Song Na, Xia Huili, Xie Yaoru, Guo Shuaikang, Zhou Rong, Shangguan Lingling, Zhuang Kun, Zhang Huiyan, An Feiran, Zheng Xueyun, Yao Lan, Yang Shihui, Chen Xiong, Dai Jun
Cooperative Innovation Center of Industrial Fermentation (Ministry of Education & Hubei Province), Key Laboratory of Fermentation Engineering (Ministry of Education), National "111" Center for Cellular Regulation and Molecular Pharmaceutics, Hubei Key Laboratory of Industrial Microbiology, School of Life and Health Sciences, Hubei University of Technology, Wuhan, Hubei, 430068, PR China.
College of Biological and Food Engineering, Huanghuai University, Zhumadian, 463000, PR China.
Synth Syst Biotechnol. 2024 Nov 26;10(1):294-306. doi: 10.1016/j.synbio.2024.11.007. eCollection 2025.
Tyrosol is an important component of pharmaceuticals, nutraceuticals, and cosmetics, and their biosynthetic pathways are currently a hot research topic. d-Erythrose 4-phosphate is a key precursor for the biosynthesis of tyrosol in . Hence, the flux of d-Erythrose 4-phosphate determined the yield of tyrosol synthesis. In this study, we first obtained an strain S19 with a tyrosol yield of 247.66 mg/L by metabolic engineering strategy. To increase the production of d-Erythrose 4-phosphate, highly active phosphoketolase BA-C was obtained by bioinformatics combined with tyrosol yield assay. The key residue sites 183, 217, and 320 were obtained by molecular docking, kinetic simulation, and tyrosol yield verification. After mutation, the highly efficient phosphoketolase BA-C was obtained, with a 37.32 % increase in enzyme activity. The tyrosol production of strain S26 with BA-C increased by 43.05 % than strain S25 with BA-C and increased by 151.19 % compared with the strain S19 without phosphoketolase in a 20 L fermenter. The mining and modification of phosphoketolase will provide strong support for the de novo synthesis of aromatic compounds.
酪醇是药物、营养保健品和化妆品的重要成分，其生物合成途径是当前的一个热门研究课题。4-磷酸-D-赤藓糖是酪醇生物合成的关键前体。因此，4-磷酸-D-赤藓糖的通量决定了酪醇合成的产量。在本研究中，我们首先通过代谢工程策略获得了一株酪醇产量为247.66 mg/L的菌株S19。为了提高4-磷酸-D-赤藓糖的产量，通过生物信息学结合酪醇产量测定获得了高活性磷酸酮醇酶BA-C。通过分子对接、动力学模拟和酪醇产量验证获得了关键残基位点183、217和320。突变后，获得了高效磷酸酮醇酶BA-C，酶活性提高了37.32%。在20 L发酵罐中，带有BA-C的菌株S26的酪醇产量比带有BA-C的菌株S25提高了43.05%，与没有磷酸酮醇酶的菌株S19相比提高了151.19%。磷酸酮醇酶的挖掘和改造将为芳香族化合物的从头合成提供有力支持。
