Pseudomonas aeruginosa ZS1 isolate derived from petroleum sludge suitable for bioremediation

机译：一种新型的鼠李糖脂生产的绿脓杆菌铜绿假单胞菌ZS1分离物适用于生物修复的石油污泥

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Petroleum pollutants emulsified by biosurfactants facilitate uptake and biodegradation by environmental microbes. In this report, we show the characterization of an indigenous surfactant-producing crude-oil-eating microbe isolated from petroleum-sludge in Zhoushan islands, China, where one of the national strategic petroleum reservoirs is located. We examined biosurfactant activities using surface tension analysis on mixed culture originated from oil-sludge microbes. In parallel, dynamics of individual microbial populations in cultures were monitored using the terminal fluorescence labeled (TFL)-RFLP method. Biosurfactant activity was found to be associated with a dominant microbial species designated as ZS1 (Zhou-Shan isolate 1). Cell-free supernatant from the ZS1 culture exhibited 100% emulsification index against crude oil and reduces surface tension to 26.5 mN/m. Sequence-based analysis suggested that the ZS1 belongs to the group of Pseudomonas aeruginosa. LC–MS/MS analysis indicated that the rhamnolipids produced by the ZS1 consisted of 7 monorhamnolipid and 11 dirhamnolipid homologues (RL7-11), two of which were novel. Maximum yield of rhamnolipids in shake-flask ZS1 culture could reach 44 g/l. Furthermore, we showed that ZS1 was capable of growing in MS medium supplemented with 1% crude oil as sole carbon source, in which cell mass increase coincided with the formation of crude-oil emulsion. Half of the crude oil was consumed by the ZS1 in 12 days. Taken together, our results show that ZS1 produces high level of rhamnolipids that effectively emulsify crude oil accelerating its uptake and degradation. Hence, it is suitable for bioremediation of petroleum pollutants in Zhoushan coastal areas.
机译：被生物表面活性剂乳化的石油污染物促进了环境微生物的吸收和生物降解。在本报告中，我们显示了从中国舟山群岛的石油污泥中分离出的一种本土生产表面活性剂的原油吞噬微生物的特征，该岛是国家战略石油储备库之一。我们对源自油泥微生物的混合培养物进行了表面张力分析，从而研究了生物表面活性剂的活性。同时，使用末端荧光标记（TFL）-RFLP方法监测培养物中单个微生物种群的动态。已发现生物表面活性剂的活性与指定为ZS1的优势微生物物种有关（Zhou-Shan分离株1）。 ZS1培养物中无细胞的上清液对原油表现出100％的乳化指数，并将表面张力降低至26.5 mN / m。基于序列的分析表明ZS1属于铜绿假单胞菌。 LC-MS / MS分析表明，ZS1产生的鼠李糖脂由7个单鼠李糖脂和11个鼠李糖脂同系物（RL7-11）组成，其中两个是新奇的。摇瓶ZS1培养物中鼠李糖脂的最大产量可达到44 g / l。此外，我们表明ZS1能够在添加1％原油作为唯一碳源的MS培养基中生长，其中细胞质量的增加与原油乳液的形成相吻合。 ZS1在12天内消耗了一半的原油。两者合计，我们的结果表明ZS1产生高水平的鼠李糖脂，可有效地乳化原油，加速其吸收和降解。因此，适用于舟山沿海地区石油污染物的生物修复。

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