CN108060200B - Preparation method of ACE inhibitory peptide composition - Google Patents

Preparation method of ACE inhibitory peptide composition Download PDF

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CN108060200B
CN108060200B CN201810031313.1A CN201810031313A CN108060200B CN 108060200 B CN108060200 B CN 108060200B CN 201810031313 A CN201810031313 A CN 201810031313A CN 108060200 B CN108060200 B CN 108060200B
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林晓婕
何志刚
林晓姿
李维新
梁璋成
任香芸
苏昊
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Fujian Academy Of Agricultural Sciences Agricultural Product Processing Research Institute
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Abstract

The invention provides a preparation method of ACE inhibitory peptide, which comprises the steps of 1) drying and crushing red yeast yellow wine lees at 55-65 ℃; 2) adding 8-12 times of water by weight, heating in water bath at 45-55 ℃, adjusting the pH value to 8-10, and then adding 0.5-1.5 wt% of protease complex for hydrolysis, wherein the protease complex comprises alkaline protease, neutral protease and papain, and the weight ratio of the protease complex to the protease complex is 520-: 250-280: 160-200; 3) inactivating enzyme of the hydrolyzed suspension at a temperature of more than 90 ℃ to terminate the hydrolysis reaction; 4) centrifuging the suspension after enzyme deactivation, collecting supernatant 5) purifying and removing impurities from the supernatant to obtain the ACE inhibitory peptide composition. According to the scheme, the red yeast yellow wine lees are subjected to mixed enzyme enzymolysis, a high-activity natural ACE inhibitory peptide composition can be prepared from the red yeast yellow wine lees, the ACE inhibitory rate of the composition can reach more than 85%, and the peptide content reaches 16-23 mg/mL. The method is simple and easy to operate, and is suitable for large-scale industrial production.

Description

Preparation method of ACE inhibitory peptide composition
Technical Field
The invention relates to the field of food processing, in particular to a preparation method of an ACE inhibitory peptide composition.
Background
With the social development and the improvement of the production and living standards of people, the average age of people is increased, the dietary structure is changed, and the salt intake is increased, so that the prevalence rate of hypertension and hypertensive heart damage is increased. The hypertension is not only an independent disease, but also an important risk factor of cerebral apoplexy, coronary heart disease, renal failure and eyeground pathological changes, and patients with hypertension are often accompanied with chronic diseases such as diabetes and the like, so that the effective prevention and control of the hypertension have important significance on human health.
The antihypertensive peptide is called angiotensin I Converting Enzyme (ACE) inhibitory peptide, and is a short-chain polypeptide substance which is separated from food protein and has the effect of remarkably reducing blood pressure. The antihypertensive peptides derived from food are generally obtained by hydrolyzing proteins by protease under mild conditions, have high edible safety, and have the common outstanding advantages of only having the effect of reducing blood pressure of patients with hypertension and having no effect of reducing blood pressure of normal blood pressure patients, so that the phenomenon of excessive blood pressure reduction is avoided. ACE inhibitory peptides from different raw materials comprise different amino acids and sequences. The Wuqiang and the like adopt a distributed enzymolysis process: firstly carrying out enzymolysis by alkaline protease and then carrying out enzymolysis by papain, and preparing ACE inhibitory peptide by utilizing the gonad of the abalone, wherein the ACE inhibitory rate reaches 78.05%; korean Fei and the like adopt alkaline protease and optimize the enzymolysis conditions of the alkaline protease, and prepare ACE inhibitory peptide by utilizing soybean protein, wherein the ACE inhibitory rate reaches 84.1%; zhang Wei and the like adopt alkaline protease and optimize the enzymolysis conditions of the alkaline protease, and prepare ACE inhibitory peptide by utilizing peanut protein, wherein the ACE inhibitory rate reaches 72.78%; zhengxung and the like adopt pepsin and a response surface method to optimize enzymolysis conditions, and prepare ACE inhibitory peptide by utilizing porcine hemoglobin, wherein the ACE inhibitory rate reaches 70.09%.
