阅读说明：本技术 一种金枪鱼肽、其提取方法及作为降压剂的应用 (Tuna peptide, extraction method thereof and application of tuna peptide as antihypertensive agent ) 是由 方诩 韩丽娟 贾洋洋 于 2020-05-27 设计创作，主要内容包括：本发明具体涉及一种金枪鱼肽、其提取方法及作为降压剂的应用。金枪鱼肽是一种具有良好的保健效果的成分,从金枪鱼加工产品的下脚料中提取金枪鱼肽是一种具有显著经济意义的资源利用方式。传统制备方法提取的金枪鱼肽往往伴随纯度、感官不良等缺陷。本发明提供了一种同时采用微生物和复合蛋白酶进行发酵和酶解的处理方法,将多肽充分的从金枪鱼生产的下脚料蛋白酶解液中充分的释放,增加提取效率,同时结合树脂吸附,同时具有无腥味、环境友好、提取率高,纯度高等优点。(The invention particularly relates to tuna peptide, an extraction method thereof and application of the tuna peptide as a blood pressure reducing agent. Tuna peptide is a component with good health care effect, and extraction of the tuna peptide from leftovers of processed products of the tuna is a resource utilization mode with remarkable economic significance. Tuna peptide extracted by the traditional preparation method is often accompanied with the defects of poor purity, poor sense and the like. The invention provides a treatment method for fermentation and enzymolysis by simultaneously adopting microorganisms and compound protease, which sufficiently releases polypeptide from leftover protein enzymolysis liquid produced by tuna, increases the extraction efficiency, is combined with resin adsorption, and has the advantages of no fishy smell, environmental friendliness, high extraction rate, high purity and the like.)

1. The extraction method of the tuna peptide is characterized by comprising the steps of adding compound protease and microorganisms into the tuna, fermenting and performing enzymolysis simultaneously to obtain absorption liquid, and purifying the absorption liquid to prepare the tuna peptide.

2. The method for extracting tuna peptide according to claim 1, comprising the steps of:

(1) adding water into tuna for homogenizing to obtain homogenate;

(2) diluting the homogenate by adopting a culture solution until the concentration of a solid matter is 8-12%, adding compound protease, and inoculating microorganisms for fermentation and enzymolysis to obtain an enzymolysis solution;

(3) sterilizing and centrifuging the enzymolysis liquid to obtain absorption liquid, purifying the absorption liquid by resin to obtain tuna peptide eluent, and concentrating and drying the tuna peptide.

3. The method for extracting tuna peptide according to claim 2, wherein in the step (1), the tuna is tuna meat or leftovers of the tuna processing;

or the homogenizing time is 0.8-1.2 hours.

4. The method for extracting tuna peptide according to claim 2, wherein in the step (2), the culture solution is a culture solution required for culturing microorganisms;

or the compound protease is a commonly used additive substance in the field of food processing, and is a protease for hydrolyzing animal and vegetable proteins; preferably, neutral as well as acidic proteases;

or the concentration of the compound protease is 2-5%.

5. The method for extracting tuna peptide according to claim 2, wherein the microorganism is a bacterium that produces protease;

or the fermentation and enzymolysis time is 44-50 h;

or the fermentation and enzymolysis temperature is 25-35 ℃.

6. The method for extracting tuna peptide according to claim 2, wherein the sterilization time is 10-60 min;

or the centrifugation parameter is 5000-.

7. The method for extracting tuna peptide according to claim 2, wherein in the step (3), the resin is a macroporous adsorption resin;

or the loading amount of the resin is equal to the weight of the resin;

or the amount of the eluent is 6-7 times of the weight of the resin;

or the elution flow rate is controlled to be 1-1.2 bed volumes per hour;

or the eluent is 75% ethanol solution;

or desorbing tuna peptide in the resin by using ethanol solution with the concentration of more than 30 percent.

8. The method for extracting tuna peptide according to claim 2, wherein the concentration is carried out under vacuum at a temperature of 80 ℃ or lower and under a vacuum degree of-0.08 Mpa or lower;

or concentrating until the specific gravity of the concentrated paste is 1.1-1.15, and then carrying out spray drying to obtain the tuna peptide.

