Partial purification and properties of S-adenosyl-l-methionine: (S)-tetrahydroprotoberberinecis-N-methyltransferase from suspension-cultured cells of Eschscholtzia and Corydalis

  title={Partial purification and properties of S-adenosyl-l-methionine: (S)-tetrahydroprotoberberinecis-N-methyltransferase from suspension-cultured cells of Eschscholtzia and Corydalis},
  author={Martina Rueffer and Gitta Zumstein and Meinhart H. Zenk},
Abstract An enzyme has been found in different species of isoquinoline alkaloid-producing plant cell cultures which specifically N -methylates certain ( S )-tetrahydroprotoberberine alkaloids such as ( S )-canadine and ( S )-stylopine at the expense of S -adenosyl- l -methionine (SAM). It was partially purified (90-fold from Eschscholtzia californica cell suspension cultures and characterized. The enzyme has a pH optimum of 8.9, a temperature optimum at 40° and a M r of about 78 000 ± 10%. The… Expand
Purification and properties of 1-hydroxycanthin-6-one: S-adenosyl-l-methionine methyltransferase from Ailanthus altissima cell cultures
Abstract An S -adenosyl- l -methionine (SAM) requiring methyltransferase responsible for the methylation of 1-hydroxycanthin-6-one has been isolated from a series of cell culture lines of AilanthusExpand
Partial Purification and Characterization of S-Adenosyl-L- Methionine:Anthranilic Acid N-Methyltransferase from Ruta Cell Suspension Cultures
The properties, the substrate specificity and the Michaelis-Menten constants of the purified enzyme were determined and it was found that the anthranilic acid specific N-methyltransferase is strongly inhibited by S-adenosyl-L-homocysteine. Expand
Molecular Cloning and Characterization of Tetrahydroprotoberberine cis-N-Methyltransferase, an Enzyme Involved in Alkaloid Biosynthesis in Opium Poppy*
S-Adenosyl-l-methionine:tetrahydroprotoberberine cis-N-methyltransferase (EC catalyzes the conversion of (S)-stylopine to the quaternary ammonium alkaloid, (S)-cis-N-methylstylopine, as aExpand
Structure–function studies of tetrahydroprotoberberine N-methyltransferase reveal the molecular basis of stereoselective substrate recognition
The insights into substrate recognition and catalysis provided here form a sound basis for the rational engineering of NMT enzymes for chemoenzymatic synthesis and metabolic engineering. Expand
Isolation and Characterization of Reticuline N-Methyltransferase Involved in Biosynthesis of the Aporphine Alkaloid Magnoflorine in Opium Poppy*
The isolation and characterization of a phylogenetically related yet functionally distinct N-methyltransferase (NMT) from opium poppy that primarily accepts 1-benzylisoquinoline and aporphine substrates possessing a tertiary amine is reported. Expand
Difference between Enzymatic and Chemical N-methylations of Protoberberine-Type Alkaloid, Dependent on the Stereoisomer of (-)-N-methyl-7,8,13,13a-tetrahydroberberinium Salt.
A possible relation between the stereostructure of (−)-(13aS)-tetrahydroberberine (1) and its enzymatic/chemical N-methylation, an important biosynthetic reaction to isoquinoline alkaloids in plants,Expand
Developmental and inducible accumulation of gene transcripts involved in alkaloid biosynthesis in opium poppy.
Opium poppy (Papaver somniferum) produces a large number of benzylisoquinoline alkaloids, including morphine and sanguinarine, derived from tyrosine via the branch-point intermediate (S)-reticuline.Expand
Enzymic 12-hydroxylation and 12-O-methylation of dihydrochelirubine in dihydromacarpine formation by Thalictrum bulgaricum
This discovery leads to the complete sequence of macarpine biosynthesis which has thus far been the longest secondary product pathway completely clarified at the enzymic level. Expand
Chapter 5 The Biotransformation of Protoberberine Alkaloids by Plant Tissue Cultures
The biochemical processes in the conversion of the protoberberines into the benzophenanthridines occur similarly in intact plants and callus tissues. Expand
Heterologous expression of alkaloid biosynthetic genes--a review.
The biosynthesis at the enzyme level of at-least two tetrahydrobenzylisoquinoline alkaloids, the benzophenanthridine alkaloid sanguinarine in the California poppy, Eschscholtzia californica, and the bisbenzymunine in barberry, Berberis stolonifera, has been elucidated in detail. Expand


Purification and properties of (S)-tetrahydroprotoberberine oxidase from suspension-cultured cells of Berberis wilsoniae.
This catalyst was responsible for the conversion of (S)-tetrahydrocolumbamine to the key intermediate, columbamine, in the metabolic pathway leading to berberine, jatrorrhizine and palmatine. Expand
S-adenosyl-l-methionine: (S) -7,8,13, 14-tetrahydroberberine--n-methyltransferase, a branch point enzyme in the biosynthesis of benzophenanthridine and protopine alkaloids.
The enzyme which transfers the CH3-group of S-adenosylmethionine to the nitrogen atom of (S)-tetrahydroberberine and (S)-stylopine is found to occur in a number of plant cell cultures originatingExpand
Purification and characterization of the berberine bridge enzyme from berberis beaniana cell cultures
The berberine bridge-forming enzyme (BBE) has been found in 66 samples taken from differentiated plants and from cell suspension cultures and was shown to be homogeneous by gel electrophoresis. Expand
S-Adenosylmethionine synthetase from Escherichia coli.
Adenosylmethionine (AdoMet) synthetase has been purified to homogeneity from Escherichia coli and studies with the phosphorothioate derivatives of ATP in the presence of Mg2+, Mn2+, or Co2+ indicate that a divalent ion is bound to the nucleotide during the reaction and provide information on the stereochemistry of the metal-nucleotide binding site. Expand
Enzymatic formation of protopines by a microsomal cytochrome P-450 system of Corydalis vaginans
Abstract A microsomal cytochrome P-450-NADPH dependent enzyme which hydroxylates stereo- and regiospecifically carbon atom 14 of (S)- cis -N- methyltetrahydroprotoberberines has been discovered in aExpand
Biosynthesis of Benzo[c]phenanthridine Alkaloids Sanguinarine, Chelirubine and Macarpine
The biosynthesis of the benzo[c]phenanthridine alkaloids was investigated in a cell suspension culture of Macleaya cordata (papaveraceae). Feeding experiments define the biosynthetic pathwayExpand
Formation of both methylenedioxy groups in the alkaloid (S)-stylopine is catalyzed by cytochrome P-450 enzymes
Abstract Two highly stereospecific microsomal cytochrome P-450 NADPH dependent enzymes have been discovered and characterized which are responsible for the methylenedioxy group formation fromExpand
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
This assay is very reproducible and rapid with the dye binding process virtually complete in approximately 2 min with good color stability for 1 hr with little or no interference from cations such as sodium or potassium nor from carbohydrates such as sucrose. Expand
One step enzymatic synthesis of dihydrosanguinarine from protopine
Abstract A microsomal cytochrome P-450 NADPH dependent enzyme which hydroxylates carbon-6 of protopine has been discovered and characterized; the monooxygenase reaction leads to 6-hydroxyprotopineExpand
Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4
Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products. Four majorExpand