Amino acid sterol ester and preparation method thereof

Abstract

The invention discloses amino acid sterol ester and a preparation method thereof. Belongs to the technical field of natural grease product structure modification. The preparation method comprises the following steps: by taking amino acid and sterol as reaction raw materials, adding a reaction solvent and a molecular sieve, fully stirring at normal temperature, then adding lipase and providing microwave to assist an esterification reaction, thereby obtaining the amino acid sterol ester. The method is rapid, efficient and simple and convenient to operate, and the obtained amino acid sterol ester does not need to be further purified and has the advantages of high bioavailability and the like.

Description

Technical Field

[0001] The present invention relates to the technical field of structural modification of natural oil products, and more particularly to an amino acid sterol ester and a preparation method thereof. Background Art

[0002] Phytosterols are widely used in food and medicine due to their antioxidant properties, ability to inhibit the development and progression of certain cancers, anti-inflammatory and antipyretic properties, disease prevention, growth promotion, and health enhancement. However, their low solubility in both aqueous solutions and oils significantly limits their practical applications. Numerous studies have shown that esterification of phytosterols with fatty acids to form phytosterol esters significantly increases the fat solubility of phytosterols / phytosterol esters, making them more easily incorporated into oils, foods, or pharmaceutical products.

[0003] Amino acids are among the many bioactive macromolecules that form the body's structure. They are the building blocks for cell construction and tissue repair. They are also used by humans to produce antibody proteins to combat bacterial and viral infections. They are the primary building blocks for sperm and egg production and an essential precursor for the synthesis of neurotransmitters. They also provide energy for bodily and brain activity, and are the source of all life.

[0004] Microwave-assisted reactions utilize microwave radiation to accelerate chemical reactions, increasing reaction rates and selectivity while minimizing side reactions and byproducts. When exposed to microwave radiation, molecules absorb the microwave energy and convert it into heat. This energy absorption and conversion process promotes chemical reactions.

[0005] At present, there are no reports on the production of amino acid sterol esters by enzyme-catalyzed, microwave-assisted esterification of plant sterols with amino acids.

[0006] In summary, how to provide a method for preparing amino acid sterol esters is an urgent problem that those skilled in the art need to solve. Summary of the Invention

[0007] In view of this, the present invention provides an amino acid sterol ester and a preparation method thereof.

[0008] In order to achieve the above object, the present invention adopts the following technical solutions:

[0009] The first object of the present invention is to provide an amino acid sterol ester.

[0010] An amino acid sterol ester, the structural formula of which is as follows:

[0011]

[0012] Here, R represents an amino acid side chain.

[0013] Further, R represents C(CH3)H-CH2-CH2-C(CH3)HC(CH3)2H, C(CH3)H-CH=CH-C(CH3)HC(CH3)2H, C(CH3)H-CH=CH-C(C2H5)HC(CH3)2H, C(CH3)H-CH=CH-C(C2H5)HC(CH3)2H;

[0014] R' represents H, CH3, C(CH3)2H, C(CH3)HCH2CH3, CH2-CH2-S-CH3, C9H8N, CH2OH, C7H7O, CH2SH, CH2-C6H5, CH2-C( =O)-NH2, CH2-CH2-C(=O)-NH2, C(OH)H-CH3, CH2-COOH, CH2-CH2-COOH, CH2-CH2-CH2-CH2-NH2, C4H5N2.

[0015] The second object of the present invention is to provide a method for preparing amino acid sterol esters.

[0016] The above-mentioned method for preparing amino acid sterol esters comprises adding amino acids, sterols, reaction solvents and molecular sieves into a reactor, and carrying out an esterification reaction under the catalysis of lipase and the assistance of microwaves to obtain amino acid sterol esters.

[0017] Furthermore, the amino acid is glycine, alanine, valine, leucine, isoleucine, methionine, tryptophan, serine, tyrosine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine or histidine.

[0018] Furthermore, the sterol is phytosterol.

[0019] Furthermore, the sterol is sitosterol or stigmasterol.