9. Tuna peptide produced by the method for extracting tuna peptide according to any one of claims 1 to 8.

10. The tuna peptide of claim 9 for use as a hypotensive agent, wherein the tuna peptide is used as a hypotensive agent comprising the steps of applying the tuna peptide as a hypotensive active ingredient to prepare a hypotensive product; preferably, the application mode comprises the step of using the tuna peptide as an active ingredient and an auxiliary ingredient together for preparing the blood pressure reducing product, and also comprises the step of using the tuna peptide and other ingredients with the blood pressure reducing effect or the auxiliary blood pressure reducing effect together for preparing the blood pressure reducing product; further preferably, the blood pressure lowering product comprises a blood pressure lowering medicine and a blood pressure lowering health care product.

Technical Field

The invention belongs to the technical field of protein polypeptide extraction methods, and particularly relates to tuna peptide, an extraction method of the tuna peptide and application of the tuna peptide as an active component for reducing blood pressure in preparation of a blood pressure reducing agent.

Background

The information in this background section is only for enhancement of understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.

The content of crude protein in the back muscle of the tuna finfish is 26.2%. The fat content is only 0.2 percent, and the food is a high-protein and low-fat health food; the protein of the back muscle of tuna finns was evaluated according to the model of essential amino acid requirement recommended by FAO/WHO in 1973 (adult model), and the back muscle contains 41.31% of amino acids in terms of flavor-developing amino acids and abundant mineral elements such as potassium, phosphorus, sodium, magnesium, calcium, zinc, selenium, etc. The muscle of the back of the tuna finfish is a high-quality raw material with rich nutrition and delicious taste.

At present, tuna is sold mainly in a fresh mode and processed into foods such as cans and seasonings. However, a large amount of high protein leftovers such as fish heads, viscera, fish skin and the like are generated in the deep processing process of the tuna, thereby causing resource waste. Therefore, the full utilization of the high-protein leftovers is beneficial to the reutilization of resources, and simultaneously, the profit of enterprises is increased to a certain extent.

The existing research shows that the tuna peptide has good health care effects, such as reducing fat in blood and improving the regeneration capacity of liver cells; the anserine in the tuna body has the effects of decomposing lactic acid and accelerating the discharge of uric acid, and is expected to be applied to gout patients. Therefore, the method for producing the polypeptide by using the tuna leftovers becomes a better resource utilization mode.

At present, the extraction method of tuna peptide mainly comprises an enzymolysis process, and a crude extract of the peptide is obtained by combining a centrifugation and alcohol precipitation method. However, the inventor believes that the purity of the tuna peptide obtained by the existing extraction method is not ideal, and the tuna peptide is often accompanied by fishy smell, so that the subsequent treatment difficulty is increased, and the market value is seriously influenced.

Disclosure of Invention

Aiming at the research background, the invention provides an optimized extraction method of tuna peptides, and the tuna peptides extracted by the method belong to small molecular peptides, have higher purity, no peculiar smell and low difficulty in subsequent processing, and are products with high economic value.

Based on the technical effects, the invention provides the following technical scheme:

according to the first aspect of the invention, the extraction method of the tuna peptide comprises the steps of adding compound protease and microorganisms into the tuna, simultaneously performing fermentation and enzymolysis to obtain absorption liquid, and purifying the absorption liquid to prepare the tuna peptide.

The invention provides an extraction method, and in a preferred technical scheme, the extraction method is used for purifying through resin. According to the research of the invention, the compound protease and the microorganism are added into the tuna together for fermentation and enzymolysis, the tuna extract prepared by the method can effectively eliminate peculiar smell, the molecular weight of the tuna peptide is obviously reduced, and the subsequent resin purification difficulty is relatively reduced. The tuna peptide obtained by the method overcomes the defects of peculiar smell and low purity of the existing product, and the product also has good activity of reducing blood pressure through verification, and is expected to have good effect when being applied to clinical medicines or health care products.

In a second aspect of the present invention, the tuna peptide prepared by the method for extracting tuna peptide of the first aspect is provided.

In a third aspect of the invention, there is provided the use of a tuna peptide according to the second aspect as a hypotensive agent.

The research of the invention proves that the tuna peptide has good ACE inhibitory activity, and is expected to be used as an antihypertensive active ingredient and auxiliary materials or be applied to preparation of medicines and health care products in combination with other antihypertensive active ingredients.

The beneficial effects of one or more technical schemes are as follows:

the research result provides a preparation method for fermenting and hydrolyzing the tuna raw material by simultaneously adopting the compound protease and the microorganism for the first time, the culture method can fully dissolve the tuna peptide from the raw material, the recovery efficiency is good, and the prepared tuna peptide has high purity, no bad smell and higher market value.

