CN114015737B - Preparation method and application of corn glutamine peptide with intestinal barrier protection function - Google Patents
Preparation method and application of corn glutamine peptide with intestinal barrier protection function Download PDFInfo
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
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- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
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- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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Abstract
The invention provides a preparation method of corn glutamine peptide with intestinal barrier protection function, which comprises the following preparation steps: carrying out first enzymolysis on zein by using alkaline protease to obtain a first enzymolysis liquid; and (3) carrying out second enzymolysis on the first enzymolysis liquid by using composite protease to obtain a second enzymolysis liquid, and drying to obtain the corn glutamine peptide. The results of the examples show that the corn glutamine peptide can efficiently release the glutamine in the corn protein, and the release amount of the glutamine accounts for 75.00% -97.50% of the glutamine in the original corn protein. Can remarkably improve the weight loss of mice, reduce the disease activity index, relieve the colon shortening, reduce the content of pro-inflammatory factors IL-1 beta, IL-6 and TNF-alpha in the colon, increase the content of anti-inflammatory cytokines IL-10 and have a certain protection effect on intestinal barriers.
Description
Technical Field
The invention belongs to the technical field of application of active peptides, and particularly relates to a preparation method and application of corn glutamine peptide with an intestinal barrier protection function.
Background
In recent years, various chronic diseases caused by intestinal tracts have a tendency to develop frequently and highly, and factors causing intestinal diseases are various, but they are fundamentally because of problems in intestinal barrier function. Abnormal intestinal barrier function can cause digestive absorption dysfunction, slow growth, disease resistance reduction and increased susceptibility to pathogenic microorganisms of nutrient substances, thereby causing various diseases such as colitis, irritable bowel disease, diarrhea, constipation and the like.
Glutamine (gin) is often used clinically as an enteral nutrient to protect the intestinal barrier, but its use is severely limited by the low solubility and ease of decomposition of gin monomers. In recent years, the research shows that the glutamine binding peptide not only maintains a plurality of physiological functions and nutrition of Gln, but also has the advantages of good water solubility, low sensitization, good absorbability, strong stability and the like, and can be used as a substitute for stabilizing Gln.
There are mainly two pathways for the preparation of glutamine peptides: firstly, the peptides are synthesized by a chemical method, and the peptides are mainly dipeptides, namely alanylglutamine (Ala-Gln) and glycylglutamine (Gly-Gln), which exist mainly in the form of injection liquid medicaments; secondly, the protease is used for controlling and hydrolyzing plant proteins such as wheat gluten to obtain peptide segments rich in glutamine. However, the chemical synthesis method has potential safety hazards such as high price and difficult complete removal of organic reagents; the reaction condition of the protease for hydrolyzing the gluten powder to prepare the glutamine peptide is mild, no toxic and harmful components are generated, but the glutamine content in the enzymolysis product is relatively low.
Disclosure of Invention
In view of the above, the present invention provides a method for preparing corn glutamine peptide with intestinal barrier protection function. The corn glutamine peptide prepared by the preparation method has high glutamine content, can release glutamine in corn protein with high efficiency, and has the intestinal barrier protection function, wherein the glutamine release amount accounts for 75.00% -97.50% of the glutamine in the original corn protein.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a preparation method of corn glutamine peptide with intestinal barrier protection function, which comprises the following preparation steps:
carrying out first enzymolysis on zein by using alkaline protease to obtain a first enzymolysis liquid;
and (3) carrying out second enzymolysis on the first enzymolysis liquid by using composite protease to obtain a second enzymolysis liquid, and drying to obtain the corn glutamine peptide.
Preferably, the alkaline protease is used in an amount of 1000 to 1500U/g zein when the first enzymolysis is performed.
Preferably, the temperature of the first enzymolysis is 60-70 ℃, the time of the first enzymolysis is 2-3 h, and the pH value of the first enzymolysis is 7.5-8.5.
Preferably, the amount of the complex protease used in the second enzymolysis is 100-300U/g zein.
Preferably, the temperature of the second enzymolysis is 45-55 ℃, the time of the second enzymolysis is 2-3 h, and the pH value of the second enzymolysis is 6.5-7.5.
