CN112626155B

CN112626155B - Preparation method of pea peptide

Info

Publication number: CN112626155B
Application number: CN202010516108.1A
Authority: CN
Inventors: 王倩
Original assignee: Nanjing Fenchem Biological Technology Co ltd
Current assignee: Nanjing Fenchem Biological Technology Co ltd
Priority date: 2020-06-09
Filing date: 2020-06-09
Publication date: 2022-02-18
Anticipated expiration: 2040-06-09
Also published as: CN112626155A

Abstract

The invention provides a preparation method of pea peptide with low bitter taste, small enzyme consumption and high peptide yield, belonging to the technical field of deep processing of peas. According to the invention, firstly, the structure of pea protein is expanded through the pretreatment of high-pressure steam sterilization, so that the enzymolysis is facilitated, the bitter value of an enzymolysis solution is greatly reduced through the composite enzymolysis of Alcalase protease and protease P, the finally obtained peptide powder has low bitter taste, and the total enzyme consumption is lower than that of the prior art. The optimal action pH value of the protease P is close to that of Alcalase protease, so that excessive increase of salt in the peptide powder is avoided, and the finally prepared peptide powder aqueous solution is close to neutral. The pea peptide powder prepared by the method has the protein content of more than or equal to 90 percent, the peptide yield reaches 73 percent, and the content of the generated free amino acid is lower than 15 percent. The pea peptide powder prepared by the invention can be used for nourishment, health food and formula food for special medicine.

Description

Preparation method of pea peptide

Technical Field

The invention belongs to the technical field of deep processing of peas, and particularly relates to a preparation method of pea peptide.

Background

Pea protein is a new protein in recent years, has low price and low sensitivity, and is a better essential amino acid source. At present, the utilization of peas in food in China is mainly concentrated on making vermicelli by using starch in the peas, and pea protein is mainly used as feed, so that the added value is low. The pea peptide is an enzymolysis product of pea protein and is one direction for deep processing of the pea protein. Researches report that the pea peptide has various functions such as antioxidation, anti-inflammation, blood pressure reduction and the like. In addition, Chinese patent 'a preparation method of pea peptide and application thereof in the whitening field' (publication number: CN109055466A) discloses application of pea peptide in the whitening field (including application in food and cosmetics); chinese patent 'preparation method of pea peptide with prebiotic effect and application of the pea peptide in food' (publication number: CN108220371A) discloses a preparation method of pea peptide with prebiotic effect; chinese patent 'pea peptide nutriment and preparation method thereof' (publication number: CN101731444A) discloses a pea peptide nutriment and a preparation method thereof.

The bitter taste of peptides is however an important obstacle for the consumer to reduce their acceptance. Therefore, reducing or even eliminating the bitter taste characteristics of food protein hydrolysates is a desirable goal of the food industry. The bitter taste removing method reported at present is a selective separation method, a covering method, a protein-like reaction debittering method and a biological debittering method. For example, Chinese patent "a debitterized edible fungus enzymatic hydrolysate and a preparation method thereof" (publication No. CN109480255A) discloses a method for debitterizing edible fungus enzymatic hydrolysate by using activated carbon adsorption and beta-cyclodextrin embedding, belonging to a selective separation method. However, many of the hydrophobic peptides and amino acids in the enzymatic hydrolysate are essential amino acids, and thus, the use of the selective separation method inevitably results in loss of proteins, and the nutritional value and biological activity of the enzymatic hydrolysate are reduced. The biological debittering method is to debitter the enzymolysis product by using microorganisms and protease. Microbial debittering typically utilizes some enzyme system produced by the microbe to reduce bitterness. Pea protein is difficult to digest, and because anti-nutritional factors and high-level fiber substances exist, the problems of high addition amount of enzyme preparations and low enzymolysis efficiency in the current report of pea protein enzymolysis research are still solved.

Chinese patent 'preparation method and application of pea protein peptide' (publication number: CN106260498A) discloses that pea protein is extruded and crushed firstly, then an enzyme promoter and neutral protease are added to hydrolyze the pea protein to obtain the pea peptide, the extrusion pretreatment improves the enzymolysis efficiency of the pea protein, but the invention also has the problem that the addition quality of the neutral protease is too high (10-12% of the weight of the pea protein powder). For another example, in a Chinese patent 'preparation method of pea peptide with prebiotic effect and application of the pea peptide in food' (publication number: CN108220371A), the pea peptide is subjected to enzymolysis through three enzyme distributions, the protein content of the pea peptide obtained in example 3 is 90% and the polypeptide content is 80%, the polypeptide is relatively pure, but the total enzyme consumption is large, and the cost is not reduced favorably.

