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通过调整表达策略和增加 GPP 供应来高效生物合成 (-)-芳樟醇。

Efficient Biosynthesis of -(-)-Linalool through Adjusting the Expression Strategy and Increasing GPP Supply in .

机构信息

Jiangsu Provincial Key Laboratory for the Chemistry and Utilization of Agro-Forest Biomass, Nanjing Forestry University, Nanjing 210037, China.

Jiangsu Key Laboratory of Biomass-Based Green Fuels and Chemicals, Nanjing Forestry University, Nanjing 210037, China.

出版信息

J Agric Food Chem. 2020 Aug 5;68(31):8381-8390. doi: 10.1021/acs.jafc.0c03664. Epub 2020 Jul 22.

Abstract

-(-)-linalool is widely used in pharmaceutical, agrochemical, and fragrance industries. However, plant extraction furnishes only limited and unstable -(-)-linalool yields that do not satisfy market demand. Therefore, a sustainable yet efficient and productive method is urgently needed. To induce the -(-)-linalool biosynthesis pathway in , we expressed several heterologous (3)-linalool synthases (LISs) and then chose a suitable LIS from (bLIS) for further study. The bLIS expression was markedly elevated by using optimized ribosomal binding sites and protein fusion tags. To increase the geranyl diphosphate content, we tested various alterations in prenyltransferases and their mutants. The final strain accumulated 100.1 and 1027.3 mg L -(-)-linalool under shake flask and fed-batch fermentation conditions, respectively. The latter is the highest reported -(-)-linalool yield to date. This work could lay theoretical and empirical foundations for engineering terpenoid pathways and optimizing other metabolic pathways.

摘要

(-)-芳樟醇在制药、农化和香料工业中被广泛应用。然而,植物提取只能提供有限且不稳定的 (-)-芳樟醇产量,无法满足市场需求。因此,迫切需要一种可持续且高效、高产的方法。为了在 中诱导 (-)-芳樟醇生物合成途径,我们表达了几种异源 (3)-芳樟醇合酶 (LIS),然后选择了一种合适的来自 (bLIS) 进行进一步研究。通过优化核糖体结合位点和蛋白融合标签,bLIS 的表达明显提高。为了增加香叶基二磷酸的含量,我们测试了各种烯丙基转移酶及其突变体的改变。最终的菌株在摇瓶和分批补料发酵条件下分别积累了 100.1 和 1027.3 mg/L 的 (-)-芳樟醇,后者是迄今为止报道的最高 (-)-芳樟醇产量。这项工作为萜类化合物途径的工程和其他代谢途径的优化奠定了理论和经验基础。

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