CN110772458A - Vaccinium myrtillus enzyme prepared by using compound biological enzyme and preparation method and application thereof - Google Patents
Vaccinium myrtillus enzyme prepared by using compound biological enzyme and preparation method and application thereof Download PDFInfo
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Abstract
The invention belongs to the field of preparation of enzymes and application of the enzymes in cosmetics, and particularly relates to a vaccinium myrtillus enzyme prepared by using compound biological enzyme and a preparation method and application of the vaccinium myrtillus enzyme. Adding sugar into the vaccinium myrtillus after enzymolysis, and sealing and fermenting the vaccinium myrtillus by using mixed bacteria to obtain vaccinium myrtillus ferment; wherein the composite bacteria are Saccharomyces cerevisiae, Lactobacillus plantarum and Bifidobacterium. The invention explores a new process for fermenting the cowberry enzyme, has simple process, fully releases nutrient substances and active ingredients in the cowberry, completely dissolves soluble substances, and is suitable for being added into cosmetics. The cosmetics can be conventional preparations such as facial masks, astringent or emulsion, and the skin care effect is obviously enhanced after the cowberry fruit enzyme is added into the essence.
Description
Technical Field
The invention belongs to the field of preparation of enzymes and application of the enzymes in cosmetics, and particularly relates to a vaccinium myrtillus enzyme prepared by using compound biological enzyme and a preparation method and application of the vaccinium myrtillus enzyme.
Background
Vaccinium, a plant of the genus Vaccinium of the family Ericaceae, comprising: more than 400 varieties of vaccinium myrtillus, vaccinium americanum, vaccinium sibiricum and the like are preferred, and vaccinium myrtillus produced in Sweden, Finland, Norway and the like is the best. The average weight of the cowberry fruit is 0.5-2.5g, the fruit has beautiful color, fine pulp, extremely small seeds and edible rate of 100 percent. The cowberry contains rich bioactive substances, has extremely strong medicinal value and nutrition and health care functions besides being eaten, and is listed as one of five kinds of human health foods by the international food and agriculture organization. The cowberry contains amino acid, vitamin, anthocyanin, flavone, organic selenium, fruit acid and other special nutrient components which are incomparable with any plant, and has more excellent antioxidant activity than common plants including blueberries. At present, people have higher and higher attention on the cowberry, and related industries are gradually increased. The development market of the blueberry deep-processed product is wide, and the deep processing will become the key development direction of the blueberry in future.
Plant enzymes are a functional product produced by fermentation techniques using biotechnology. The Edward Howeer doctor discovered the enzyme and key codes of human life and health in the eighties of the last century, and the enzyme attracts attention of people all over the world. The cowberry fruit ferment mainly contains anthocyanin, pectin, flavonoid, vitamins, minerals, etc. with health promotion function. Especially contains rich anthocyanin, has multiple physiological effects of scavenging free radicals, resisting inflammation, improving immunity, delaying aging, resisting cancer and the like, and can be developed into various health care products and cosmetics. The lead, mercury, arsenic, hormone, fluorescent agent whitening agent and the like are added to the current cosmetics on the market to a certain extent, the cosmetics have certain moisturizing and whitening functions but have side effects on human bodies, and the current cosmetics basically have moisturizing and whitening functions and do not have health care functions.
Disclosure of Invention
Aiming at the problem of side effect of additives in cosmetics on the market to human bodies, the invention aims to provide a vaccinium myrtillus enzyme prepared by utilizing compound biological enzyme and a preparation method and application thereof.
In order to realize the purpose, the invention is realized by the following technical scheme:
a Vaccinium myrtillus enzyme prepared by compound biological enzyme is prepared by performing enzymolysis on Vaccinium myrtillus, adding sugar, and sealing and fermenting with mixed bacteria to obtain Vaccinium myrtillus enzyme; wherein the composite bacteria are Saccharomyces cerevisiae, Lactobacillus plantarum and Bifidobacterium.
The strains used in the invention can be purchased through various commercial approaches, for example, the strains can be purchased through China Industrial microbial culture Collection center (CICC), and the strains purchased through commercial channels are all suitable for the invention.
