阅读说明：本技术 一种水果酵素及其制备方法 (Fruit enzyme and preparation method thereof ) 是由 耿燕 史劲松 段文慧 张晓娟 曹刚刚 贾友彬 李宣衡 于 2021-07-13 设计创作，主要内容包括：本发明公开了一种水果酵素及其制备方法。本发明采用乌梅果肉,利用酵母菌、乳酸菌、芽孢杆菌、醋酸菌对乌梅进行协同发酵。酵母菌利用自身酶系统将乌梅中的糖水解成单糖,并加入乳酸菌与酵母菌协同发酵,能够对单糖有更加充分的利用,产生乳酸并分泌具有杀菌功能的乳酸菌素来抑制有害菌的生长,同时,乳酸菌本身所产生的酸性代谢产物,维持了酒精发酵环境的偏酸性。在醋酸发酵过程中添加了芽孢杆菌,芽孢杆菌发酵后产生更多酚类物质,增强发酵液抗氧化活性。同时也添加了乳酸菌,乳酸菌的对乌梅进一步发酵,为发酵微生物提供生长繁殖可利用的必需氨基酸和各种维生素,促进发酵微生物的生长与繁殖,也能提高发酵酶系对乌梅的发酵能力。(The invention discloses a fruit ferment and a preparation method thereof. The method adopts the dark plum pulp, and utilizes saccharomycetes, lactic acid bacteria, bacillus and acetic acid bacteria to carry out synergistic fermentation on the dark plum. The saccharomycete hydrolyzes sugar in the dark plum into monosaccharide by using an enzyme system of the saccharomycete, adds the lactic acid bacteria and the saccharomycete for synergistic fermentation, can utilize the monosaccharide more fully, generates lactic acid and secretes the lactic acid bacteria element with a sterilization function to inhibit the growth of harmful bacteria, and simultaneously, acidic metabolites generated by the lactic acid bacteria maintain the acidity of an alcohol fermentation environment. The bacillus is added in the acetic fermentation process, and more phenolic substances are generated after the bacillus is fermented, so that the antioxidant activity of the fermentation liquor is enhanced. Meanwhile, lactic acid bacteria are added, and further fermentation of the dark plum by the lactic acid bacteria provides necessary amino acids and various vitamins which can be utilized by growth and reproduction for fermentation microorganisms, promotes the growth and reproduction of the fermentation microorganisms, and can also improve the fermentation capacity of the fermentation enzyme system to the dark plum.)

1. The fermentation method of the fruit ferment is characterized by comprising the following steps:

s1, raw material pretreatment: cleaning mume fructus, removing core, pulping to obtain pulp, and performing enzymolysis to obtain enzymolysis pulp;

s2, alcoholic fermentation: adding saccharomyces cerevisiae into the enzymolysis fruit pulp obtained in the step S1, fermenting for 20-30h at 28-30 ℃, supplementing 0.5-2% of glucose, adding lactic acid bacteria, stirring once every 0.5-2 days, and fermenting in a sealed manner for 5-7 days;

s3, acetic fermentation: inoculating lactobacillus and bacillus into the fermentation liquor obtained in the step S2, heating to 35-37 ℃, fermenting for 2-4 days, inoculating acetic acid bacteria, and performing mixed fermentation for 4-6 days at 30-35 ℃ to obtain fermented vinegar liquid, wherein the stirring speed in the whole acetic acid fermentation process is 180-200 r/min;

s4, filtering, blending, sterilizing and canning the fermented vinegar liquid obtained in the step S3 to obtain the fruit enzyme.

2. The fermentation method according to claim 1, wherein in the step S2, the Saccharomyces cerevisiae is Saccharomyces cerevisiae CICC1012, and the lactic acid bacteria are Lactobacillus fermentum LFE02 with a preservation number of CGMCC No. 12934.

3. The fermentation method according to claim 1, wherein the amount of the inoculated Saccharomyces cerevisiae in the step of S2 is 10¹⁰-10¹²Per kg; the inoculation amount of the lactobacillus is 10¹²-10¹⁴One per kg.

4. The fermentation method according to claim 1, wherein in the step S3, the lactic acid bacteria are one or more of Lactobacillus fermentum with a collection number of CICC21829, Lactobacillus plantarum AR-307 with a collection number of CGMCC No.18388, and Lactobacillus plantarum P-2 with a collection number of CGMCC No. 18389; the bacillus is Bacillus belgii with the preservation number of CICC 24434; the acetic acid bacteria is Pasteur acetic acid bacteria G3-2 with the preservation number of CGMCC No. 12930.

5. The fermentation method according to claim 1, wherein the amount of acetic acid bacteria inoculated in the step S3 is 10⁸-10¹⁰Per kg; the inoculation amount of the lactic acid bacteria is 10¹²-10¹⁵Per kg; the inoculation amount of the bacillus is 10⁸-10¹⁰One per kg.

6. The fermentation method of claim 1, wherein in the step S1, the enzymolysis is performed by using cellulase and pectinase to respectively hydrolyze the pulp, and after the enzymolysis, the two hydrolyzed pulp are mixed according to a ratio of 5-8:3-4 to obtain a mixed hydrolyzed pulp.

7. The fermentation method according to claim 6, wherein the cellulase and pectinase are added in an amount of 0.4-0.6% by mass of the pulp.

8. The fermentation method according to claim 6, wherein the enzymolysis temperature is 45-50 ℃ and the enzymolysis time is 2-2.5 h.

9. The fermentation method as claimed in claim 1, wherein in the step S4, the filtering is performed by diatomite, the blending is performed by adding honey, and the sterilization is performed by 140-160MPa high-pressure instantaneous sterilization for homogeneous sterilization.

