CN111903957A - Application of a kind of plant protease hydrolyzate as polypeptide surfactant - Google Patents

Abstract

The invention relates to the application of a plant protease hydrolyzate as a polypeptide surfactant, and belongs to the technical field of light chemical industry. The application is to use a plant protease hydrolyzate with a surface tension of 25mN/m to 45mN/m when the protein mass fraction is 0.4% to 1% in a solution or suspension state as a polypeptide surfactant, and the surface of the polypeptide is applied. Active agents can be used as food flavorings, protein drinks, meat products, or cosmetic emulsifiers. Compared with the emulsification of protein, the emulsification of polypeptide surfactants is stronger. Because of its small molecular weight, peptides are easily absorbed by human skin, hair and intestinal tract, so they have nutritional effects; peptide surfactants also have the functions of moisturizing, anti-oxidation, lowering blood pressure, lowering blood lipids, and antibacterial, which are extremely safe. It is a multifunctional biosurfactant with good properties and biocompatibility. It can be used in cosmetics, medicines, food and other products that come into direct contact with the human body.

Description

Application of a kind of plant protease hydrolyzate as polypeptide surfactant

technical field

The invention relates to the application of a plant protease hydrolyzate as a polypeptide surfactant, and belongs to the technical field of light chemical industry.

Background technique

Biosurfactants are natural surfactants produced by microorganisms, animals and plants. Compared with chemically synthesized surfactants, the raw materials of biosurfactants are renewable biomass resources, which have the advantages of easy degradation, high biocompatibility and high safety, and are especially suitable for products in direct contact with the human body, such as cosmetics, pharmaceuticals and food, etc.

The hydrophobic end of biosurfactants generally has a long fatty acid chain, the hydrophilic end is generally sugars, cyclic peptides, amino acids, phosphoric acid carboxylic acids or alcohols, and the molecular weight is generally 500Da to 1500Da. Biosurfactants can be divided into glycolipids, phospholipids, fatty acids, lipopeptides and lipoproteins according to their chemical structures. Western developed countries started production and application in the middle of the 20th century, while research in my country started late and basically stayed in the laboratory stage. With the continuous development and progress of human society, people's awareness of environmental protection and safety is generally enhanced, and the development, production and application of biosurfactants will become an inevitable trend.

The production methods of biosurfactants include microbial (bacteria, yeast and some filamentous fungi) fermentation, enzymatic synthesis and natural biological extraction. Biosurfactants synthesized by microbial fermentation, various glycolipid biosurfactants such as rhamnolipid, microbial fermentation sugar and trehalolipid, the microbial fermentation biosurfactant prepared by Bacterococcus has been used as a humectant by Kao Co., Ltd. in Japan For cosmetics, the trade name is Sofina. Enzymatic biosynthesis can be used for the synthesis of biosurfactants such as monoglycerides, glycolipids, phospholipids and alkyl glycosides. In recent years, the combination of enzyme synthesis and microbial fermentation has become a development trend. The natural biological extraction method can directly extract biosurfactants from natural biological raw materials. Biosurfactants such as phospholipids and lecithins exist in natural biological raw materials such as egg yolk or soybean. Raw material restrictions.

Biosurfactants can be widely used in the petroleum industry, washing cosmetics, pharmaceuticals, food, agriculture and the environment. Used in the petroleum industry, it can reduce the viscosity of oil, improve the recovery of crude oil, enhance the fluidity of oil in oil pipelines, and realize the demulsification and dehydration of oil and the bioremediation of oil-contaminated soil. Used in washing cosmetics, it has high surface activity, strong emulsifying ability, good foaming, good biocompatibility, non-allergic, digestible and high safety. It is often used as emulsifier, penetrant, detergent, wetting agent, Bactericides, antioxidants and cosmetic additives such as hair dyes. Biosurfactants are often used as solubilizers, suspending agents, antioxidants, penetrants and preservatives in pharmacy, which can reduce the surface tension between the two phases of the solvent and the insoluble drug, and liquefy the two immiscible substances into a phase. Soluble emulsion or suspension, phospholipids with strong emulsifying ability in biosurfactant can be used as emulsifier for intravenous injection. Biosurfactants can be used as food additives in the food industry, such as emulsifiers, stabilizers, wetting agents, foaming agents and thickeners, etc., to change the consistency, viscosity, taste and freshness of food, etc. Biosurfactants In the process of human digestion, it can be decomposed, absorbed or eliminated from the body, and has no side effects on the human body. The application of biosurfactants in the agricultural field is also gradually increasing. Its emulsification and permeability can promote the transmission and dispersion of water in compost particles, and its moisturizing properties can slow down the evaporation of water in the compost and maintain the activity of microorganisms. Biosurfactants can also be used as pesticides and animal feed additives. In environmental engineering, the desorption of biosurfactants promotes the degradation of organic matter in sewage and reduces the degradation time. Biosurfactants can also remove various pollutants and heavy metals, and have good environmental compatibility.

