CN117243370A

CN117243370A - Western Mei Jiaosu and preparation method thereof

Info

Publication number: CN117243370A
Application number: CN202311318653.XA
Authority: CN
Inventors: 赵文竹; 杨曼妮; 颜林悦之; 于志鹏
Original assignee: Hainan University; Sanya Research Institute of Hainan University
Current assignee: Hainan University; Sanya Research Institute of Hainan University
Priority date: 2023-10-11
Filing date: 2023-10-11
Publication date: 2023-12-19

Abstract

The invention relates to the technical field of food processing, in particular to a western Mei Jiaosu and a preparation method thereof. The method comprises the following steps: preparing western Mei Yun slurry; enzymolysis: adding cellulase, carrying out water bath for 3-4 hours at 35-40 ℃, and inactivating enzyme; saccharifying for 0.5-1 h by adding saccharifying enzyme, inactivating enzyme and cooling; regulating pH value to 3.5-4.5, adding amylase and papain mixed enzyme, heating in water bath at 55-65 deg.c for 2.5-3.5 hr, inactivating enzyme, filtering and sterilizing; fermentation: according to the volume ratio of the feed liquid of 3:7, adding water, inoculating saccharomycetes, adding white granulated sugar and ascorbic acid, uniformly mixing, and carrying out primary fermentation at the temperature of 27-32 ℃ for 4-6 hours; inoculating lactobacillus plantarum, and performing secondary fermentation at the temperature of 33-37 ℃ for 8-12 hours; filtering with 0.45 μm filter membrane, and packaging. The content of functional components in the product is increased by optimizing the preparation process, so that the flavor and the nutritional value are improved.

Description

Western Mei Jiaosu and preparation method thereof

Technical Field

The invention relates to the technical field of food processing, in particular to a western Mei Jiaosu and a preparation method thereof.

Background

With the acceleration of the current life rhythm and the increase of life pressure, the work and rest laws and eating habits of people are changed. Food intake of "three high" (high sugar, high fat, high calorie) is increasing, but food intake contributing to intestinal peristalsis, such as dietary fiber-rich food intake is greatly reduced, and constipation is becoming increasingly common. Meanwhile, with age, constipation problems in the elderly are becoming more common, as well as physical activity is reduced and a variety of chronic diseases are involved. Typical symptoms of constipation include dry stool, difficulty in defecation, prolonged defecation time, and the like. Since the long-term dependence of drugs for constipation may affect the normal functions of the intestinal tract and may cause adverse reactions such as diarrhea, abdominal pain, nausea, dehydration, etc., people start worrying about the method of drugs for constipation. Under such a background, more and more researchers are working on developing novel functional foods with the effect of relaxing bowel so as to meet the demands of people for more safely solving the constipation problem.

The prune is a low calorie fruit, and contains abundant vitamin C, vitamin K, vitamin A, dietary fiber, antioxidant substances, and various minerals such as potassium, magnesium, and calcium. In addition, prune is also rich in plant compounds such as polyphenols and carotenoids. Functionally, prunes have multiple benefits. Firstly, the soluble fiber is abundant, which is helpful for stimulating intestinal peristalsis and preventing constipation. Second, the antioxidant substances, such as vitamin C and polyphenols, help neutralize free radicals, reduce the adverse effects of oxidative stress on physical health, and thereby enhance the immune system function. In addition, the potassium element contained in the blood pressure regulator can regulate blood pressure, and the vitamin K can participate in the blood coagulation process, so that the blood pressure regulator is beneficial to maintaining cardiovascular health.

The ferment product is prepared by taking fresh vegetables, fruits and other plants as raw materials through the processes of juicing, extracting and the like. Then adding fermentation strain for fermentation, the ferment contains rich saccharide, organic acid and other bioactive components, and important enzymes, which are composed of amino acids and have special bioactivity. Enzymes exist in all living animals and plants, and are important to maintain normal body functions, promote food digestion, repair tissues and other vital activities. However, the current ferment products have some disadvantages, such as insufficient nutrition ingredients, low content of functional ingredients, and the like. In addition, many ferment products in the prior art also adopt natural fermentation, which leads to the doping of mixed bacteria and unstable properties of the fermented products; and the fermentation period of some ferment products is too long, the industrial production efficiency is low, and the market demand can not be met.

