Preparation method for preparing kojic acid dipalmitate through compound enzyme method

Abstract

The present invention relates to a preparation method for preparing kojic acid dipalmitate through a compound enzyme method, wherein kojic acid and palmitic acid are placed into an anhydrous organic solvent, a compound lipase enzyme inhibitor adopted as an esterification reaction catalyst is added, and the kojic acid and the palmitic acid are esterified to generate the kojic acid dipalmitate. According to the present invention, a variety of compound lipases are adopted as the catalyst to prepare the kojic acid dipalmitate, the requirement on the reaction equipment is reduced, the product treatment cost after the reaction is substantially reduced, the advantages of mildness, high efficiency, environmental protection, low cost, high raw material conversion rate and the like are provided, the reaction steps are reduced, the production cost is reduced, and the high industrial application and economic values are provided.

Description

technical field

[0001] The invention belongs to the technical field of biochemical industry, and in particular relates to a method for preparing kojic acid dipalmitate by a compound enzyme method. Background technique

[0002] Kojic acid is a colorless, odorless, safe and non-toxic tyrosinase inhibitor, which can be used as a preservative, preservative, and color-protecting agent in food. Kojic acid has a strong inhibitory effect on the formation of melanin in human skin. Kojic acid inactivates tyrosinase to chelate its essential copper ions, and inhibits the interconversion from dopa chromium to DHICA (5,6- dihydroxyindole-2-carboxylic acid). L-ascorbic acid and its derivatives, including MAP, reduce melanin intermediates and prevent the oxidation chain reaction from tyrosine / DOPA to various points in melanin. Therefore, kojic acid can be produced for the treatment of freckles , age spots, pigmentation whitening cosmetics, deeply loved by consumers. However, because koji...

Examples

Embodiment 1

[0030] Pour 20ml of acetone liquid into a 50ml conical flask, dissolve 0.2 gram of kojic acid and 0.72 gram of palmitic acid in 20ml of acetone, then add 0.05 gram of lipase Lipozyme RMIM (derived from Rhizomucor miehei), 0.05 gram Novozyme 435 (immobilized lipase from Candida antarctica), 1 g color-changing silica gel. After sealing, put it in a shaker at 40°C and 170rpm to react for 28h. After the reaction was completed, centrifuge at 12000 rpm for 10 min to remove enzyme and solid particles, and perform chromatographic detection and analysis. The conversion rate of the substrate was 94%. The function of using color-changing silica gel during the reaction is to remove the moisture in the reaction system.

Embodiment 2

[0032] Pour 15ml of petroleum ether-tetrahydrofuran mixed solvent (volume ratio: petroleum ether: tetrahydrofuran = 1:2) into a 30ml conical flask, dissolve 0.1 gram of kojic acid and 0.5 gram of palmitic acid in this mixed solvent, and finally add to this solution Add 0.01 g of immobilized cells containing Bacillus subtilis lipase, 0.02 g of lipase A (from Candida antarctica), and 0.5 g of discolored silica gel. After sealing, place it in a shaker at 35°C and 170rpm to react for 45h. After the reaction was completed, the reaction was centrifuged at 12000 rpm for 10 minutes to remove enzymes and solid particles. Chromatographic detection and analysis showed that the conversion rate of the substrate was 87%. The function of using color-changing silica gel during the reaction is to remove the moisture in the reaction system.

Embodiment 3

[0034] Pour 60ml of dichloromethane-1,4-dioxane-ether mixed solvent (dichloromethane:1,4-dioxane:ether=1:4:2 by volume) into a 100ml Erlenmeyer flask, 2 grams of kojic acid and 12 grams of palmitic acid were dissolved in the mixed solvent, and finally, 0.001 gram of Rhizopus oryzae lipase, 0.001 gram of porcine pancreatic lipase and 3.0 grams of molecular sieves were added to the solution. After sealing, put it in a shaker at 60°C and 170rpm to react for 60h. After completion of the reaction, centrifuge at 12000 rpm for 10 min to remove enzymes and solid particles, and perform chromatographic detection and analysis. The conversion rate of the substrate is 62%. The function of using color-changing silica gel during the reaction is to remove the moisture in the reaction system.

technical field

[0001] The invention belongs to the technical field of biochemical industry, and in particular relates to a method for preparing kojic acid dipalmitate by a compound enzyme method. Background technique

[0002] Kojic acid is a colorless, odorless, safe and non-toxic tyrosinase inhibitor, which can be used as a preservative, preservative, and color-protecting agent in food. Kojic acid has a strong inhibitory effect on the formation of melanin in human skin. Kojic acid inactivates tyrosinase to chelate its essential copper ions, and inhibits the interconversion from dopa chromium to DHICA (5,6- dihydroxyindole-2-carboxylic acid). L-ascorbic acid and its derivatives, including MAP, reduce melanin intermediates and prevent the oxidation chain reaction from tyrosine / DOPA to various points in melanin. Therefore, kojic acid can be produced for the treatment of freckles , age spots, pigmentation whitening cosmetics, deeply loved by consumers. However, because koji...

Examples

Embodiment 1

[0030] Pour 20ml of acetone liquid into a 50ml conical flask, dissolve 0.2 gram of kojic acid and 0.72 gram of palmitic acid in 20ml of acetone, then add 0.05 gram of lipase Lipozyme RMIM (derived from Rhizomucor miehei), 0.05 gram Novozyme 435 (immobilized lipase from Candida antarctica), 1 g color-changing silica gel. After sealing, put it in a shaker at 40°C and 170rpm to react for 28h. After the reaction was completed, centrifuge at 12000 rpm for 10 min to remove enzyme and solid particles, and perform chromatographic detection and analysis. The conversion rate of the substrate was 94%. The function of using color-changing silica gel during the reaction is to remove the moisture in the reaction system.

Embodiment 2

[0032] Pour 15ml of petroleum ether-tetrahydrofuran mixed solvent (volume ratio: petroleum ether: tetrahydrofuran = 1:2) into a 30ml conical flask, dissolve 0.1 gram of kojic acid and 0.5 gram of palmitic acid in this mixed solvent, and finally add to this solution Add 0.01 g of immobilized cells containing Bacillus subtilis lipase, 0.02 g of lipase A (from Candida antarctica), and 0.5 g of discolored silica gel. After sealing, place it in a shaker at 35°C and 170rpm to react for 45h. After the reaction was completed, the reaction was centrifuged at 12000 rpm for 10 minutes to remove enzymes and solid particles. Chromatographic detection and analysis showed that the conversion rate of the substrate was 87%. The function of using color-changing silica gel during the reaction is to remove the moisture in the reaction system.

Embodiment 3

[0034] Pour 60ml of dichloromethane-1,4-dioxane-ether mixed solvent (dichloromethane:1,4-dioxane:ether=1:4:2 by volume) into a 100ml Erlenmeyer flask, 2 grams of kojic acid and 12 grams of palmitic acid were dissolved in the mixed solvent, and finally, 0.001 gram of Rhizopus oryzae lipase, 0.001 gram of porcine pancreatic lipase and 3.0 grams of molecular sieves were added to the solution. After sealing, put it in a shaker at 60°C and 170rpm to react for 60h. After completion of the reaction, centrifuge at 12000 rpm for 10 min to remove enzymes and solid particles, and perform chromatographic detection and analysis. The conversion rate of the substrate is 62%. The function of using color-changing silica gel during the reaction is to remove the moisture in the reaction system.

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