Disclosure of Invention
Therefore, it is desirable to provide a method for preparing a highly active natural ACE inhibiting peptide composition. To achieve the above object, the inventors provide a method for preparing an ACE inhibitory peptide composition, comprising the steps of:
1) drying red yeast yellow wine lees at 55-65 deg.C, and pulverizing;
2) adding 8-12 times of water into the dried and crushed red yeast yellow wine lees, uniformly stirring, and heating to 45-55 ℃ to obtain a suspension; adjusting pH of the suspension to 8-10, adding 0.5-1.5 wt% protease complex, and hydrolyzing at 45-55 deg.C and pH 8-10 for 2-3 hr; the protease complex comprises alkaline protease, neutral protease and papain, and the weight ratio of the alkaline protease to the neutral protease is 520-560: 250-280: 160-200;
3) inactivating enzyme of the hydrolyzed suspension at a temperature of more than 90 ℃ to terminate the hydrolysis reaction;
4) centrifuging the suspension after enzyme deactivation, collecting supernatant,
5) purifying and removing impurities from the supernatant to obtain the ACE inhibitory peptide composition.
Preferably, in the step 3), the pH value of the suspension is adjusted to 8.4-8.6, protease with the weight percentage of 0.5-1.5% is added into the suspension for hydrolysis, the hydrolysis temperature is controlled at 50 ℃, and the hydrolysis time is 3 hours.
Preferably, the mass ratio of the alkaline protease to the neutral protease to the papain is 547:268: 185.
Preferably, the content of the peptide in the ACE inhibitory peptide composition is 16-23 mg/mL.
Different from the prior art, the scheme of carrying out mixed enzyme enzymolysis on the red yeast yellow wine lees provided by the technical scheme can prepare a high-activity natural ACE inhibitory peptide composition from the red yeast yellow wine lees, the ACE inhibitory rate of the composition can reach more than 85%, and the peptide content reaches 16-23 mg/mL. The method is simple and easy to operate, and is suitable for large-scale industrial production. Meanwhile, the composition has rich nutrition and good delicate flavor and sweet taste, and can be used for developing high-grade flavor seasonings and used as a functional food additive. Meanwhile, the composition can be absorbed by the body to a high degree, and can be widely applied to the fields of functional foods and medical biology as a crude product after further purification.
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FIG. 1 is a graph showing the effect of different protease enzymatic digestions on ACE inhibition;
FIG. 2 is a graph of the response of the enzyme mixture design test;
FIG. 3 is a contour plot of a mixed enzyme design assay;
fig. 4 is a graph showing the effect of various factors on ACE inhibition.
Detailed Description
To explain technical contents, structural features, and objects and effects of the technical solutions in detail, the following detailed description is given with reference to the accompanying drawings in conjunction with the embodiments.
Food-borne ACE inhibitory peptides overview ACE inhibitory peptides, angiotensin converting enzyme inhibitory peptides are polypeptide substances having Angiotensin Converting Enzyme (ACE) inhibitory activity, and the amino acid sequences and the peptide chain lengths of the polypeptides are different, but all have similar functions, namely, the function of lowering blood pressure, and are also called antihypertensive peptides.
The red yeast yellow wine is an important brewed wine in Fujian province, and the main byproduct of yellow wine production is red yeast yellow wine lees which are solid matters left after fermented mash is squeezed and separated to remove wine liquid. The red rice yellow wine can produce 20-30% of distiller's grains in the production process, and can be used as food processing material. The red yeast yellow wine lees in the embodiment is purchased from Fujiangningde yellow wine industry, and the protein content is 38.2 percent and the water content is 10.3 percent through detection.
In this embodiment, the formula for calculating the ACE inhibition ratio of the I-sample is:
I＝(Sa-Co)/Sa×100％
wherein, I is the ACE inhibition rate of the sample; sa-peak area of hippuric acid in control group; co-area of hippuric acid peak in test group. The HPLC chromatographic conditions are as follows: zorbax SB-C18 analytical column; the column temperature is 30 ℃; mobile phase: 30% acetonitrile (containing 0.05% acetic acid); flow rate: 1.0 mL/min; the sample injection volume is 5 mu L; the detection wavelength is 228 nm.
