Production of free fatty acids (FFAs) and derivatives from renewable non-food biomass by microbial fermentation is of great interest. Here, we report the development of engineered Rhodococcus opacus strains producing FFAs, fatty acid ethyl esters (FAEEs) and long-chain hydrocarbons (LCHCs). Culture conditions were optimized to produce 82.9 g l⁻¹ of triacylglycerols from glucose, and an engineered strain with acyl-coenzyme A (CoA) synthetases deleted, overexpressing three lipases with lipase-specific foldase produced 50.2 g l⁻¹ of FFAs. Another engineered strain with acyl-CoA dehydrogenases deleted, overexpressing lipases, foldase, acyl-CoA synthetase and heterologous aldehyde/alcohol dehydrogenase and wax ester synthase produced 21.3 g l⁻¹ of FAEEs. A third engineered strain with acyl-CoA dehydrogenases and alkane-1 monooxygenase deleted, overexpressing lipases, foldase, acyl-CoA synthetase and heterologous acyl-CoA reductase, acyl-ACP reductase and aldehyde deformylating oxygenase produced 5.2 g l⁻¹ of LCHCs. Metabolic engineering strategies and engineered strains developed here may help establish oleaginous biorefinery platforms for the sustainable production of chemicals and fuels.

通过微生物发酵从可再生的非食物生物质中生产游离脂肪酸（FFA）和衍生物非常受关注。在这里，我们报告了工程化的不育红球菌菌株的产生，所述菌株产生了FFA，脂肪酸乙酯（FAEE）和长链烃（LCHC）。优化培养条件以从葡萄糖产生82.9 g l ^-1的三酰基甘油，并删除了带有酰基辅酶A（CoA）合成酶的工程菌株，过表达具有脂肪酶特异性折叠酶的三种脂肪酶产生了50.2 g l ^-1的FFA。另一个带有酰基辅酶A脱氢酶的工程菌株被删除，过表达脂肪酶，折叠酶，酰基辅酶A合成酶和异源醛/醇脱氢酶和蜡酯合酶产生21.3 g l ^-1FAEEs。删除了带有酰基辅酶A脱氢酶和烷烃1单加氧酶的第三个工程菌株，过表达脂肪酶，折叠酶，酰基辅酶A合成酶和异源酰基辅酶A还原酶，酰基辅酶ACP还原酶和醛脱甲酰加氧酶产生了5.2 g ^-1的LCHCs。这里开发的代谢工程策略和工程菌株可能有助于建立油质生物精炼平台，以可持续地生产化学品和燃料。