Disclosure of Invention

The invention provides a western Mei Jiaosu and a preparation method thereof for solving the problems.

The first aim of the invention is to provide a preparation method of the Western Mei Jiaosu, which comprises the following steps:

s1, preparing western Mei Yun slurry: cleaning fresh prune, removing cores, cutting into blocks, soaking in ascorbic acid solution for 8-15 min after cutting into blocks, pulping to obtain prune Mei Yun pulp;

s2, enzymolysis: adding cellulase into the western Mei Yun pulp prepared in the step S1, carrying out water bath for 3-4 hours at the temperature of 35-40 ℃, boiling and inactivating enzyme; adding saccharifying enzyme, saccharifying at 60-70 deg.c for 0.5-1 hr, boiling to deactivate enzyme and cooling; regulating the pH value to 3.5-4.5, adding amylase and papain mixed enzyme, heating in a water bath at 55-65 ℃ for 2.5-3.5 h, boiling to inactivate enzyme, filtering out excessive water and sterilizing;

s3, fermenting: according to the volume ratio of the feed liquid of 3: adding water, and uniformly mixing to obtain the to-be-fermented western Mei Jiang; inoculating saccharomycetes, respectively adding 3-5% of white granulated sugar and 0.1-0.2% of ascorbic acid into the to-be-fermented western Mei Jiang, uniformly mixing, and carrying out primary fermentation at the temperature of 27-32 ℃ for 4-6 hours; inoculating lactobacillus plantarum, and performing secondary fermentation at the temperature of 33-37 ℃ for 8-12 hours;

s4, packaging: filtering with 0.45 μm filter membrane after fermentation, and packaging the liquid to obtain the final product of western Mei Jiaosu.

Preferably, the cellulase, saccharifying enzyme, alpha-amylase and papain are added in the step S2 in an amount of 0.8-1.2%, 0.1-0.3%, 0.5-1% and 0.4-0.8% of the mass of the prune homogenate, respectively.

Preferably, the inoculation amount of the saccharomycetes and the lactobacillus plantarum in the step S3 is 3-5% and 2-4% of the mass of the prune pulp to be fermented.

Preferably, the inoculation amount of the saccharomycetes and the lactobacillus plantarum in the step S3 is 4% and 3% of the mass of the prune pulp to be fermented.

Preferably, the fermentation temperature of the primary fermentation is 31 ℃; the fermentation temperature of the secondary fermentation was 34 ℃.

Preferably, the fermentation time of the primary fermentation is 5 hours; the fermentation time of the secondary fermentation is 10 hours.

Preferably, the mass ratio of amylase to papain is 5:3.

preferably, the saccharomycetes are pretreated before being added, and the pretreatment is as follows: adding ultrapure water into a certain amount of saccharomycetes according to the volume ratio of 1:100, uniformly mixing, and carrying out water bath for 18-25 min at the temperature of 23-27 ℃.

A second object of the present invention is to provide a western Mei Jiaosu prepared by the method for preparing a western Mei Jiaosu.

Compared with the prior art, the invention has the following beneficial effects:

the invention fully utilizes the functional advantages of the prune and carries out compound fermentation with a specific fermentation strain, thereby developing the prune Mei Jiaosu. In addition, the content of functional components in the product is increased by optimizing the preparation process, so that the flavor and the nutritional value of the product are improved, and the actual effect of the product in the health field is improved.

The method has the advantages that the prune is taken as a main raw material, and beneficial ingredients in the prune are fully released and converted through enzymolysis, fermentation and other processes, so that the activity and the functionality of enzyme products are improved, the safety of the products is researched, and no harmful effect on human bodies is ensured under reasonable dosage; the development of the product aims at combining health and functions and developing an innovative functional product. The product has the functional benefits of promoting intestinal health, improving constipation and the like, and the flavor and the nutritional value of the product are enhanced by improving the content of nutritional ingredients and functional ingredients in the product. Compared with the prior art, the invention has the advantages of uniqueness and remarkable advantages.

Drawings

Fig. 1 is a flowchart of a preparation method of western Mei Jiaosu provided according to an embodiment of the present invention.

