阅读说明：本技术 一种酶法制备Omega-3脂肪酸磷脂的方法 (Method for preparing Omega-3 fatty acid phospholipid by enzyme method ) 是由 刘汝萃 范书琴 张建全 马良 杨业坤 张延秀 闫亚兵 孙盼盼 于 2021-01-20 设计创作，主要内容包括：本发明公开了一种酶法制备Omega-3脂肪酸磷脂的方法,包括以下步骤：将乙酯型鱼油、磷脂、复合磷脂酶加入反应釜中进行反应,在反应过程中不断从反应釜底部充入氮气,抽出产生的乙醇液体。本发明利用复合磷脂酶转化乙酯型鱼油和磷脂生成Omega-3脂肪酸磷脂,原料易得,反应条件温和,副产物少,易于后期的纯化提取,对环境污染小,适合广泛应用。(The invention discloses a method for preparing Omega-3 fatty acid phospholipid by an enzyme method, which comprises the following steps: adding the ethyl ester type fish oil, the phospholipid and the compound phospholipase into a reaction kettle for reaction, continuously introducing nitrogen from the bottom of the reaction kettle in the reaction process, and extracting the generated ethanol liquid. The Omega-3 fatty acid phospholipid is generated by converting the ethyl ester type fish oil and the phospholipid by using the compound phospholipase, the raw materials are easy to obtain, the reaction condition is mild, the byproducts are few, the purification and extraction at the later stage are easy, the environmental pollution is low, and the method is suitable for wide application.)

1. A method for preparing Omega-3 fatty acid phospholipid by an enzymatic method, which is characterized by comprising the following steps:

adding the ethyl ester type fish oil, the phospholipid and the compound phospholipase into a reaction kettle for reaction, continuously introducing nitrogen from the bottom of the reaction kettle in the reaction process, and extracting the generated ethanol liquid.

2. The method for preparing Omega-3 fatty acid phospholipid by the enzyme method according to claim 1, wherein the mass ratio of the ethyl ester type fish oil, the phospholipid and the compound phospholipase is (6-10): 0.6-1.3): 0.15-0.25.

3. The enzymatic process for producing Omega-3 fatty acid phospholipids as claimed in claim 1, wherein said complex phospholipase consists of the following phospholipases: the mass ratio of the phospholipase A1 to the phospholipase B is (3-6) to (4-8).

4. The method for preparing Omega-3 fatty acid phospholipid by the enzymatic method according to claim 1, wherein the reaction temperature is 40-70 ℃, the reaction time is 30-60h, and the reaction pressure is-0.08 to-0.1 MPa.

5. The method for preparing Omega-3 fatty acid phospholipid by the enzymatic method according to claim 1, wherein the reaction kettle rotating speed is 10-25 r/min.

Technical Field

The invention belongs to the technical field of health care products, and particularly relates to a method for preparing Omega-3 fatty acid phospholipid by an enzymatic method.

Background

Compared with triglyceride type and ethyl ester type fish oil, the Omega-3 fatty acid phospholipid is easier to be absorbed by human body, and the Omega-3 fatty acid phospholipid is more stable in property, the Omega-3 polyunsaturated fatty acid is fatty acid which can not be synthesized by human body, and is necessary nutrient for human body to synthesize various hormones and endogenous substances, and the physiological function of human body can be normally operated only by supplementing oleic acid from outside of food. The Omega-3 fatty acid component is eicosapentaenoic acid (EPA, containing 5 unsaturated bonds) and docosahexaenoic acid (DHA, containing 6 unsaturated bonds) which are derived from seal, fish and other substances, and the unsaturated bonds of the two are more, so the Omega-3 fatty acid component has stronger function of regulating blood fat, promotes neutral or acid cholesterol to be discharged from excrement, inhibits the synthesis of intrahepatic lipid and lipoprotein, can reduce cholesterol, triglyceride, low-density lipoprotein (LDL) and very low-density lipoprotein (VLDL) in blood plasma, increases high-density lipoprotein (HDL), and can be used for preventing and treating cardiovascular and cerebrovascular diseases, resisting cancer and diminishing inflammation, and promoting the development of nervous system and visual system.

At present, methods for preparing Omega-3 fatty acid phospholipid include natural extraction method, chemical synthesis method and microbial fermentation method, wherein the natural extraction method has the following technical problems: the raw material containing Omega-3 fatty acid phospholipid is few, and the purification cost is high, and the chemical synthesis method has the following technical problems: the reaction conditions are severe, the number of byproducts is large, the safety cannot be guaranteed, and the microbial fermentation method has the following technical problems: the specific strains required by compounding are not well screened, and the fermentation products are more and the purification is difficult.

Therefore, the development of a preparation method of Omega-3 fatty acid phospholipid, which has the advantages of easily available raw materials, mild reaction conditions, few byproducts and easy purification, is a problem to be solved by the technical personnel in the field.

