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.
J Agric Food Chem. 2021 May 26;69(20):5663-5670. doi: 10.1021/acs.jafc.1c01101. Epub 2021 May 13.
-(-)-Linalool is widely used in the pharmaceutical, agrochemical, and fragrance industries; however, its applications are limited owing to low yield and high cost of production. To improve the production efficiency of -(-)-linalool in , three enzymes [-derived isopentenyl diphosphate isomerase, -derived geranyl diphosphate synthase, and -derived (3)-linalool synthases] were physically colocalized to synthetic complexes using synthetic protein scaffolds of GTPase-binding domain, Src homology 3, and PSD95/DlgA/Zo-1. -(-)-Linalool was produced at the highest concentration in the strain IGL114 containing a scaffold ratio of 1:1:4. By further optimizing the inducer, temperature, and glycerol concentration, the production titer of -(-)-linalool in the shake flask was increased by approximately 10 times compared with that of the scaffold-free control and was 2.78 times the previously reported yield. The production in the fermenter was about 1.5 times the previous highest production. In general, the final strain accumulated 277.8 and 1523.2 mg/L -(-)-linalool under the conditions of shake-flask and fed-batch fermentation, respectively. This study provides a foundation for the assembly of bacterial intracellular protein scaffolds.
(-)-芳樟醇广泛应用于制药、农化和香料行业；然而，由于其产量低、生产成本高，其应用受到限制。为了提高 (-)-芳樟醇在 的生产效率，使用 GTP 结合域、Src 同源 3 和 PSD95/DlgA/Zo-1 的合成蛋白支架，将三种酶[-衍生的异戊烯二磷酸异构酶、-衍生的香叶基二磷酸合酶和 -衍生的（3）-芳樟醇合酶]物理共定位到合成复合物中。在含有支架比为 1:1:4 的 IGL114 菌株中，(-)-芳樟醇的浓度最高。通过进一步优化诱导剂、温度和甘油浓度，与无支架对照相比，摇瓶中 (-)-芳樟醇的产量提高了约 10 倍，是之前报道产量的 2.78 倍。发酵罐中的产量约为之前最高产量的 1.5 倍。总的来说，在摇瓶和分批补料发酵条件下，最终菌株分别积累了 277.8 和 1523.2 mg/L 的(-)-芳樟醇。本研究为细菌细胞内蛋白支架的组装提供了基础。
