CN111528332A

CN111528332A - Bird's nest peptide and preparation method and application thereof

Info

Publication number: CN111528332A
Application number: CN202010387357.5A
Authority: CN
Inventors: 范群艳; 连建梅; 李鸿铭; 陈小玲; 谢加凤; 叶淑贤
Original assignee: Xiamen Yanzhiwu Silken Food Co ltd
Current assignee: Xiamen Yanzhiwu Silken Food Co ltd
Priority date: 2020-05-09
Filing date: 2020-05-09
Publication date: 2020-08-14

Abstract

The invention discloses an edible bird's nest peptide and a preparation method and application thereof, and belongs to the technical field of food processing. The invention discloses a preparation method of the protected cubilose peptide, which has simple and convenient process flow, maintains the original flavor of cubilose, has no obvious enzyme or peptide flavor, has soft mouthfeel and no astringent feeling, and can be used as a raw material for food or skin care products because the cubilose peptide prepared by controlling the enzymolysis process conditions has low molecular weight, is easy to absorb and has small viscosity, so the preparation method of the cubilose peptide is suitable for market popularization and application and industrial production.

Description

Bird's nest peptide and preparation method and application thereof

Technical Field

The invention belongs to the technical field of food processing, and particularly relates to a bird's nest peptide and a preparation method and application thereof.

Background

The bird's nest is used as an old nourishing treasure and a beautifying holy product, and has the functions of nourishing yin, moistening lung, nourishing and beautifying, promoting metabolism of a human body and the like. In recent years, with the increasing activity of the health food market, the bird's nest products are more and more favored by consumers. The main components of the bird's nest comprise water-soluble protein, carbohydrate, trace elements such as calcium, phosphorus, iron, sodium, potassium and the like and amino acid, wherein the protein content in the bird's nest is high, each 100 g of dry bird's nest contains about 49.9g of protein, the bird's nest mainly exists in the form of glycoprotein, protease secreted by a human body is gradually reduced along with the increase of the age, the problems of difficult digestion, low protein bioavailability and the like exist after the human body takes the protein, and the low nutrient absorption rate in the bird's nest is easily caused in the process of directly eating the bird's nest, so that the eating effect is influenced. Therefore, in order to improve the absorption rate of protein polypeptide in the cubilose, the small molecular peptide extraction technology of the cubilose has been researched at the present stage, but the flavor is not ideal, and the process is relatively complex.

The existing preparation method of the cubilose peptide mainly comprises an enzyme method and an acid method, wherein although the enzyme extraction has specificity, the cubilose peptide prepared finally is easy to have the flavor residue and the astringent feeling of the enzyme, thereby influencing the application range of the product; in addition, in the acid extraction process, the efficiency is low due to the violent reaction with protein and the complex operation process, and the phenomenon of incomplete deacidification is also caused.

Therefore, the technical problem to be solved by the technical personnel in the field is how to provide the method for preparing the cubilose peptide with simple preparation process and good flavor.

Disclosure of Invention

Aiming at the problems, the invention provides the preparation method of the cubilose peptide, the preparation process flow is simple, the operation is easy, the flavor is good, and the taste and the flavor of the cubilose after stewing are reserved; the finally prepared cubilose peptide can be directly applied to the traditional instant cubilose product, and the sialic acid content is increased, so that the nutritional value of the product is improved.

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

a preparation method of cubilose peptide specifically comprises the following steps:

(1) soaking and picking bird's nest: soaking dried nidus Collocaliae in purified water, removing impurities, mincing, and draining;

(2) stewing and grinding: adding purified water into the cubilose soaked in the step (1) for stewing, and then finely grinding to obtain cubilose pulp;

the edible bird's nest is soaked in water, and then is stewed until the edible bird's nest is transparent, soft and glutinous and has no hard core, and the powder and short strips produced in the processes of soaking and sorting the edible bird's nest are used for extracting the edible bird's nest peptide by enzymolysis, and then are added into the edible bird's nest product, so that the utilization rate of the edible bird's nest can be effectively improved.

