State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China; Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
College of Life Sciences, Hebei Agricultural University, Baoding 071000, China.
Microbiol Res. 2023 Jul;272:127390. doi: 10.1016/j.micres.2023.127390. Epub 2023 Apr 20.
Trehalose dicorynomycolates are structurally important constituents of the cell envelope in Corynebacterium glutamicum. The genes treS, treY, otsA, mytA and mytB are necessary for the biosynthesis of trehalose dicorynomycolates. In this study, the effect of biosynthesis of trehalose dicorynomycolates on L-isoleucine production in C. glutamicum has been investigated by deleting the genes treS, treY, otsA, mytA, and mytB in the L-isoleucine producing C. glutamicum WM001. L-isoleucine production was slightly improved in the mutants ΔtreY, ΔotsA, and ΔtreYA, and not improved in the single deletion mutant ΔtreS , but significantly improved in the triple deletion mutant ΔtreSYA. Deletion of mytA or mytB in ΔtreSYA could further improve L-isoleucine production. However, deletion of both mytA and mytB in ΔtreSYA significantly decreased L-isoleucine production. The final L-isoleucine producing C. glutamicum WL001 was constructed by deletion of treS, treY, otsA, and mytB, insertion of lrp, and replacement of the native promoter of ilvA with the L-isoleucine sensitive promoter P7. WL001 grew worse than the control WM001, but produced 36.1% more L-isoleucine after 72 h shake flask cultivation than WM001.
海藻糖二胞壁酸酯是谷氨酸棒杆菌细胞包膜结构的重要组成部分。treS、treY、otsA、mytA 和 mytB 基因是海藻糖二胞壁酸酯生物合成所必需的。在本研究中,通过敲除生产 L-异亮氨酸的谷氨酸棒杆菌 WM001 中的 treS、treY、otsA、mytA 和 mytB 基因,研究了海藻糖二胞壁酸酯生物合成对 L-异亮氨酸生产的影响。在突变体 ΔtreY、ΔotsA 和 ΔtreYA 中,L-异亮氨酸的产量略有提高,而在单缺失突变体 ΔtreS 中没有提高,但在三缺失突变体 ΔtreSYA 中显著提高。在 ΔtreSYA 中缺失 mytA 或 mytB 可以进一步提高 L-异亮氨酸的产量。然而,在 ΔtreSYA 中同时缺失 mytA 和 mytB 会显著降低 L-异亮氨酸的产量。通过敲除 treS、treY、otsA 和 mytB,插入 lrp,并将 ilvA 的天然启动子替换为 L-异亮氨酸敏感启动子 P7,构建了最终的 L-异亮氨酸生产谷氨酸棒杆菌 WL001。WL001 的生长状况比对照 WM001 差,但在 72 小时摇瓶培养后,比 WM001 多产生 36.1%的 L-异亮氨酸。