Disclosure of Invention

The invention aims to overcome the defects in the prior art and provides a preparation method of pea peptide with low bitter taste, small enzyme consumption and high peptide yield.

In order to solve the technical problems, the invention provides the following technical scheme:

the invention provides a preparation method of pea peptide with low bitter taste, less enzyme consumption and high peptide yield, and firstly provides pretreatment for high-pressure steam sterilization of pea protein isolate. The pretreatment method comprises the steps of stirring and mixing pea protein isolate and deionized water to obtain mixed feed liquid, and placing the mixed feed liquid into a high-pressure steam sterilization pot for constant-temperature treatment.

The invention provides a preparation method of pea peptide with low bitter taste, small enzyme dosage and high peptide yield, and also provides a method for reducing the bitter taste of the pea peptide by carrying out enzymolysis on pea protein by adopting complex enzyme on mixed feed liquid subjected to high-pressure steam sterilization pretreatment. The method comprises the steps of cooling the pretreated mixed feed liquid to room temperature, adding Alcalase protease firstly, then adding protease P, and obtaining enzymatic hydrolysate after a certain enzymolysis time. And (3) after enzymolysis is carried out for a certain time, heating the enzymolysis liquid to 90-100 ℃, obtaining supernatant after enzyme deactivation treatment and centrifugation treatment, obtaining clear enzymolysis liquid by the supernatant through filtering equipment, and packaging after spray drying.

The preparation method of the pea peptide provided by the invention has the advantages of low bitter taste, small enzyme consumption and high peptide yield, and comprises the following specific steps:

the method comprises the following steps:

step S1: the pretreatment of high-pressure steam sterilization comprises the following specific steps:

step S1-1: adding deionized water into pea protein isolate serving as a raw material according to the mass of 3-7% of the pea protein, and uniformly stirring to obtain a mixed feed liquid; step S1-2 is executed;

step S1-2: placing the mixed material liquid in a high-pressure steam sterilization pot, and carrying out constant-temperature treatment for 10-30 min; proceed to step S2;

step S2: performing compound enzyme enzymolysis, specifically as follows:

step S2-1: cooling the mixed material liquid pretreated in the step A to room temperature, adding Alcalase protease for 1-4 h, and then adding protease P for 2-7 h; step S2-2 is executed;

step S2-2: heating the enzymatic hydrolysate subjected to the enzymolysis by the complex enzyme in the step S2-1 to 90-100 ℃, carrying out enzyme deactivation treatment for 10-20 min, and carrying out centrifugal treatment for 10-30 min at the rotating speed of 8000-10000 r/min to obtain supernatant; proceed to step S3;

step S3: filtering and spray drying, which comprises the following steps:

and (4) filtering the supernatant to obtain clear enzymolysis liquid, and packaging after spray drying.

Further, in step S1-1, the protein content (dry basis) of the pea protein isolate is greater than or equal to 85%.

Further, in step S1-2, the constant temperature treatment temperature is 100-130 ℃.

Further, in step S2-1, Alcalase protease is a commercially available food grade enzyme from Novitin. Adding the protein in an amount of 900-3000U/g, adjusting the pH value to 7.5-9.0 by using 2mol/L NaOH at an action temperature of 45-60 ℃, and keeping the pH value unchanged.

Further, in step S2-1, protease P is a commercially available food grade enzyme available from Nippon Kogyo K.K. It contains both an endonuclease and an exonuclease. Before adding, enzyme deactivation of the enzymolysis liquid is not needed, the adding amount is 60-200U/g protein, the acting temperature is 40-50 ℃, and the pH value is adjusted to 7.5-8.5 by using 1mol/L HCl.

Further, in step S3, the filtration apparatus is formed by combining a 0.45 μm microfiltration membrane and a plate and frame filter.

Further, in step S3, the inlet temperature of spray drying is 170-180 ℃, and the outlet temperature is 70-90 ℃.

Compared with the prior art, the preparation method of the pea peptide provided by the invention has the following benefits:

1. firstly, the high-pressure steam sterilization pretreatment is adopted, so that the structure of the pea protein is expanded, and the enzymolysis is facilitated.

2. According to the invention, through the composite enzymolysis of Alcalase protease and protease P, the bitterness value of the enzymolysis liquid is greatly reduced, the finally obtained peptide powder has low bitterness, and the total enzyme consumption is lower than that of the prior art.

3. The optimal action pH value of the protease P is close to that of Alcalase protease, so that excessive increase of salt in the peptide powder is avoided, and the finally prepared peptide powder aqueous solution is close to neutral.