The mixed enzyme adopted by the enzymolysis is cellulase, hemicellulase, pectinase and acid protease; the respective addition amounts are based on the mass ratio of the cowberry fruit crushing mixed solution, the addition amount of the cellulase is 0.1-0.5%, the addition amount of the hemicellulase is 0.1-0.5%, the addition amount of the pectinase is 0.12-0.18%, and the addition amount of the acid protease is 0.1-0.5%.
A preparation method of a vaccinium myrtillus enzyme prepared by using compound biological enzyme comprises the steps of mixing washed and crushed vaccinium myrtillus with water, adding mixed enzyme, carrying out enzymolysis for 2-4h in a water bath at 50-60 ℃ with the pH value of 4.0-5.0, adding sugar with the mass of 14% -20% of the crushed mixture, inoculating mixed bacteria to an enzymolysis product after adjustment, and carrying out sealed fermentation to obtain the vaccinium myrtillus enzyme.
The method comprises the steps of cleaning and crushing the refrigerated fresh vaccinium myrtillus fruits, and mixing the crushed fruits with water according to the volume ratio of 1-3:0.8-1.5 for later use.
Adding mixed enzyme into the cowberry fruit crushing mixed solution, uniformly mixing, adjusting the pH value of the mixed solution to 4.0-5.0, and placing in a water bath at 50-60 ℃ for enzymolysis for 2-4 h; then adding sugar to adjust the sugar degree, adjusting the sugar degree at room temperature, and adding sugar which is 14-20% of the mass of the blueberry crushing mixed liquid; adjusting sugar degree, sterilizing, and inactivating; and (5) standby.
①, opening a cover plate, carefully wiping the detection prism by using a soft cloth, ② taking a plurality of drops of solution to be detected, placing the drops on the detection prism, slightly closing the cover plate to avoid bubbles, enabling the solution to be distributed on the surface of the prism, ③, aligning an instrument light inlet plate to a light source or a bright place, observing a visual field by eyes, rotating an eyepiece adjusting hand wheel, ④, and enabling a blue-white boundary of the visual field to be clear, wherein the scale value of the boundary is the concentration of the solution.
The mixed enzyme adopted by the enzymolysis is cellulase, hemicellulase, pectinase and acid protease; the respective addition amount is based on the mass ratio of the cowberry fruit crushing mixed solution, and the addition amount of the cellulase is 0.1-0.5 percent, preferably 0.2-0.35 percent; the adding amount of the hemicellulase is 0.1-0.5%, and the preferable adding amount is 0.2-0.35%; the addition amount of the pectinase is 0.12-0.18%, and the preferable addition amount is 0.14-0.16%; the addition amount of the acid protease is 0.1-0.5%, and the preferable addition amount is 0.2-0.35%.
The sealed fermentation comprises the following steps: respectively activating yeast Saccharomyces cerevisiae, Lactobacillus plantarum and Bifidobacterium, and culturing to middle and late logarithmic growth stage; standby;
mixing the enzymolysis product with the activated saccharomyces cerevisiae, lactobacillus plantarum and bifidobacterium, and uniformly stirring according to the mass ratio of 40-60:0.8-1.5:0.8-1.5:0.8-1.5, wherein the preferable ratio range is as follows: 48-54:1.0-1.2:1.0-1.2:1.0-1.2, and then sealing and fermenting at 25-32 ℃ for 4-7 days, and stirring once every 10h to obtain the fermentation liquor.
And filtering and sterilizing the fermentation liquor to obtain the vaccinium myrtillus ferment.
The bilberry of the present invention may be produced anywhere, preferably in europe, such as finland, sweden, etc.
The invention has the advantages that:
the bilberry enzyme is prepared by using the compound biological enzyme method to assist probiotics, contains active ingredients of procyanidine and organic acid, the content of procyanidine is far higher than that of enzymes prepared by other plants, and the organic acid contains various organic acids including citric acid, pyruvic acid, succinic acid and the like. The cowberry fruit ferment prepared by the invention not only can brighten uniform skin color and make the skin more moist, but also has the functions of eliminating free radicals and delaying aging, and is beneficial to human bodies, and the cowberry fruit ferment specifically comprises the following components:
1. the invention adopts composite biological enzyme and enzymolysis to assist beneficial microorganism fermentation, and the blueberry procyanidine is protected to the maximum extent in the fermentation process, so that the nutrient substances and effective components in the blueberry are fully released, and the soluble substances are completely dissolved; the cowberry fruit enzyme prepared by the method is in a light purple transparent liquid state, and comprises the following main effective components: procyanidin, organic acid, pectin, flavonoid, vitamins, minerals, etc. Wherein the content of procyanidin is 46-75mg/ml, the content of organic acid is 14.21-18.89mg/ml, and the types of organic acid mainly comprise lactic acid, citric acid, pyruvic acid and succinic acid.
