College of Enology, Northwest A&F University, Yangling 712100, China.
State Key Laboratory of Microbial Technology, Shandong University, Qingdao 266237, China.
ACS Synth Biol. 2024 Aug 16;13(8):2545-2554. doi: 10.1021/acssynbio.4c00306. Epub 2024 Jun 11.
is a potential workhorse for production of various value-added chemicals including terpenoids, oleo-chemicals, and enzymes from low-cost feedstocks. However, the limited genetic toolbox is hindering its metabolic engineering. In the present study, four type I and one novel type II peroxisomal targeting signal (PTS1/PTS2) were characterized and employed for limonene production for the first time in . The implant of the biosynthesis pathway into the peroxisome led to 111.5 mg/L limonene in a shake flask culture. The limonene titer was further boosted to 1.05 g/L upon dual-metabolic regulation in the cytoplasm and peroxisome, which included employing the acetoacetyl-CoA synthase NphT7, adding an additional copy of native ATP-dependent citrate lyase, etc. The final yield was 0.053 g/g glucose, which was the highest ever reported. The newly characterized PTSs should contribute to the expansion of genetic toolboxes for. The results demonstrated that could be explored for efficient production of terpenoids.
是生产各种附加值化学品的潜在主力军，包括萜类化合物、油脂化学品和酶类，可利用低成本的饲料进行生产。然而，有限的遗传工具阻碍了其代谢工程的发展。在本研究中，首次对四种 I 型和一种新型 II 型过氧化物酶体靶向信号（PTS1/PTS2）进行了特征分析，并首次将其应用于柠檬烯的生产。将生物合成途径植入过氧化物酶体后，在摇瓶培养中得到了 111.5mg/L 的柠檬烯。通过在细胞质和过氧化物酶体中进行双重代谢调控，进一步提高了柠檬烯的产量，包括使用乙酰乙酰辅酶 A 合酶 NphT7，添加额外的天然 ATP 依赖性柠檬酸裂合酶等。最终的产率为 0.053g/g 葡萄糖，这是迄今为止报道的最高产量。新鉴定的 PTS 应该有助于扩大的遗传工具包。研究结果表明可以探索用于萜类化合物的高效生产。