1. Screening test for protease:
the inventor further carried out a screening test of the red yeast yellow wine lees protease, respectively carried out enzymolysis by five proteases under respective proper action conditions (table 1), and carried out preliminary screening by taking the ACE inhibition rate as an index, and the result is shown in figure 1.
TABLE 1 conditions of enzymatic hydrolysis of proteases
Figure BDA0001546559540000041
As can be seen from figure 1, the difference of ACE inhibition rate of each yellow wine lees enzymatic hydrolysate is large, the ACE inhibition rate of the alkaline protease enzymatic hydrolysate is the highest and reaches 88.550%, and neutral protease and papain are used as secondary factors. These three enzymes were selected for mixed enzyme studies.
2. Material mixing design test:
the inventor also carries out a mixed material design test, takes the inhibition rate as an index, and optimizes the proportion combination and reaction conditions of the 3 enzymes.
2.1Scheffe simplex lattice design:
the mixed enzyme research is carried out by adopting a Scheffe simplex type grid design, the mixed material design test is shown in table 2, A, B, C respectively shows the percentage of alkaline protease, neutral protease and papain in a mixed material system, and the mixing condition A + B + C is 1. The protocol and results are shown in Table 2.
TABLE 2 compounding design test arrangement and results and predicted values
Figure BDA0001546559540000042
Figure BDA0001546559540000051
2.2 regression model establishment and analysis of variance:
and (3) performing multivariate regression fitting analysis on the response value by using Design expert8.0.5 software, wherein an incomplete cubic regression model equation is as follows:
Y＝89.630A+77.04213924B+63.879C+19.023AB+28.321AC+36.321BC+112.240ABC
the results of the anova are shown in table 3. Incomplete cubic regression model and assistance (P < 0.0001), determination coefficient R-Squared which is 0.9939, and correction determination coefficient Adj R-Squared which is 0.989 are both more than 0.9, which shows that the model can well fit each index and the mixed enzyme ratio.
TABLE 3 regression model equation analysis of variance
Figure BDA0001546559540000052
2.3 predicting the optimal mixed enzyme ratio:
the response surface and contour plots of the mixed enzyme design assay are shown in FIGS. 2 and 3. Fig. 2 shows that the slope of the response surface is very steep, indicating that the ACE inhibition rate is very sensitive to changes in the mixing ratio of the three enzymes. By optimizing by using a Design expert8.0.5 software model, the optimal mixed enzyme ratio is calculated as follows: 54.7% of alkaline protease, 26.8% of neutral protease and 18.5% of papain, and under the condition, the ACE inhibition rate is 91.990%.
2.4 optimal mixed enzyme ratio verification test:
in order to verify the accuracy of the model prediction result, the test is carried out under the process parameters of model optimization, the test is repeated for 3 times, the ACE inhibition rates are 92.833%, 92.027% and 92.990% respectively, the average value is 92.617%, the relative error xi between the measured value and the model prediction value is 0.681%, and the model fitting degree is good.
2.5 Mixed enzyme enzymolysis conditions Single factor test:
in order to determine the optimal hydrolysis conditions of the mixed enzyme, four factors and three levels L are adopted for the temperature, time, enzyme adding amount and pH value of the mixed enzyme9(34) And (4) orthogonal experimental design. The inhibition ratio was used as an index, and the design factor level scheme is shown in table 4.
TABLE 4L9(34) Orthogonal experimental design factors and levels
Figure BDA0001546559540000061
The influence of various factors of the mixed enzyme on the ACE inhibition rate of the enzymolysis liquid is shown in a graph of the influence of various factors on the ACE inhibition rate in fig. 4.