Preferably, the zein comprises a de-starched zein powder; the starch removal treatment comprises: mixing corn protein powder to be treated with alpha-amylase, and performing starch removal enzymolysis to obtain corn protein; the mass of the alpha-amylase is 0.8-1.2% of the mass of the corn gluten meal to be treated.
Preferably, the temperature of the starch removal enzymolysis is 60-70 ℃, and the time of the starch removal enzymolysis is 1.5-2.5 h.
Preferably, before drying, the method further comprises nanofiltration treatment of the second enzymolysis liquid; the nanofiltration pressure is 15-25 Bar and the temperature is 15-25 ℃.
The invention also provides the corn glutamine peptide with the intestinal barrier protection function, which is characterized in that the content of glutamine in the corn glutamine peptide is 10% -13% of the total protein, the glutamine in the corn protein can be efficiently released, the release amount of the glutamine accounts for 75.00% -97.50% of the glutamine in the original corn protein (the mass percentage of the protein in the glutamine peptide obtained in the specific implementation process of the invention is 83%).
The invention also provides application of the corn glutamine peptide in preparing food, medicine or health care product with intestinal barrier protection function.
The invention provides a preparation method of corn glutamine peptide with an intestinal barrier protection function. According to the invention, the corn protein is subjected to sequential enzymolysis by adopting alkaline protease and composite protease, so that the corn glutamine peptide is obtained, and the effective utilization of the corn protein is realized. The corn glutamine peptide prepared by the preparation method provided by the invention can efficiently release glutamine in corn protein, wherein the content of the glutamine is 10% -13% of the total protein in the corn glutamine peptide, and the content of the glutamine accounts for 75.00% -97.50% of the glutamine in the original corn protein. Can obviously reduce the content of pro-inflammatory factors IL-1 beta, IL-6 and TNF-alpha in colon tissues, increase the content of anti-inflammatory factors IL-10, improve the weight reduction of mice caused by ulcerative colitis, effectively relieve colon shortening, reduce disease activity index, lighten infiltration of inflammatory cells in colon tissues, reduce colon mucosa permeability and relieve the colonitis of the mice.
Drawings
FIG. 1 is the effect of maize glutamine peptide on DSS-induced colitis mice weight change;
FIG. 2 is the effect of maize glutamine peptide on disease activity index of DSS-induced colitis mice;
FIG. 3 is the effect of maize glutamine peptide on DSS-induced colitis mouse colon length;
FIG. 4 is the effect of maize glutamine peptide on DSS-induced colitis mouse anti-inflammatory factor IL-10;
FIG. 5 is the effect of maize glutamine peptide on DSS-induced colitis mouse pro-inflammatory factor TNF- α;
FIG. 6 is the effect of maize glutamine peptide on DSS-induced colitis mouse pro-inflammatory factor IL-6;
FIG. 7 is the effect of maize glutamine peptide on DSS-induced colitis mouse proinflammatory factor IL-1. Beta.
Detailed Description
The invention provides a preparation method of corn glutamine peptide with intestinal barrier protection function, which comprises the following steps:
carrying out first enzymolysis on zein by using alkaline protease to obtain a first enzymolysis liquid;
and (3) carrying out second enzymolysis on the first enzymolysis liquid by using composite protease to obtain a second enzymolysis liquid, and drying to obtain the corn glutamine peptide.
The invention uses alkaline protease to carry out first enzymolysis on corn protein to obtain first enzymolysis liquid. In the first enzymolysis according to the present invention, the alkaline protease is preferably used in an amount of 1000 to 1500U/g zein, more preferably 1200 to 1450U/g zein, and still more preferably 1200 to 1300U/g zein. The temperature of the first enzymolysis is preferably 60-70 ℃, and more preferably 60-65 ℃; the time of the first enzymolysis is preferably 2 to 3 hours, more preferably 2.2 to 2.8 hours; the pH of the first enzymatic hydrolysis is preferably 7.5 to 8.5, more preferably 7.6 to 8.2. In the enzymolysis process, the pH and the temperature are monitored in real time, so that the temperature and the pH are unchanged in the second enzymolysis process. The reagent used for regulating the pH value is not strictly required, and 1mol/L NaOH and/or 1mol/L HCl can be used. The source of the alkaline protease is not particularly limited, and a conventional commercial product can be adopted, and in the implementation process of the invention, the alkaline protease is novinarian alkaline protease Alcalase 2.4L which is purchased from Denmark novinary.