Detailed Description

Hereinafter, embodiments of the present invention will be described with reference to the accompanying drawings. In the following description, like modules are denoted by like reference numerals. In the case of the same reference numerals, their names and functions are also the same. Therefore, a detailed description thereof will not be repeated.

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be further described in detail with reference to the accompanying drawings and specific embodiments. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not to be construed as limiting the invention.

Strain sources: yeasts are purchased from Angel Yeast Co., ltd.: 12015704; lactobacillus plantarum was purchased from (Siemens Polymer technology Co., ltd.: HH-LP 56).

Example 1

A method for preparing western Mei Jiaosu, comprising the steps of:

s1, preparing western Mei Yun slurry: cleaning fresh prune, removing core, cutting, soaking in 0.1% ascorbic acid solution for 10min, pulping to obtain prune Mei Yun pulp;

s2, enzymolysis: adding cellulase into the western Mei Yun pulp prepared in the step S1, carrying out water bath for 3.5 hours at 38 ℃, boiling and inactivating enzyme; adding saccharifying enzyme, saccharifying at 65deg.C for 1 hr, boiling to deactivate enzyme, and cooling; adjusting pH to 4, adding amylase and papain mixed enzyme, heating in water bath at 60deg.C for 3 hr, boiling to deactivate enzyme, filtering to remove excessive water, and sterilizing; the addition amounts of cellulase, saccharifying enzyme, alpha-amylase and papain are respectively 1%, 0.2%, 1% and 0.6% of the mass of the prune homogenate;

s3, fermenting: according to the volume ratio of the feed liquid of 3: adding water, and uniformly mixing to obtain the to-be-fermented western Mei Jiang;

inoculating saccharomycetes, respectively adding the white granulated sugar with the volume fraction of 4% and the ascorbic acid with the volume fraction of 0.1% into the to-be-fermented western medicine Mei Jiang, uniformly mixing, and carrying out primary fermentation at the temperature of 31 ℃ for 5 hours;

inoculating lactobacillus plantarum, and performing secondary fermentation at 34 ℃ for 10 hours; the inoculation amount of the saccharomycetes and the lactobacillus plantarum is 4 percent and 3 percent of the mass of the prune pulp to be fermented;

the saccharomycetes are pretreated before being added, and the method is as follows: adding ultrapure water into a certain amount of saccharomycetes according to the volume ratio of 1:100, uniformly mixing, and carrying out water bath at 25 ℃ for 20min for activation;

Example 2

A method for preparing western Mei Jiaosu, comprising the steps of:

s2, enzymolysis: adding cellulase into the western Mei Yun pulp prepared in the step S1, carrying out water bath for 4.5 hours at 35 ℃, boiling and inactivating enzyme; adding saccharifying enzyme, saccharifying at 65deg.C for 1 hr, boiling to deactivate enzyme, and cooling; adjusting pH to 4.5, adding amylase and papain mixed enzyme, heating in water bath at 65deg.C for 3 hr, boiling to deactivate enzyme, filtering to remove excessive water, and sterilizing; the addition amounts of cellulase, saccharifying enzyme, alpha-amylase and papain are respectively 1%, 0.2%, 1% and 0.6% of the mass of the prune homogenate;

inoculating saccharomycetes, respectively adding the white granulated sugar with the volume fraction of 4% and the ascorbic acid with the volume fraction of 0.1% into the to-be-fermented western medicine Mei Jiang, uniformly mixing, and carrying out primary fermentation at the temperature of 27 ℃ for 6 hours;

Example 3

A method for preparing western Mei Jiaosu, comprising the steps of:

s2, enzymolysis: adding cellulase into the western Mei Yun pulp prepared in the step S1, carrying out water bath for 4.5 hours at 35 ℃, boiling and inactivating enzyme; adding saccharifying enzyme, saccharifying at 70deg.C for 0.5 hr, boiling to deactivate enzyme, and cooling; adjusting pH to 4, adding amylase and papain mixed enzyme, heating in water bath at 55deg.C for 3.5 hr, boiling to deactivate enzyme, filtering to remove excessive water, and sterilizing; the addition amounts of cellulase, saccharifying enzyme, alpha-amylase and papain are respectively 1%, 0.2%, 1% and 0.6% of the mass of the prune homogenate;