The research reports on biosurfactants are increasing, but the preparation of pure polypeptide surfactants by plant proteolysis has not been reported yet. The emulsification system of protein and polypeptide exists in human daily life. Milk is a typical protein emulsification system, and a stable system of cream and water is formed through the emulsification of casein; Various peptides formed by heating and degrading meat protein play the role of emulsifier; soy protein isolate products can also be used for emulsification of food. At present, plant protease hydrolysate is a short-chain peptide segment formed by enzymatic hydrolysis of plant protein by protease, which is mainly used for taste, nutrition and functional health care, and there is no report on emulsification. In the prior art, the invention patent application with the application number of 201510248236.1 is related to polypeptide emulsification. It introduces a preparation method of a collagen polypeptide-based surfactant. Filtration, distillation and concentration under reduced pressure, and then reacting with oleoyl chloride to obtain a peptidyl surfactant with a peptidyl group at the hydrophilic end and an oleoyl group at the hydrophobic end.

SUMMARY OF THE INVENTION

In order to overcome the defects of the prior art, the purpose of the present invention is to provide the application of a plant protease hydrolyzate as a polypeptide surfactant.

In order to achieve the purpose of the present invention, the following technical solutions are provided.

The application of a plant protease hydrolyzate as a polypeptide surfactant is as follows:

When the plant protease hydrolyzate satisfies the surface tension of 25mN/m-45mN/m when the protein mass fraction is 0.4%-1% in solution or suspension state, the plant protease hydrolyzate is used as a polypeptide surfactant.

Preferably, the polypeptide surfactant is used as a food seasoning, protein drink, meat product or cosmetic emulsifier.

The plant protease hydrolyzate is a short-chain peptide segment formed by enzymatic hydrolysis of plant protein by protease.

Preferably, the plant protease hydrolyzate is prepared by the following method, and the method steps are as follows:

(1) the pulverized vegetable protein raw material or vegetable protein extract is prepared into feed liquid with water;

(2) adjust the pH value of feed liquid to the optimum pH value of protease;

(3) adding more than one endoprotease to the feed solution to obtain a reaction solution;

(4) Stir the reaction solution until fully mixed, and enzymolysis for 2h to 18h; in the case of single protease enzymolysis, the enzymolysis temperature is the optimum temperature of the endoprotease, and when two or more proteases are enzymolyzed at the same time, the enzymolysis temperature is based on the enzymolysis time. Determine the temperature at which the degree of hydrolysis of plant protein is maximized;

(5) heat inactivating the enzyme to obtain a plant protease hydrolyzate.

In step (1):

Preferably, the vegetable protein raw material or the vegetable protein extract is crushed and sieved at 40-60 mesh.

Preferably, the mass fraction of the vegetable protein raw material or vegetable protein extract is 1% to 25% based on the total mass of water as 100%.

Preferably the vegetable protein raw material is one or more of soybean, soybean meal and peanut meal; preferably the vegetable protein extract is one or more of soybean protein concentrate, soybean protein isolate, peanut protein concentrate, peanut protein isolate, corn gluten and gluten protein .

In step (2):

The acid or base used to adjust the pH value of the feed liquid is determined according to the requirements of the field of use, for example, the acid and base allowed by relevant laws and regulations in the field of food or cosmetics.

In step (3):

Preferably, the mass fraction of the endoprotease is 0.1% to 20% based on the total mass of the vegetable protein raw material or the vegetable protein extract being 100%.

Preferably, the endoprotease is more than one of papain, bromelain, neutral protease, alkaline protease, chymotrypsin, trypsin, pepsin and Protamex protease, and Protamex protease comes from Novozymes (China) Investment Co., Ltd. ( hereinafter referred to as Novozymes).