[0020] Furthermore, the reaction solvent is ethyl acetate.

[0021] Further, the specific steps are as follows:

[0022] (1) adding amino acids and sterols into a reaction kettle, and then adding reaction solvent and molecular sieves in sequence, stirring uniformly to obtain a mixture;

[0023] (2) Transesterification reaction: lipase was added to the mixture and microwave-assisted treatment was performed. After the reaction, the mixture was cooled to 25°C.

[0024] (3) Post-treatment: The product obtained in step (2) is filtered to remove lipase, and the reaction solvent is removed by distillation under reduced pressure. After washing with water and drying, amino acid sterol esters are obtained.

[0025] Furthermore, in step (1),

[0026] The molar ratio of amino acids to sterols is 1:1 to 20;

[0027] The weight of the reaction solution is 50% of the total weight of the amino acid and sterol;

[0028] The addition amount of molecular sieve is 40-60g / L.

[0029] Furthermore, in step (2),

[0030] The mass of the lipase is 2-10% of the total mass of the amino acid, sterol, reaction solvent and molecular sieve.

[0031] Furthermore, in step (2), the parameters of the microwave-assisted treatment are 100-300 kW, 40-65° C., and a reaction time of 10-35 min.

[0032] Furthermore, in the step (3), the reaction solvent after the reduced pressure distillation treatment can be recycled.

[0033] As can be seen from the above technical solution, compared with the prior art, the present invention achieves the following beneficial effects: the present invention, through the microwave-assisted esterification method, can greatly improve the rate and conversion rate of enzymatic conversion of amino acid sterol esters. The method of the present invention has the characteristics of mild reaction conditions, rapid reaction speed, high product yield, and environmental protection. It solves the problems of low rate, low yield, and insufficient output existing in traditional methods for preparing fatty acid sterol esters, and can be used for the safe, clean, efficient, and standardized preparation of plant sterol esters. DETAILED DESCRIPTION

[0034] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0035] The reagents required for the present invention are conventional experimental reagents, purchased from commercial channels;

[0036] The experimental methods not mentioned are conventional experimental methods and will not be described in detail here.

[0037] Example 1

[0038] Preparation of glycine sitosterol ester

[0039] 7.5 g of glycine and 49.8 g of sitosterol were added to a reactor, and 28.7 g of ethyl acetate and 1.3 g of molecular sieves were added in sequence and stirred to obtain a mixture. 2.6 g of lipase was added to the mixture, and microwave assistance was provided at 200 kW and 60° C. for 25 min, the reaction was stopped, and the mixture was naturally cooled to 25° C. The resulting product was filtered to remove the lipase, and the reaction solvent (ethyl acetate) was removed by vacuum distillation. After washing and drying, 44.6 g of glycine sitosterol ester (structural formula as follows) was obtained. The purity of glycine sitosterol ester was 91.4% by high-resolution liquid chromatography-mass spectrometry (HPLC-MS / MS), and the total yield of the reaction (the purity of glycine sitosterol ester multiplied by the mass of the obtained glycine sitosterol ester, and then divided by the theoretical yield of glycine sitosterol ester) was 86.5%. (Detected by HPLC-MS / MS).

[0040]

[0041] Example 2

[0042] Preparation of Alanine Sitosterol Ester

[0043] 8.9 g of alanine and 49.8 g of sitosterol were added to a reactor, and 29.4 g of ethyl acetate and 1.6 g of molecular sieves were added in sequence and stirred to obtain a mixture. 2.7 g of lipase was added to the mixture, and microwave assistance was provided. The reaction was carried out at 280 kW and 65 ° C for 20 minutes, and the reaction was stopped and naturally cooled to 25 ° C. The resulting product was filtered to remove the lipase, and the reaction solvent (ethyl acetate) was removed by vacuum distillation. After washing and drying, 46.1 g of alanine sitosterol ester (structural formula as follows) was obtained. The product purity was 93.8% by high-resolution liquid chromatography-mass spectrometry, and the total reaction yield (the purity of alanine sitosterol ester multiplied by the mass of the obtained alanine sitosterol ester, and then divided by the theoretical yield of alanine sitosterol ester) was 89.1%.

