Method for preparing acyl-L-carnitine and/or L-carnitine

Abstract

The invention discloses a method for preparing acyl-L-carnitine and / or L-carnitine, which comprises the following steps: 1) adding crotonbetaine and carnitine anhydrase to an aqueous solution containing [Bmim]PF6 ionic liquid, and reacting at 30-45 DEG C for 2-12h; 2) removing the carnitine anhydrase and water content in the reaction system in step 1); and 3) adding lipase and acyl donors to the reaction system in step 2), and reacting at 30-80 DEG C for 2-24h to obtain a reaction system containing the acyl-L-carnitine. The method for preparing acyl-L-carnitine has the following advantages: (1) the problem of low conversion rate in L-carnitine synthesis using crotonbetaine as a precursor is solved; (2) the problems on key technologies of product separation and purification in the L-carnitine synthesis using the crotonbetaine as the precursor are solved; and (3) the acyl-L-carnitine can be prepared. The acyl-L-carnitine can be used as the final product, and also can be hydrolyzed into L-carnitine.

Description

technical field

[0001] The present invention relates to a method for preparing acyl-L-carnitine and / or L-carnitine. Background technique

[0002] L-carnitine is widely used in pharmaceutical industry, food industry and feed industry. At present, the methods for producing L-carnitine mainly include extraction, chemical synthesis and biocatalysis. Among them, chemical synthesis and biocatalysis are of commercial value. Compared with the chemical synthesis method, the biocatalysis method has the characteristics of environmental friendliness, low cost, and high product safety, and has a better development prospect.

[0003] The synthesis of L-carnitine from croton betaine under the catalysis of carnitine dehydratase is one of the important ways to prepare L-carnitine, and it is also the main method to produce L-carnitine by biocatalysis. However, the method for preparing L-carnitine with croton betaine as a precursor has the following problems: (1) The conversion rate of the ...

Examples

Embodiment 1

[0032] Embodiment 1, preparation palmitoyl-L-carnitine

[0033] 1) Mix 20ml of [Bmim]PF 6 Ionic liquid (purchased from Henan Lihua Pharmaceutical Co., Ltd.), 1.5mmol croton betaine (Fan Liqiang, Yuan Qinsheng. Microbial conversion method-a new idea of ​​DL-carnitine untwisting [J]. Medicinal Chemistry, 1999, 34 (5): 335-337), 100mg KNO 3 , 200 mg of carnitine dehydratase and 1 ml of pH6.5 phosphate buffer solution were placed in a 125 ml round bottom flask, and reacted for 3 hours at 40° C. and 120 rpm on a vacuum rotary evaporator to obtain L-carnitine.

[0034] The reaction at this stage is the hydration reaction of L-carnitine from croton betaine catalyzed by carnitine dehydratase.

[0035] 2) Centrifuge the reaction solution of the above step 1) at 4200rpm for 1min to remove the carnitine dehydratase in the reaction system, place the separated carnitine dehydratase in 1ml of pH 6.5 phosphate buffer solution for the hydration reaction of the next cycle ; Then keep the re...

Embodiment 2

[0043] Embodiment 2, preparation palmitoyl-L-carnitine (the process of step 2-4 is not repeated)

[0044] 1) Mix 20ml of [Bmim]PF 6 Ionic liquid (purchased from Henan Lihua Pharmaceutical Co., Ltd.), 1.5mmol croton betaine (Fan Liqiang, Yuan Qinsheng. Microbial conversion method-a new idea of ​​DL-carnitine untwisting [J]. Medicinal Chemistry, 1999, 34 (5): 335-337), 100mg KNO 3 , 200mg of carnitine dehydratase and 1ml of pH6.5 phosphate buffer solution were placed in a 125ml round bottom flask, and reacted for 12h on a vacuum rotary evaporator at 30°C and 120rpm to obtain L-carnitine.

[0045] The reaction at this stage is the hydration reaction of L-carnitine from croton betaine catalyzed by carnitine dehydratase.

[0046] 2) Centrifuge the reaction solution in the above step 1) at 500rpm for 30min to remove the carnitine dehydratase in the reaction system, place the separated carnitine dehydratase in 1ml of pH 6.5 phosphate buffer solution for the hydration reaction of the ...

Embodiment 3

[0052] Embodiment 3, preparation of palmitoyl-L-carnitine (the process of steps 2-4 is repeated once)

[0053] 1) Mix 20ml of [Bmim]PF 6 Ionic liquid (purchased from Henan Lihua Pharmaceutical Co., Ltd.), 1.5mmol croton betaine (Fan Liqiang, Yuan Qinsheng. Microbial conversion method-a new idea of ​​DL-carnitine untwisting [J]. Medicinal Chemistry, 1999, 34 (5): 335-337), 50mg fumaric acid, 200mg carnitine dehydratase and 1ml pH6.5 phosphate buffer solution were placed in a 125ml round bottom flask, and reacted for 2h at 45°C and 120rpm on a vacuum rotary evaporator to obtain L- carnitine.

