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Optimization of fatty alcohol biosynthesis pathway for selectively enhanced production of C12/14 and C16/18 fatty alcohols in engineered Escherichia coli

机译:优化脂肪醇生物合成途径以选择性提高工程化大肠杆菌中C12 / 14和C16 / 18脂肪醇的产量

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Background With the increasing stress from oil price and environmental pollution, aroused attention has been paid to the microbial production of chemicals from renewable sources. The C12/14 and C16/18 alcohols are important feedstocks for the production of surfactants and detergents, which are widely used in the most respected consumer detergents, cleaning products and personal care products worldwide. Though bioproduction of fatty alcohols has been carried out in engineered E. coli, several key problems have not been solved in earlier studies, such as the quite low production of C16/18 alcohol, the lack of optimization of the fatty alcohol biosynthesis pathway, and the uncharacterized performance of the engineered strains in scaled-up system. Results We improved the fatty alcohol production by systematically optimizing the fatty alcohol biosynthesis pathway, mainly targeting three key steps from fatty acyl-acyl carrier proteins (ACPs) to fatty alcohols, which are sequentially catalyzed by thioesterase, acyl-coenzyme A (CoA) synthase and fatty acyl-CoA reductase. By coexpression of thioesterase gene BTE, acyl-CoA synthase gene fadD and fatty acyl-CoA reductase gene acr1, 210.1?mg/L?C12/14 alcohol was obtained. A further optimization of expression level of BTE, fadD and acr1 increased the C12/14 alcohol production to 449.2?mg/L, accounting for 75.0% of the total fatty alcohol production (598.6?mg/L). In addition, by coexpression of thioesterase gene ‘tesA, acyl-CoA synthase gene fadD and fatty acyl-CoA reductase gene FAR, 101.5?mg/L?C16/18 alcohol was obtained, with C16/18 alcohol accounting for 89.2% of the total fatty alcohol production. Conclusions To our knowledge, this is the first report on selective production of C12/14 and C16/18 alcohols by microbial fermentation. This work achieved high-specificity production of both C12/14 and C16/18 alcohols. The encouraging 598.6?mg/L of fatty alcohols represents the highest titer reported so far. In addition, the 101.5?mg/L 89.2%?C16/18 alcohol suggests an important breakthrough in C16/18 alcohol production. A more detailed optimization of the expression level of fatty alcohol biosynthesis pathway may contribute to a further improvement of fatty alcohol production.
机译:背景技术随着来自油价和环境污染的压力越来越大,引起了人们对由可再生资源化学生产微生物的关注。 C12 / 14和C16 / 18醇是生产表面活性剂和清洁剂的重要原料,已广泛用于全球最受尊敬的消费性清洁剂,清洁产品和个人护理产品中。尽管已在工程化的大肠杆菌中进行了脂肪醇的生物生产,但早期研究尚未解决一些关键问题,例如C16 / 18醇的生产量非常低,脂肪醇生物合成途径的优化不足以及工程菌在放大系统中的未表征的性能。结果我们通过系统地优化脂肪醇的生物合成途径来改善脂肪醇的生产,主要针对从脂肪酰基-酰基载体蛋白(ACP)到脂肪醇的三个关键步骤,这些步骤依次被硫酯酶,酰基辅酶A(CoA)合酶催化和脂肪酰基辅酶A还原酶。通过硫酯酶基因BTE的共表达,得到酰基辅酶A合酶基因fadD和脂肪酰基辅酶A还原酶基因acr1,得到210.1μmg/L≤C12/ 14的醇。 BTE,fadD和acr1表达水平的进一步优化使C12 / 14酒精产量增加至449.2?mg / L,占脂肪醇总产量(598.6?mg / L)的75.0%。另外,通过硫酯酶基因tesA,酰基辅酶A合酶基因fadD和脂肪酰基辅酶A还原酶基因FAR的共表达,得到101.5μg/L≤C16/ 18醇,其中C16 / 18醇占89.2%。总脂肪醇产量。结论据我们所知,这是关于通过微生物发酵选择性生产C12 / 14和C16 / 18醇的第一份报告。这项工作实现了C12 / 14和C16 / 18醇的高特异性生产。令人鼓舞的598.6mg / L脂肪醇是迄今为止报道的最高滴度。此外,101.5?mg / L 89.2%?C16 / 18醇表明C16 / 18醇生产有重要突破。脂肪醇生物合成途径的表达水平的更详细的优化可能有助于脂肪醇生产的进一步改善。

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