4. The pea peptide powder prepared by the method has the protein content of more than or equal to 90 percent, the peptide yield reaches 73 percent, the peptide segment content with the relative molecular mass of less than 1000 accounts for 80.79 percent, and the content of the generated free amino acid is less than 15 percent.

Drawings

FIG. 1 is a graph showing the relative molecular mass distribution of comparative example 1 and examples 1 to 3 in the method for preparing pea peptide with low bitterness, small enzyme consumption and high peptide yield according to the present invention.

Detailed Description

The following detailed description of embodiments of the invention is provided in connection with the accompanying drawings and the detailed description.

Specific examples of the present invention are defined as follows:

1. the protein content is measured by adopting a Kjeldahl method;

2. peptide assay GB/T22492-;

3. the determination of the peptide yield is defined as the ratio of the peptide content to the content of the raw material protein;

4. the bitterness value was determined as follows: 5 sensory evaluators were invited to participate and the training was first carried out for 1h, during which time the evaluators were asked to evaluate the various concentrations of quinine solution. Quinine concentrations were 0.1, 0.2, 0.4, 0.6, 0.8, 1mg/100mL, respectively, corresponding bitterness scores were: not bitter (1), slightly bitter (2), moderately bitter (3), very bitter (4), and extremely bitter (5). The test sample (10mg/mL) was placed in numbered cups at room temperature and randomly assigned to the panelists (the evaluation contained duplicates of the test sample, but with different numbers). And evaluating the sample by the evaluation group, comparing with the standard substance, and scoring the sample if the bitterness intensity is consistent. Mineral water was used to rinse thoroughly each sample before tasting it. The score for each sample was averaged and reported as the bitterness value of the sample.

Example 1, the preparation method of pea peptide with low bitter taste, small enzyme consumption and high peptide yield provided by the invention is adopted, and the relative molecular mass distribution of the pea peptide powder obtained by the preparation method is shown in figure 1.

The method comprises the following specific steps:

step A1: preparing pea protein isolate powder into mixed feed liquid with the protein concentration of 5% by using deionized water, and uniformly stirring; go to step A2;

step A2: placing the mixed liquid in a high pressure steam sterilization pot, setting the temperature at 100 deg.C, and treating at constant temperature for 20 min; go to step A3;

step A3: cooling the heat-treated mixed feed liquid to room temperature, adding Alcalase (2160U/g protein) at 55 deg.C, adjusting pH to 8.5 with 2mol/L NaOH, maintaining pH unchanged, and performing enzymolysis for 3 hr; adding protease P (120U/g protein), adjusting pH to 8.0 with 1mol/L HCl, and performing enzymolysis for 6 h; go to step A4;

step A4: heating the enzymolysis solution to 95 deg.C, inactivating enzyme for 15min, centrifuging at 8000r/min for 15min to obtain supernatant; go to step A5;

step A5: and (3) filtering the supernatant to obtain clear enzymolysis liquid, and packaging after spray drying.

Example 2, the preparation method of pea peptide with low bitter taste, small enzyme consumption and high peptide yield provided by the invention is adopted, and the relative molecular mass distribution of the pea peptide powder obtained by the preparation method is shown in figure 1.

The method comprises the following specific steps:

step B1: preparing pea protein isolate powder into mixed feed liquid with the protein concentration of 5% by using deionized water, and uniformly stirring; go to step B2;

step B2: placing the mixed liquid in a high pressure steam sterilization pot, setting the temperature at 121 ℃, and carrying out constant temperature treatment for 20 min; go to step B3;

step B3: cooling the heat-treated mixed material liquid to room temperature, adding Alcalase (3000U/g protein), adjusting the pH value to 8.5 with 2mol/L NaOH at the temperature of 55 ℃, keeping the pH value unchanged, and performing enzymolysis for 3 h; adding protease P (120U/g protein), adjusting pH to 8.0 with 1mol/L HCl, and performing enzymolysis for 4 h; go to step B4;

step B4: heating the enzymolysis solution to 95 deg.C, inactivating enzyme for 15min, centrifuging at 8000r/min for 15min to obtain supernatant; go to step B5;

step B5: and (3) filtering the supernatant to obtain clear enzymolysis liquid, and packaging after spray drying.

Example 3, the preparation method of pea peptide with low bitter taste, small enzyme consumption and high peptide yield provided by the invention is adopted, and the relative molecular mass distribution of the pea peptide powder obtained by the preparation method is shown in figure 1.