The suppression rate tends to increase and decrease with increasing temperature. When the temperature is 50 ℃, the ACE inhibition rate of the enzymolysis liquid is the highest. This is probably because the proper temperature promotes the interaction between the catalytic group of the enzyme and the substrate, increases the enzymolysis probability, and increases the content of short peptides in the enzymolysis liquid, thereby increasing the ACE inhibition rate. When the temperature exceeds 50 ℃, the enzyme denaturation is increased at high temperature, the activity is reduced, and the enzymolysis degree is reduced.
The inhibition rate rises firstly and then falls with the time extension, the protein is gradually degraded into short peptide with ACE inhibitory activity in the initial stage of enzymolysis, and the short peptide is further degraded into peptide and amino acid with smaller molecular weight with the time extension of enzymolysis, so that the inhibitory activity is reduced.
As the amount of the enzyme added increases, the inhibition rate increases and then becomes gentle. When the enzyme adding amount is 1%, the inhibition rate reaches the highest value. The reason may be that when a small amount of enzyme is added, the amount of enzyme is insufficient to completely bind to the substrate, the rate of proteolysis is low, and the inhibitory peptide produced is small. With the increase of the enzyme adding amount, the enzyme is completely combined with the substrate to reach a saturated state, the enzymolysis rate cannot be increased by continuously adding the enzyme, and the inhibition rate tends to be flat.
The inhibition rate tends to increase and decrease with increasing pH, and the inhibition rate reaches a maximum value at pH 8.5. This is probably because, at pH8.5, each enzyme maintains a high activity, which facilitates further degradation of the enzymatic hydrolysate into short peptides of smaller molecular weight, thereby increasing the inhibition rate. Higher or lower pH inhibited the activity of each of the other enzymes except the pH-optimum enzyme.
Mixed enzyme enzymolysis condition orthogonal test: on the basis of the single-factor test, the mixed enzyme enzymolysis condition orthogonal test is carried out, and the test results are shown in the table.
TABLE 5 results and analysis of orthogonal experiments
Figure BDA0001546559540000071
It can be seen that the time and pH are first and second order factors that influence the inhibition rate. The analysis of variance is shown in Table 6.
TABLE 6 ANOVA TABLE
Figure BDA0001546559540000072
As can be seen from the table, the ACE inhibition rate of the enzymolysis product is extremely obvious (P is less than 0.01) by the time, the pH and the temperature. The main and secondary sequence of the influence of each factor of enzymolysis on the ACE inhibition rate of an enzymolysis product is that B is more than D and more than A is more than C, namely, time is more than pH and more than temperature, and enzyme adding amount is more than. The optimum conditions of enzymolysis are as follows: the A2B3C3D2 is the most preferable combination of A2B3C1D2, namely, the temperature is 50 ℃, the time is 3h, the enzyme adding amount is 0.5 percent and the pH is 8.5, considering that the difference of the enzyme adding amount C is not significant and the enzyme adding amount directly influences the production cost.
2.6 macroporous resin purification impurity removal test:
soaking a certain amount of resin in absolute ethyl alcohol for 24h, and then washing the resin with absolute ethyl alcohol until no absorption exists at 220nm for later use.
Column conditions were as follows: glass column: 2.6X 40 cm; bed Volume (BV): 180 mL; resin: DA 201-C; sample loading concentration: 20 mg/mL; the sample loading flow rate is 1/2 BV/h; the loading volume is 50 ml. Washing with distilled water at a flow rate of 1BV/h until the conductivity is basically unchanged; the elution flow rate is 1BV/h with 80% ethanol.
Example 1
A preparation method of an ACE inhibitory peptide composition comprises the following steps:
1) drying red yeast yellow wine lees at 60 ℃, and crushing for later use;
2) adding 10 times of water by weight into the red yeast yellow wine lees dried and crushed in the step 1), uniformly stirring, and heating in a water bath to 50 ℃ to obtain a suspension; adjusting the pH value of the suspension to 8.5, adding 1 weight percent of protease compound for hydrolysis, controlling the hydrolysis temperature to be 50 ℃ and the hydrolysis time to be 3 hours; the protease compound comprises alkaline protease, neutral protease and papain in a weight ratio of 547:268: 185;
3) inactivating enzyme of the hydrolyzed suspension at 90 deg.C for 10min, and terminating hydrolysis reaction;
4) centrifuging the suspension after enzyme deactivation at 10000r/min for 15 minutes, collecting supernatant,
5) and (3) carrying out macroporous resin column chromatography on the supernatant, washing the supernatant by using distilled water to remove salt, sugar and macromolecular impurities, eluting the supernatant by using 80% ethanol, and collecting eluent which is the ACE inhibitory peptide composition.