In the present invention, the zein may be obtained by purchase or may be prepared by itself. When the invention prepares the corn protein by itself, the corn protein powder is preferably used as the raw material. The corn protein powder is a main byproduct of corn starch produced by corn kernels through a wet method, the protein content of the corn protein powder is about 60%, and the glutamine content of the corn protein powder accounts for about 8.3% of total protein of corn. The source of the corn gluten meal is not particularly limited, and the corn gluten meal is obtained by adopting a conventional commercial product, and in the specific embodiment of the invention, the corn gluten meal is purchased from zizihahal Longjingfengfeng biotechnology limited company.
The invention preferably carries out extrusion treatment on the corn gluten meal to obtain the extruded corn gluten meal. The extrusion temperature of the present invention is preferably 160 to 180 ℃, more preferably 165 to 175 ℃, and still more preferably 170 ℃. The invention has no strict requirement on the extrusion time, and ensures that the water content of the extruded corn protein powder is 10-20wt%. The present invention is not critical to the manner of extrusion, and preferably a twin screw extruder is used.
The invention preferably carries out starch removal treatment on the extruded corn protein powder, and the corn protein is obtained after enzyme deactivation treatment. The starch removing treatment comprises the following steps: and mixing the extruded corn protein with alpha-amylase for enzymolysis. The mass of the alpha-amylase is preferably 0.8-1.2% of the mass of the extruded corn gluten meal, and more preferably 0.9-1.0%. The enzymolysis temperature is preferably 60-70 ℃, and more preferably 60-65 ℃; the time for the enzymolysis is preferably 1.5 to 2.5 hours, more preferably 1.8 to 2.0 hours. The temperature of the enzyme deactivation treatment is preferably 90-110 ℃, and more preferably 95-100 ℃; the time for the enzyme deactivation treatment is preferably 15 to 25 minutes, more preferably 18 to 20 minutes. The separation mode is preferably centrifugation, and the rotation speed of the centrifugation is preferably 7500 r/min-8500 r/min, more preferably 7800 r/min-8200 r/min; the time for the centrifugation is preferably 5 to 15 minutes, more preferably 8 to 10 minutes. The invention preferably carries out drying treatment on the precipitate obtained by centrifugal treatment to obtain the zein. The drying temperature of the present invention is preferably 60 to 70℃and more preferably 62 to 65 ℃. The drying mode is not critical, and an oven is preferably used.
The invention utilizes the alpha-amylase to carry out enzymolysis to squeeze the corn protein powder, so that the starch in the corn protein powder can be removed, the purity of the obtained corn protein is high, the space structure is more fully exposed, and the enzymolysis is easy.
After the first enzymolysis liquid is obtained, the invention carries out second enzymolysis on the first enzymolysis product by utilizing compound protease to obtain the second enzymolysis liquid. In the second enzymolysis, the amount of the composite protease is preferably 100 to 300U/g zein, more preferably 150 to 250U/g zein, and even more preferably 200 to 220U/g zein. The temperature of the second enzymolysis is preferably 45-55 ℃, and more preferably 48-52 ℃; the time of the second enzymolysis is preferably 2 to 3 hours, more preferably 2.2 to 2.8 hours; the pH of the second enzymatic hydrolysis is preferably 6.5 to 7.5, more preferably 6.8 to 7.2. The reagent used for regulating the pH value is not strictly required, and 1mol/L NaOH and/or 1mol/L HCl can be used. The source of the compound protease is not particularly limited, and a conventional commercial product can be adopted, and in the implementation process of the invention, the compound protease is North-vitamin complex protease Protamex which is purchased from Denmark North-vitamin company.
In the invention, the first enzymolysis liquid is preferably pretreated before the second enzymolysis is carried out on the first enzymolysis liquid by utilizing the compound protease, and the temperature and the pH of the first enzymolysis liquid are adjusted to the temperature and the pH required by the second enzymolysis. In the enzymolysis process, the pH and the temperature are monitored in real time, so that the temperature and the pH are unchanged in the second enzymolysis process.