[0044]

[0045] Example 3

[0046] Preparation of Threonine Stigmasterol Ester

[0047] 11.9 g of threonine and 41.3 g of stigmasterol were added to a reactor, and 26.6 g of ethyl acetate and 1.2 g of molecular sieves were added in sequence and stirred to obtain a mixture. 2.4 g of lipase was added to the mixture, and microwave assistance was provided. The reaction was carried out at 220 kW and 60 ° C for 20 min, and the reaction was stopped and naturally cooled to 25 ° C. The resulting product was filtered to remove the lipase, and the reaction solvent (ethyl acetate) was removed by vacuum distillation. After washing and drying, 48.9 g of threonine stigmasterol ester was obtained. The product purity was 95.5% by high-resolution liquid chromatography-mass spectrometry, and the total reaction yield (the purity of threonine stigmasterol ester multiplied by the mass of the obtained threonine stigmasterol ester, and then divided by the theoretical yield of threonine stigmasterol ester) was 90.9%.

[0048]

[0049] The various embodiments in this specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the various embodiments can be referenced to each other.

[0050] The above description of the disclosed embodiments is intended to enable one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be readily apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention is not limited to the embodiments shown herein but is intended to conform to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. An amino acid sterol ester, characterized in that Its structural formula is as follows: Among them, R represents C(CH3)H-CH2-CH2-C(CH3)HC(CH3)2H, C(CH3)H-CH=CH-C(CH3)HC(CH3)2H, C(CH3)H-CH=CH-C(C2H5)HC(CH3)2H, C(CH3)H-CH2-CH2-C(C2H5)HC(CH3)2H; R' represents H, CH3, C(CH3)2H, C(CH3)HCH2CH3, CH2-CH2-S-CH3, C9H8N, CH2OH, C7H7O, CH2SH, CH2-C6H5, CH2-C( =O)-NH2, CH2-CH2-C(=O)-NH2, C(OH)H-CH3, CH2-COOH, CH2-CH2-COOH, CH2-CH2-CH2-CH2-NH2, C4H5N2.

2. The method for preparing the amino acid sterol ester according to claim 1, wherein Amino acid, sterol, reaction solvent and molecular sieve are added into a reactor, and an esterification reaction is carried out under the catalysis of lipase and the assistance of microwave to obtain amino acid sterol ester.

3. The preparation method according to claim 2, wherein The amino acid is glycine, alanine, valine, leucine, isoleucine, methionine, tryptophan, serine, tyrosine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine or histidine.

4. The preparation method according to claim 2, wherein The sterols are phytosterols.

5. The preparation method according to claim 2, wherein The reaction solvent is ethyl acetate.

6. The preparation method according to claim 2, wherein The specific steps are as follows: (1) adding amino acids and sterols into a reaction kettle, and sequentially adding reaction solvent and molecular sieves, stirring uniformly to obtain a mixture; (2) Transesterification reaction: lipase was added to the mixture and microwave-assisted treatment was performed. After the reaction, the mixture was cooled to 25°C. (3) Post-treatment: The product obtained in step (2) is filtered to remove lipase, and the reaction solvent is removed by distillation under reduced pressure. After washing with water and drying, amino acid sterol esters are obtained.

7. The preparation method according to claim 6, wherein In the step (1), The molar ratio of amino acids to sterols is 1:1 to 20; The weight of the reaction solution is 50% of the total weight of the amino acid and sterol; The addition amount of molecular sieve is 40-60g / L.

8. The preparation method according to claim 6, wherein In the step (2), The mass of the lipase is 2-10% of the total mass of the amino acid, sterol, reaction solvent and molecular sieve.

9. The preparation method according to claim 6, wherein In the step (2), the parameters of the microwave-assisted treatment are 100-300 kW, 40-65° C., and a reaction time of 10-35 min.

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