阅读说明：本技术 一种海参肽的制备方法 (Preparation method of sea cucumber peptide ) 是由 闫云霞 于 2020-06-24 设计创作，主要内容包括：本发明涉及一种海参肽的制备方法,其包括：S1、选取新鲜海参,预处理备用；S2、向预处理后的海参中加入2～6倍纯净水,研磨磨碎,制匀浆,依次加入脂肪酶、中性蛋白酶,氨肽酶进行酶解；S3、固液分离,将所得液体进行脱脂纯化,吸附脱色得澄清液；S4、澄清液过离子交换树脂脱钠、再浓缩干燥。本发明方法生产周期短、成本低、对环境无污染,而且所得产品安全性高,纯度高、颜色浅、口感好。所得产品可广泛应用于食品、特殊医学用配方食品、保健品等领域。(The invention relates to a preparation method of sea cucumber peptide, which comprises the following steps: s1, selecting fresh sea cucumbers, and pretreating for later use; s2, adding 2-6 times of purified water into the pretreated sea cucumber, grinding, preparing homogenate, and sequentially adding lipase, neutral protease and aminopeptidase for enzymolysis; s3, carrying out solid-liquid separation, carrying out degreasing purification on the obtained liquid, and carrying out adsorption decoloration to obtain a clear liquid; and S4, passing the clear liquid through ion exchange resin to remove sodium, and then concentrating and drying. The method has the advantages of short production period, low cost, no environmental pollution, high safety of the obtained product, high purity, light color and good taste. The obtained product can be widely applied to the fields of food, formula food for special medicine, health care products and the like.)

1. The preparation method of the sea cucumber peptide is characterized by comprising the following steps:

s1, selecting fresh sea cucumbers, and pretreating for later use;

s2, adding 2-6 times of purified water into the pretreated sea cucumber, grinding, preparing homogenate, and sequentially adding lipase, neutral protease and aminopeptidase for enzymolysis;

s3, carrying out solid-liquid separation, carrying out degreasing purification on the obtained liquid, and carrying out adsorption decoloration to obtain a clear liquid;

and S4, passing the clear liquid through ion exchange resin to remove sodium, and then concentrating and drying.

2. The method of claim 1, wherein in step S1, the pretreatment method is: selecting fresh sea cucumber, dissecting to remove viscera, shearing into small pieces less than 0.5cm, and cleaning with purified water for use.

3. The method of claim 1, wherein the homogenate is prepared in step S2 by: adding 1-3 times of purified water into the pretreated sea cucumber, and grinding the sea cucumber into slurry by using a grinder; and adding 1-3 times of purified water into the slurry, and uniformly stirring to form homogenate.

4. The method according to claim 1, wherein the homogenate is heated to 40 ℃ to 50 ℃ in step S2, lipase in an amount of 0.1% to 0.3% by mass and neutral protease in an amount of 1% to 3% by mass of fresh sea cucumber are added and the mixture is kept for 40min to 60min, and aminopeptidase in an amount of 0.3% to 0.5% by mass and the mixture is kept at 90min to 150 min.

5. The method according to claim 1, wherein in step S2, after the enzymatic hydrolysis, the temperature of the enzymatic hydrolysate is raised to 105 ℃, and the enzymatic hydrolysis is maintained for 5-10 min to inactivate the enzyme.

6. The production method according to claim 1, wherein in step S3, the solid-liquid separation equipment used is: filtering by a horizontal gong centrifuge at a centrifuge speed of 3400r/min to obtain liquid, and degreasing and purifying by a three-phase centrifuge.

7. The method of claim 1, wherein in step S3, the adsorption decoloring method is: adsorbing and decoloring by using active carbon, wherein the using amount of the active carbon is 0.5-5 percent of that of the fresh sea cucumber, the time is 30-60 min, and filtering by using a pledge type plate and frame filter.

8. The method according to claim 1, wherein in step S4, the equipment used for the concentration and drying is: a double-effect falling film thickener and a pressure type spray dryer.