In step (5):

Preferably, the enzyme is inactivated at 100°C for 5 min to 10 min.

beneficial effect

The invention provides the application of a plant protease hydrolyzate as a polypeptide surfactant. By optimizing the selection of plant protease hydrolysis conditions, the protein content of the enzymolyzate in a solution or suspension state is 0.4% to 1%. The surface tension is 25mN/m～45mN/m, which can be used as a polypeptide surfactant. Compared with the emulsification of protein, the emulsification of polypeptide surfactants is stronger. Because of its small molecular weight, peptides are easily absorbed by human skin, hair and intestinal tract, so they have nutritional effects; peptide surfactants also have the functions of moisturizing, anti-oxidation, lowering blood pressure, lowering blood lipids, and antibacterial, which are extremely safe. It is a multifunctional biosurfactant with good properties and biocompatibility. It can be used in cosmetics, medicines, food and other products that come into direct contact with the human body.

Detailed ways

The present invention will be described in detail below in conjunction with specific embodiments, but not as a limitation on the patent of the present invention.

In the following examples:

The degree of hydrolysis was determined by formaldehyde titration;

The protein content was determined by the Kjeldahl method.

Example 1

(1) Measure 100mL of water and heat it to 50°C in a 250mL beaker, add 10g of soybean protein isolate (Shandong Jiahua Oil Co., Ltd.), and prepare a feed solution;

(2) adjusting the pH of the feed liquid with a concentration of 2M NaOH solution is 7.5, and stirring until soybean protein isolate reaches a dissolved state;

(3) 0.3 g of alkaline protease (Novozymes) was added to obtain a reaction solution;

(4) The reaction solution was stirred at 60 rpm to 200 rpm until fully mixed, and enzymatically hydrolyzed at 50°C for 3 hours;

(5) heating to 100° C. and maintaining the enzyme activity for 5 minutes to obtain a soybean polypeptide enzymatic hydrolysate.

The soybean polypeptide enzymatic hydrolysate obtained in this example was detected, and the results were as follows:

The degree of hydrolysis of the soybean polypeptide enzymatic hydrolysis solution was determined to be 17%; the soybean polypeptide enzymatic hydrolysis solution was diluted with water to become an aqueous solution with a protein content of 1%, and its surface tension was detected to be 35 mN/m.

Example 2

(1) Measure 100mL of distilled water and heat it to 50°C in a 250mL beaker, add 5g of soybean protein isolate (Shandong Yuwang Food Co., Ltd.), and prepare a feed liquid;

(2) adjust the pH of the feed liquid to be 7.0 with the NaOH solution of 2M, stir until soybean protein isolate reaches the dissolved state;

(3) add papain (Nanning Pangbo Biological Engineering Co., Ltd.) 0.05g to obtain reaction solution;

(4) The reaction solution was stirred at 60 rpm to 200 rpm until fully mixed, and enzymatically hydrolyzed at 50°C for 6 hours;

(5) heating to 100° C. and maintaining the enzyme activity for 5 minutes to obtain a soybean polypeptide enzymatic hydrolysate.

The soybean polypeptide enzymatic hydrolysate obtained in this example was detected, and the results were as follows:

The degree of hydrolysis of the soybean polypeptide enzymatic hydrolysis solution was determined to be 14%; the soybean polypeptide enzymatic hydrolysis solution was diluted with water to become an aqueous solution with a protein content of 1%, and its surface tension was detected to be 38 mN/m.

Example 3

(1) Measure 100mL of distilled water in a 250mL beaker, heat to 55°C, add 10g of peanut protein isolate (Henan Liangjian Technology Co., Ltd.), and prepare a feed solution;

(2) adjusting the pH of the feed liquid with a concentration of 2M hydrochloric acid (HCl) solution is 6.0, and stirring until the peanut protein isolate reaches a dissolved state;

(3) add Protamex protease (novozymes) 0.05g to obtain reaction solution;

(4) The reaction solution was stirred at 60 rpm to 200 rpm until fully mixed, and enzymatically hydrolyzed at 55°C for 4 hours;

(5) heating to 100° C. and maintaining the enzyme activity for 5 minutes to obtain a peanut polypeptide enzymatic hydrolysate.

The peanut polypeptide enzymolysis solution prepared in the present embodiment is detected, and the results are as follows:

The degree of hydrolysis of the peanut polypeptide enzymolysis solution was determined to be 18%; the peanut polypeptide enzymolysis solution was diluted with water to become an aqueous solution with a protein content of 1%, and its surface tension was 32 mN/m.

