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Neutral Lipid Biosynthesis in Engineered Escherichia coli: Jojoba Oil-Like Wax Esters and Fatty Acid Butyl Esters

机译:工程化大肠杆菌中的中性脂质生物合成:霍霍巴油样蜡酯和脂肪酸丁酯

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摘要

Wax esters are esters of long-chain fatty acids and long-chain fatty alcohols which are of considerable commercial importance and are produced on a scale of 3 million tons per year. The oil from the jojoba plant (Simmondsia chinensis) is the main biological source of wax esters. Although it has a multitude of potential applications, the use of jojoba oil is restricted, due to its high price. In this study, we describe the establishment of heterologous wax ester biosynthesis in a recombinant Escherichia coli strain by coexpression of a fatty alcohol-producing bifunctional acyl-coenzyme A reductase from the jojoba plant and a bacterial wax ester synthase from Acinetobacter baylyi strain ADP1, catalyzing the esterification of fatty alcohols and coenzyme A thioesters of fatty acids. In the presence of oleate, jojoba oil-like wax esters such as palmityl oleate, palmityl palmitoleate, and oleyl oleate were produced, amounting to up to ca. 1% of the cellular dry weight. In addition to wax esters, fatty acid butyl esters were unexpectedly observed in the presence of oleate. The latter could be attributed to solvent residues of 1-butanol present in the medium component, Bacto tryptone. Neutral lipids produced in recombinant E. coli were accumulated as intracytoplasmic inclusions, demonstrating that the formation and structural integrity of bacterial lipid bodies do not require specific structural proteins. This is the first report on substantial biosynthesis and accumulation of neutral lipids in E. coli, which might open new perspectives for the biotechnological production of cheap jojoba oil equivalents from inexpensive resources employing recombinant microorganisms.
机译:蜡酯是长链脂肪酸和长链脂肪醇的酯,它们在商业上具有重要意义,年产量为300万吨。来自霍霍巴植物(Simmondsia chinensis)的油是蜡酯的主要生物来源。尽管它具有多种潜在应用,但由于其价格高昂,荷荷巴油的使用受到限制。在这项研究中,我们描述了通过重组表达荷荷巴油中产生脂肪醇的双功能酰基辅酶A还原酶和来自不动杆菌Baylyi菌株ADP1的细菌蜡酯合酶的表达,在重组大肠杆菌菌株中建立异源蜡酯的生物合成。脂肪酸的酯化和脂肪酸的辅酶A硫酯。在油酸酯的存在下,产生了霍霍巴油状蜡酯,例如棕榈基油酸酯,棕榈基棕榈油酸酯和油基油酸酯,总计约达10%。细胞干重的1%。除蜡酯外,在油酸酯的存在下意外地观察到了脂肪酸丁酯。后者可归因于培养基成分Bacto胰蛋白present中存在的1-丁醇溶剂残留。重组大肠杆菌中产生的中性脂质被积累为胞浆内包裹体,表明细菌脂质体的形成和结构完整性不需要特定的结构蛋白。这是有关大肠杆菌中中性脂质大量生物合成和积累的第一份报告,这可能为利用重组微生物从廉价资源中廉价生产霍霍巴油当量的生物技术生产开辟新的前景。

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