Technical Field

The invention belongs to the technical field of deep processing of marine products, and particularly relates to a preparation method of sea cucumber peptide.

Background

The sea cucumber recorded in the compendium of materia medica supplement is characterized by the effects of tonifying kidney channel, benefiting marrow and essence, eliminating debility and saliva, controlling urination, strengthening yang and generating white pulse: has high nutritive value and health promotion effect. The traditional sea cucumber products comprise salted sea cucumbers, half-dried sea cucumbers, freeze-dried sea cucumbers and the like, and have the problems of troublesome use and low protein absorption rate. The traditional sea cucumber products are usually eaten in a flaky and random manner, and have the problems of more eating, no absorption and less eating effect.

With the improvement of bioengineering technology in recent years, some enterprises gradually enter the sea cucumber deep processing industry, and also relate to the production and processing of sea cucumber peptides, but the products are dark in color, heavy in fishy smell and poor in taste, and are relatively limited in the use process, and the terminal products are generally capsules and tablets, so that the safety ratio is relatively poor and the use is relatively troublesome.

Disclosure of Invention

Technical problem to be solved

In order to solve the above problems in the prior art, the present invention provides a method for preparing sea cucumber peptide.

(II) technical scheme

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

a preparation method of sea cucumber peptide comprises the following steps:

s1, selecting fresh sea cucumbers, and pretreating for later use;

s2, adding 2-6 times of purified water into the pretreated sea cucumber, grinding, preparing homogenate, and sequentially adding lipase, neutral protease and aminopeptidase for enzymolysis;

s3, carrying out solid-liquid separation, carrying out degreasing purification on the obtained liquid, and carrying out adsorption decoloration to obtain a clear liquid;

and S4, passing the clear liquid through ion exchange resin to remove sodium, and then concentrating and drying.

The preparation method as described above, preferably, in step S1, the pretreatment method is: selecting fresh sea cucumber, dissecting to remove viscera, shearing into small pieces less than 0.5cm, and cleaning with purified water for use.

The preparation method as described above, preferably, in step S2, the process of preparing the homogenate is: adding 1-3 times of purified water into the pretreated sea cucumber, and grinding the sea cucumber into slurry by using a grinder; and adding 1-3 times of purified water into the slurry, and uniformly stirring to form homogenate.

In the preparation method, preferably, in step S2, the homogenate is heated to 40 ℃ to 50 ℃, lipase accounting for 0.1% -0.3% of the mass of the fresh sea cucumber and neutral protease accounting for 1% -3% of the mass of the fresh sea cucumber are added, the mixture is kept for 40min to 60min, and then aminopeptidase accounting for 0.3% -0.5% of the mass of the fresh sea cucumber is added, and the temperature is kept for 90min to 150 min.

In the preparation method as described above, preferably, in step S2, after the enzymolysis, the temperature of the enzymolysis liquid is raised to 105 ℃, and the enzymolysis liquid is kept for 5min to 10min to inactivate the enzyme.

In the above-described production method, preferably, in step S3, the solid-liquid separation equipment is: filtering by a horizontal gong centrifuge at a centrifuge speed of 3400r/min to obtain liquid, and degreasing and purifying by a three-phase centrifuge.

The preparation method as described above, preferably, in step S3, the adsorption decoloring method is: adsorbing and decoloring by using active carbon, wherein the using amount of the active carbon is 0.5-5 percent of that of the fresh sea cucumber, the time is 30-60 min, and filtering by using a pledge type plate and frame filter.

In the above-described production method, preferably, in step S4, the equipment used for the concentration and drying is: a double-effect falling film thickener and a pressure type spray dryer.

