Lipase catalysis method for synthesis of astaxanthin succinate

A technology of succinate and astaxanthin, applied in the biological field, can solve the problems of loss of biological activity, decreased activity, instability of astaxanthin, etc., and achieve the effects of easy digestion and absorption, mild reaction conditions, and good application potential

Inactive Publication Date: 2015-04-15
UNIV OF SCI & TECH BEIJING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, because astaxanthin is extremely unstable, it is easy to lose its biological activity under conditions such as light, heat and radiation, and the cell wall of Phaffia rhodozyme is difficult to break
Extracting astaxanthin from Phaffia rhodozyma cells requires severe cell wall breaking methods such as acid cooking, then solvent extraction, solution concentration (rotary evaporation method), and then purification by means of crystallization and chromatography. The instability of xanthin is easy to degrade during the separation and purification process, resulting in low yield of final astaxanthin
[0004] During the storage and processing of astaxanthin products, due to the high instability of astaxanthin, its activity is greatly reduced and its use effect is affected

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] 2L of Phaffia rhodozyme fermentation broth was directly crushed with a high-pressure homogenizer, and the crushed Phaffia rhodozyme cells were recovered by centrifugation. After freeze-drying, they were mixed with isopropanol and n-hexane (V:V=2: 1) Carry out astaxanthin extraction, the solid-liquid ratio is 15mL / g. The extracted solution was subjected to rotary evaporation to obtain the crude extract of astaxanthin. Take 1.2 mL of the crude extract, add 3.8 mL of reaction solvent DMSO and 0.2 g of yeast lipase, and add succinic anhydride to make the molar ratio of succinic anhydride to astaxanthin 1500 times. / min) for 8 hours, the conversion rate of astaxanthin was 53.70%, of which the content of monoester in astaxanthin succinate was 82.14%, and the content of diester was 17.86%.

Embodiment 2

[0020] 2L of Phaffia rhodozyme fermentation broth was directly crushed with a high-pressure homogenizer, and the crushed Phaffia rhodozyme cells were recovered by centrifugation. After freeze-drying, they were mixed with isopropanol and n-hexane (V:V=2: 1) Carry out astaxanthin extraction, the solid-liquid ratio is 15mL / g. The extracted solution was subjected to rotary evaporation to obtain the crude extract of astaxanthin. Take 1.2mL of crude extract, add 3.8mL of reaction solvent DMF and 0.2g of yeast lipase, add succinic anhydride, the molar ratio of succinic anhydride to astaxanthin is 1500 times, and react in a constant temperature shaker (160 r / min) at 35°C After 6 hours, the conversion rate of astaxanthin was 25.96%, and the generated astaxanthin succinate was all astaxanthin succinate monoester.

Embodiment 3

[0022] 2L of Phaffia rhodozyme fermentation broth was directly crushed with a high-pressure homogenizer, and the crushed Phaffia rhodozyme cells were recovered by centrifugation. After freeze-drying, they were mixed with isopropanol and n-hexane (V:V=2: 1) Carry out astaxanthin extraction, the solid-liquid ratio is 15mL / g. The extracted solution was subjected to rotary evaporation to obtain the crude extract of astaxanthin. Take 1.2mL of crude extract, add reaction solvent DMSO2.4mL and 0.2g yeast lipase, add succinic anhydride, the molar ratio of succinic anhydride to astaxanthin is 1500 times, 45℃ constant temperature shaker (160 r / min) for 24h , the conversion rate of astaxanthin is 86.50%, of which the monoester content of astaxanthin succinate is 68.01%, and the content of diester is 31.99%.

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Abstract

The invention belongs to the field of biotechnology and relates to a directly to a lipase catalysis method for synthesis of astaxanthin succinate by directly using Phaffia rhodozyma as raw material through separation and coupling reaction. The specific method is as below: breaking Phaffia rhodozyma cells by a high-pressure homogenization method, centrifuging, recovering, freezing, drying, adding a mixed solution of isopropanol / hexane for astaxanthin extraction, subjecting the extraction liquid to rotary evaporation and concentration, directly adding reaction solvents of lipase, succinic anhydride and DMF at a temperature of 20-50 DEG C for esterification, so as to synthesize astaxanthin succinate. The method has important application value and prospect in solving the problem of high unstability of astaxanthin, and production of stable efficient antioxidant astaxanthin succinate.

Description

technical field [0001] The invention belongs to the field of biotechnology, and provides a method for preparing astaxanthin succinate by using Phaffia rhodozyma cells as raw materials and adopting a lipase-catalyzed separation and reaction coupling. This method is useful for solving the high instability of astaxanthin It has important application value and prospect in the production of efficient and stable antioxidant astaxanthin succinate. Background technique [0002] Astaxanthin (3,3'-dihydroxy-4,4'-diketo-beta-carotene) is a red carotenoid found in the shell of langoustines, oysters and salmon, which is intracellular and can Combined with protein to form green and blue. Astaxanthin is the strongest antioxidant in nature. Its antioxidant capacity is 1000 times that of vitamin E. It can effectively remove oxygen free radicals in cells, enhance cell regeneration, remove aging cells and maintain body balance. It has the functions of protecting skin, promoting Hair growt...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P23/00
Inventor 尹春华韩烨李卉冯卓闫海刘晓璐许倩倩吕乐
Owner UNIV OF SCI & TECH BEIJING
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