The gene encoding polyneuridine aldehyde esterase of monoterpenoid indole alkaloid biosynthesis in plants is an ortholog of the alpha/betahydrolase super family

Eur J Biochem. 2000 Mar;267(5):1397-406. doi: 10.1046/j.1432-1327.2000.01136.x.

Abstract

The biosynthesis of the anti-arrhythmic alkaloid ajmaline is catalysed by more than 10 specific enzymes. In this multistep process polyneuridine aldehyde esterase (PNAE) catalyses a central reaction by transforming polyneuridine aldehyde into epi-vellosimine, which is the immediate precursor for the synthesis of the ajmalane skeleton. PNAE was purified from cell suspension cultures of Rauvolfia serpentina. The N-terminal sequence and endoproteinase LysC fragments of the purified protein were used for primer design and for the amplification of specific PCR products leading to the isolation of PNAE-encoding cDNA from a R. serpentina library. The PNAE cDNA was fused with a C-terminal His-tag, expressed in Escherichia coli and purified to homogeneity using Ni-affinity chromatography. The pure enzyme shows extraordinary substrate specificity, completely different to other esterases. Sequence alignments indicate that PNAE is a new member of the alpha/beta hydrolase super family.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkaloids / biosynthesis*
  • Alkaloids / chemistry
  • Amino Acid Sequence
  • Base Sequence
  • Carboxylic Ester Hydrolases / genetics*
  • Cloning, Molecular
  • DNA, Complementary
  • Indoles / chemistry
  • Molecular Sequence Data
  • Plants / enzymology*
  • Sequence Homology, Amino Acid
  • Terpenes / chemistry

Substances

  • Alkaloids
  • DNA, Complementary
  • Indoles
  • Terpenes
  • Carboxylic Ester Hydrolases

Associated data

  • GENBANK/AF178576