The method comprises the following specific steps:

step C1: preparing pea protein isolate powder into mixed feed liquid with the protein concentration of 5% by using deionized water, and uniformly stirring; go to step C2;

step C2: placing the mixed liquid in a high pressure steam sterilization pot, setting the temperature at 121 ℃, and carrying out constant temperature treatment for 20 min; go to step C3;

step C3: cooling the heat-treated mixed feed liquid to room temperature, adding Alcalase (2160U/g protein) at 55 deg.C, adjusting pH to 8.5 with 2mol/L NaOH, maintaining pH unchanged, and performing enzymolysis for 3 hr; adding protease P (120U/g protein), adjusting pH to 8.0 with 1mol/L HCl, and performing enzymolysis for 6 h; go to step C4;

step C4: heating the enzymolysis solution to 95 deg.C, inactivating enzyme for 15min, centrifuging at 9000r/min for 15min to obtain supernatant; go to step C5;

step C5: and (3) filtering the supernatant to obtain clear enzymolysis liquid, and packaging after spray drying.

In comparative example 1, the preparation method of pea peptide with low bitter taste, small enzyme consumption and high peptide yield provided by the invention is adopted, and the relative molecular mass distribution of the pea peptide powder obtained by the preparation method is shown in figure 1.

The method comprises the following specific steps:

step D1: preparing pea protein isolate powder into mixed feed liquid with the protein concentration of 5% by using deionized water, and uniformly stirring; go to step D2;

step D2: placing the mixed liquid in a high pressure steam sterilization pot, and standing at normal temperature for 20 min; go to step D3;

step D3: cooling the heat-treated mixed feed liquid to room temperature, adding Alcalase (2160U/g protein) at 55 deg.C, adjusting pH to 8.5 with 2mol/L NaOH, maintaining pH unchanged, and performing enzymolysis for 3 hr; adding protease P (120U/g protein), adjusting pH to 8.0 with 1mol/L HCl, and performing enzymolysis for 6 h; go to step D4;

step D4: heating the enzymolysis solution to 95 deg.C, inactivating enzyme for 15min, centrifuging at 8000r/min for 15min to obtain supernatant; go to step D5;

step D5: and (3) filtering the supernatant to obtain clear enzymolysis liquid, and packaging after spray drying.

Indexes of the pea peptide powder obtained in the comparative example 1 and the examples 1 to 3 were measured, and the results are shown in table 1.

TABLE 1 comparison of index detection results of pea peptide powders obtained in comparative example 1 and examples 1 to 3

The pea peptide powders obtained in comparative example 1 and examples 1 to 3 were examined to have a relative molecular mass distribution as shown in fig. 1.

The above embodiments and examples are specific supports for the technical idea of the method for preparing pea peptide with low bitter taste, small enzyme consumption and high peptide yield, and the protection scope of the invention should not be limited thereby, and any equivalent changes or equivalent modifications made on the basis of the technical scheme according to the technical idea of the invention still belong to the protection scope of the technical scheme of the invention.

Claims

1. A preparation method of pea peptide is characterized by comprising the following steps: the method comprises the following steps:

step S1-1: preparing pea protein isolate powder into mixed feed liquid with the protein concentration of 5% by using deionized water, uniformly stirring, and transferring to the step S1-2;

step S1-2: placing the mixed liquid in a high pressure steam sterilization pot, setting the temperature at 121 deg.C, and treating at constant temperature for 20 min; proceed to step S2;

step S2: performing compound enzyme enzymolysis, specifically as follows:

step S2-1: cooling the mixed material liquid pretreated in the step S1 to room temperature, firstly adding 2160U/g protein Alcalase protease, adjusting the pH value to 8.5 with 2mol/L NaOH at 55 ℃, keeping the pH value unchanged, and carrying out enzymolysis for 3 h; then adding protease P of 120U/g protein, adjusting the pH value to 8.0 by using 1mol/L HCl, and carrying out enzymolysis for 6 h; step S2-2 is executed;

step S2-2: heating the enzymolysis liquid after the enzymolysis of the compound enzyme in the step S2-1 to 95 ℃, inactivating the enzyme for 15min, and centrifuging at the rotating speed of 9000r/min for 15min to obtain a supernatant; proceed to step S3;

step S3: filtering and spray drying, which comprises the following steps:

2. The method for preparing a pea peptide according to claim 1, wherein: the protein content of the pea protein isolate in the step S1-1 is more than or equal to 85 percent on a dry basis.

3. The method for preparing a pea peptide according to claim 1, wherein: in the step S3, the filtering device is formed by combining a microfiltration membrane of 0.45 μm and a plate-and-frame filter.

4. The method for preparing a pea peptide according to claim 1, wherein: in the step S3, the inlet temperature of spray drying is 170-180 ℃, and the outlet temperature is 70-90 ℃.

5. A pea peptide product or a pea peptide-containing product prepared by the method for preparing pea peptide according to any one of claims 1 to 4.