2. The invention utilizes yeast, lactobacillus plantarum and bifidobacterium to mix and ferment the vaccinium myrtillus to produce the ferment, breaks through the traditional natural fermentation mode and the pure strain fermentation process, and explores a new process for fermenting the vaccinium myrtillus ferment. The enzyme prepared by the method is simple in process and is suitable for being added into cosmetics. The fermentation speed can be increased by adding the activating microbial inoculum and keeping the fermentation temperature at 25-32 ℃, and the fermentation can be more uniform by stirring once every 10 hours.
3. The cowberry fruit ferment provided by the invention has the effects of improving and brightening skin color, and is specifically characterized in that the skin color is uniform, and the glossiness is improved. Sensory tests of the facial mask added with the cowberry enzyme show that the cowberry enzyme has exact anti-aging effect, and after the facial mask is applied to various parts of the face, including the forehead, the periphery of eyes and the cheeks, the skin elasticity value is improved, and the water content is obviously improved.
Description of the drawings:
fig. 1 is a graph of skin care effect test results of cowberry enzyme provided in the embodiment of the present invention, wherein a is forehead, B is eye periphery, and C is cheek.
Detailed Description
The present invention is described in further detail with reference to the following detailed description, but the scope of the present invention is not limited thereto.
Example 1:
(1) preparing materials: cleaning and crushing the refrigerated vaccinium myrtillus, and mixing the crushed vaccinium myrtillus and clean water according to the volume ratio of 2:1 to obtain a crushed vaccinium myrtillus mixed solution for later use.
(2) Activation of mixed strains: respectively activating yeast Saccharomyces cerevisiae (YPD culture medium), Lactobacillus plantarum (LB culture medium) and Bacillus bifidus (LB culture medium), culturing to middle and late logarithmic growth period, collecting thallus, washing with sterile physiological saline, and removing supernatant.
(3) Biological enzymolysis: adding mixed enzyme into the crushed cowberry mixed solution, wherein the addition amount of cellulase is 0.1% of the crushed cowberry mixed solution, the addition amount of hemicellulase is 0.1% of the crushed cowberry mixed solution, the addition amount of pectinase is 0.12% of the crushed cowberry mixed solution, the addition amount of acid protease is 0.1% of the crushed cowberry mixed solution; then adjusting the pH value of the mixed solution to 4.0, and placing the mixed solution in a water bath at 50 ℃ for enzymolysis for 2 h.
(4) And (3) sugar degree adjustment: adding white sugar directly into the mixture after enzymolysis to adjust sugar degree, and measuring sugar degree at room temperature with a handheld sugar degree meter to adjust sugar concentration to 14%.
(5) Sterilization and enzyme deactivation: sterilizing the mixture of cowberry juice by pasteurization, sterilizing at 85 deg.C for 20min, and rapidly cooling in ice water.
(6) Adding mixed bacteria into the sterilized cowberry fruit juice mixture, and directly mixing and stirring the mixture, the saccharomyces cerevisiae, the lactobacillus plantarum and the bifidobacterium uniformly according to the volume ratio of 50:1:1: 1.
(7) And (3) sealing fermentation: mixing, sealing, fermenting at 25 deg.C for 4 days, and stirring once every 10 hr to obtain fermentation liquid.
(8) And (3) filtering and sterilizing: firstly, filtering the fermentation liquor by using gauze to remove filter residues, and then carrying out ultrafiltration sterilization to obtain the vaccinium myrtillus ferment.
The obtained ferment is detected by high performance liquid chromatography, the content of procyanidine in the prepared cowberry ferment is 51mg/ml, the content of organic acid is 15.3mg/ml, and the cowberry ferment contains abundant organic acid by HPLC organic acid analysis, so that the organic acid types are confirmed to be mainly lactic acid, citric acid, pyruvic acid and succinic acid.