Example 4

(1) Measure 100mL of distilled water in a 250mL beaker, heat to 37°C, add 8g of gluten (crude protein content 83%, Shandong Guanxian Ruixiang Biotechnology Development Co., Ltd.) to prepare a feed solution;

(2) adjust the pH of the feed liquid to be 2.0 with a concentration of 10M HCl, and stir until the gluten powder reaches a mixed state;

(3) finally add 0.1g pepsin (Sichuan Deyang City Biochemical Products Co., Ltd.) to obtain reaction solution;

(4) The reaction solution was stirred at 60 rpm to 200 rpm until fully mixed, and enzymatically hydrolyzed at 37°C for 10 hours;

(5) heating to 100° C. and maintaining the enzyme activity for 5 minutes to obtain a gluten protein polypeptide enzymatic hydrolyzate solution.

Detecting the gluten protein polypeptide enzymatic hydrolysate prepared in this example, the results are as follows:

The degree of hydrolysis of the glutenin polypeptide enzymolysis solution was determined to be 16%; the glutenin polypeptide enzymolysis solution was diluted with water to become an aqueous solution with a protein content of 1%, and its surface tension was detected to be 40mN/m.

Example 5

(1) 20g of soybean polypeptide enzymolysis solution obtained in Example 1, 15g soybean oil, 15g chicken oil, 150g chicken breast enzymolysis solution (protein content 10% (w/w)), 10g salt were added into a 250mL cup, Heat to boiling to obtain a mixed solution;

(2) The mixed solution was placed in an emulsifying homogenizer (Zhejiang Chint Electric Co., Ltd., model TDCC2-0.5) and homogenized at 4000 r/min to obtain a white soup stock emulsion.

Example 6

(1) 10g of soybean polypeptide enzymolysis solution obtained in Example 3 and 3g of dried walnuts, 2g of dried almonds, 5g of dried peanut kernels, 20g of white granulated sugar, 160g of water were boiled in a stainless steel small pot for 5 minutes to obtain a mixture;

(2) Transfer the mixture to a Joyoung multifunctional food processor and beat at high speed for 3 minutes to obtain a white protein milk drink.

Example 7

(1) 2Kg of beef, 0.7Kg of tallow are minced with an orifice plate of 8mm in diameter, and 0.8Kg of chicken breast is minced with an orifice plate of 8mm in diameter to obtain minced meat material;

(2) the minced meat material is beaten into mashed meat with a masher, add 0.18Kg salt, 0.5g sodium nitrite, 1Kg ice water high-speed beating for 1 minute, then add the embodiment 1 that is cooled to 4 ℃ and obtain Soybean polypeptide enzymolysis solution 1Kg, 0.3Kg soybean protein, 40g carrageenan, 120g sugar, 25g monosodium glutamate, 1g I+G, 15g white pepper, 1.5Kg ice water, beat at high speed for 1 minute, add 1.5Kg ice water, add modified Corn starch 1Kg continue to beat evenly;

(3) Pour the meat slurry into the nylon casing, and then punch in at both ends after tightening;

(4) Cooking: Cook at 85℃～90℃ for 50 minutes～60 minutes.

Example 8

The soybean polypeptide enzymolysis solution 8mL obtained in Example 2, 8mL of walnut oil, 2g of myristate isopropanol, 6g of white oil, 2g of lanolin, and 5g of glycerol were placed in an emulsifying homogenizer (Zhejiang Chint Electric Co., Ltd., Model TDCC2-0.5) is homogenized at 3000r/min to obtain a uniform moisturizing cream lotion, which is applied to the skin and has skin care functions such as moisturizing, water retention and nutrition.

Claims (10)

1. The application of a plant protein zymolyte as a polypeptide surfactant is characterized in that: when the surface tension of the plant protein zymolyte is 25 mN/m-45 mN/m when the protein mass fraction is 0.4% -1% in a solution or suspension state, the plant protein zymolyte is applied as a polypeptide surfactant, and the plant protein zymolyte is a short-chain peptide segment formed by the enzymolysis of plant protein through protease.

2. The use of a plant protein hydrolysate as a polypeptide surfactant according to claim 1, wherein the plant protein hydrolysate is selected from the group consisting of: the polypeptide surfactant is used as an emulsifier for food seasonings, protein beverages, meat products or cosmetics.

