The gene encoding polyneuridine aldehyde esterase of monoterpenoid indole alkaloid biosynthesis in plants is an ortholog of the alpha/betahydrolase super family.
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.
Dogru
E
E
Lehrstuhl für Pharmazeutische Biologie, Institut für Pharmazie, Johannes Gutenberg-Universität Mainz.
Warzecha
H
H
Seibel
F
F
Haebel
S
S
Lottspeich
F
F
Stöckigt
J
J
D000470:Alkaloids / Q000096:biosynthesis* / Q000737:chemistry; D000595:Amino Acid Sequence; D001483:Base Sequence; D002265:Carboxylic Ester Hydrolases / Q000235:genetics*; D003001:Cloning, Molecular; D018076:DNA, Complementary; D007211:Indoles / Q000737:chemistry; D008969:Molecular Sequence Data; D010944:Plants / Q000201:enzymology*; D017386:Sequence Homology, Amino Acid; D013729:Terpenes / Q000737:chemistry
D000470:Alkaloids; D018076:DNA, Complementary; D007211:Indoles; D013729:Terpenes; D002265:Carboxylic Ester Hydrolases