Characterization of a Cytosolic Acyl-Activating Enzyme Catalyzing the Formation of 4-Methylvaleryl-CoA for Pogostone Biosynthesis in Pogostemon Cablin

Plant Cell Physiol. 2021 Dec 3;62(10):1556-1571. doi: 10.1093/pcp/pcab111.

Abstract

Pogostone, a compound with various pharmaceutical activities, is a major constituent of the essential oil preparation called Pogostemonis Herba, which is obtained from the plant Pogostemon cablin. The biosynthesis of pogostone has not been elucidated, but 4-methylvaleryl-CoA (4MVCoA) is a likely precursor. We analyzed the distribution of pogostone in P. cablin using gas chromatography-mass spectrometry (GC-MS) and found that pogostone accumulates at high levels in the main stems and leaves of young plants. A search for the acyl-activating enzyme (AAE) that catalyzes the formation of 4MVCoA from 4-methylvaleric acid was launched, using an RNAseq-based approach to identify 31 unigenes encoding putative AAEs including the PcAAE2, the transcript profile of which shows a strong positive correlation with the distribution pattern of pogostone. The protein encoded by PcAAE2 was biochemically characterized in vitro and shown to catalyze the formation of 4MVCoA from 4-methylvaleric acid. Phylogenetic analysis showed that PcAAE2 is closely related to other AAE proteins in P. cablin and other species that are localized to the peroxisomes. However, PcAAE2 lacks a peroxisome targeting sequence 1 (PTS1) and is localized in the cytosol.

Keywords: Pogostemon cablin; 4-Methylvaleric acid •; 4-Methylvaleryl-CoA; Acyl-activating enzyme; Branched-chain fatty acid; Pogostone.

MeSH terms

  • Amino Acid Sequence
  • Coenzyme A Ligases / chemistry
  • Coenzyme A Ligases / genetics*
  • Coenzyme A Ligases / metabolism
  • Oils, Volatile / metabolism*
  • Phylogeny
  • Plant Proteins / chemistry
  • Plant Proteins / genetics*
  • Plant Proteins / metabolism
  • Pogostemon / genetics*
  • Pogostemon / metabolism
  • Sequence Alignment

Substances

  • Oils, Volatile
  • Plant Proteins
  • Pogostone
  • Coenzyme A Ligases