(3) First biological enzymolysis: adjusting the concentration and the pH of the cubilose pulp prepared in the step (2), then adding neutral protease into the prepared cubilose pulp, stirring for enzymolysis, inactivating the enzyme, cooling, and sieving by a 100-200-mesh sieve to obtain a first enzymolysis liquid;

and the enzyme deactivation and cooling process after enzymolysis comprises the following steps: and continuously heating the enzymolysis liquid to above 85 ℃ to start enzyme deactivation for 15-20 min, cooling to below 50 ℃ after enzyme deactivation is finished, and sieving to obtain the first enzymolysis liquid.

(4) Homogenizing and filtering: homogenizing and roughly filtering the first enzymolysis liquid prepared in the step (3) to obtain enzymolysis homogenized liquid;

(5) and (3) second biological enzymolysis: adjusting the concentration and pH of the enzymatic hydrolysis homogeneous liquid prepared in the step (4), then adding compound protease into the prepared enzymatic hydrolysis homogeneous liquid, stirring for enzymatic hydrolysis, inactivating enzyme, and cooling to obtain a second enzymatic hydrolysis liquid;

(6) refining and spray drying: and (4) carrying out fine filtration and RO membrane reverse osmosis concentration on the secondary enzymolysis liquid prepared in the step (5), and then carrying out spray drying to obtain the cubilose peptide.

Specifically, the secondary enzymolysis liquid is subjected to fine filtration treatment by a 0.1-5 mu m microporous membrane to obtain clear and transparent fine filtration liquid of the edible bird's nest; and then carrying out reverse osmosis concentration on the fine bird's nest filtrate by an RO (reverse osmosis) membrane for 2-7 times, and carrying out spray drying, wherein the spray inlet temperature is 130-180 ℃, the outlet temperature is 70-100 ℃, and the pressure is 30-40 Mpa, and the sprayed powder is the bird's nest peptide.

In addition, in the biological enzymolysis process, the stirring speed in the enzymolysis tank needs to be kept at 30-70 r/min, so that the bird's nest pulp is in a stirring state in the biological enzymolysis process, the enzymolysis is uniform, and the enzymolysis efficiency is improved.

The preparation method of the cubilose peptide disclosed and protected by the invention is simple and convenient in process flow, and the cubilose peptide prepared by the process flow and the control conditions keeps the original flavor of cubilose, has no obvious enzyme taste or peptide taste, and is soft in taste and free of astringent feeling.

Furthermore, because the bird's nest peptide prepared by the invention has better taste and flavor, the bird's nest peptide can be directly used after being concentrated or can be freeze-dried into powder in vacuum to be used as a health ingredient added into other foods besides being prepared into peptide powder by spray drying.

Preferably, in the step (1), the mass of the added dry cubilose is 1-8% of that of the purified water, the soaking time is 30-60 min, and the soaking temperature is 50-60 ℃.

Preferably, in the step (2), the amount of the purified water added is 20-60 times of the mass of the cubilose, and the stewing time is 0.5-1 h; and grinding the stewed cubilose for 2-3 times by using a colloid mill, and controlling the fineness of cubilose pulp particles within 10 mu m.

The invention is characterized in that the colloid fine grinding is carried out before the biological enzymolysis of the cubilose, and the fineness of the size of the cubilose pulp after the colloid fine grinding is controlled to increase the contact surface area of the cubilose and the biological enzyme, improve the enzymolysis efficiency, shorten the enzymolysis time and reduce the energy consumption.

Preferably, in the step (3), the mass concentration of the cubilose pulp before the first biological enzymolysis is 1-8%, and the pH value is 6.0-8.0.

Further, the adding mass of the neutral protease is 0.02-0.07% of the mass of the dry cubilose, the enzymolysis temperature is 50-60 ℃, and the enzymolysis time is 30-60 min.

Preferably, in the step (4), the homogenization pressure is 20-35 MPa, the homogenization temperature is 40-80 ℃, and the enzymatic hydrolysis homogenization liquid is obtained after rough filtration by a 300-mesh filter screen.

Preferably, in the step (5), the mass concentration of the enzymatic homogeneous liquid before the second biological enzymolysis is 1-8%, and the pH value is 6.0-8.0.