Detailed Description

It is to be understood that the following detailed description is exemplary and is intended to provide further explanation of the invention as claimed. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the invention. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.

As introduced in the background art, aiming at the defects in the prior art, the invention provides an extraction method of tuna peptide, the tuna peptide prepared by the extraction method and application.

According to the first aspect of the invention, the extraction method of the tuna peptide comprises the steps of adding compound protease and microorganisms into the tuna, simultaneously performing fermentation and enzymolysis to obtain absorption liquid, and purifying the absorption liquid to prepare the tuna peptide.

Preferably, the method for extracting tuna peptide specifically comprises the following steps:

(1) adding water into tuna for homogenizing to obtain homogenate;

(2) diluting the homogenate by adopting a culture solution until the concentration of a solid matter is 8-12%, adding compound protease, and inoculating microorganisms for fermentation and enzymolysis to obtain an enzymolysis solution;

(3) sterilizing and centrifuging the enzymolysis liquid to obtain absorption liquid, purifying the absorption liquid by resin to obtain tuna peptide eluent, and concentrating and drying the tuna peptide.

In the step (1), it is further preferable that the tuna is tuna meat or leftovers of the tuna in the processing process, and specifically includes high-protein leftovers such as fish head, viscera, fish skin and the like.

Further preferably, the homogenization time is 0.8 to 1.2 hours.

In the step (2), it is further preferable that the culture solution is a culture solution required for culturing microorganisms; specifically, the culture medium is glucose culture medium.

More preferably, the compound protease is an additive commonly used in the field of food processing, is a protease for hydrolyzing animal and vegetable proteins, and further is a neutral and acidic protease. The compound protease adopted by the invention is a compound product of common protease, for example, papain, neutral protease, trypsin or protease obtained by fermenting other strains.

Further preferably, the concentration of the compound protease is 2-5% (mass fraction).

Further preferably, the microorganism is a protease-producing bacterium; in a specific embodiment, the microorganism is candida tropicalis.

Further preferably, the fermentation and enzymolysis time is 44-50 h.

Further preferably, the fermentation and enzymolysis temperature is 25-35 ℃.

Further preferably, the sterilization time is 10-60 min.

Further preferably, the centrifugation parameter is 5000-.

In the step (3), it is further preferable that the resin is a macroporous adsorption resin.

More preferably, the loading amount of the resin is equal to the weight of the resin.

More preferably, the amount of the elution liquid is 6 to 7 times the weight of the resin.

Further preferably, the elution flow rate is controlled to be 1-1.2 bed volumes per hour.

Further preferably, the eluent is 30% -75% ethanol solution.

The research of the invention finds that when the concentration of the eluent is in the concentration range, the higher elution recovery rate can be kept, and when the concentration reaches 75%, the highest elution recovery rate is achieved; when the ethanol concentration further increased, the elution efficiency started to decrease instead.

Further preferably, 75% ethanol solution is used for desorbing tuna peptide in the resin.

Further preferably, the concentration adopts vacuum decompression concentration, the concentration temperature is less than or equal to 80 ℃, and the concentration vacuum degree is less than or equal to-0.08 Mpa.

Preferably, the concentrated extract is concentrated to a specific gravity of 1.1-1.15, and then spray drying is carried out to obtain the tuna peptide.

In a second aspect of the present invention, the method for extracting tuna peptide or obtaining tuna peptide according to the first aspect is provided.

In a third aspect of the invention, there is provided the use of a tuna peptide according to the second aspect as a hypotensive agent.

The application of the tuna peptide as the antihypertensive agent comprises the step of applying the tuna peptide as the antihypertensive active ingredient to the preparation of the antihypertensive product, preferably, the application mode comprises the step of applying the tuna peptide as the active ingredient and auxiliary ingredients together to the preparation of the antihypertensive product, and the step of applying the tuna peptide and other ingredients with the antihypertensive effect or the auxiliary antihypertensive effect together to the preparation of the antihypertensive product.

Further preferably, the blood pressure lowering product comprises a blood pressure lowering medicine and a blood pressure lowering health care product.

In order to make the technical solutions of the present invention more clearly understood by those skilled in the art, the technical solutions of the present invention will be described in detail below with reference to specific embodiments.