In the preparation method of the sea cucumber peptide, grinding and lipase hydrolysis are adopted, so that the structure of sea cucumber cells can be easily destroyed, the release of active ingredients in the cells can be further promoted, the extraction of the sea cucumber peptide is facilitated, and the product yield is improved. Meanwhile, the link of high-temperature cooking in production is avoided, and the problem of deep color caused by partial browning reaction is reduced. A large number of experimental researches show that when the added amount of the lipase is less than 0.1 percent of the fresh sea cucumber, the lipase cannot effectively remove fat, the product has slight fishy smell, the yield is reduced, when the used amount is more than 0.3 percent, the effect is not obviously changed, and the use amount is preferably 0.1 to 0.3 percent to save the cost. The using amount of the neutral protease is less than 1 percent of the fresh sea cucumber, the product yield is greatly reduced, the obtained sea cucumber peptide is more than 1000Kd, the product purity is reduced, and when the using amount is more than 3 percent, the sea cucumber peptide is excessively hydrolyzed, so the preferable using amount is 1 to 3 percent of the fresh sea cucumber.

After the macromolecular protein is subjected to enzymolysis, hydrophobic amino acid at the tail end of a peptide chain is exposed, and bitter taste is generated when the macromolecular protein contacts taste buds. In the invention, after the endo neutral protease is used for hydrolysis, the bitter groups at two ends of the peptide bond are cut off by exo-aminopeptidase, so that the product has better taste and fully meets the requirements of food and health-care products on the taste of the raw materials. Researches show that the usage amount of the exo-aminopeptidase is 0.3-0.5% of the fresh sea cucumber, the bitter taste can be effectively removed, the usage amount is less than 0.3%, the bitter taste is remained, and when the usage amount is more than 0.5%, excessive hydrolysis can occur, so that the free amino acid in the product is increased, the taste of the product is influenced, and the biological activity of the product is weakened. Therefore, the amount of exoaminopeptidase used is preferably 0.3% to 0.5%.

The main salty taste of the product is glutamic acid sodium salt, and after the product is decolorized, sodium ions in the product are removed by using ion exchange resin, so that sodium in the product can be effectively removed, and the taste of the product is improved.

(III) advantageous effects

The invention has the beneficial effects that:

in the preparation method of the sea cucumber peptide, provided by the invention, lipase, endo-neutral protease and exo-aminopeptidase are adopted for enzymolysis, solid-liquid separation, degreasing and purification, decoloration treatment, concentration and drying are carried out, and the high-purity sea cucumber peptide is obtained. The obtained sea cucumber peptide has the advantages of molecular weight less than 1000Kd, high activity, strong oxidation resistance, no fishy smell, extremely low sodium content and high zinc content. The product obtained by the method can be widely applied to the fields of food, formula food for special medicine, health-care products and the like.

Detailed Description

For better explanation of the present invention and for ease of understanding, the present invention will be described in detail below by way of specific embodiments, and commercially available products may be used as the enzyme preparation used.

Example 1

A preparation method of sea cucumber peptide specifically comprises the following steps:

step 1, selecting 10 kilograms of fresh sea cucumbers, dissecting and removing internal organs, shearing into small blocks less than 0.5cm, and cleaning with purified water for later use;

step 2, adding 20 kg of purified water into the sea cucumber treated in the step 1, and grinding the small sea cucumber blocks into slurry by using a grinder;

step 3, adding 20 kilograms of purified water into the slurry, stirring uniformly to form homogenate, heating to 45 ℃, adding 20 grams of lipase (100000U/g) and 100 grams of endo-neutral protease (200000U/g), keeping for 60min, then adding 30 grams of exoaminopeptidase (30000U/g), keeping for 120min, finally heating to 105 ℃, keeping for 5min, and inactivating enzyme;

step 4, filtering the enzyme-inactivated solution by using a horizontal gong centrifuge (the centrifugal speed is 3400r/min), and degreasing and purifying the filtrate by using a three-phase centrifuge to obtain a purified solution;

step 5, adding 1 kg of active carbon into the purified solution for adsorption and decoloration for 40min, and filtering by using a mass-press plate-and-frame filter to obtain clear filtrate;

step 6, removing sodium from the clear filtrate through ion exchange resin, and then further concentrating by using a double-effect concentrator;

and 7, drying the concentrated solution at 80 ℃ by using spray drying to obtain the sea cucumber peptide.