Further, the compound protease comprises the following proteases in percentage by mass:

0.04-0.3% of neutral protease, 0.05-0.4% of alkaline protease, 0.1-0.5% of flavourzyme and 0.1-0.5% of trypsin;

and the protease is based on the mass ratio of the dry cubilose, the enzymolysis temperature is 50-60 ℃, and the enzymolysis time is 4-10 h.

The invention also claims the bird's nest peptide prepared by the method and the application of the bird's nest peptide in bird's nest food and skin care products.

The cubilose peptide has low molecular weight and low viscosity, and is suitable for being applied to instant cubilose food and skin care products.

According to the technical scheme, compared with the prior art, the invention discloses and provides the cubilose peptide and the preparation method and the application thereof, and the cubilose peptide has the following excellent effects:

(1) the invention adopts the processes of primary biological enzymolysis, homogenization, filtration, secondary biological enzymolysis, fine filtration, concentration and spray drying, utilizes the biotechnology to effectively improve the utilization rate of the bird's nest, not only extracts the nutrition and the essence of the bird's nest, but also changes macromolecular bird's nest into small molecular peptide, improves the absorption and the utilization rate of the bird's nest in a human body, and simultaneously avoids the problems of bad smell, poor solubility, bad color and the like existing in the prior enzymolysis technology by the process treatment and the compound enzyme optimization.

(2) The preparation method of the cubilose peptide disclosed by the invention is simple and convenient in process flow, the cubilose peptide prepared by the process flow and the control conditions keeps the original flavor of cubilose, has no obvious enzyme taste or peptide taste, is soft in taste and has no astringent feeling, and the cubilose peptide is low in molecular weight and small in viscosity and can be suitable for cubilose foods and skin care products, so that the preparation method of the cubilose peptide disclosed by the invention is suitable for popularization and application in the market.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.

FIG. 1 is a molecular weight chromatogram of the bird's nest peptide prepared in example 1 of the present invention.

FIG. 2 is a molecular weight chromatogram of the bird's nest peptide prepared in comparative example 1 of the present invention.

FIG. 3 is a graph comparing the protein content of different enzymatic hydrolyses according to the invention.

FIG. 4 is a graph showing the comparison of the degree of hydrolysis of the bird's nest hydrolyzed by different enzymes according to the present invention.

FIG. 5 is a graph showing comparison of sialic acid hydrolyzed by different enzymes of the present invention.

FIG. 6 is a graph showing the effect of different amounts of flavourzyme added on hydrolysis molecular weight according to the present invention.

FIG. 7 is a graph showing the effect of different amounts of alkaline protease of the present invention on hydrolysis molecular weight.

FIG. 8 is a graph showing the effect of different amounts of neutral protease of the present invention on hydrolysis molecular weight.

FIG. 9 is a graph showing the effect of different amounts of trypsin according to the present invention on hydrolysis molecular weight.

Detailed Description

The present invention will be described more clearly and completely with reference to the accompanying drawings and technical solutions provided in the embodiments of the present invention, and it is to be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the 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.

It should be understood that the processing equipment or devices not specifically mentioned in the following examples are conventional in the art; all pressure values and ranges refer to absolute pressures.

Furthermore, it is to be understood that one or more method steps mentioned in the present invention does not exclude that other method steps may also be present before or after the combined steps or that other method steps may also be inserted between these explicitly mentioned steps, unless otherwise indicated. Moreover, unless otherwise indicated, the numbering of the various method steps is merely a convenient tool for identifying the various method steps, and is not intended to limit the order in which the method steps are arranged or the scope of the invention in which the invention may be practiced, and changes or modifications in the relative relationship may be made without substantially changing the technical content.

The embodiment of the invention discloses a method for preparing cubilose peptide with simple preparation process and good flavor, which comprises the following steps:

1) cleaning: soaking dry edible bird's nest with medical-grade purified water at 50-60 ℃ for 30-60 min, rubbing, cleaning, draining, adding 20-60 times of medical-grade purified water, and stewing for 30-60 min until the edible bird's nest is transparent, soft and glutinous and has no hard core;

2) grinding: grinding the stewed cubilose for 2-3 times by using a colloid mill, and controlling the fineness of cubilose pulp particles within 10 mu m to obtain cubilose pulp;