Example 2:
the difference from the embodiment 1 is that:
biological enzymolysis: adding mixed enzyme into the cowberry fruit crushing mixed solution, wherein the addition amount of cellulase is 0.25%, the addition amount of hemicellulase is 0.25%, the addition amount of pectinase is 0.14%, and the addition amount of acid protease is 0.25%; then adjusting the pH value of the mixed solution to 4.0, and placing the mixed solution in a water bath at the temperature of 55 ℃ for enzymolysis for 3 hours.
And (3) sugar degree adjustment: and (3) directly adding white granulated sugar into the mixture after enzymolysis to adjust the sugar degree, and measuring the sugar degree by using a handheld sugar degree instrument at room temperature to adjust the sugar concentration to 16%.
And (3) sealing fermentation: mixing, sealing, fermenting at 28 deg.C for 5 days, and stirring once every 10 hr to obtain fermentation liquid.
The obtained ferment is subjected to high performance liquid chromatography detection, the content of procyanidine in the prepared cowberry fruit ferment is 75mg/ml, the content of organic acid is 18.89mg/ml, and the types of the organic acid mainly comprise lactic acid, citric acid, pyruvic acid and succinic acid through HPLC organic acid type analysis.
Example 3:
the difference from the embodiment 1 is that:
biological enzymolysis: adding mixed enzyme into the cowberry fruit crushing mixed solution, wherein the addition amount of cellulase is 0.4%, the addition amount of hemicellulase is 0.4%, the addition amount of pectinase is 0.16%, and the addition amount of acid protease is 0.4%; then adjusting the pH value of the mixed solution to 4.0, and placing the mixed solution in a water bath at the temperature of 55 ℃ for enzymolysis for 3 hours.
And (3) sugar degree adjustment: adding white sugar directly into the mixture after enzymolysis to adjust sugar degree, and measuring sugar degree at room temperature with a handheld sugar degree meter to adjust sugar concentration to 18%.
And (3) sealing fermentation: mixing, sealing, fermenting at 30 deg.C for 6 days, and stirring once every 10 hr to obtain fermentation liquid.
The obtained ferment is detected by high performance liquid chromatography, the content of procyanidine in the prepared cowberry fruit ferment is 59mg/ml, the content of organic acid is 15.5mg/ml, and the variety of the organic acid mainly comprises lactic acid, citric acid, pyruvic acid and succinic acid through HPLC organic acid type analysis.
Example 4:
the difference from the embodiment 1 is that:
biological enzymolysis: adding mixed enzyme into the cowberry fruit crushing mixed solution, wherein the addition amount of cellulase is 0.5%, the addition amount of hemicellulase is 0.5%, the addition amount of pectinase is 0.18%, and the addition amount of acid protease is 0.5%; then adjusting the pH value of the mixed solution to 4.0, and placing the mixed solution in a water bath at 60 ℃ for enzymolysis for 4 h.
And (3) sugar degree adjustment: adding white sugar directly into the mixture after enzymolysis to adjust sugar degree, and measuring sugar degree at room temperature with a handheld sugar degree meter to adjust sugar concentration to 20%.
And (3) sealing fermentation: mixing, sealing, fermenting at 32 deg.C for 7 days, and stirring once every 10 hr to obtain fermentation liquid.
Detecting the obtained ferment by high performance liquid chromatography, wherein the content of procyanidine in the prepared cowberry fruit ferment is 55mg/ml, and the content of organic acid is 15.8mg/ml, and analyzing the obtained ferment by HPLC organic acid types to determine that the enzyme mainly comprises lactic acid, citric acid, pyruvic acid and succinic acid.
Comparative example 1:
(1) preparing materials: cleaning and crushing the refrigerated vaccinium myrtillus, and mixing the crushed vaccinium myrtillus and clean water according to the volume ratio of 2:1 to obtain a crushed vaccinium myrtillus mixed solution for later use.
(2) Activation of mixed strains: respectively activating Saccharomyces cerevisiae (YPD culture medium) and Lactobacillus plantarum (LB culture medium), culturing to middle and late logarithmic growth period, collecting thallus, washing with sterile normal saline, and removing supernatant.