The column conditions were as follows: glass column: 2.6X 40 cm; bed Volume (BV): 180 mL; resin: DA 201-C; sample loading concentration: 20 mg/mL; the sample loading flow rate is 1/2 BV/h; the loading volume is 50 ml. The resin is required to be soaked in absolute ethyl alcohol for 24 hours and then packed into a column, and the chromatographic column is used when the chromatographic column is required to be washed by absolute ethyl alcohol until no absorption exists at a position of 220 nm. Washing with distilled water as eluent at flow rate of 1BV/h until the conductivity is basically unchanged; then eluting with 80% ethanol at the flow rate of 1 BV/h.
The ACE inhibitory peptide composition prepared in example 1 has an ACE inhibitory rate of 92.814% and a peptide content of 22.4 mg/mL. In the examples, peptides were detected as GBT 22492-.
Example 2
A preparation method of an ACE inhibitory peptide composition comprises the following steps:
1) drying red yeast yellow wine lees at 60 ℃, and crushing for later use;
2) adding 10 times of water by weight into the red yeast yellow wine lees dried and crushed in the step 1), uniformly stirring, and heating in a water bath to 45 ℃ to obtain a suspension; adjusting the pH value of the suspension to 8.5, adding 1 weight percent of protease compound for hydrolysis, controlling the hydrolysis temperature to 45 ℃ and the hydrolysis time to 2 hours; the protease compound comprises alkaline protease, neutral protease and papain in a weight ratio of 520:250: 160;
3) inactivating enzyme of the hydrolyzed suspension at 100 ℃ for 10min, and terminating the hydrolysis reaction;
4) centrifuging the suspension after enzyme deactivation at 10000r/min for 15 minutes, collecting supernatant,
5) and (3) carrying out macroporous resin column chromatography on the supernatant, washing the supernatant by using distilled water to remove salt, sugar and macromolecular impurities, eluting the supernatant by using 80% ethanol, and collecting eluent which is the ACE inhibitory peptide composition.
The column conditions were as follows: glass column: 2.6X 40 cm; bed Volume (BV): 180 mL; resin: DA 201-C; sample loading concentration: 20 mg/mL; the sample loading flow rate is 1/2 BV/h; the loading volume is 50 ml. The resin is required to be soaked in absolute ethyl alcohol for 24 hours and then packed into a column, and the chromatographic column is used when the chromatographic column is required to be washed by absolute ethyl alcohol until no absorption exists at a position of 220 nm. Washing with distilled water as eluent at flow rate of 1BV/h until the conductivity is basically unchanged; then eluting with 75% ethanol at the flow rate of 1 BV/h.
The ACE inhibitory peptide composition prepared in example 2 has an ACE inhibitory rate of 87.159% and a peptide content of 18.73 mg/mL. In the examples, peptides were detected as GBT 22492-.
Example 3
A preparation method of an ACE inhibitory peptide composition comprises the following steps:
1) drying red yeast yellow wine lees at 55 ℃, and crushing for later use;
2) adding water in an amount which is 12 times the weight of the red yeast yellow wine lees dried and crushed in the step 1), uniformly stirring, and heating in a water bath to 55 ℃ to obtain a suspension; adjusting the pH value of the suspension to 8.5, adding 0.5 wt% of protease complex for hydrolysis, controlling the hydrolysis temperature to 55 ℃ and the hydrolysis time to 2 hours; the protease compound comprises alkaline protease, neutral protease and papain, and the weight ratio of the alkaline protease to the neutral protease to the papain is 560:280: 200;
3) inactivating enzyme of the hydrolyzed suspension at 90 deg.C for 10min, and terminating hydrolysis reaction;
4) centrifuging the suspension after enzyme deactivation at 12000r/min for 10min, collecting supernatant,
5) and (3) carrying out macroporous resin column chromatography on the supernatant, washing the supernatant by using distilled water to remove salt, sugar and macromolecular impurities, eluting the supernatant by using 80% ethanol, and collecting eluent which is the ACE inhibitory peptide composition.