3) first biological enzymolysis: pouring the cubilose pulp into an enzymolysis tank, adding purified water, wherein the material-liquid ratio is (1-8): 100, adjusting the pH value to 6.0-8.0, starting stirring, rotating at 30-70 rpm (keeping the cubilose pulp in a stirring state), heating to 50-60 ℃, adding neutral protease (0.02-0.07 percent of the dry cubilose amount) for enzymolysis for 30-60 min, and controlling the temperature at 50-60 ℃ in the constant temperature process;

4) enzyme deactivation and filtration: after the first biological enzymolysis is finished, continuously heating the enzymolysis liquid to more than 85 ℃ and starting enzyme deactivation for 15-20 min, cooling to below 50 ℃ after enzyme deactivation, and sieving with a 100-200-mesh sieve to obtain first enzymolysis liquid;

5) homogenizing: passing the first enzymolysis liquid through a homogenizer for 1-2 times, homogenizing at 20-35 Mpa and 40-80 ℃, and coarse-filtering with a 300-mesh filter screen to obtain a homogenized liquid;

6) and (3) second biological enzymolysis: introducing the homogenized solution into an enzymolysis tank, performing constant volume to 1-8% of dry cubilose concentration by using medical-grade purified water, adjusting the pH value to 6.0-8.0, starting stirring at a rotating speed of 30-70 rpm (keeping the cubilose pulp in a stirring state), heating to raise the temperature of materials in the tank to 50-60 ℃, adding neutral protease (0.04-0.3% of the dry cubilose), alkaline protease (0.05-0.4% of the dry cubilose weight), flavourzyme (0.1-0.5% of the dry cubilose weight) and trypsin (0.1-0.5% of the dry cubilose weight) to perform secondary enzymolysis, wherein the enzymolysis time is 4-10 h, the temperature is controlled to 50-60 ℃ in a constant temperature process, and then performing enzyme deactivation and cooling to obtain secondary enzymolysis solution;

7) fine filtering: secondary fine filtering the secondary enzymolysis liquid by a 0.1-5 mu m microporous filter membrane to obtain clear and transparent fine filtering liquid of the bird's nest;

8) concentration and spray drying: and (3) concentrating the fine filtrate by reverse osmosis of an RO membrane by 2-7 times, and performing spray drying at the spray inlet temperature of 130-180 ℃, the spray outlet temperature of 70-100 ℃ and the pressure of 30-40 Mpa to spray out powder to obtain the bird's nest peptide.

The excellent results of the method for preparing the bird's nest peptide disclosed and protected by the invention are further illustrated below with reference to specific examples, but the content of the invention is not limited to the following examples.

Example 1

A preparation method of cubilose peptide comprises the following steps:

1) soaking dried nidus Collocaliae with medical grade purified water at 50 deg.C for 40min, mincing, cleaning, draining, adding 30 times of medical grade purified water, and stewing for 30min until the nidus Collocaliae is transparent, soft and glutinous and has no hard core;

2) grinding the stewed bird's nest with colloid for 2 times, and controlling the fineness of bird's nest pulp particles within 10 μm to obtain bird's nest pulp;

3) pouring the cubilose pulp into an enzymolysis tank, adding medical-grade purified water until the concentration of dry cubilose is 5%, adjusting the pH value to 7.5, starting stirring at the rotating speed of 40rpm, heating to 50 ℃ and then adding 0.03% of neutral protease for carrying out primary enzymolysis for 40min, and controlling the temperature in the constant temperature process to 50 ℃;

4) after the first enzymolysis is finished, continuously heating the enzymolysis liquid to 85 ℃ to start enzyme deactivation for 15min, cooling to 45 ℃ after the enzyme deactivation is finished, and filtering by 200 meshes to obtain the first enzymolysis liquid;

5) passing the first enzymolysis solution through a homogenizer for 2 times, homogenizing at 45 deg.C under 25Mpa, and coarse-filtering with 300 mesh filter screen to obtain homogenized solution;

6) introducing the homogenized solution into an enzymolysis tank, performing constant volume with medical grade purified water until the concentration of dry nidus Collocaliae is 2.5%, adjusting pH value to 7.5, starting stirring at 40rpm, heating to 50 deg.C, adding neutral protease 0.05%, alkaline protease 0.3%, flavourzyme 0.1%, and trypsin 0.1%, performing second enzymolysis for 6h, controlling temperature at constant temperature to 50 deg.C, inactivating enzyme, and cooling to obtain second enzymolysis solution;

