CN111363767A

CN111363767A - Preparation method of phosphatidylserine for food

Info

Publication number: CN111363767A
Application number: CN202010321012.XA
Authority: CN
Inventors: 孙荣; 刘建民; 任明; 张雷
Original assignee: Shandong Huishilai Biotechnology Co ltd
Current assignee: Shandong Huishilai Biotechnology Co ltd
Priority date: 2020-04-22
Filing date: 2020-04-22
Publication date: 2020-07-03

Abstract

The invention discloses a preparation method of phosphatidylserine for food, which comprises the following steps: mixing lecithin and embedding agent, stirring with water, embedding, adding L-serine and CaCl₂Phospholipase D and acetic acid buffer solution are mixed evenly for enzyme conversion; and after the conversion is finished, adding an embedding agent into the reaction solution, cooling to separate out a precipitate after the embedding treatment, and collecting the precipitate to obtain a water-soluble phosphatidylserine product. The invention has the beneficial effects that: the problem of organic solvent residue in the phosphatidylserine product is solved, and the application safety of the product in the food field is improved; the dispersion state of the substrate in the water phase is improved, the water consumption of a conversion system is reduced, the problem that lecithin in the water phase is hydrolyzed is effectively solved, the generation of by-product phosphatidic acid is reduced, and the product quality is improved; improves the dispersibility and solubility of the phosphatidylserine in food, and expands the application fields of the product in the aspects of special medical food, health-care food, common food and the like.

Description

Preparation method of phosphatidylserine for food

Technical Field

The invention relates to the technical field of biological enzyme catalysis, in particular to a preparation method of phosphatidylserine for food.

Background

Phosphatidylserine (PS) is a member of the phospholipid family, is one of the important components of cell membrane phospholipids, and has important regulatory effects on many cellular metabolic processes. In recent years, animal experiments and clinical application experiments prove that phosphatidic acid serine can improve cognitive ability, improve Alzheimer's disease, treat depression, and relieve mental stress and physical fatigue.

The national health council added PS to the new resources food catalogue at 21/10/2010, allowing addition as new resources to food.

The preparation mainly comprises a solvent extraction method and an enzyme conversion method. The extraction method mainly extracts lecithin from animal and plant cells, and the extraction sources mainly comprise brains and internal organs of animals. The solvent extraction method is simple to operate and large in treatment capacity, once is a main production method of PS, but because the extraction method consumes a large amount of organic solvent, the environment is burdened, and more importantly, along with the large-scale occurrence of mad cow disease, the safety of extracting PS from animal brains faces severe examination. Compared with an extraction method, the enzyme conversion method for preparing the PS is safer, low-carbon and environment-friendly. In this context, the enzymatic conversion to prepare PS has become a hot spot in the current research.

The enzyme method for producing PS takes soybean lecithin as a raw material and generates PS through enzyme conversion. Due to the hydrophobic property of lecithin, enzyme conversion is mostly carried out in a water/organic phase bidirectional system, but a PS organic solvent obtained by conversion production in the bidirectional system is difficult to completely remove, so that the organic solvent residue is dangerous, and the application of PS in food is influenced.

In this context, the production of PS by enzymatic conversion in the pure aqueous phase is being investigated by those skilled in the art. Since lecithin is a hydrophobic substance, in order to ensure the dispersibility of lecithin in an aqueous phase, a specific homogenization process is required, or excessive water is used for dispersing lecithin, so as to improve the contact between a substrate and an enzyme, but the excessive exposure of lecithin in the aqueous phase can cause a substrate hydrolysis reaction, so that the content of phosphatidic acid serving as a byproduct in a reaction system is too high, and the product quality is influenced.

Disclosure of Invention

In order to solve the technical problems, the invention provides the preparation method of the phosphatidylserine for the food, the whole conversion process is free from the intervention of an organic solvent, the problem of organic solvent residue in the product is thoroughly solved, the safety of the product is improved, the water phase in a reaction system is reduced, and the generation of a by-product phosphatidic acid is reduced.

The specific invention can be realized by the following technical method:

the operation steps are as follows:

(1) mixing lecithin and embedding agent, stirring with water, embedding, adding L-serine and CaCl₂Phospholipase D, mix with acetic acid buffer solution, carry on the enzyme conversion.

(2) And after the conversion is finished, adding a certain amount of embedding agent into the reaction solution, cooling to separate out a precipitate after the embedding treatment, and collecting the precipitate to obtain the water-soluble phosphatidylserine product.

The lecithin is soybean lecithin (PC is more than or equal to 98%);

the embedding agent is as follows: one or more of cyclodextrin, maltodextrin and porous starch, wherein the addition amount of the embedding agent is 1-2 times of that of lecithin;

the water adding amount is 2-5 times (w/v) of that of lecithin;

the embedding treatment comprises the following steps: one of ultrasonic treatment, homogenization and heating and stirring;

the ultrasonic wave: the power is 100-;

the homogenization is as follows: the homogenizing pressure is 600-800 Pa;

the heating and stirring: the temperature is 70-80 ℃, the stirring speed is 100-;

the addition amount of the L-serine is 1.5 to 3 times of that of lecithin, and CaCl is added₂The adding amount is 0.1-0.3 times of lecithin, the adding amount of phospholipase is 20-50U/g lecithin, and the adding amount of acetic acid buffer solution is 1-2 times (W/V) of lecithin;

the concentration of the acetic acid buffer solution is 0.1mol/L, and the pH value is 5.5-6;

the phospholipase D is as follows: one of commercially available phospholipase D and phospholipase D fermentation broth;

the enzyme conversion conditions are as follows: at the temperature of between 33 and 38 ℃ and at the speed of 100r/min, and transforming for 5 to 10 hours;

the cooling temperature and time are as follows: standing for 12-24h at 4 ℃;

the collected precipitate is as follows: and (4) centrifuging and collecting.

