Development of a Genetic System for Marinobacter atlanticus CP1 (sp. nov.), a Wax Ester Producing Strain Isolated From an Autotrophic Biocathode
Development of a Genetic System for Marinobacter atlanticus CP1 (sp. nov.), a Wax Ester Producing Strain Isolated From an Autotrophic Biocathode
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DOI：
10.3389/fmicb.2018.03176
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发表时间：
2018-12-21
影响因子：
5.2
通讯作者：
Glaven, Sarah M.
中科院分区：
文献类型：
--
作者：
Bird, Lina J.;Wang, Zheng;Glaven, Sarah M.
在这里，我们报告了一个海洋杆菌遗传系统的开发。菌株CP1，以前从生物毒素MCL群落中分离出来，被证明能氧化铁并以阴极生物膜的形式生长。对CP1的16S rRNA基因大大亚基的序列分析以及与部分保守蛋白的比较表明，CP1与海洋杆菌HP15和海洋杆菌属的亲缘关系最近。ES.042。在电子DNA-DNA杂交中，使用基因组到基因组距离计算器(GGDC)预测CP1是一种新的海洋细菌，在这里被描述为大西洋海洋杆菌。CP1能与供体菌株WM3064接合转化到质粒DNA中，在没有抗生素选择的情况下，绿色荧光蛋白(GFP)的结构性表达是稳定的。定向双缺失致突变水华支原体脂肪酰辅酶A还原酶(AcrB)和脂肪醛还原酶(FARA)基因的同源物导致蜡酯产量的损失，然而，这两个基因的单缺失突变导致回收的蜡酯总量增加。为CP1提供的基因工具将使我们能够进一步探索用于生物技术应用的蜡酯合成，以及进一步努力了解CP1在生物催化MCL群落中的作用。
Here, we report on the development of a genetic system for Marinobacter sp. strain CP1, previously isolated from the Biocathode MCL community and shown to oxidize iron and grow as a cathodic biofilm. Sequence analysis of the small and large subunits of the 16S rRNA gene of CP1, as well as comparison of select conserved proteins, indicate that it is most closely related to Marinobacter adhaerens HP15 and Marinobacter sp. ES.042. In silico DNA-DNA hybridization using the genome-to-genome distance calculator (GGDC) predicts CP1 to be a new species of Marinobacter described here as Marinobacter atlanticus. CP1 is competent for transformation with plasmid DNA using conjugation with Escherichia coli donor strain WM3064 and constitutive expression of green fluorescent protein (GFP) is stable in the absence of antibiotic selection. Targeted double deletion mutagenesis of homologs for the M. aquaeoli fatty acyl-CoA reductase (acrB) and fatty aldehyde reductase (farA) genes resulted in a loss of production of wax esters; however, single deletion mutants for either gene resulted in an increase in total wax esters recovered. Genetic tools presented here for CP1 will enable further exploration of wax ester synthesis for biotechnological applications, as well as furthering our efforts to understand the role of CP1 within the Biocathode MCL community.