7) performing secondary fine filtration treatment on the secondary enzymolysis liquid by using a 1-micron microporous filter membrane to obtain clear and transparent bird's nest fine filtration liquid;

8) concentrating the fine filtrate by reverse osmosis of RO membrane to 5 times, spray drying at inlet temperature of 135 deg.C, outlet temperature of 80 deg.C and pressure of 30Mpa, and spraying powder to obtain nidus Collocaliae peptide.

Example 2:

a preparation method of cubilose peptide comprises the following steps:

1) soaking dried nidus Collocaliae with medical grade purified water at 60 deg.C for 60min, mincing, cleaning, draining, adding 40 times of medical grade purified water, and stewing for 50min until the nidus Collocaliae is transparent, soft and glutinous and has no hard core;

2) grinding the stewed bird's nest with colloid for 3 times, and controlling the fineness of bird's nest pulp particles within 10 μm to obtain bird's nest pulp;

3) pouring the cubilose pulp into an enzymolysis tank, adding medical-grade purified water until the concentration of dry cubilose is 8%, adjusting the pH value to 7.0, starting stirring at a rotation speed of 50rpm, heating to 60 ℃ in the tank, adding 0.07% of neutral protease, performing primary enzymolysis for 50min, and controlling the temperature in the constant temperature process to 60 ℃;

4) after the first enzymolysis is finished, continuously heating the enzymolysis liquid to more than 85 ℃ to start enzyme deactivation for 20min, cooling to 40 ℃ after enzyme deactivation, and filtering by 150 meshes to obtain first enzymolysis liquid;

5) passing the first enzymolysis solution through a homogenizer for 2 times, homogenizing at 33Mpa and 50 deg.C, and coarse-filtering with 300 mesh filter screen to obtain homogenized solution;

6) introducing the homogenized solution into an enzymolysis tank, performing constant volume with medical grade purified water until the concentration of dry nidus Collocaliae is 5.0%, adjusting pH value to 7.0, starting stirring at a rotation speed of 50rpm, heating to 60 deg.C, adding neutral protease 0.3%, alkaline protease 0.08%, flavourzyme 0.5%, and trypsin 0.4%, performing second enzymolysis for 10h, controlling temperature at constant temperature of 60 deg.C, inactivating enzyme, and cooling to obtain second enzymolysis solution;

7) secondary fine filtration treatment is carried out on the secondary enzymolysis liquid by a 0.5 mu m microporous membrane to obtain clear and transparent bird's nest fine filtration liquid;

8) concentrating the fine filtrate by reverse osmosis of RO membrane to 7 times, spray drying at spray inlet temperature of 175 deg.C, outlet temperature of 100 deg.C and pressure of 40Mpa, and spraying powder to obtain nidus Collocaliae peptide.

Comparative example 1:

a preparation method of cubilose peptide comprises the following steps:

1) soaking dried nidus Collocaliae in water at room temperature for 30min, and stewing in 20 times of water for 30min until the nidus Collocaliae is transparent, soft and glutinous and has no hard core;

3) pouring cubilose pulp into an enzymolysis tank, adding water to a certain amount until the concentration of the dry cubilose is 2.5%, adjusting the pH value to 7.0, starting stirring, rotating at 50rpm, heating to 55 ℃ and then adding 0.4% of neutral protease, 0.5% of animal protease, 0.05% of flavor protease and 0.6% of trypsin for enzymolysis, wherein the enzymolysis time is 6h, and the temperature in the constant temperature process is controlled to be 55 ℃;

4) after enzymolysis, continuously heating the enzymolysis liquid to more than 85 ℃ to start enzyme deactivation for 20min, cooling to 40 ℃ after enzyme deactivation, and filtering by 150 meshes to obtain the enzymolysis liquid;

5) secondary fine filtration treatment is carried out on the enzymatic hydrolysate by a microporous membrane with the diameter of 5 mu m, so as to obtain clear and transparent fine filtration liquid of the edible bird's nest;

6) concentrating the fine filtrate by reverse osmosis of RO membrane by 10 times, spray drying at spray inlet temperature of 120 deg.C, outlet temperature of 101 deg.C and pressure of 40Mpa, and spraying powder to obtain nidus Collocaliae peptide.

