数字农科院2.0

Improved methyl supply for 5-methyltetrahydrofolate production in E. coli through a novel C1 transfer pathway

文献类型： 外文期刊

作者： Wen Liu;Jing Guo;Wei Lu;Tao Cheng;Yiting Li;Mo Xian;Rubing Zhang

作者机构：

关键词： 5-MTHF;Escherichia coli;Metabolic engineering;Methyl supply

期刊名称： Microbial Cell Factories

ISSN： 1475-2859

年卷期： 2025 年 24 卷 1 期

页码：

收录情况： SCIE（2025版）

摘要： Background: L-5-Methyltetrahydrofolate (5-MTHF) is the sole biologically active form of folate present in human blood and serves as an essential nutritional supplement. While microbial biosynthesis of 5-MTHF offers a sustainable alternative to chemical synthesis, its low yield limits industrial potential. Results: In this study, strategies for improving the methyl supply combined with engineering the tetrahydrofolate (THF) synthetic pathway were employed in E. coli to increase 5-MTHF production. First, a new exogenous C1 pathway was introduced to improve the intracellular methyl supply through acetyl-CoA breakdown. High expression of key rate-limiting genes folE, folP and purU enhanced metabolic flux of THF pathway, resulting in a 5-MTHF titer of 1.075 mg/L during shake-flask fermentation. A subsequent increase in 5-MTHF production was achieved by knocking out the metE gene, which is involved in the consumption of 5-MTHF. The best engineered strain, M3012, produced 8.2 mg/L 5-MTHF in a 5 L bioreactor via fed-batch fermentation, which presented the highest 5-MTHF titer to date. Conclusion: We successfully engineered E. coli by introducing a novel exogenous C1 metabolic pathway to augment the methyl donor pool essential for the biosynthesis of 5-MTHF. Further metabolic optimizations, including the enhancement of the THF precursor flux and the elimination of competing degradation pathways, developed a recombinant strain with significantly increased yield, which paves the way for industrial production of 5-MTHF.

分类号：

  • 相关文献

[1]Polysialic acid biosynthesis and production in Escherichia coli: current state and perspectives. Lin, Bai-Xue,Tao, Yong,Qiao, Yu,Shi, Bo.

[2]Synergetic Fermentation of Glucose and Glycerol for High-Yield N-Acetylglucosamine Production in Escherichia coli. Kaikai Wang,Xiaolu Wang,Huiying Luo,Yaru Wang,Yuan Wang,Tao Tu,Huoqing Huang,Bin Yao,Xiaoyun Su,Jie Zhang,Xing Qin,Yingguo Bai. 2022

[3]Metabolic Engineering of Escherichia coli for Efficient l-Isoleucine Production based on the Citramalate Pathway. Zhang, Qiquan,Wang, Yuheng,Wang, Xiaolu,Bai, Yingguo,Wang, Yaru,Wang, Yuan,Tu, Tao,Qin, Xing,Su, Xiaoyun,Yao, Bin,Luo, Huiying,Liu, Xu,Huang, Huoqing,Zhang, Jie. 2025

[4]一株大肠埃希菌烈性噬菌体ΦTRI-1的分离及特性分析. 高苗,杨金广,孙航军,刘旭,刘伟,王凤龙. 2015

[5]Progress of vitamin E metabolic engineering in plants. Chen, Shuangyan,Li, Hongjie,Liu, Gongshe. 2006

[6]Down-regulation of crambe fatty acid desaturase and elongase in Arabidopsis and crambe resulted in significantly increased oleic acid content in seed oil. Li, Xueyuan,Fan, Jing,Zhu, Li-Hua,Mei, Desheng,Liu, Qing,Singh, Surinder,Green, Allan,Zhou, Xue-Rong,Mei, Desheng,Liu, Qing,Singh, Surinder,Zhou, Xue-Rong,Fan, Jing.

[7]Bioactive compounds in functional buckwheat food. Shao, Ji-Rong,Zhang, Zhan-Lu,Zhou, Mei-Liang,Tang, Yi-Xiong,Shao, Ji-Rong,Zhang, Zhan-Lu,Xue, Wen-Tong,Zhang, Zhan-Lu,Zhou, Mei-Liang,Wu, Yan-Min,Tang, Yu,Li, Fa-Liang.

