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Metabolic engineering of plant alkaloid biosynthesis.
Plant alkaloids, one of the largest groups of natural products, provide many pharmacologically active compounds. Several genes in the biosynthetic pathways for scopolamine, nicotine, and berberine
Molecular Characterization of theS-Adenosyl-l-methionine:3′-Hydroxy-N-methylcoclaurine 4′-O-Methyltransferase Involved in Isoquinoline Alkaloid Biosynthesis in Coptis japonica *
TLDR
Characterization of the substrate saturation and product inhibition kinetics indicated that 4′-OMT follows an ordered Bi Bi mechanism, whereas 6-O-methyltransferase follows a Ping-Pong BiBi mechanism.
Molecular Cloning and Characterization of CoclaurineN-Methyltransferase from Cultured Cells of Coptis japonica *
TLDR
Recombinant CNMT was purified to homogeneity, and enzymological characterization confirmed that CoptisCNMT has quite broad substrate specificity, i.e. not only for 6-O-methylnorlaudanosoline and norreticuline but also for 6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline.
Overexpression of Coptis japonica norcoclaurine 6-O-methyltransferase overcomes the rate-limiting step in Benzylisoquinoline alkaloid biosynthesis in cultured Eschscholzia californica.
TLDR
Investigation of the effect of the overexpression of putative rate-limiting step enzymes in benzylisoquinoline alkaloid biosynthesis found that a 6OMT-specific gene is missing and 4OMT catalyzes the 6O-methyltransferase reaction with low activity in California poppy, which supports the notion that the 6OMt reaction is important for alkaloids biosynthesis in this plant species.
Molecular cloning of columbamine O-methyltransferase from cultured Coptis japonica cells.
TLDR
The result clearly indicated that EST analysis was useful for isolating the candidate gene in a relatively well-characterized biosynthetic pathway and the relationship between the structure and substrate recognition of the O-methyltransferases involved in isoquinoline alkaloid biosynthesis, and a reconsideration of the biosynthesis pathway to palmatine are discussed.
Knockdown of berberine bridge enzyme by RNAi accumulates (S)-reticuline and activates a silent pathway in cultured California poppy cells
TLDR
C cultured California poppy cells are established, in which berberine bridge enzyme was knocked down by RNA interference, to accumulate the important key intermediate reticuline, and the potential application of RNAi technology in metabolic modification and the flexibility of plant secondary metabolism is discussed.
Transient RNA Silencing of Scoulerine 9-O-Methyltransferase Expression by Double Stranded RNA in Coptis japonica Protoplasts
TLDR
The transient RNAi of endogenous biosynthetic genes involved in isoquinoline alkaloid biosynthesis in Coptis japonica protoplasts is reported and substantial silencing of SMT gene expression was obtained by the introduction of these SMT dsRNAs.
In Vivo Bioconversion of Tetrahydroisoquinoline by Recombinant Coclaurine N-Methyltransferase
TLDR
The possibility of converting 6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline into its N-methylated product using transgenic Escherichia coli, which expressed recombinant coclaurine N- methyltransferase, without the addition of a methyl-group donor is examined.
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