Short-chain dehydrogenase/reductase catalyzing the final  step of noscapine biosynthesis is localized to laticifers in  opium poppy.
The final step in the biosynthesis of the phthalideisoquinoline alkaloid noscapine involves a purported dehydrogenation of the narcotinehemiacetal keto moiety. A short-chain dehydrogenase/reductase (SDR), designated noscapine synthase (NOS), that catalyzes dehydrogenation of narcotinehemiacetal to noscapine was identified in opium poppy and functionally characterized. The NOS gene was isolated using an integrated transcript and metabolite profiling strategy and subsequently expressed in Escherichia coli. Noscapine synthase is highly divergent from other characterized members of the NADPH-dependent SDR superfamily involved in benzylisoquinoline alkaloid metabolism, and it exhibits exclusive substrate specificity for narcotinehemiacetal. Kinetic analyses showed that NOS exhibits higher catalytic efficiency with NAD+ as the cofactor compared with NADP+. Suppression of NOS transcript levels in opium poppy plants subjected to virus-induced gene silencing resulted in a corresponding reduction in the accumulation of noscapine and an increase in narcotinehemiacetal levels in the latex. Noscapine and NOS transcripts were detected in all opium poppy organs, but both were most abundant in stems. Unlike other putative biosynthetic genes clustered in the opium poppy genome, and their corresponding proteins, NOS transcripts and the cognate enzyme were abundant in latex, indicating that noscapine metabolism is completed in a distinct cell type compared with the rest of the pathway.
Xue
X
Chen
Peter J
PJ
Facchini
D001483:Base Sequence; D055162:Biocatalysis; D002851:Chromatography, High Pressure Liquid; D017931:DNA Primers; D017343:Genes, Plant; D007700:Kinetics; D008025:Ligases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009665:Noscapine / Q000378:metabolism*; D009892:Opium / Q000378:metabolism*; D010088:Oxidoreductases / Q000378:metabolism*; D010207:Papaver / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D053719:Tandem Mass Spectrometry
D017931:DNA Primers; D009892:Opium; D009665:Noscapine; D010088:Oxidoreductases; D008025:Ligases