Comparative analysis and results

The molecular weights of the bird's nest peptides prepared in the examples 1-2 and the comparative example 1 are respectively compared, and the comparison results are shown in the attached figures 1-2, the following tables 1 and 2.

Table 1 example 1 molecular weight data

Table 2 comparative example 1 molecular weight data

As can be seen from FIGS. 1 to 2 and tables 1 and 2, in example 1, the proportion of molecular weight < 8000Da is > 60%, and the proportion of molecular weight < 1000Da is > 40%; in comparative example 1, the proportion of molecular weight > 10000 was > 70%. Therefore, the enzymolysis degree of the bird's nest peptide prepared by adopting the secondary enzymolysis disclosed by the

embodiments

1 and 2 of the invention is obviously superior to that of the bird's nest peptide prepared by adopting the single enzymolysis degree in the comparative example 1.

In addition, in order to further verify the non-obvious nature of the biological enzymolysis process in the preparation method of the cubilose peptide disclosed and protected by the application compared with the prior art, the inventor also performs the following experiments:

experiment 1: the preferred method of the biological enzyme species is,

1) soaking dried nidus Collocaliae with medical grade purified water at 50 deg.C for 40min, mincing, cleaning, draining, adding 30 times of medical grade purified water, and stewing for 35min until the nidus Collocaliae is transparent, soft and glutinous and has no hard core;

3) passing the cubilose pulp through a homogenizer for 1-2 times, homogenizing under the pressure of 20-35 Mpa and at the temperature of 40-80 ℃, and coarsely filtering through a 300-mesh filter screen to obtain homogenized liquid;

4) respectively adding 0.05-0.2% of pepsin, 0.05-0.2% of alkaline protease, 0.05-0.2% of papain, 0.05-0.2% of trypsin, 0.05-0.2% of flavourzyme and 0.04-0.2% of neutral protease in the step 3) to measure hydrolysis conditions and enzyme activity;

5) respectively adding 0.1-0.25% of test pepsin, 0.1-0.25% of alkaline protease, 0.1-0.25% of papain, 0.1-0.25% of trypsin, 0.1-0.25% of flavourzyme and 0.1-0.25% of neutral protease into the supernatant in step 3), and testing the dissolution rate of the bird's nest protein by using Coomassie brilliant blue and Kjeldahl nitrogen;

6) 0.15 to 0.35 percent of pepsin, 0.15 to 0.35 percent of alkaline protease, 0.15 to 0.35 percent of papain, 0.15 to 0.35 percent of trypsin, 0.15 to 0.35 percent of flavourzyme and 0.15 to 0.35 percent of neutral protease are respectively added in the step 3) to test the influence on the degree of hydrolysis of the cubilose protein;

7) respectively adding 0.25-0.4% of pepsin, 0.25-0.4% of alkaline protease, 0.25-0.4% of papain, 0.25-0.4% of trypsin, 0.25-0.4% of flavourzyme and 0.25-0.4% of neutral protease into the mixture obtained in step 3), testing the hydrolysis release effect of the sialic acid in the cubilose, and detecting the free sialic acid in the hydrolyzed supernatant;

8) adding pepsin 0.35-0.55%, alkaline protease 0.35-0.55%, papain 0.35-0.55%, trypsin 0.35-0.55%, flavourzyme 0.35-0.55%, and neutral protease 0.35-0.55% into 3), respectively, testing the molecular weight change of hydrolyzed polypeptide, and detecting the supernatant after the bird's nest is hydrolyzed. The screening results are shown in the following table 3 and the attached figures 3-5, and the results show that the oligopeptide content, the protein content and the free sialic acid content of the supernatant are higher after the hydrolysis of trypsin, alkaline protease and flavourzyme.

9) Combining 4), 5), 6), 7) and 8) screening results, and selecting four biological enzymes of neutral protease, alkaline protease, flavourzyme and trypsin as complex enzymes to serve as biological enzymes for enzymolysis of the bird's nest peptide.