After the second enzymatic hydrolysate is obtained, the invention preferably carries out enzyme deactivation treatment on the second enzymatic hydrolysate, and the supernatant is obtained by centrifugation. The temperature of the enzyme deactivation treatment is preferably 90-110 ℃, and more preferably 95-100 ℃; the time for the enzyme deactivation treatment is preferably 15 to 25 minutes, more preferably 18 to 20 minutes. The separation mode is preferably centrifugation, and the rotation speed of the centrifugation is preferably 3000 r/min-5000 r/min, more preferably 3200 r/min-4800 r/min, and even more preferably 3500-4000 r/min; the time for the centrifugation is preferably 15 to 25 minutes, more preferably 18 to 20 minutes.
The invention preferably carries out nanofiltration treatment and drying treatment on the supernatant to obtain the corn glutamine peptide. The pressure of the nanofiltration is preferably 15 to 25Bar, and more preferably 18 to 20Bar; the temperature of the nanofiltration is preferably 15 to 25 ℃, and more preferably 18 to 20 ℃. The invention has no strict requirement on the nanofiltration time, and ensures that the volume of the solution obtained after nanofiltration is 1/3-2/3 of the volume of the supernatant.
According to the invention, the corn protein is subjected to sequential enzymolysis by adopting alkaline protease and composite protease, so that corn glutamine peptide is obtained, effective utilization of the corn protein is realized, the glutamine content in the obtained corn glutamine peptide is 6% -7.8% of the total protein, the glutamine release amount is 75.00% -97.50% of the glutamine in the original corn protein, and efficient release of the glutamine in the raw materials is realized.
The invention also provides application of the corn glutamine peptide in preparing food, medicine or health care product with intestinal barrier protection function. The corn glutamine peptide prepared by the invention can efficiently release glutamine in corn protein, the release amount of the glutamine accounts for 75.00% -97.50% of that of the original corn protein, the content of pro-inflammatory factors IL-1 beta, IL-6 and TNF-alpha in colon tissues can be obviously reduced, the content of anti-inflammatory factors IL-10 is increased, the weight reduction of colonitis mice is improved, the colonic shortening is relieved, the disease activity index is reduced, the infiltration of inflammatory cells in colon tissues is reduced, the permeability of colonic mucosa is reduced, the colonic inflammation of the mice is relieved, and the effects of relieving acute enteritis and chronic enteritis and protecting intestinal barrier function are achieved.
The technical solutions of the present invention will be clearly and completely described in the following in connection with the embodiments of the present invention. It will be apparent that the described embodiments are only some, but not all, embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1
The preparation of the corn glutamine peptide comprises the following steps:
(1) Pretreatment of raw materials: adjusting the water content of 200g of corn gluten meal (the protein content is about 60 percent, wherein the glutamine content accounts for 8.3 percent of the total protein of the corn) to 15 percent, and extruding and puffing at 170 ℃ by a double screw extruder; drying at room temperature after puffing, crushing and sieving with a 60-mesh sieve to obtain 200g of extruded corn protein powder.
(2) Performing starch removal pretreatment on the extruded corn protein powder: the extruded corn protein powder in the step (1) is prepared into a suspension with the concentration of 10% (m/v) by using 0.02mol/L phosphate buffer with the pH of 6.0, and alpha-amylase with the mass of 1% of the extruded corn protein powder is added for hydrolysis for 2 hours at the temperature of 65 ℃ to remove starch. After the reaction is finished, enzyme is inactivated for 20min at 100 ℃, centrifugation is carried out for 10min at 8000r/min, the precipitate is taken and dried in a baking oven at 60-70 ℃, and is crushed and sieved by a 60-mesh sieve, thus obtaining 150g of pretreated corn protein powder (namely corn protein).
(3) Protease hydrolysis: preparing pretreated corn protein powder in the step (2) into suspension with the substrate concentration of 10% (m/v), adding 195000U (namely 1300U/g protein) alkaline protease, adjusting the pH value of the solution to 8.0, and hydrolyzing at 60 ℃ for 2.5 hours to obtain a first enzymolysis liquid;
(4) Adjusting the pH value of the first enzymolysis liquid to 7.0, adding 30000U (namely 200U/g protein) compound protease, and hydrolyzing at 50 ℃ for 2.5 hours to obtain a second enzymolysis liquid;
(5) Placing the second enzymolysis liquid into a boiling water bath with the temperature of 100 ℃ to inactivate enzymes for 20min, centrifuging for 15min at 4000r/min, and taking supernatant;
(6) Desalting the supernatant with nanofiltration membrane with molecular weight cut-off of 300Da, wherein the nanofiltration parameters are as follows: the volume of the solution is 3L, the operating pressure is 20Bar, the temperature is 20 ℃, the volume is concentrated to 1/2 after nanofiltration is finished, and the corn glutamine peptide of 90g is obtained through freeze drying.