Example 4

The optimization experiment of the fermentation strain is specifically as follows:

1. fermenting western Mei Jiaosu with yeast

1. Single factor experiment

Single-factor experiments of the preparation of the Western Mei Jiaosu are carried out by taking saccharomycetes as fermentation strains and taking inoculum sizes (1%, 2%, 3%, 4% and 5%), fermentation temperatures (23 ℃, 27 ℃, 31 ℃, 35 ℃ and 39 ℃) and feed-liquid ratios (1:7, 2:7, 3:7, 4:7 and 5:7) as variables, wherein the experimental conditions are shown in table 1;

TABLE 1 Yeast fermentation Process Single factor test survey Table

The specific operation is as follows:

s3, fermenting: according to the volume ratio of the feed liquid of 3: adding water, and uniformly mixing to obtain the to-be-fermented western Mei Jiang; inoculating saccharomycetes, respectively adding white granulated sugar with the volume fraction of 4% and ascorbic acid with the volume fraction of 0.1% into the to-be-fermented western medicine Mei Jiang, uniformly mixing, and fermenting;

s4, packaging: after the fermentation, the mixture was filtered through a 0.45 μm filter membrane to obtain Western Mei Jiaosu.

Determining the total acid content and superoxide dismutase activity of each group;

(1) Total acid determination

The determination is carried out according to the NaOH titration method in GB 12456-2021 determination of total acids in food safety national Standard food; the total acid content of the liquid edible plant ferment is more than or equal to 1.2g/100g according to the specification of T/CBFIA 08003-2017 edible plant ferment.

(2) Superoxide dismutase activity determination

The enzyme solution was centrifuged at 8000r/min for 10min, and the enzyme activity was measured using a superoxide dismutase kit.

As a result, the results of total acid content and superoxide dismutase activity were best when the fermentation conditions were respectively 4% inoculum size, 31℃fermentation temperature, and 3:7 feed liquid ratio, while the other conditions were unchanged.

2. Response surface optimization

Based on the single factor investigation result, the response surface optimization is carried out by taking the superoxide dismutase activity as an index according to the Box-Behnken principle (table 2).

TABLE 2 response surface test factors and levels for Yeast fermentation processes

The experimental result of the response surface shows that the optimal fermentation conditions are as follows: yeast inoculation amount is 4%, fermentation temperature is 31 ℃, and feed-liquid ratio is 3:7. The preparation of the Western Mei Jiaosu is carried out under the optimal condition, the fermentation is carried out for 25 hours after inoculation, the total acid content is 1.80g/kg after the Western Mei Jiaosu is measured, and the superoxide dismutase (SOD) activity is 409.52U/g.

2. Fermenting western Mei Jiaosu with Lactobacillus plantarum

1. Single factor experiment

Single-factor experiments of the preparation of the western Mei Jiaosu are carried out by taking saccharomycetes as fermentation strains and taking the addition amount (2%, 4% and 6%) of white granulated sugar, the fermentation temperature (29 ℃, 33 ℃ and 37 ℃) and the feed-liquid ratio (2:7, 3:7 and 4:7) as variables, and the experimental conditions are shown in table 3;

TABLE 3 Single factor test survey Table for Lactobacillus plantarum fermentation process

The specific operation is as follows:

s3, fermenting: according to the volume ratio of the feed liquid of 3: adding water, and uniformly mixing to obtain the to-be-fermented western Mei Jiang; inoculating lactobacillus plantarum, respectively adding white granulated sugar with the volume fraction of 4% and ascorbic acid with the volume fraction of 0.1% into the to-be-fermented western medicine Mei Jiang, uniformly mixing, and fermenting;

(1) Total acid determination

(2) Superoxide dismutase activity determination

The results showed that the total acid content and superoxide dismutase activity were best when the fermentation conditions were respectively white granulated sugar addition 4%, fermentation temperature 33 ℃, feed ratio 3:7, and other conditions were unchanged.

2. Response surface optimization

Based on the single factor investigation result, the response surface optimization is carried out by taking the superoxide dismutase activity as an index according to the Box-Behnken principle (table 4).