Disclosure of Invention

In view of the above, the present invention provides an enzymatic method for preparing Omega-3 fatty acid phospholipids.

In order to achieve the purpose, the invention adopts the following technical scheme:

a method for preparing Omega-3 fatty acid phospholipid by an enzymatic method, which comprises the following steps:

adding the ethyl ester type fish oil, the phospholipid and the compound phospholipase into a reaction kettle for reaction, continuously introducing nitrogen from the bottom of the reaction kettle in the reaction process, and extracting the generated ethanol liquid.

The invention has the beneficial effects that: the Omega-3 fatty acid phospholipid is generated by converting the ethyl ester type fish oil and the phospholipid by using the compound phospholipase, the raw materials are easy to obtain, the reaction condition is mild, the byproducts are few, the purification and extraction at the later stage are easy, the environmental pollution is low, and the method is suitable for wide application.

Furthermore, the mass ratio of the ethyl ester type fish oil, the phospholipid and the enzyme is (6-10): (0.6-1.3): 0.15-0.25).

The beneficial effects of the further technical scheme are that: under the proportion, the ethyl ester type fish oil and the phospholipid have the largest effect to react, and the content of the omega-3 fatty acid phospholipid is the highest.

Further, the compound phospholipase consists of the following phospholipases: the mass ratio of the phospholipase A1 to the phospholipase B is (3-6) to (4-8).

Furthermore, the reaction temperature is 40-70 ℃, the reaction time is 30-60h, and the reaction pressure is-0.08 to-0.1 MPa.

The beneficial effects of the further technical scheme are that: ethanol generated by the reaction under the process condition is discharged at any time along with the reaction, the reaction efficiency is improved,

further, the rotating speed of the reaction kettle is 10-25 r/min.

The beneficial effects of the further technical scheme are that: under the condition of the rotating speed, the reaction substrates can be promoted to be fully mixed uniformly, and the reaction can be carried out at the lipase interface.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1

A method for preparing Omega-3 fatty acid phospholipid by an enzymatic method, comprising the following steps:

adding 6g of ethyl ester type fish oil, 1.3g of phospholipid and 0.25g of compound phospholipase, wherein 0.1g of phospholipase A and 0.15g of phospholipase B into a reaction kettle for reaction, the reaction temperature is 70 ℃, the reaction time is 30h, the reaction pressure is-0.1 MPa, the rotating speed of the reaction kettle is 25r/min, continuously filling nitrogen from the bottom of the reaction kettle in the reaction process, and extracting the generated ethanol liquid to obtain a product, wherein the content of phospholipid type fatty acid in the obtained product is 80%, and the content of Omega-3 fatty acid phospholipid is 60%.

Example 2

A method for preparing Omega-3 fatty acid phospholipid by an enzymatic method, comprising the following steps:

adding 8g of ethyl ester type fish oil, 0.8g of phospholipid and 0.2g of compound phospholipase into a reaction kettle for reaction, wherein the phospholipase A is 0.1g and the phospholipase B is 0.1g, the reaction temperature is 50 ℃, the reaction time is 50 hours, the reaction pressure is-0.085 MPa, the rotating speed of the reaction kettle is 20r/min, continuously filling nitrogen into the bottom of the reaction kettle in the reaction process, and extracting the generated ethanol liquid to obtain a product, wherein the content of phospholipid type fatty acid in the product is 75 percent, and the content of Omega-3 fatty acid phospholipid is 63 percent.

Example 3

A method for preparing Omega-3 fatty acid phospholipid by an enzymatic method, comprising the following steps:

adding 7g of ethyl ester type fish oil, 0.8g of phospholipid and 0.18g of compound phospholipase, wherein the phospholipase A is 10.108g and the phospholipase B is 0.072g into a reaction kettle for reaction, the reaction temperature is 60 ℃, the reaction time is 40h, the reaction pressure is-0.09 MPa, the rotating speed of the reaction kettle is 15r/min, continuously filling nitrogen from the bottom of the reaction kettle in the reaction process, and extracting the generated ethanol liquid. The phospholipid type fatty acid content of the obtained product was 78%, wherein the content of Omega-3 fatty acid phospholipid was 62%.

Example 4

A method for preparing Omega-3 fatty acid phospholipid by an enzymatic method, comprising the following steps:

10g of ethyl ester type fish oil, 0.6g of phospholipid and 0.15g of compound phospholipase, wherein 0.105g of phospholipase A and 0.045g of phospholipase B are added into a reaction kettle for reaction, the reaction temperature is 40 ℃, the reaction time is 60 hours, the reaction pressure is-0.08 MPa, the rotating speed of the reaction kettle is 10r/min, nitrogen is continuously filled from the bottom of the reaction kettle in the reaction process, and the produced ethanol liquid is extracted. The phospholipid type fatty acid content of the obtained product was 77%, wherein the content of Omega-3 fatty acid phospholipid was 62%.

The description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.