(3) Biological enzymolysis: adding mixed enzyme into the cowberry fruit crushing mixed solution, wherein the addition amount of cellulase is 0.4%, the addition amount of hemicellulase is 0.4%, the addition amount of pectinase is 0.16%, and the addition amount of acid protease is 0.4%; then adjusting the pH value of the mixed solution to 4.0, and placing the mixed solution in a water bath at the temperature of 55 ℃ for enzymolysis for 3 hours.
(4) And (3) sugar degree adjustment: and (3) directly adding white granulated sugar into the mixture after enzymolysis to adjust the sugar degree, and measuring the sugar degree by using a handheld sugar degree instrument at room temperature to adjust the sugar degree to 16%.
(5) Sterilization and enzyme deactivation: sterilizing the mixture of cowberry juice by pasteurization, sterilizing at 85 deg.C for 20min, and rapidly cooling in ice water.
(6) Adding mixed bacteria into the treated cowberry fruit juice mixture, and directly stirring the mixture, the saccharomyces cerevisiae, the lactobacillus plantarum and the bifidobacterium uniformly according to the proportion of 50:1: 1.
(7) And (3) sealing fermentation: mixing, sealing, fermenting at 28 deg.C for 5 days, and stirring once every 10 hr to obtain fermentation liquid.
(8) And (3) filtering and sterilizing: firstly, filtering the fermentation liquor by using gauze to remove filter residues, and then carrying out ultrafiltration sterilization to obtain the vaccinium myrtillus ferment.
Detecting the obtained ferment by high performance liquid chromatography, wherein the content of procyanidine in the prepared cowberry ferment is 49.8mg/ml, and the content of organic acid is 14.5mg/ml, and analyzing the obtained ferment by HPLC organic acid types to determine that the cowberry ferment contains lactic acid, citric acid and pyruvic acid and hardly contains succinic acid.
Comparative example 2:
the difference from the embodiment 1 is that:
activation of mixed strains: respectively activating Saccharomyces cerevisiae (YPD culture medium), Lactobacillus plantarum (LB culture medium) and Bifidobacterium (LB culture medium), culturing to middle and late logarithmic growth period, collecting thallus, washing with sterile normal saline, and removing supernatant.
Biological enzymolysis: adding cellulase in an amount of 0.4% and pectase in an amount of 0.16% into the mixture. Adjusting the pH value of the mixed solution to 4.0, and placing the mixed solution in a water bath at the temperature of 55 ℃ for enzymolysis for 3 hours.
Directly inoculating the saccharomyces cerevisiae, the lactobacillus plantarum and the bifidobacterium prepared in the step (2) into the treated cowberry fruit juice mixture according to the proportion of 50:1:1:1, and uniformly stirring.
Through detection, the prepared cowberry fruit ferment contains procyanidine, the content of organic acid is 13.85mg/ml, and the types of the organic acid mainly comprise lactic acid, citric acid, pyruvic acid and succinic acid.
Detecting the obtained ferment by high performance liquid chromatography, wherein the content of procyanidine in the prepared cowberry fruit ferment is 40.25mg/ml, the content of organic acid is 10.25mg/ml, and the blueberry fruit ferment is determined to comprise lactic acid, citric acid, pyruvic acid and succinic acid by HPLC organic acid type analysis.
Comparative example 3:
the difference from the embodiment 1 is that:
activation of mixed strains: respectively activating Saccharomyces cerevisiae (YPD culture medium) and Lactobacillus plantarum (LB culture medium), culturing to middle and late logarithmic growth period, collecting thallus, washing with sterile normal saline, and removing supernatant.
Biological enzymolysis: adding cellulase in an amount of 0.4% and pectase in an amount of 0.16% (w/w) into the mixture. Adjusting the pH value of the mixed solution to 4.0, and placing the mixed solution in a water bath at the temperature of 55 ℃ for enzymolysis for 3 hours.
And (3) directly inoculating the saccharomyces cerevisiae and the lactobacillus plantarum prepared in the step (2) into the treated cowberry fruit juice mixture according to the ratio of 50:1:1, and uniformly stirring.