TABLE 4 response surface test factors and levels of Lactobacillus plantarum fermentation Process

The experimental result of the response surface shows that when lactobacillus plantarum is used for fermentation, the optimal fermentation conditions are as follows: white granulated sugar addition amount is 4%, fermentation temperature is 33 ℃, and feed-liquid ratio is 3:7. The preparation of the Western Mei Jiaosu is carried out under the optimal condition, the fermentation is carried out for 15 hours after inoculation, the total acid content is measured to be 0.87g/kg by Western Mei Jiaosu, and the superoxide dismutase (SOD) activity is 485.22U/g.

3. Single factor experiment using mixed strain fermentation process

1. Experimental methods and conditions

Taking saccharomycetes and lactobacillus plantarum as fermentation strains at the same time, taking the results as reference, setting the inoculation amount of the saccharomycetes and the lactobacillus plantarum as quantification, respectively taking the inoculation sequence, the fermentation temperature and the fermentation time as variables to carry out single-factor experiments for preparing the Western Mei Jiaosu, and setting experimental conditions of a control group (independently inoculated saccharomycetes and independently inoculated lactobacillus plantarum) as shown in Table 5;

table 5 single factor test survey table for mixed strain fermentation process

2. Measurement index

(1) Total acid determination

The determination is carried out according to the NaOH titration method in GB 12456-2021 determination of total acids in food safety national Standard food. The total acid content of the liquid edible plant ferment is more than or equal to 1.2g/100g according to the specification of T/CBFIA 08003-2017 edible plant ferment.

(2) Superoxide dismutase activity determination

3. Western Mei Jiaosu active ingredient and functional study

(1) Crude polysaccharide content

Determining the ferment protein content by using a Bradford protein concentration determination kit; the free amino acid content of the ferment is determined by adopting a Soxhibao amino acid content detection kit (BC 1570).

(2) Monosaccharide composition of polysaccharide

And respectively weighing appropriate amounts of mannose, ribose, rhamnose, glucuronic acid, galacturonic acid, anhydrous glucose, galactose, xylose, arabinose and fucose reference substances, and preparing standard substance stock solutions with different mass concentrations. 1mL of the sample and 2mL of trifluoroacetic acid (2 mol/L) are placed in a 10mL headspace bottle, hydrolyzed for 8h at 110 ℃ to volatilize the trifluoroacetic acid, and then 2mL of purified water is added for redissolution.

PMP derivatization: mu.L of the sample solution, 250. Mu.L of 0.6mol/L NaOH, 500. Mu.L of 0.4mol/L PMP-methanol solution were placed in a 5mL EP tube and reacted at 70℃for 1 hour. Cooling in cold water for 10min, adding 500 μL of 0.3mol/L HCl and 1mL of chloroform, mixing, centrifuging at 3000r/min for 10min, collecting supernatant, repeating for three times, and mixing the supernatants.

Chromatographic conditions: chromatographic column: xtime (C18 4.6X100 mm,5 μm); column temperature: 30 ℃; flow rate: 1.0mL/min; detection wavelength: 250nm; sample injection amount: 20. Mu.L; mobile phase: 0.05mol/L potassium dihydrogen phosphate solution (pH adjusted to 6.70 with sodium hydroxide solution) -acetonitrile=83-17.

(3) Composition of free amino acids

And respectively weighing a proper amount of standard substances of aspartic acid, glutamic acid, serine, glycine, histidine, arginine, threonine, alanine, proline, tyrosine, valine, methionine, isoleucine, leucine, phenylalanine and lysine, and dissolving the standard substances by using 0.1mol/L HCl solution to prepare standard substance stock solutions with different mass concentrations. 3mL of ferment, 3mL of concentrated hydrochloric acid and 4mL of concentrated hydrochloric acid (6 mol/L) are taken to be placed in a hydrolysis tube, 3-4 drops of phenol are added to be mixed evenly, sealing is carried out, hydrolysis is carried out in a 110 ℃ electrothermal blowing drying box for 22h, cooling is carried out to room temperature, then the hydrolysis liquid is taken, ultrapure water is used for constant volume in a 50mL volumetric flask, 1mL of the hydrolysis liquid is taken to be placed in an ammonia blowing instrument for nitrogen blowing after being mixed evenly, and 1mL of hydrochloric acid (0.1 mol/L) is added for redissolution after the hydrolysis liquid is dried.