The column conditions were as follows: glass column: 2.6X 40 cm; bed Volume (BV): 180 mL; resin: DA 201-C; sample loading concentration: 20 mg/mL; the sample loading flow rate is 1/2 BV/h; the loading volume is 50 ml. The resin is required to be soaked in absolute ethyl alcohol for 24 hours and then packed into a column, and the chromatographic column is used when the chromatographic column is required to be washed by absolute ethyl alcohol until no absorption exists at a position of 220 nm. Washing with distilled water as eluent at flow rate of 1BV/h until the conductivity is basically unchanged; then eluting with 85% ethanol at the flow rate of 1 BV/h.
The ACE inhibitory peptide composition prepared in example 3 had an ACE inhibitory rate of 90.904% and a peptide content of 20.89 mg/mL. In the examples, peptides were detected as GBT 22492-.
The inventor finds out in research and analysis of amino acids in the red yeast yellow wine lees ACE inhibitory peptide that: the mass fractions of the unnecessary amino acids in the red yeast yellow wine lees are respectively 10.90 percent of aspartic acid, 14.30 percent of glutamic acid, 5.42 percent of serine, 2.36 percent of histidine, 4.97 percent of arginine, 6.04 percent of glycine, 5.37 percent of proline and 5.32 percent of alanine. The mass fraction and the nutrition score of the essential amino acids in the red yeast yellow wine lees ACE inhibitory peptide are shown in the table 1. The total mass fraction of 18 amino acids in the red yeast yellow wine lees ACE inhibitory peptide is 88.91%, and the first limiting amino acid is tryptophan and accounts for 0.75% of the total amino acids. The essential amino acid in the red yeast yellow wine lees ACE inhibitory peptide accounts for 41.30% of the total amino acid. The mass fraction of the essential amino acids is 8.36 percent of leucine at most. Table 1 shows that, in addition to methionine, cysteine and tryptophan, each essential amino acid CS is large and has a composition close to that of the corresponding essential amino acid of standard egg protein. Lysine is the first limiting amino acid of most grains, while lysine AAS is higher in monascus yellow wine lees ACE inhibitory peptide, probably because of microbial metabolism during monascus yellow wine fermentation. Therefore, the red yeast yellow wine lees ACE inhibitory peptide has better nutrition balance than the five-cereal coarse cereals. The EAAI, BV and NI of the red yeast yellow wine lees ACE inhibitory peptide are 76.52, 71.70 and 74.99 respectively, which shows that the red yeast yellow wine lees ACE inhibitory peptide can be absorbed by organisms to a higher degree. The SRCAA is 79.03, which shows that the dispersion of various essential amino acids in the red yeast yellow wine lees ACE inhibitory peptide deviating from the mode amino acid is small, the nutritive value is relatively high, and the red yeast yellow wine lees ACE inhibitory peptide can be determined to be a high-quality protein source. The total mass fraction of the umami amino acids (aspartic acid, glutamic acid, phenylalanine, valine and leucine) and the glycine in the red yeast yellow wine lees ACE inhibitory peptide reaches 53.50%. Has good delicate flavor and sweet flavor, can be used as a functional food additive, and has good taste and consumer acceptance.
Table 7 amino acid composition in red grain and score
Figure BDA0001546559540000111
Figure BDA0001546559540000121
Amino acid analysis: the amino acid content was determined by the method outlined in the determination of amino acids in GB/T5009.124-2003 food.
