Regulating malonyl-CoA metabolism via synthetic antisense RNAs for enhanced biosynthesis of natural products

Yaping Yang, Yuheng Lin, Lingyun Li, Robert J. Linhardt, Yajun Yan

Research output: Contribution to journalArticlepeer-review

158 Scopus citations

Abstract

Malonyl-CoA is the building block for fatty acid biosynthesis and also a precursor to various pharmaceutically and industrially valuable molecules, such as polyketides and biopolymers. However, intracellular malonyl-CoA is usually maintained at low levels, which poses great challenges to efficient microbial production of malonyl-CoA derived molecules. Inactivation of the malonyl-CoA consumption pathway to increase its intracellular availability is not applicable, since it is usually lethal to microorganisms. In this work, we employ synthetic antisense RNAs (asRNAs) to conditionally down-regulate fatty acid biosynthesis and achieve malonyl-CoA enrichment in Escherichia coli. The optimized asRNA constructs with a loop-stem structure exhibit high interference efficiency up to 80%, leading to a 4.5-fold increase in intracellular malonyl-CoA concentration when fabD gene expression is inhibited. Strikingly, this strategy allows the improved production of natural products 4-hydroxycoumarin, resveratrol, and naringenin by 2.53-, 1.70-, and 1.53-fold in E. coli, respectively. In addition, down-regulation of other fab genes including fabH, fabB, and fabF also leads to remarkable increases in 4-hydroxycoumarin production. This study demonstrates a novel strategy to enhance intracellular malonyl-CoA and indicates the effectiveness of asRNA as a powerful tool for use in metabolic engineering.

Original languageEnglish
Pages (from-to)217-226
Number of pages10
JournalMetabolic Engineering
Volume29
DOIs
StatePublished - 1 May 2015
Externally publishedYes

Keywords

  • 4-hydroxycoumarin
  • Antisense RNA
  • Fatty acid biosynthesis
  • Malonyl-CoA
  • Naringenin
  • Resveratrol

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