PITC derivatization: and (3) after the sample solution and the standard substance solution are derived, passing through a microporous filter membrane of 0.22 mu m to obtain the sample solution and the standard substance solution.

Chromatographic conditions: chromatographic column: venusil AA column (C18.6X105 mm,5 μm); ultraviolet detector wavelength: 254nm; column temperature: 35 ℃; sample injection amount: 20. Mu.L; flow rate: 1mL/min; mobile phase: a is 0.1mol/L sodium acetate-acetonitrile (volume ratio is 97:3) solution (pH 6.5 is adjusted by glacial acetic acid), and B is 80% acetonitrile solution.

(4) Organic acid composition

And weighing a proper amount of standard substances of malic acid, ascorbic acid, lactic acid, succinic acid and citric acid, and preparing standard substance stock solutions with different mass concentrations. 5mL of the solution was aspirated and centrifuged at 3000r/min for 15min. The supernatant was removed and filtered through a 0.22 μm aqueous filter, and the filtrate was subjected to HPLC.

Chromatographic conditions: chromatographic column: LG column (C18 4.6X250 mm,5 μm); mobile phase: a: methanol, B:20mmol/L disodium hydrogen phosphate (phosphoric acid adjusts pH to 2.5) -methanol (99:1); sample injection amount: 10. Mu.L; flow rate: 0.8mL/min; column temperature: 40 ℃; ultraviolet detector wavelength: 210nm.

(5) Effects on cell viability

Collecting cells in good growth state and logarithmic phase, and adjusting cell density to 3×10 ⁴ Inoculating 100 mu L of each well into a 96-well plate, and placing the 96-well plate into constant temperature CO ₂ Culturing in an incubator for 24 hours, and discarding the old culture solution after the cells grow on the wall. The ferment group and the control group are respectively provided with 5 administration concentrations, the administration concentrations are respectively 0, 50, 100, 150, 200 and 250 mu L/mL by adopting a fold dilution method and using a culture solution containing 10% bovine serum, and a blank control group is simultaneously provided, each concentration is respectively provided with 3 parallel compound holes, and the administration time is respectively 48 hours. After the administration time is reached, the drug-containing culture medium is sucked and removed, each compound well is washed 1-2 times by PBS, and then 10 mu L of MTT solution is added into each compound well after 100 mu L of 10% bovine serum culture solution is added into each compound well again; meanwhile, bubbles in the compound wells are avoided when the culture solution and the MTT are added, and OD value reading is influenced. Incubate for 4h in incubator and then determine Optical Density (OD) with an enzyme-labeled instrument at 490nm wavelength.

The cell viability calculation formula is as follows: cell viability (%) = (experimental OD value-control well OD value)/(control OD value-control well OD value) ×100%.

(6) Effects on cell cycle

The concentration was set at 2X 10 ⁵ individual/mL cell suspensions were seeded into 6-well plates at 37 ℃,5% co ₂ Is cultured in a cell culture incubator for 24 hours. The old culture solution is discarded, and the enzyme group and the control group are respectively prepared into 0, 50, 100 and 150 mu L/mL of drug-containing culture solution to be administered to each pore plate cell for 24 hours, and a blank control group is arranged, and each concentration is arranged in parallel to 3. Transferring old medicated culture solution in 6-well plate into centrifuge tube, digesting cells with pancreatin, adding new culture solution to stop digestion, transferring blown-down cells into centrifuge tube which has previously collected old culture solution, centrifuging at 1000rmp for 3-5min, and discarding supernatant. Adding ice-bath precooled PBS to resuspend cells, centrifuging, discarding supernatant, then adding ice-bath precooled 70% ethanol to fix for 24h, centrifuging, and precipitating cells. 0.5mL of propidium iodide staining solution is added to each sample tube, cells are slowly resuspended, and the cells are detected by a flow cytometer after being subjected to light-shielding warm bath for 30min at 37 ℃.