The analysis shows that the ACE inhibitory peptide composition prepared from red yeast yellow wine lees has high absorption and utilization degree by organisms, has rich nutrition, good delicate flavor and sweet flavor, can be used as a functional food additive, and has high acceptable degree of mouthfeel for consumers.
It is noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or terminal that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or terminal. Without further limitation, an element defined by the phrases "comprising … …" or "comprising … …" does not exclude the presence of additional elements in a process, method, article, or terminal that comprises the element. Further, herein, "greater than," "less than," "more than," and the like are understood to exclude the present numbers; the terms "above", "below", "within" and the like are to be understood as including the number.
It should be noted that, although the above embodiments have been described herein, the invention is not limited thereto. Therefore, based on the innovative concepts of the present invention, the technical solutions of the present invention can be directly or indirectly applied to other related technical fields by making changes and modifications to the embodiments described herein, or by using equivalent structures or equivalent processes performed in the content of the present specification and the attached drawings, which are included in the scope of the present invention.

Claims (4)

1. A preparation method of an ACE inhibitory peptide composition is characterized by comprising the following steps:
1) drying red yeast yellow wine lees at 55-65 deg.C, and pulverizing;
2) adding 8-12 times of water into the dried and crushed red yeast yellow wine lees, uniformly stirring, and heating to 45-55 ℃ to obtain a suspension; adjusting pH of the suspension to 8-10, adding 0.5-1.5 wt% protease complex, and hydrolyzing at 45-55 deg.C and pH 8-10 for 2-3 hr; the protease complex comprises alkaline protease, neutral protease and papain, and the weight ratio of the alkaline protease to the neutral protease is 520-560: 250-280: 160-200;
3) inactivating enzyme of the hydrolyzed suspension at a temperature of more than 90 ℃ to terminate the hydrolysis reaction;
4) centrifuging the suspension after enzyme deactivation, and collecting supernatant;
5) purifying and removing impurities from the supernatant to obtain the ACE inhibitory peptide composition.
2. The method for preparing the ACE inhibitory peptide composition according to claim 1, wherein in the step 2), the pH of the suspension is adjusted to 8.4-8.6, and the suspension is hydrolyzed by adding 0.5-1.5 wt% of protease, wherein the hydrolysis temperature is controlled to 50 ℃ and the hydrolysis time is controlled to 3 hours.
3. The method for preparing the ACE inhibitory peptide composition according to claim 1, wherein the mass ratio of the alkaline protease to the neutral protease to the papain is 547:268: 185.
4. The method for preparing the ACE inhibitory peptide composition of claim 1, wherein the peptide content of the ACE inhibitory peptide composition is 16-23 mg/mL.
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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007054933A3 (en) * 2005-11-10 2007-12-06 Rimonest Ltd Methods and compositions employing pomegranate extracts and fementation products of statin-producing fungi
CN105002249A (en) * 2015-08-14 2015-10-28 湖北工业大学 Method for preparing ACE (angioensin I converting enzyme) inhibitory peptides through liquid state fermentation of monascus
CN105524966A (en) * 2016-02-26 2016-04-27 天津现代职业技术学院 Method for preparing ACE inhibitory peptides through bean pulp enzymolysis
CN105646646A (en) * 2016-03-17 2016-06-08 湖北工业大学 Purification method of monascus thallus ACE (Angioensin I Converting Enzyme) inhibitory peptide

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007054933A3 (en) * 2005-11-10 2007-12-06 Rimonest Ltd Methods and compositions employing pomegranate extracts and fementation products of statin-producing fungi
CN105002249A (en) * 2015-08-14 2015-10-28 湖北工业大学 Method for preparing ACE (angioensin I converting enzyme) inhibitory peptides through liquid state fermentation of monascus
CN105524966A (en) * 2016-02-26 2016-04-27 天津现代职业技术学院 Method for preparing ACE inhibitory peptides through bean pulp enzymolysis
CN105646646A (en) * 2016-03-17 2016-06-08 湖北工业大学 Purification method of monascus thallus ACE (Angioensin I Converting Enzyme) inhibitory peptide

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