TH9858B - The process of producing planuotol using enzymes geranylgeraniol-18-hydroxylase - Google Patents

Abstract

DC60 (27/01/40) The invention is the production of plaunotol which has medicinal value from the precursor. geranylgeraniol which is a substance that is cheap and easy to synthesize using the enzyme geranyl- geraniol-18-hydroxylase This enzyme was the first found in this invention in leaves. of the croton plant, a plant found in Thailand The discovery of this enzyme led to the a new method for obtaining plaunotol using enzymatic conversion methods. Study some properties of enzymes. geranylgeraniol-18-hydroxylase which was found to be The enzyme contained in 20,000 g microsomal fraction with high heat resistance, good catalytic efficiency at pH 5 and high specificity to geranyl- geraniol substrate. in medicine from the precursor geranylgeraniol which is a substance that is cheap and easy to synthesize using the enzyme geranyl- geraniol-18-hydroxylase This enzyme was the first found in this invention in leaves. of the croton plant, a plant found in Thailand The discovery of this enzyme led to the a new method for obtaining plaunotol using enzymatic conversion methods. Study some properties of enzymes. geranylgeraniol-18-hydroxylase which was found to be Enzyme contained in 20,000 g microsomal fraction with high heat resistance, good catalytic efficiency at pH 5 and high specificity to geranyl- geraniol substrate.

Claims (6)

1. Process of preparation Geraniol-Eighteen-Hydroxylase Enzyme (geranylgeraniol-18-hydroxylase) that is specific to the hydroxylation of the geraniol geraniol layer at the eighteen-position carbon to obtain plaunotol, where the process consists of The following steps: 1.1 Extraction of enzyme extracts. geranylgeraniol-18-hydroxylase From the leaves of the little pula (Croton sublyratus) with buffer solution containing the extraction method by adding buffer solution for use. In the extraction of the powder from the leaves of the little Crao leaves, which was crushed in the cotyledon, which was cooled with liquid nitrogen and stirred into a suspension solution, which, when filtered, the extract of the enzyme was obtained.1.2 Enzyme separation geranylgeraniol-18-hydroxylase From the extract obtained from 1.1 with different centrifugation, that is, centrifugate the extracted extract at an initial strength of 3000 g to obtain a liquid supernatant layer that when centrifuged. The second time, at 20,000 g, gave the pellet a section called the enzyme garden. The pre-purified geranylgeraniol-18-hydroxylase was induced by boiling at 100 ° C for 30 minutes. 2. Process according to claim 1 The extraction buffer has the following composition: 2.1 Trizine-sodium hydroxide at pH 7.8 2.2 beta-mercaptoethanol 2.3 sucrose 2.4 EDTA. (EDTA 2.5 DTT (DTT) 2.6 Magnesium Chloride 2.7 BSA (BSA) 3.eranylgeraniol-18-hydroxylase enzyme The purer process obtained by claim 1 is a particle size of 20 to 40 nanometers with the highest hydroxylactic acidity at pH 5. Hydroxylation goranylgoraniol at the 18 position carbon provides pelanitol And increased activity by the use of coenzyme 4. Enzyme according to claim 3 can be activated by heating at 100 ํ C for 30 minutes. 5. Process for production of non-toxic substances using enzymes geranylgeraniol-18-hydroxylase according to claim 3, add a hydroxyl group at the carbon position 18 of the substrate. geranylgeraniol In conditions where the manufacturing process is at pH 5, the enzyme is co-enzyme. The enzyme was activated by heating it at 100 ํ C for 30 minutes before the hydroxylation. 6. Production process of pelanotol according to claim 5, the co-enzyme used is NADPH (NADPH).