[0054] The reaction at this stage is the hydration reaction of L-carnitine from croton betaine catalyzed by carnitine dehydratase.

[0055] 2) Centrifuge the reaction solution of the above step 1) at 10000rpm for 0.5min to remove the carnitine dehydratase in the reaction system, place the separated carnitine dehydratase in 1ml of pH 6.5 phosphate buffer solution for the next cycle of hyd...

technical field

[0001] The present invention relates to a method for preparing acyl-L-carnitine and / or L-carnitine. Background technique

[0002] L-carnitine is widely used in pharmaceutical industry, food industry and feed industry. At present, the methods for producing L-carnitine mainly include extraction, chemical synthesis and biocatalysis. Among them, chemical synthesis and biocatalysis are of commercial value. Compared with the chemical synthesis method, the biocatalysis method has the characteristics of environmental friendliness, low cost, and high product safety, and has a better development prospect.

[0003] The synthesis of L-carnitine from croton betaine under the catalysis of carnitine dehydratase is one of the important ways to prepare L-carnitine, and it is also the main method to produce L-carnitine by biocatalysis. However, the method for preparing L-carnitine with croton betaine as a precursor has the following problems: (1) The conversion rate of the ...

Examples

Embodiment 1

[0032] Embodiment 1, preparation palmitoyl-L-carnitine

[0033] 1) Mix 20ml of [Bmim]PF 6 Ionic liquid (purchased from Henan Lihua Pharmaceutical Co., Ltd.), 1.5mmol croton betaine (Fan Liqiang, Yuan Qinsheng. Microbial conversion method-a new idea of ​​DL-carnitine untwisting [J]. Medicinal Chemistry, 1999, 34 (5): 335-337), 100mg KNO 3 , 200 mg of carnitine dehydratase and 1 ml of pH6.5 phosphate buffer solution were placed in a 125 ml round bottom flask, and reacted for 3 hours at 40° C. and 120 rpm on a vacuum rotary evaporator to obtain L-carnitine.

[0034] The reaction at this stage is the hydration reaction of L-carnitine from croton betaine catalyzed by carnitine dehydratase.

[0035] 2) Centrifuge the reaction solution of the above step 1) at 4200rpm for 1min to remove the carnitine dehydratase in the reaction system, place the separated carnitine dehydratase in 1ml of pH 6.5 phosphate buffer solution for the hydration reaction of the next cycle ; Then keep the re...

Embodiment 2

[0043] Embodiment 2, preparation palmitoyl-L-carnitine (the process of step 2-4 is not repeated)

[0044] 1) Mix 20ml of [Bmim]PF 6 Ionic liquid (purchased from Henan Lihua Pharmaceutical Co., Ltd.), 1.5mmol croton betaine (Fan Liqiang, Yuan Qinsheng. Microbial conversion method-a new idea of ​​DL-carnitine untwisting [J]. Medicinal Chemistry, 1999, 34 (5): 335-337), 100mg KNO 3 , 200mg of carnitine dehydratase and 1ml of pH6.5 phosphate buffer solution were placed in a 125ml round bottom flask, and reacted for 12h on a vacuum rotary evaporator at 30°C and 120rpm to obtain L-carnitine.

[0045] The reaction at this stage is the hydration reaction of L-carnitine from croton betaine catalyzed by carnitine dehydratase.

[0046] 2) Centrifuge the reaction solution in the above step 1) at 500rpm for 30min to remove the carnitine dehydratase in the reaction system, place the separated carnitine dehydratase in 1ml of pH 6.5 phosphate buffer solution for the hydration reaction of the ...

Embodiment 3

[0052] Embodiment 3, preparation of palmitoyl-L-carnitine (the process of steps 2-4 is repeated once)

[0053] 1) Mix 20ml of [Bmim]PF 6 Ionic liquid (purchased from Henan Lihua Pharmaceutical Co., Ltd.), 1.5mmol croton betaine (Fan Liqiang, Yuan Qinsheng. Microbial conversion method-a new idea of ​​DL-carnitine untwisting [J]. Medicinal Chemistry, 1999, 34 (5): 335-337), 50mg fumaric acid, 200mg carnitine dehydratase and 1ml pH6.5 phosphate buffer solution were placed in a 125ml round bottom flask, and reacted for 2h at 45°C and 120rpm on a vacuum rotary evaporator to obtain L- carnitine.

[0054] The reaction at this stage is the hydration reaction of L-carnitine from croton betaine catalyzed by carnitine dehydratase.

[0055] 2) Centrifuge the reaction solution of the above step 1) at 10000rpm for 0.5min to remove the carnitine dehydratase in the reaction system, place the separated carnitine dehydratase in 1ml of pH 6.5 phosphate buffer solution for the next cycle of hyd...

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