S-tetrahydroprotoberberineoxidase, process for producing the same and use thereof
AssigneeConsortium fur Elektrochemische Industrie GmbH
InventorsManfred Amann, Naotaka Nagakura, Meinhart H. Zenk
Life of this patent
Estimating the term…
Abstract
The invention relates to the enzyme S-tetrahydroprotoberberineoxidase, which oxidizes selectively S-tetrahydroprotoberberines and S-1-benzyl-1,2,3,4-tetrahydroisoquinoline in the presence of oxygen, resulting in the corresponding protoberberines and 1-benzyl-3,4-dihydroisoquinolines. The invention also relates to a process of preparing S-tetrahydroprotoberberineoxidase by extraction from certain plant materials. The oxidation products of several compounds find use in the pharmaceutical industries.
Claims19
The compound s-tetrahydroproberberineoxidase having the following parameters:
- (a) molar weight:
- 100,000.+-.10% as determined by gel filtration;
- (b) isoelectric point is 5.7;
- (c) catalytic activity occurs at 40.degree. C.;
- (d) catalytic activity occurs at pH of 6-10;
- (e) K.sub.M value for R,S-corypalmine at 40.degree. C. and pH 8.9 is 1.3 .mu.Moles;
- (f) Intensity maximum of the fluorescense-emission spectrum is 520 nm at an excitation wavelength of 450 nm;
- (g) complete and reversible inhibition is exhibited in the presence of a member selected from the group consisting of acetylacetone and morin;
- and
- (h) prosthetic group flavin.
A method for producing S-tetrahydroprotoberberineoxidase having the following parameters:
- (a) molar weight:
- 100,000.+-.10% as determined by gel filtration;
- (b) isoelectric point is 5.7;
- (c) catalytic activity occurs at 40.degree. C.;
- (d) catalytic activity occurs at pH of 6-10;
- (e) K.sub.M value for R,S-corypalmine at 40.degree. C. and pH 8.9 is 1.3 .mu.Moles;
- (f) intensity maximum of the fluoroescense-emission spectrum is 520 nm at an excitation wavelength of 450 nm;
- (g) complete and reversible inhibition is exhibited in the presence of a member selected from the group consisting of acetylacetone and morin;
- and
- (h) prosthetic group flavin, comprising the step of extracting said S-tetrahydroprotoberberineoxidase from a plant material from a member of a family selected from the group consisting of Papaveraceae, Berberidaceae, Menispermaceae, Annonaceae, Ranunculaceae, and a combination thereof.
A method for the enzymatic oxidation of a compound of the general formula:
- ##STR2## with S-configuration, wherein R.sub.1, R.sub.2 R.sub.5, R.sub.6 and R.sub.7 are independently elected from the group consisting of a hydrogen atom and hydroxy, alkyl and alkoxy groups;
- R.sub.3 is a member selected from the group consisting of hydrogen and an --OH group;
- R.sub.4 stands for hydrogen;
- and
- R.sub.8 stands for a member selected from the group consisting of hydrogen and an alkyl group with 1-3 carbon atoms;
- comprising the step of treating said compound with S-tetrahydroproberberineoxidase having the following parameters:
- (a) molar weight:
- 100,000.+-.10% as determined by gel filtration;
- (b) isoelectric point is 5.7;
- (c) catalytic activity occurs at 40.degree. C.;
- (d) cataltyic activity occurs at pH of 6-10;
- (e) K.sub.M value for R,S-corypalmine at 40.degree. C. and pH 8.9 is 1.3 .mu.Moles;
- (f) intensity maximum of the fluoroescense-emission spectrum is 520 nm at an excitation wavelength of 450 nm;
- (g) completed and reversible inhibition is exhibited in the presence of a member selected from the group consisting of acetylacetone and morin;
- and
- (h) prosthetic group flavin.
An enzymatically acting composition comprising as an active substance an effective amount of S-tetrahydroprotoberberineoxidase having the following parameters:
- (a) molar weight:
- 100,000.+-.10% as determined by gel filtration;
- (b) isoelectric point is 5.7;
- (c) catalytic activity occurs at 40.degree. C.;
- (d) catalytic activity occurs at pH of 6-10;
- (e) K.sub.M value for R,S-corypalmine at 40.degree. C. and pH 8.9 is 1.3 .mu.Moles;
- (f) intensity maximum of the fluoroescense-emission spectrum is 520 nm at an excitation wavelength of 450 nm;
- (g) complete and reversible inhibition is exhibited in the presence of a member selected from the group consisting of acetylacetone and morin;
- and
- (h) prosthetic group flavin.
Dependencies are inferred from the claim text.