(7) Effects on apoptosis

Taking human lung cancer NCl-H1299 cells with good growth condition and logarithmic phase, and adjusting concentration to 2×10 ⁵ Cell suspension of individual/mL, human lung cancer NCI-H1299 cells were seeded at appropriate concentration in 6-well plates, then incubated at constant temperature CO ₂ Culturing in an incubator for 24 hours, and discarding old culture solution after the cells grow on the wall. The ferment group and the control group are respectively dosed for 48 hours according to 0, 50, 100 and 150 mu L/mL, meanwhile, a blank control group is arranged, 3 compound holes are arranged at each concentration, the ferment group and the control group are cultured for 48 hours, samples are processed according to the instruction book of an Annexin V-FITC/PI detection kit, and are analyzed by a flow cytometer, so that the apoptosis rate is calculated.

4. Western plum ferment safety index determination

(1) Microorganism index

The detection is carried out according to methods provided in GB 4789.4-2016 salmonella test for food safety national standard food microbiology, GB 4789.10-2016 staphylococcus aureus test for food safety national standard food microbiology, GB 4789.15-2016 mould and yeast count for food safety national standard food microbiology test, GB 4789.2-2022 total colony count determination for food safety national standard food microbiology test, GB 4789.3-2016 coliform count for food safety national standard food microbiology test.

(2) Heavy metal content

The determination was carried out according to the method provided in GB 5009.268-2016 "determination of Multi-elements in food safety national Standard food".

3. Results

(1) Fermenting by solely using saccharomycetes: 24 hours after fermentation, the content of the crude polysaccharide is measured to be 90mg/g; after 48 hours, the crude polysaccharide content was 140mg/g; glucose accounts for 55% of the polysaccharide, and fructose accounts for 30% of the polysaccharide; galactose accounts for 15% of the polysaccharide; lysine accounts for 10% of the total amino acids, phenylalanine accounts for 8% of the total amino acids; lactic acid content is 2g/L, malic acid content is 1.8g/L;10% fermentation product resulted in 85% cell viability and 20% fermentation product resulted in 70% cell viability; after fermentation product treatment, cells enter G1 phase for delay, and the duration of S phase is increased; the expression of apoptosis marker Bax is increased and the expression of apoptosis marker Bcl-2 is decreased.

(2) The lactobacillus plantarum is adopted independently for fermentation: 24 hours after fermentation, the content of the crude polysaccharide is 80mg/g, and after 48 hours, the content of the crude polysaccharide is 120mg/g; glucose accounts for 50% of the polysaccharide, and fructose accounts for 25% of the polysaccharide; galactose accounts for 8% of the polysaccharide; lysine accounts for 9% of the total amino acids, and phenylalanine accounts for 5% of the total amino acids; lactic acid content is 1.5g/L, malic acid content is 1.2g/L;10% fermentation product resulted in 80% cell viability and 20% fermentation product resulted in 60% cell viability; after fermentation product treatment, cells enter G2 phase for delay, and S phase duration is reduced; the expression of apoptosis marker Bax is increased and the expression of apoptosis marker Bcl-2 is decreased.

(3) Fermenting by mixed bacteria fermentation process

The fermentation process with the best effect is as follows: inoculating 4% yeast, adding 4% white granulated sugar and 0.1% ascorbic acid, mixing, and fermenting for 5 hr; inoculating 3% lactobacillus plantarum, fermenting for 10 hours, and fermenting for 15 hours; after 24 hours from the end of fermentation, the content of the crude polysaccharide is determined to be 100mg/g, and after 48 hours, the content of the crude polysaccharide is determined to be 150mg/g; glucose accounts for 60% of the polysaccharide, and fructose accounts for 35% of the polysaccharide; galactose accounts for 20% of the polysaccharide; lysine accounts for 12% of the total amino acids, phenylalanine accounts for 11% of the total amino acids; lactic acid content is 3.7g/L, malic acid content is 2.5g/L;10% fermentation product resulted in a cell viability of 95% and 20% fermentation product resulted in a cell viability of 75%; after fermentation product treatment, cells enter G1 phase for delay, and the duration of S phase is increased; apoptosis markers Bcl-2 and Caspase-3 were decreased. The group 7 has poor effect, and can be unstable due to natural fermentation in the early stage or bring mixed bacteria, saccharomycetes, lactobacillus plantarum and the like to form competition, so that the natural fermentation process is abandoned, and a specific strain is selected to be inoculated in the fermentation, so that the fermentation process is more controllable.

