Columbamine, the central intermediate in the late stages of protoberberine biosynthesis

A specific methyltransferase which in the presence of S-adenosylmethionine [SAM] converts columbamine to palmatine has been demonstrated to occur in cell cultures. The enzyme acts only on the quaternary alkaloid as substrate, not on its tetrahydro derivative, which fact is in sharp contrast to previous claims

Canadine synthase from Thalictrum tuberosum cell cultures catalyses the formation of the methylenedioxy bridge in berberine synthesis

An enzyme system catalysing the formation of the methylenedioxy bridge at ring A of (S)-canadine [ = (S)-tetrahydroberberine] from (S)-tetrahydrocolumbamine has been detected in microsomal preparations from different Ranunculaceae and Berberidaceae cell cultures. The cytochrome P-450 enzyme complex has been partly characterized from a protoberberine alkaloid producing Thalictrum tuberosum L. cell line. The enzyme complex consisting of a microsomal associated oxidase with a cytochrome P-450 reductase has a pH optimum at pH 8.5 and a temperature optimum of 40°. The apparent Km values are 33 μM for NADPH and 11.5 μM for tetrahydrocolumbamine

The formation of corydaline and related alkaloids in Corydalis cava in vivo and in vitro

Kein Inhaltsverz

Alternative final steps in berberine biosynthesis in Coptis japonica cell cultures

In Coptis japonica cell cultures an alternative pathway has been discovered which leads from (S)-tetrahydrocolumbamine via (S)-canadine to berberine. The two enzymes involved have been partially purified. (S)-Tetrahydrocolumbamine is stereospecifically transformed into (S)-canadine under formation of the methylenedioxy bridge in ring A. This new enzyme was named (S)-canadine synthase. (S)-Canadine in turn is stereospecifically dehydrogenated to berberine by an oxidase, (S)-canadine oxidase (COX), which was partially purified (25-fold). This enzyme has many physical properties in common with the already known (S)-tetrahydroprotoberberine oxidase from Berberis but grossly differs from the latter enzyme in its cofactor requirement (Fe) and its substrate specificity. Neither (S)-norreticuline nor (S)-scoulerine serves as substrate for the Coptis enzyme, while both substrates are readily oxidized by the Berberis enzyme. The four terminal enzymes catalyzing the pathway from (S)-reticuline to berberine are housed in Berberis as well as in Coptis in smooth vesicles with a density of =1.14 g/ml. These vesicles have been enriched and characterized by electron microscopy