Example 2

A preparation method of sea cucumber peptide specifically comprises the following steps:

step 1, selecting 10 kilograms of fresh sea cucumbers, dissecting and removing internal organs, shearing into small blocks less than 0.5cm, and cleaning with purified water for later use;

step 2, adding 15 kg of purified water into the sea cucumbers, and grinding the small sea cucumber blocks into slurry by a grinder;

step 3, adding 25 kilograms of purified water into the slurry, stirring uniformly to form homogenate, heating to 40 ℃, adding 15 grams of lipase (100000U/g) and 200 grams of endo-neutral protease (200000U/g), keeping for 50 minutes, then adding 40 grams of exoaminopeptidase (30000U/g), keeping for 100 minutes, finally heating to 105 ℃, keeping for 7 minutes, and inactivating the enzyme;

step 4, filtering the enzyme-inactivated solution by using a horizontal gong centrifuge (the centrifugal speed is 3400r/min), and degreasing and purifying the filtrate by using a three-phase centrifuge to obtain a purified solution;

and 5, adding 0.8 kg of active carbon into the purified solution, adsorbing and decoloring for 50min, and filtering by using a press type plate-and-frame filter to obtain clear filtrate.

Step 6, removing sodium from the clear filtrate through ion exchange resin, and then further concentrating the filtrate by using a double-effect concentrator;

and 7, drying the concentrated solution by spray drying to obtain the sea cucumber peptide.

Example 3

A preparation method of sea cucumber peptide specifically comprises the following steps:

step 1, selecting 10 kilograms of fresh sea cucumbers, dissecting and removing internal organs, shearing into small blocks less than 0.5cm, and cleaning with purified water for later use;

step 2, adding 25 kg of purified water into the sea cucumbers, and grinding the small sea cucumber blocks into slurry by using a grinder;

step 3, adding 10 kilograms of purified water into the slurry, stirring uniformly to form homogenate, heating to 48 ℃, adding 23 grams of lipase (100000U/g) and 250 grams of endo-neutral protease (200000U/g), keeping for 45 minutes, then adding 45 grams of exoaminopeptidase (30000U/g), keeping for 90 minutes, finally heating to 105 ℃, keeping for 10 minutes to inactivate enzyme;

step 4, filtering the enzyme-inactivated solution by using a horizontal gong centrifuge (the centrifugal speed is 3400r/min), and degreasing and purifying the filtrate by using a three-phase centrifuge to obtain a purified solution;

and 5, adding 1.5 kg of active carbon into the purified solution, adsorbing and decoloring for 30min, and filtering by using a press type plate-and-frame filter to obtain clear filtrate.

Step 6, removing sodium from the clear filtrate through ion exchange resin, and then further concentrating the filtrate by using a double-effect concentrator;

and 7, drying the concentrated solution by spray drying to obtain the sea cucumber peptide.

Comparative example 1

The procedure is as in example 1, except that no lipase is added.

Comparative example 2

The procedure is as in example 1, except that no aminopeptidase is added.

Comparative example 3

The procedure of example 2 was followed except that the decolorization by adsorption on activated carbon was not performed and sodium removal by ion exchange resin was not performed.

Detecting the proportion of protein hydrolysate with relative molecular mass less than 1000u by high performance gel filtration chromatography, measuring to obtain sea cucumber peptide with molecular mass less than 1000Da, and determining product yield, sodium content, and product color and taste, wherein the result is shown in Table 1.

TABLE 1 test results

The product yield is the mass percentage of the obtained dried sea cucumber peptide and the fresh sea cucumber, the protein content of the fresh sea cucumber is about 6%, and the obtained product yield shows that the sea cucumber peptide obtained by the method has higher yield and effectively improves the economic benefit. The detection results show that the enzyme preparation used in the method of the invention can prepare enzymolysis products with almost all molecular weights less than 1000Kd, and the obtained products have high yield. The prepared product is very easy to dissolve in water, can be directly eaten after being brewed, and has no sediment.

The prepared product can be directly eaten, and can also be matched with other foods to be eaten to prepare products with special purposes such as health care products and the like.