Efficient Biosynthesis of R-(-)-Linalool through Adjusting the Expression Strategy and Increasing GPP Supply in Escherichia coli.,Journal of Agricultural and Food Chemistry

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Efficient Biosynthesis of R-(-)-Linalool through Adjusting the Expression Strategy and Increasing GPP Supply in Escherichia coli.
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2020-07-13 , DOI: 10.1021/acs.jafc.0c03664
Xun Wang _{1,

2,

3} , Jing Wu _{1,

2,

3} , Jiaming Chen _{1,

2,

3} , Longjie Xiao _{1,

2,

3} , Yu Zhang _{1,

2,

3} , Fei Wang _{1,

2,

3} , Xun Li _{1,

2,

3}

Affiliation

R-(−)-linalool is widely used in pharmaceutical, agrochemical, and fragrance industries. However, plant extraction furnishes only limited and unstable R-(−)-linalool yields that do not satisfy market demand. Therefore, a sustainable yet efficient and productive method is urgently needed. To induce the R-(−)-linalool biosynthesis pathway in Escherichia coli, we expressed several heterologous (3R)-linalool synthases (LISs) and then chose a suitable LIS from Streptomyces clavuligerus (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^–1R-(−)-linalool under shake flask and fed-batch fermentation conditions, respectively. The latter is the highest reported R-(−)-linalool yield to date. This work could lay theoretical and empirical foundations for engineering terpenoid pathways and optimizing other metabolic pathways.

中文翻译：

R-（-）-Linalool的有效生物合成，通过调整表达策略和增加GPP在大肠杆菌中的供应。

R -（-）-芳樟醇广泛用于制药，农业化学和香料工业。然而，植物提取仅提供了有限且不稳定的R -（-）-芳樟醇产量，不能满足市场需求。因此，迫切需要一种可持续而高效的生产方法。为了诱导大肠杆菌中的R -（-）-芳樟醇生物合成途径，我们表达了几种异源（3 R）-芳樟醇合酶（LISs），然后从棒状链霉菌中选择了合适的LIS（bLIS）有待进一步研究。通过使用优化的核糖体结合位点和蛋白质融合标签，bLIS表达显着升高。为了增加二磷酸香叶基酯的含量，我们测试了异戊二烯基转移酶及其突变体的各种变化。在摇瓶和分批补料发酵条件下，最终菌株分别积累了100.1和1027.3 mg L ^–1R -（-）-芳樟醇。后者是迄今为止报道的最高R -（-）-芳樟醇收率。这项工作可以为工程类萜途径和优化其他代谢途径奠定理论和经验基础。

更新日期：2020-08-05

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