Example 2
The difference is that the alkaline protease in the step (3) is enzymatically decomposed for 2 hours as in example 1.
Example 3
The difference is that the alkaline protease in the step (3) is enzymatically decomposed for 3 hours as in example 1.
Example 4
The difference is that the complex protease of step (4) is enzymatically hydrolyzed for 2 hours as in example 1.
Example 5
The difference is that the complex protease of step (4) is enzymatically hydrolyzed for 3 hours as in example 1.
Comparative example 1
The procedure is as in example 1, except that the complex protease is not used for the enzymolysis.
Comparative example 2
The same as in example 1, except that the alkaline protease was not used for the enzymolysis.
Comparative example 3
The procedure is as in example 1, except that the complex proteolytic enzyme is first carried out and then the alkaline proteolytic enzyme is used.
Test example 1
The glutamine content in the corn glutamine peptides obtained in example 1 and comparative examples 1 to 3 was measured, respectively. Specific: the corn glutamine peptides obtained in example 1 and comparative examples 1 to 3 were divided into two equal mass samples to be tested, wherein one sample was subjected to acid hydrolysis after two hours of protection with bis- (trifluoroacetoxy) -iodobenzene (BTI), the other sample was directly subjected to acid hydrolysis, and the glutamic acid content was measured after the hydrolysis, and the difference between the two equal mass samples was the glutamine content in the corresponding treatment mode.
The results of the examination showed that the content of glutamine in the corn glutamine peptide obtained in example 1 was 12% of the total protein mass, the content of glutamine in the corn glutamine peptide obtained in comparative example 1 was 8.79% of the total protein mass, the content of glutamine in the corn glutamine peptide obtained in comparative example 2 was 8.29% of the total protein mass, and the content of glutamine in the corn glutamine peptide obtained in comparative example 3 was 9.99% of the total protein mass. The corn glutamine peptides obtained in comparative examples 1 to 3 all had a glutamine content lower than that of the present invention, indicating that the alkaline protease and the complex protease of the present invention were high in glutamine content obtained by the sequential hydrolysis of zein.
Test example 2
Corn glutamine peptide has effect of relieving symptoms of mice with DSS-induced intestinal barrier damage.
1. 60 healthy male Kunming mice (purchased from Gibbs laboratory animal technology Co., ltd.) of 6-8 weeks old were randomly divided into 6 groups of 10 animals each: control, model, maize glutamine peptide (APH) low (100 mg/kg.d), medium (300 mg/kg.d), high (500 mg/kg.d) and glutamine (Gln) groups.
2. And (3) establishing a model: mice were gavaged for 14 days, with low, medium, and high doses of corn glutamine peptide (corn glutamine peptide) of example 1, control and model groups were gavaged with equal amounts of saline, and 6 groups were fed with normal water (purchased from Shanghai Biotechnology Co., ltd.). Experiments were started on day 8, specific: the model group and the corn glutamine peptide group were free to drink 3% aqueous Dextran Sodium Sulfate (DSS) (purchased from biotechnology (Shanghai) Co., ltd.) and the control group was fed with water normally. The specific groupings and modes of administration are shown in Table 1.
Table 1 animal experiment group and mode of administration
3. During the modeling period, each group of mice was monitored for changes in body weight and changes in Disease Activity Index (DAI) for each group of mice, and the DAI scoring details are given in table 2 below.
TABLE 2DAI scoring rules
The results of the weight change of the mice in each group are shown in Table 3 and FIG. 1, and the results of the Disease Activity Index (DAI) change of the mice in each group are shown in Table 4 and FIG. 2, wherein the ordinate of FIG. 2 shows the sum of the scores of weight loss, fecal viscosity and fecal occult blood, for example, score 7 in FIG. 2 may indicate that weight loss is 10% and score 2, fecal viscosity is mucoid and score 3, fecal occult blood is perianal blood and score 2, and the score is 7.