TABLE 6 safety index determination results of prune ferment

TABLE 7 results of measurement of the metal content of prune ferment

The measurement results of the safety indexes of the prune ferment are shown in table 6; the heavy metal content results are shown in table 7; from the table, the western Mei Jiaosu fermented by the fermentation process meets the national standard, and the safety is good.

The invention has the advantages that:

(1) Innovations in raw material selection

The prune contains rich vitamins, minerals, dietary fibers, antioxidants and unique chemical compositions such as pectin, anthocyanin and the like, and the components endow the product with excellent biological activity and functionality together, so that a rich foundation is provided for the development of prune ferment.

(2) Innovations in process optimization

The preparation method has the advantages that the processes of enzymolysis, fermentation and the like are skillfully combined, the beneficial components in the prune are released to the greatest extent, the defect of low release rate of the active components in the traditional ferment in the processing process is overcome, the bioactive components are reserved to the greatest extent by optimizing the process parameters, and the effectiveness of the functions of the product is ensured.

(3) Combination of biological Activity and safety

The bioactive components of the western Mei Fuhan anthocyanin, polyphenol and the like have the characteristics of antioxidation, anti-inflammatory, anti-aging and the like. Combining these bioactive ingredients with the enzyme product imparts a broader functional benefit to the product and ensures safety, thereby increasing its acceptance in the marketplace.

(4) Application of multiple functional benefits

The prune ferment combines multiple functional benefits such as promoting intestinal health, improving constipation, enhancing immune system function, maintaining cardiovascular health, etc., highlighting the comprehensive health benefits of the prune Mei Jiaosu and increasing the attractiveness and market competitiveness of the product. Not only satisfies the basic functions of the traditional ferment products, but also solves the health problems brought by modern life style in a targeted way.

In conclusion, the development of the prune ferment brings new value and prospect to the field of ferment products.

It should be appreciated that various forms of the flows shown above may be used to reorder, add, or delete steps. For example, the steps described in the present disclosure may be performed in parallel, sequentially, or in a different order, provided that the desired results of the technical solutions of the present disclosure are achieved, and are not limited herein.

The above embodiments do not limit the scope of the present invention. It will be apparent to those skilled in the art that various modifications, combinations, sub-combinations and alternatives are possible, depending on design requirements and other factors. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the scope of the present invention.

Claims

1. A method for preparing the western Mei Jiaosu, which is characterized by comprising the following steps:

2. A method of preparing a west Mei Jiaosu according to claim 1, wherein: the addition amounts of the cellulase, the saccharifying enzyme, the alpha-amylase and the papain in the step S2 are respectively 0.8-1.2%, 0.1-0.3%, 0.5-1% and 0.4-0.8% of the mass of the prune homogenate.

3. A method of preparing a west Mei Jiaosu according to claim 2, wherein: the inoculation amount of the saccharomycetes and the lactobacillus plantarum in the step S3 is 3-5% and 2-4% of the mass of the prune pulp to be fermented.

4. A process for preparing a compound Mei Jiaosu as claimed in claim 3, wherein: the inoculation amount of the saccharomycetes and the lactobacillus plantarum in the step S3 is 4% and 3% of the mass of the prune pulp to be fermented.

5. The method for preparing the western Mei Jiaosu according to claim 4, wherein: the fermentation temperature of the primary fermentation is 31 ℃; the fermentation temperature of the secondary fermentation is 34 ℃.

6. The method for preparing the western Mei Jiaosu according to claim 5, wherein: the fermentation time of the primary fermentation is 5 hours; the fermentation time of the secondary fermentation is 10 hours.

7. The method for preparing the western Mei Jiaosu of claim 6, wherein: the mass ratio of the amylase to the papain is 5:3.

8. the method for preparing the western Mei Jiaosu of claim 7, wherein: the saccharomycetes are pretreated before being added, and the saccharomycetes are specifically as follows: adding ultrapure water into a certain amount of saccharomycetes according to the volume ratio of 1:100, uniformly mixing, and carrying out water bath for 18-25 min at the temperature of 23-27 ℃.

9. A process for preparing a form of west Mei Jiaosu as claimed in claim 1, wherein said process comprises preparing said west Mei Jiaosu.