3. The use of a plant protein hydrolysate as claimed in claim 1 or 2 as a polypeptide surfactant, wherein: the plant protein zymolyte is prepared by the following method:

(1) preparing the crushed plant protein raw material or plant protein extract into feed liquid by using water;

(2) adjusting the pH value of the feed liquid to the optimal pH value of the protease;

(3) adding more than one endoprotease into the feed liquid to obtain reaction liquid;

(4) stirring the reaction solution until the reaction solution is fully mixed, and performing enzymolysis for 2-18 h; when single protease is used for enzymolysis, the enzymolysis temperature is the optimal temperature of endoprotease, and when more than two proteases are used for enzymolysis simultaneously, the enzymolysis temperature is determined according to the temperature at which the hydrolysis degree of the plant protein is maximum within the enzymolysis time;

(5) heating to inactivate enzyme to obtain plant protein zymolyte.

4. The use of a plant protein hydrolysate as a polypeptide surfactant according to claim 3, wherein the plant protein hydrolysate is selected from the group consisting of: in the step (1): the plant protein raw material or the plant protein extract is crushed and sieved in 40-60 meshes.

5. The use of a plant protein hydrolysate as a polypeptide surfactant according to claim 3, wherein the plant protein hydrolysate is selected from the group consisting of: in the step (1): the mass fraction of the plant protein raw material or the plant protein extract is 1 to 25 percent based on the total mass of the water as 100 percent;

in the step (3): the mass fraction of the endoprotease is 0.1-20% based on 100% of the total mass of the plant protein raw material or the plant protein extract.

6. The use of a plant protein hydrolysate as a polypeptide surfactant according to claim 3, wherein the plant protein hydrolysate is selected from the group consisting of: in the step (1): the vegetable protein material is more than one of soybean, soybean meal and peanut meal; the vegetable protein extract is more than one of soybean protein concentrate, soybean protein isolate, peanut protein concentrate, peanut protein isolate, zein and glutelin;

in the step (3): the endoprotease is one or more of papain, bromelain, neutral protease, alkaline protease, chymotrypsin, trypsin, pepsin and Protamex protease from Novesin (China) investment Limited.

7. The use of a plant protein hydrolysate as a polypeptide surfactant according to claim 3, wherein the plant protein hydrolysate is selected from the group consisting of: in the step (5): keeping the enzyme inactivation for 5min to 10min at the temperature of 100 ℃.

8. The use of a plant protein hydrolysate as a polypeptide surfactant according to claim 3, wherein the plant protein hydrolysate is selected from the group consisting of: in the step (1): the plant protein raw material or the plant protein extract is crushed and sieved in 40 meshes to 60 meshes;

the mass fraction of the plant protein raw material or the plant protein extract is 1 to 25 percent based on the total mass of the water as 100 percent;

the vegetable protein material is more than one of soybean, soybean meal and peanut meal; the vegetable protein extract is one or more of soybean protein concentrate, soybean protein isolate, peanut protein concentrate, peanut protein isolate, zein and glutelin.

9. The use of a plant protein hydrolysate as a polypeptide surfactant according to claim 3, wherein the plant protein hydrolysate is selected from the group consisting of: in the step (3): the mass fraction of the endoprotease is 0.1-20 percent based on 100 percent of the total mass of the plant protein raw material or the plant protein extract;

the endoprotease is one or more of papain, bromelain, neutral protease, alkaline protease, chymotrypsin, trypsin, pepsin and Protamex protease from Novesin (China) investment Limited.

10. The use of a plant protein hydrolysate as a polypeptide surfactant according to claim 3, wherein the plant protein hydrolysate is selected from the group consisting of: in the step (1): the plant protein raw material or the plant protein extract is crushed and sieved in 40 meshes to 60 meshes;

the mass fraction of the plant protein raw material or the plant protein extract is 1 to 25 percent based on the total mass of the water as 100 percent;

the vegetable protein material is more than one of soybean, soybean meal and peanut meal; the vegetable protein extract is more than one of soybean protein concentrate, soybean protein isolate, peanut protein concentrate, peanut protein isolate, zein and glutelin;

in the step (3):

the mass fraction of the endoprotease is 0.1-20 percent based on 100 percent of the total mass of the plant protein raw material or the plant protein extract;

the endoprotease is more than one of papain, bromelain, neutral protease, alkaline protease, chymotrypsin, trypsin, pepsin and Protamex protease, and the Protamex protease is from Novesin (China) investment Limited;

in the step (5): keeping the enzyme inactivation for 5min to 10min at the temperature of 100 ℃.