TABLE 3 rate of weight change of mice in each group during molding
TABLE 4 Disease Activity Index (DAI) for groups of mice during modeling
As can be seen from tables 3-4 and FIGS. 1-2, different doses of corn glutamine peptide significantly improved weight loss in mice and decreased disease activity index compared to the model group.
Mice from each group were gavaged for 14 days in the manner of Table 1, and after 12 hours of last feeding, the mice were sacrificed, the colon length of each group was measured, and colon tissues were taken to detect cytokines such as TNF- α, IL-1β, IL-6, and IL-10, and the detection results are shown in Table 5 and FIGS. 3 to 7.
TABLE 5 colon length and inflammatory factor content in mice 14 days after gastric lavage
As can be seen from table 5 and figures 3-7, different doses of corn glutamine peptide can alleviate colonic shortening compared to the model group. After DSS treatment, the content of pro-inflammatory cytokines IL-1 beta, IL-6 and TNF-alpha in colon tissues is increased to respectively reach 1.30ng/g colon tissue, 1.13ng/g colon tissue and 6.32ng/g colon tissue, and the content of anti-inflammatory cytokine IL-10 is reduced to reach 2.25ng/g colon tissue. Compared with the model group, after being treated by corn glutamine peptide with different dosages, the content of pro-inflammatory factors IL-1 beta, IL-6 and TNF-alpha in colon can be obviously reduced, and the content of anti-inflammatory cytokine IL-10 can be increased. In particular to a corn glutamine peptide high dose group, which reduces pro-inflammatory cytokines IL-1 beta, IL-6 and TNF-alpha by 19.23 percent, 27.88 percent and 33.76 percent respectively, increases anti-inflammatory cytokine IL-10 by 66.81 percent, which is equivalent to the level of a normal group, is superior to that of a Gln group, and shows that the corn glutamine peptide with a certain dose can effectively relieve DSS-induced intestinal barrier damage, has a certain protective effect on the intestinal barrier, enables the intestinal barrier to recover to the normal level, and has the effect superior to that of the single Gln.
According to the embodiment, the corn glutamine peptide prepared by the preparation method can efficiently release glutamine in corn protein, effectively improve weight loss of mice, reduce disease activity index, relieve colon shortening, reduce the content of pro-inflammatory factors IL-1 beta, IL-6 and TNF-alpha in colon, increase the content of anti-inflammatory cytokine IL-10, effectively relieve DSS-induced intestinal barrier injury, and have a certain protection effect on intestinal barrier.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (7)
1. The application of corn glutamine peptide in preparing medicine for treating ulcerative colitis;
the preparation method of the corn glutamine peptide comprises the following steps:
carrying out first enzymolysis on zein by using alkaline protease to obtain a first enzymolysis liquid;
carrying out second enzymolysis on the first enzymolysis liquid by using compound protease Protamex to obtain second enzymolysis liquid, and drying to obtain corn glutamine peptide;
the temperature of the first enzymolysis is 60-70 ℃, the time is 2-3 h, and the pH value is 7.5-8.5;
the temperature of the second enzymolysis is 45-55 ℃, the time is 2-3 h, and the pH value is 6.5-7.5.
2. The use according to claim 1, wherein the alkaline protease is used in an amount of 1000 to 1500U/g zein when the first enzymatic hydrolysis is performed.
3. The use according to claim 1, wherein the amount of the complex protease Protamex used in the second enzymatic hydrolysis is 100-300U/g zein.
4. The use according to claim 1, wherein the zein comprises a de-starched zein powder; the starch removal treatment comprises: mixing corn protein powder to be treated with alpha-amylase, and performing starch removal enzymolysis to obtain corn protein; the mass of the alpha-amylase is 0.8% -1.2% of the mass of the corn gluten meal to be treated.
5. The use according to claim 4, wherein the temperature of the starch-removing enzymolysis is 60-70 ℃ and the time of the starch-removing enzymolysis is 1.5-2.5 h.
6. The use according to claim 1, further comprising nanofiltration of the second enzymatic hydrolysate prior to drying; the pressure of the nanofiltration is 15-25 Bar, and the temperature is 15-25 ℃.
7. The use according to any one of claims 1 to 6, wherein the mass of glutamine in the corn glutamine peptide is 10% to 13% of the total protein mass in the corn glutamine peptide.
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