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在红球菌 PD630 中重新布线中性脂质生产以从头合成蜡酯。

Rewiring neutral lipids production for the de novo synthesis of wax esters in Rhodococcus opacus PD630.

机构信息

Instituto de Biociencias de la Patagonia (INBIOP), Universidad Nacional de la Patagonia San Juan Bosco y CONICET, Km 4-Ciudad Universitaria 9000, Comodoro Rivadavia, Chubut, Argentina.

Instituto de Biociencias de la Patagonia (INBIOP), Universidad Nacional de la Patagonia San Juan Bosco y CONICET, Km 4-Ciudad Universitaria 9000, Comodoro Rivadavia, Chubut, Argentina.

出版信息

J Biotechnol. 2017 Oct 20;260:67-73. doi: 10.1016/j.jbiotec.2017.09.009. Epub 2017 Sep 14.

Abstract

Rhodococcus opacus PD630 accumulates significant amounts of triacylglycerols (TAG), but is not able to de novo synthesize wax esters (WE) from structural unrelated carbon sources, such as gluconate. In this study, strain PD630 was engineered to produce WE by heterologous expression of maqu_2220 gene, which encodes a fatty acyl-CoA reductase for the production of fatty alcohols in Marinobacter hydrocarbonoclasticus. Recombinant cells produced ca. 46% of WE and 54% of TAG (of total WE+TAG) from gluconate compared with the wild type, which produced 100% of TAG. Cell growth was not affected by the heterologous expression of MAQU_2220. Several saturated and monounsaturated WE species were produced by cells, with C18:C16, C16:C16 and C16:C18 as main species. The fatty acid composition of WE fraction in PD630maqu_2220 was enriched with C, C, whereas C, C and C predominated in the TAG fraction. Significant amounts of WE and TAG were accumulated by PD630maqu_2220 from whey, an inexpensive waste material from dairy industries, without affecting cell biomass production. This is the first report on WE synthesis by R. opacus from gluconate, which demonstrates that lipid metabolism of this bacterium is flexible enough to assimilate heterologous components for the production of new lipid derivatives with industrial interest.

摘要

红平红球菌 PD630 能够积累大量三酰基甘油(TAG),但无法从头合成结构上不相关的碳源(如葡萄糖酸盐)的蜡酯(WE)。在这项研究中,通过异源表达 Marinobacter hydrocarbonoclasticus 中编码脂肪酸酰基辅酶 A 还原酶的 maqu_2220 基因,对 PD630 菌株进行了工程改造,以生产 WE。与野生型相比,重组细胞从葡萄糖酸盐中产生了约 46%的 WE 和 54%的 TAG(总 WE+TAG),而野生型则产生了 100%的 TAG。异源表达 MAQU_2220 对细胞生长没有影响。细胞产生了几种饱和和单不饱和的 WE 物质,主要物种为 C18:C16、C16:C16 和 C16:C18。WE 馏分中 MAQU_2220 的脂肪酸组成富含 C、C,而 TAG 馏分中 C、C 和 C 占优势。PD630maqu_2220 可以从乳清(乳制品工业的廉价废物)中积累大量的 WE 和 TAG,而不会影响细胞生物量的产生。这是红平红球菌首次从葡萄糖酸盐中合成 WE 的报道,证明了该细菌的脂质代谢足够灵活,可以吸收异源成分,生产具有工业价值的新脂质衍生物。

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