Food and Fermentation Industries >

2023 , Vol. 49 >Issue 23: 16 - 24

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.035600

Metabolic engineering of Escherichia coli for 8-prenylkaempferol synthesis

  • ZHAO Wanying ,
  • ZHOU Jingwen ,
  • HOU Ying
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  • 1(College of Bioengineering, Tianjin University of Science and Technology, Tianjin 300000, China)
    2(College of Bioengineering, Jiangnan University, Wuxi 214122, China)

Received date: 2023-03-23

  Revised date: 2023-04-14

  Online published: 2024-01-02

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Abstract

8-Prenylkaempferol (8-pk) is a flavonoid with excellent anti-inflammatory and anticancer pharmacological activities. At present, the biosynthesis of 8-pk is mainly obtained from kaempferol (Kae) by the catalysis of plant-derived prenyltransferases (PTs). However, plant-derived PTs contain signal peptides, which greatly hinder their heterologous expression and efficient catalysis in prokaryotes. In addition, the insufficient supply of the precursor molecule dimethylallyl pyrophosphate (DMAPP) in the catalytic pathway severely limits the biosynthesis yield of 8-pk. In this study, Escherichia coli was used as a chassis cell, further enhancing the supply of precursor DMAPP and truncating the signal peptide of prenyltransferase for 8-pk efficient synthesis. Firstly, epimedium derived prenyltransferase (EkF8DT) was expressed in E. coli BL21(DE3) and IUP pathway was introduced to increase the supply of DMAPP, with an 8-pk titer of 2.14 mg/L. Subsequently, the structure of EkF8DT was predicted by AlphaFold2. Subsequently, the structure of EkF8DT was predicted by AlphaFold2 and the highest 8-pk yield of 6.46 mg/L was achieved when the N-end truncation of EkF8DT was truncated by 60 amino acids. By fusing the protein tag to the N-end truncation of EkF8DT and optimizing fermentation conditions a titer of 24.28 mg/L was achieved. Finally, 8-pk titer of 44.33 mg/L was achieved in a 5 L fermenter, which is the higher level reported. This study provides an effective strategy for 8-pk biosynthesis.

Key words： 8-prenylkaempferol; prenyltransferases; IUP pathway; signal peptide truncation; protein tags

Cite this article

ZHAO Wanying , ZHOU Jingwen , HOU Ying . Metabolic engineering of Escherichia coli for 8-prenylkaempferol synthesis[J]. Food and Fermentation Industries, 2023 , 49(23) : 16 -24 . DOI: 10.13995/j.cnki.11-1802/ts.035600

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