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芽孢杆菌属中生物表面活性剂的检测:基因与产物鉴定

Detection of biosurfactants in Bacillus species: genes and products identification.

作者信息

Płaza G, Chojniak J, Rudnicka K, Paraszkiewicz K, Bernat P

机构信息

Department of Environmental Microbiology, Institute for Ecology of Industrial Areas, Katowice, Poland.

Laboratory of Gastroimmunology, Department of Immunology and Infectious Biology, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, Poland.

出版信息

J Appl Microbiol. 2015 Oct;119(4):1023-34. doi: 10.1111/jam.12893. Epub 2015 Sep 4.

Abstract

AIM

To screen environmental Bacillus strains for detection of genes encoding the enzymes involved in biosurfactant synthesis and to evaluate their products e.g. surfactin, iturin and fengycin.

MATERIALS AND RESULTS

The taxonomic identification of isolated from the environment Bacillus strains was performed by Microgene ID Bacillus panel and GEN III Biolog system. The polymerase chain reaction (PCR) strategy for screening of genes in Bacillus strains was set up. Liquid chromatography-mass spectrometry (LC-MS/MS) method was used for the identification of lipopeptides (LPs). All studied strains exhibited the presence of srfAA gene and produced surfactin mostly as four homologues (C13 to C16). Moreover, in 2 strains (KP7, T'-1) simultaneous co-production of 3 biosurfactants: surfactin, iturin and fengycin was observed. Additionally, it was found out that isolate identified as Bacillus subtilis ssp. subtilis (KP7), beside LPs co-production, synthesizes surfactin with the efficiency much higher than other studied strains (40·2 mg l(-1) ) and with the yield ranging from 0·8 to 8·3 mg l(-1) .

CONCLUSION

We showed that the combined methodology based on PCR and LC-MS/MS technique is an optimal tool for the detection of genes encoding enzymes involved in biosurfactant synthesis as well as their products, e.g. surfactin, iturin and fengycin. This approach improves the screening and the identification of environmental Bacillus co-producing biosurfactants-stimulating and facilitating the development of this area of science.

SIGNIFICANCE AND IMPACT OF THE STUDY

The findings of this work will help to improve screening of biosurfactant producers. Discovery of novel biosurfactants and biosurfactants co-production ability has shed light on their new application fields and for the understanding of their interactions and properties.

摘要

目的

筛选环境芽孢杆菌菌株,以检测编码参与生物表面活性剂合成的酶的基因,并评估其产物,如表面活性素、伊枯草菌素和丰原素。

材料与结果

通过Microgene ID芽孢杆菌鉴定板和GEN III Biolog系统对从环境中分离的芽孢杆菌菌株进行分类鉴定。建立了用于筛选芽孢杆菌菌株中基因的聚合酶链反应(PCR)策略。采用液相色谱-质谱联用(LC-MS/MS)方法鉴定脂肽(LP)。所有研究菌株均显示存在srfAA基因,且产生的表面活性素大多为四种同系物(C13至C16)。此外,在2株菌株(KP7、T'-1)中观察到同时共产生3种生物表面活性剂:表面活性素、伊枯草菌素和丰原素。另外,发现鉴定为枯草芽孢杆菌枯草亚种(KP7)的分离株,除了共产生LP外,合成表面活性素的效率远高于其他研究菌株(40.2 mg l(-1)),产量范围为0.8至8.3 mg l(-1)。

结论

我们表明,基于PCR和LC-MS/MS技术的联合方法是检测编码参与生物表面活性剂合成的酶及其产物(如表面活性素、伊枯草菌素和丰原素)的基因的最佳工具。这种方法改进了对共产生生物表面活性剂的环境芽孢杆菌的筛选和鉴定,刺激并促进了该科学领域的发展。

研究的意义和影响

这项工作的发现将有助于改进生物表面活性剂生产者的筛选。新型生物表面活性剂的发现及其共生产能力为其新的应用领域以及理解它们的相互作用和性质提供了线索。

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