[8]Soybean transcription factor GmMYBZ2 represses catharanthine biosynthesis in hairy roots of Catharanthus roseus. Zhou, Mei-Liang,Shao, Ji-Rong,Zhou, Mei-Liang,Wu, Yan-Min,Tang, Yi-Xiong,Hou, Hong-Li,Zhu, Xue-Mei.

[9]Production and metabolic engineering of bioactive substances in plant hairy root culture. Zhou, Mei-Liang,Shao, Ji-Rong,Zhou, Mei-Liang,Tang, Yi-Xiong,Wu, Yan-Min,Zhou, Mei-Liang,Zhu, Xue-Mei.

[10]An protocol for genetic transformation of Catharanthus roseus by Agrobacterium rhizogenes A4. Zhou, Mei-Liang,Wu, Yan-Min,Tang, Yi-Xiong,Zhou, Mei-Liang,Shao, Ji-Rong,Zhu, Xue-Mei.

[11]Genome-wide identification of genes involved in raffinose metabolism in Maize. Zhou, Mei-Liang,Zhang, Qian,Zhou, Ming,Shao, Ji-Rong,Zhou, Mei-Liang,Sun, Zhan-Min,Tang, Yi-Xiong,Wu, Yan-Min,Zhu, Xue-Mei.

[12]Metabolic engineering of microorganisms to produce omega-3 very long-chain polyunsaturated fatty acids. Gong, Yangmin,Wan, Xia,Jiang, Mulan,Hu, Chuanjiong,Hu, Hanhua,Huang, Fenghong.

[13]Trehalose Metabolism-Related Genes in Maize. Zhou, Mei-Liang,Zhang, Qian,Shao, Ji-Rong,Zhou, Mei-Liang,Sun, Zhan-Min,Liu, Bo-Xin,Zhang, Kai-Xuan,Tang, Yi-Xiong,Wu, Yan-Min,Chen, Li-Hui,Zhu, Xue-Mei. 2014

[14]Carotenoids in Staple Cereals: Metabolism, Regulation, and Genetic Manipulation. Shengnan Zhai, Xianchun Xia and Zhonghu He. 2016

[15]Direct conversion of carbon dioxide to glucose using metabolically engineered Cupriavidus necator. Xiaolu Wang,Huiying Luo,Yaru Wang,Yuan Wang,Tao Tu,Xing Qin,Xiaoyun Su,Huoqing Huang,Yingguo Bai,Bin Yao,Jie Zhang. 2022

[16]Engineering a universal and efficient platform for terpenoid synthesis in yeast. Ma Y.,Zu Y.,Huang S.,Stephanopoulos G.. 2023

[17]Production of N-acetylglucosamine from carbon dioxide by engineering Cupriavidus necator H16. Xiaolu Wang,Fangfang Chang,Tingting Wang,Huiying Luo,Xiaoyun Su,Tao Tu,Yuan Wang,Yingguo Bai,Xing Qin,Honglian Zhang,Yaru Wang,Bin Yao,Huoqing Huang,Jie Zhang. 2023

[18]Progress And Perspective Of Biosynthetic P.latform For Higher-Order Biofuels. Su, HF, Lin, JF, Tan, FR. 2017

[19]Development of homozygous transgenic Atropa belladonna plants with glyphosate resistance and high-yield scopolamine using metabolic engineering. Qiaozhuo Zhang,Mengjiao Liang,Yuanyuan Liu,Chunxian Yang,Junlan Zeng,Jianbo Qin,Xiaozhong Lan,Min Lin,Min Chen,Jin Wang,Zhihua Liao. 2021

[20]Metabolic engineering of Pichia pastoris for myo-inositol production by dynamic regulation of central metabolism. Zhang, Qiquan,Wang, Xiaolu,Luo, Huiying,Wang, Yaru,Wang, Yuan,Tu, Tao,Qin, Xing,Su, Xiaoyun,Huang, Huoqing,Yao, Bin,Bai, Yingguo,Zhang, Jie. 2022

作者其他论文 更多>>