Through detection, the content of procyanidine in the prepared cowberry fruit ferment is 45.86mg/ml, the content of organic acid is 13.55mg/ml, the types of the organic acid mainly comprise lactic acid and citric acid, the content of pyruvic acid is less, and succinic acid is hardly detected.
The obtained ferment is subjected to high performance liquid chromatography detection, the content of procyanidine in the prepared cowberry fruit ferment is 36.25mg/ml, the content of organic acid is 8.45mg/ml, and the cowberry fruit ferment mainly comprises lactic acid and citric acid, has low pyruvic acid content and hardly detects succinic acid through HPLC organic acid type analysis.
Application example
The enzyme prepared by the above embodiment and the enzyme obtained by the comparative example are used for testing the application effect of the enzyme in cosmetics:
first, test method
1. Test grouping
The test is divided into 5 groups and 5 groups of test groups (the blueberry ferment of comparative example 1, comparative example 2, comparative example 3, example 2 and example 1 is added into the prepared facial mask essence respectively and is respectively used as the test groups 1, 2, 3, 4 and 5, the addition amount is 0.80%), and the control sample is the facial mask essence prepared in advance.
2. Test method
Test method 1: double-blind, random and contrast tests were selected, 100 volunteers were selected, 20 per group, and the prepared test samples 1-5 were used for area contrast tests for observation of effects. The left face tested the control sample, the right face tested the test samples (1) - (5), used nightly, and the use was terminated for 15 days. Before and after use, artificial observation and scoring are adopted for comparison, the observation result is recorded in a scoring system of 0-10 points, and the higher the score is, the better the effect is. The scoring results were used to analyze the effect of bilberry enzyme in the serum.
The test method 2: the blueberry enzyme of example 2 was applied to a cosmetic mask (0.80% enzyme was added to the mask essence), and the mask was subjected to sensory tests. 20 volunteers are selected, a skin tester is used for performing instrument skin test on the tested person, data test is respectively performed on the forehead, the periphery of the eye and the cheek before and after the wild Vaccinium myrtillus seaweed mask is used, the test indexes comprise elasticity, oil content and water content, and the test is repeated for 3 times.
3. Test results (see fig. 1 and table 1).
TABLE 1 test samples (1) - (5) for skin lightening effect test results
According to test results, the cowberry fruit ferment prepared by the method has an obvious effect of brightening the skin color. The obtained enzyme was pale purple and transparent. The biological complex enzyme decomposes cellulose, pectin, polysaccharide, pigment and the like in the cowberry fruit pulp, so that the contents of active ingredients such as procyanidine, organic acid, vitamin and the like in the prepared enzyme are increased, and the quality and the yield of the cowberry enzyme are improved. Application tests show that the essence added with the cowberry fruit ferment has the effects of brightening the uniform skin color and improving the skin elasticity and water content compared with the common essence;
meanwhile, according to the technical scheme, the specific strain is adopted at a certain sugar content, and the ferment obtained by decomposing the specific enzyme is compared with the ferment obtained by a comparative example, so that the ferment obtained by the invention has a more obvious effect of brightening the uniform skin color, and the ferment obtained by the embodiment of the invention has a remarkable effect, and especially, when the addition amount of cellulase is controlled to be 0.25%, the addition amount of hemicellulase is 0.25%, the addition amount of pectinase is 0.14%, the addition amount of acid protease is 0.25%, and meanwhile, the addition amount of sugar is controlled to be 16%, the blueberry ferment with the optimal quality can be obtained. Under the condition, the contents and types of procyanidine and organic acid of the obtained enzyme are the highest, so that the effect of brightening and homogenizing skin color is the best.
And the results of data testing by a skin tester before and after 20 volunteers randomly selected from 100 volunteers by using the enzyme of example 2 of the present invention are shown (see fig. 1), it can be seen from fig. 1 that the indexes of elasticity, moisture and oil content of the face before and after the enzyme mask of example 2 of the present invention is used are obviously changed, wherein the elasticity coefficient and the moisture coefficient of the skin of the face mask using wild vaccinium myrtillus seaweed are obviously increased, and the skin oil content coefficient is greatly reduced. Therefore, the mask essence prepared by the blueberry enzyme disclosed by the invention has the effects of obviously improving the skin elasticity value and the water content, and no one of all volunteers has symptoms such as skin allergy and the like.