The invention has the beneficial effects that:

1. the method for preparing the phosphatidylserine by pure water phase inversion has the advantages that no organic solvent is introduced in the whole system and the production process, the problem of organic solvent residue in the phosphatidylserine product is solved, and the safety of the product in the food field is improved.

According to the invention, the embedding agent is added into the enzyme conversion system to embed the lecithin, so that the dispersion state of the substrate in the water phase is improved, the water consumption of the conversion system is reduced, the problem that the lecithin in the water phase is hydrolyzed is effectively solved, the generation of by-product phosphatidic acid is reduced, and the product quality is improved.

The product phosphatidylserine after the conversion is further embedded to obtain a water-soluble product, so that the dispersibility and the solubility of the phosphatidylserine in food are improved, and the application fields of the product in the aspects of special medical food, health-care food, common food and the like are expanded.

DETAILED DESCRIPTION OF EMBODIMENT (S) OF INVENTION

The present invention is further illustrated by the following examples.

Example 1

Weighing 50g lecithin (PC is more than or equal to 98%) and 50g cyclodextrin, adding into 150ml pure water, stirring at 100r/min for 5h in 70 deg.C water bath, adding 50g L-serine and CaCl₂5g, phospholipase D1000U and 50ml of acetic acid buffer solution with the pH value of 5.5, uniformly stirring, converting at 35 ℃ for 5 hours at 150r/min, adding 50g of cyclodextrin after the conversion is finished, carrying out ultrasonic treatment at 100KHz for 30 minutes, cooling at 4 ℃ and standing for 12 hours, centrifuging and collecting precipitates to obtain the water-soluble phosphatidylserine.

Example 2

Weighing 50g lecithin (PC is more than or equal to 98 percent) and 100g cyclodextrin, adding into 200ml pure water, stirring uniformly, homogenizing at 800Pa, adding 100g L-serine and CaCl₂5g, phospholipase D1000U, 100ml of acetic acid buffer solution with the pH value of 5.5, uniformly stirring, converting at 35 ℃, 200r/min for 8h, adding 50g of cyclodextrin after the conversion is finished, homogenizing at 800Pa, cooling at 4 ℃, standing for 24h, centrifuging and collecting precipitates to obtain the water-soluble phosphatidylserine.

Example 3

2 parts of 50g lecithin (PC is more than or equal to 98%) are weighed respectively, 100g of cyclodextrin is added into one part, and no cyclodextrin is added into the other part, other operation steps are the same as example 1, the conversion rate of the lecithin is examined, and the result is shown in the following table:

TABLE 1 Effect of lecithin embedding on conversion

	Conversion rate of lecithin	Phosphatidic acid production rate
			Adding cyclodextrin	75%	5%
Without addition of cyclodextrin	45%	15%

Claims

1. A preparation method of phosphatidylserine for food is characterized by comprising the following steps:

(1) mixing lecithin and embedding agent, stirring with water, embedding, adding L-serine and CaCl₂Phospholipase D and acetic acid buffer solution are mixed evenly for enzyme conversion;

(2) and after the conversion is finished, adding an embedding agent into the reaction solution, cooling to separate out a precipitate after the embedding treatment, and collecting the precipitate to obtain the water-soluble phosphatidylserine product.

2. The method for preparing phosphatidylserine for food according to claim 1, wherein in the steps (1) and (2), the embedding agent is: one or more of cyclodextrin, maltodextrin and porous starch, wherein the addition amount of the embedding agent is 1-2 times of that of lecithin, and the embedding treatment comprises the following steps: ultrasonic treatment, homogenizing, heating and stirring.

3. The method for preparing phosphatidylserine for food according to claim 2, wherein the ultrasonic condition is: the power is 100-: the homogenizing pressure is 600-800Pa, and the heating and stirring conditions are as follows: the temperature is 70-80 ℃, the stirring speed is 100-.

4. The process according to claim 1, wherein the amount of L-serine added is 2-5 times (w/v) that of lecithin, and the amount of CaCl is 1.5-3 times that of lecithin in steps (1) and (2)₂The adding amount is 0.1-0.3 time of lecithin, the adding amount of phospholipase is 20-50U/g lecithin, the adding amount of acetate buffer is 1-2 times (w/v) of lecithin, the concentration of acetate buffer is 0.1mol/L, the pH is 5.5-6, and the enzyme conversion conditions are as follows: at the temperature of between 33 and 38 ℃ and at the speed of 100 and 200r/min, and converting for 5 to 10 hours, wherein the cooling is as follows: standing for 12-24h at 4 ℃, wherein the collected precipitate is as follows: and (4) centrifuging and collecting.