TABLE 3 molecular weight ratio of different enzymatically hydrolyzed polypeptides

Experiment 2: preferred method of dosing biological enzymes

A preparation method of cubilose peptide comprises the following steps:

3) pouring the cubilose pulp into an enzymolysis tank, adding medical-grade purified water until the concentration of the dry cubilose is 5%, adjusting the pH to 7.0, starting stirring at the rotating speed of 40rpm, and heating until the temperature of the materials in the tank is 50 ℃.

4) Adding neutral protease, alkaline protease, flavourzyme and trypsin in the step 3) for compounding, performing a four-factor three-level orthogonal experiment, and performing enzymolysis for 8 hours to obtain an enzymolysis solution.

5) Through carrying out orthogonal test on the step 4), as can be seen from tables 4 and 5, the biological enzymes affecting the molecular weight of the bird's nest polypeptide are trypsin, alkaline protease, flavourzyme and neutral protease in a primary and secondary sequence, and 0.04-0.3% of neutral protease, 0.05-0.4% of alkaline protease, 0.1-0.5% of flavourzyme and 0.1-0.5% of trypsin are selected as the optimal mixture ratio of the bird's nest complex enzyme for enzymolysis. Wherein, the influence graphs of different addition amounts of different protease species on the hydrolysis molecular weight are shown in fig. 6-9, and the influence graphs of different addition amounts of the flavourzyme, the alkaline protease, the neutral protease and the trypsin on the hydrolysis molecular weight are shown in sequence.

TABLE 4 orthogonal experiment factor horizon

Level of	Flavourzyme protease	Alkaline protease	Neutral protease	Trypsin
						1	0.1％	0.1％	0.1％	0.1％
2	0.3％	0.3％	0.3％	0.3％
					3	0.5％	0.5％	0.5％	0.5％

TABLE 5L₉(3⁴) Results of orthogonal experiments

The previous 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.

Claims

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

2. The method for preparing cubilose peptide according to claim 1, wherein in the step (1), the mass of the added dry cubilose is 1-8% of the mass of the purified water, the soaking time is 30-60 min, and the soaking temperature is 50-60 ℃.

3. The preparation method of the cubilose peptide according to claim 1, wherein in the step (2), the amount of purified water is 20-60 times of the mass of the cubilose, and the stewing time is 0.5-1 h; and grinding the stewed cubilose for 2-3 times by using a colloid mill, and controlling the fineness of cubilose pulp particles within 10 mu m.

4. The method for preparing cubilose peptide according to claim 1, wherein in the step (3), the mass concentration of cubilose pulp before first biological enzymolysis is 1-8%, and the pH value is 6.0-8.0;

the adding mass of the neutral protease is 0.02-0.07% of the mass of the dry cubilose, the enzymolysis temperature is 50-60 ℃, and the enzymolysis time is 30-60 min.

5. The method for preparing cubilose peptide according to claim 1, wherein in the step (4), the homogenization pressure is 20-35 MPa, the homogenization temperature is 40-80 ℃, and the enzymolysis homogenization liquid is obtained after rough filtration through a 300-mesh filter screen.

6. The method for preparing cubilose peptide according to claim 1, wherein in the step (5), the mass concentration of the enzymatic hydrolysis homogeneous liquid before the second biological enzymolysis is 1-8%, and the pH value is 6.0-8.0.

7. The method for preparing the cubilose peptide according to claim 1 or 6, wherein the compound protease consists of the following proteases in percentage by mass:

8. The method for preparing cubilose peptide according to claim 1, wherein in the step (6), the aperture of the secondary fine filtration of the enzymatic hydrolysate is 0.1-5 μm, the reverse osmosis concentration multiple of an RO membrane is 2-7 times, the inlet temperature of spray drying is 130-180 ℃, the outlet temperature is 70-100 ℃, and the pressure is 30-40 Mpa.

9. An edible bird's nest peptide, characterized in that the edible bird's nest peptide is prepared by the method of any one of claims 1 to 8.

10. Use of the bird's nest peptide prepared by the method of any one of claims 1-8 or the bird's nest peptide of claim 9 in bird's nest food and skin care products.