Comprehensive analysis shows that the ferment is rich in procyanidin, organic acid and vitamins, and the combined action of the procyanidin, the organic acid and the vitamins is visible, application tests show that the essence added with the blueberry ferment has the effects of brightening the uniform skin color and improving the skin elasticity and the water content compared with the common essence, and the blueberry ferment prepared by the method is excellent in quality and suitable for being added into cosmetics.
Claims (8)
1. A vaccinium myrtillus ferment prepared by utilizing compound biological enzyme is characterized in that: adding sugar into the vaccinium myrtillus after enzymolysis, and sealing and fermenting the vaccinium myrtillus by using mixed bacteria to obtain vaccinium myrtillus ferment; wherein the composite bacteria are Saccharomyces cerevisiae, Lactobacillus plantarum and Bifidobacterium.
2. The bilberry enzyme prepared by using the complex bio-enzyme according to claim 1, wherein: the mixed enzyme adopted by the enzymolysis is cellulase, hemicellulase, pectinase and acid protease; the respective addition amounts are based on the mass ratio of the cowberry fruit crushing mixed solution, the addition amount of the cellulase is 0.1-0.5%, the addition amount of the hemicellulase is 0.1-0.5%, the addition amount of the pectinase is 0.12-0.18%, and the addition amount of the acid protease is 0.1-0.5%.
3. The method for preparing Vaccinium myrtillus ferment prepared by complex biological enzyme according to claim 1, which comprises the following steps: mixing the cleaned and crushed Vaccinium myrtillus with water, adding mixed enzyme, performing enzymolysis in water bath with pH value of 4.0-5.0 and 50-60 deg.C for 2-4h, adding sugar with quality of 14-20% of the crushed mixture, inoculating mixed bacteria to the enzymolysis product, and sealing and fermenting to obtain the Vaccinium myrtillus ferment.
4. The method for preparing Vaccinium myrtillus ferment by using composite biological enzyme as claimed in claim 3, wherein: the method comprises the steps of cleaning and crushing the refrigerated fresh vaccinium myrtillus fruits, and mixing the crushed fruits with water according to the volume ratio of 1-3:0.8-1.5 for later use.
5. The method for preparing Vaccinium myrtillus ferment by using composite biological enzyme as claimed in claim 3, wherein: adding mixed enzyme into the cowberry fruit crushing mixed solution, uniformly mixing, adjusting the pH value of the mixed solution to 4.0-5.0, and placing in a water bath at 50-60 ℃ for enzymolysis for 2-4 h; then adding sugar to adjust the sugar degree, adjusting the sugar degree at room temperature, and adding sugar which is 14-20% of the mass of the blueberry crushing mixed liquid; adjusting sugar degree, sterilizing, and inactivating; and (5) standby.
6. The method for preparing Vaccinium myrtillus ferment using complex bio-enzyme according to claim 3 or 5, wherein: the mixed enzyme adopted by the enzymolysis is cellulase, hemicellulase, pectinase and acid protease; the respective addition amounts are based on the mass ratio of the cowberry fruit crushing mixed solution, the addition amount of the cellulase is 0.1-0.5%, the addition amount of the hemicellulase is 0.1-0.5%, the addition amount of the pectinase is 0.12-0.18%, and the addition amount of the acid protease is 0.1-0.5%.
7. The method for preparing Vaccinium myrtillus ferment by using composite biological enzyme as claimed in claim 3, wherein: the sealed fermentation comprises the following steps: respectively activating yeast Saccharomyces cerevisiae, Lactobacillus plantarum and Bifidobacterium, and culturing to middle and late logarithmic growth stage; standby;
and mixing the enzymolysis product with activated saccharomyces cerevisiae, lactobacillus plantarum and bifidobacterium, uniformly stirring according to the mass ratio of 40-60:0.8-1.5:0.8-1.5:0.8-1.5, sealing, fermenting for 4-7 days at the temperature of 25-32 ℃, and stirring once every 10 hours to obtain the fermentation liquor.
8. The method of preparing Vaccinium myrtillus ferment using complex bio-enzyme according to claim 7, wherein: and filtering and sterilizing the fermentation liquor to obtain the vaccinium myrtillus ferment.
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