10. A fruit ferment prepared by the fermentation method according to any one of claims 1 to 9.

Technical Field

The invention relates to a fruit ferment and a preparation method thereof, and belongs to the technical field of biological fermentation.

Background

The enzyme is a functional fermentation product obtained by fermenting animals, plants or fungi with microorganisms. The edible plant enzyme defined by the national standard enzyme product classification guide (QB/T5324) is an enzyme product which is prepared by taking plants capable of being used for food processing as main raw materials, adding or not adding auxiliary materials and carrying out microbial fermentation and contains specific bioactive components and can be eaten by human beings, so that a large number of plant fermentation products formed by microbial fermentation in the market belong to the field of edible enzymes, which is a provision from the aspects of industrial application and industrial practice and is beneficial to the development and development of enzyme industry. However, this also provides the possibility for many low-quality enzyme products to enter the market. For example, simple fermentation with a single lactic acid bacterium, yeast, acetic acid bacterium, or the like, although it is formally in accordance with the concept of enzyme, or improved in terms of taste and function, these products do not fully embody the technical connotation of enzyme products.

In recent years, more and more researches on flavone polyphenol in food are carried out, and the flavone polyphenol is an important functional substance. The flavone polyphenol is widely present in various plants and fruits, has strong free radical scavenging ability, and also has antibacterial, anti-inflammatory, immunity enhancing, cardiovascular improving, liver protecting, anti-tumor, and anticancer effects. The dark plum also contains polyphenol flavonoids, which is one of the important reasons that dark plum has the effects of resisting oxidation, resisting bacteria, diminishing inflammation, protecting liver, resisting tumors and the like. Therefore, the flavone polyphenol can also be used as an important index for evaluating the functionality of the ferment.

At present, the research on dark plum enzyme products is less, and an invention patent with the patent application number of CN107890094A discloses a fruit enzyme powder, but the invention only utilizes dark plum juice, which may cause the loss of some functional components in dark plum and can not fully exert the functions of dark plum. The fermentation process of the invention is simple, and the raw materials are not sufficiently fermented, so that the produced nutrient functional substances are greatly reduced. The invention patent with the patent application number of CN109908305A discloses a fruit and vegetable enzyme and a processing technology thereof. However, the invention only utilizes the lactic acid bacteria agent, and the dark plum is only one component in the formula, so the contribution degree of the dark plum to the product efficacy is not clear. At present, a ferment fermentation method capable of fully utilizing dark plums does not exist.

The traditional enzyme product is rooted in the traditional fermentation and is of a multi-bacterium mixed ecological fermentation type, so the fermentation form and the process thereof fully embody the action of environmental functional microorganisms and follow the principle of natural selection, wherein a plurality of ecological microorganisms have a long-term competitive, alternating and evolving fermentation process, and in the process, plant raw materials are fully transformed and utilized for multiple rounds. Pure fermentation, lactic acid bacteria and yeast, can not achieve good effect under single action. Even if multi-bacterium composite fermentation is carried out, an open environment is not simply adopted for fermentation, but a multi-bacterium synergistic mechanism is needed to be known, the characteristics of raw materials and the target of fermentation conversion are known, specific functional microorganisms are screened on the basis, a composite fermentation mode is needed to be researched, and a multi-bacterium composite fermentation process is determined by carrying out necessary detection and process analysis on characteristic products formed by fermentation in the fermentation process.

Disclosure of Invention

In order to solve the problems, the invention provides a fermentation method of fruit ferment, which obviously improves the content of organic acid and the content of total flavone polyphenol in the ferment.

The first purpose of the present invention is to provide a fermentation method of fruit ferment, comprising the following steps:

s1, raw material pretreatment: cleaning mume fructus, removing core, pulping to obtain pulp, and performing enzymolysis to obtain enzymolysis pulp;

s2, alcoholic fermentation: adding saccharomyces cerevisiae into the enzymolysis fruit pulp obtained in the step S1, fermenting for 20-30h at 28-30 ℃, supplementing 0.5-2% of glucose, adding lactic acid bacteria, stirring once every 0.5-2 days, and fermenting in a sealed manner for 5-7 days;

s3, acetic fermentation: inoculating lactobacillus and bacillus into the fermentation liquor obtained in the step S2, heating to 35-37 ℃, fermenting for 2-4 days, inoculating acetic acid bacteria, and performing mixed fermentation for 4-6 days at 30-35 ℃ to obtain fermented vinegar liquid, wherein the stirring speed in the whole acetic acid fermentation process is 180-200 r/min;

s4, filtering, blending, sterilizing and canning the fermented vinegar liquid obtained in the step S3 to obtain the fruit enzyme.

Further, in the step S2, the Saccharomyces cerevisiae is Saccharomyces cerevisiae (CICC 1012), the Lactobacillus is Lactobacillus fermentum (LFE 02), and the preservation number is CGMCC No. 12934.

Further, in the step S2, the inoculation amount of the saccharomyces cerevisiae is 10¹⁰-10¹²Per kg; the inoculation amount of the lactobacillus is 10¹²-10¹⁴One per kg.

Further, in the step S3, the Lactobacillus is Lactobacillus fermentum (Lactobacillus fermentum) with the preservation number of CICC21829 and Lactobacillus plantarum AR-307 with the preservation number of CGMCC No.18388 and Lactobacillus plantarum P-2 with the preservation number of one or more of CGMCC No. 18389; the Bacillus is Bacillus velezensis (Bacillus velezensis), and the preservation number is CICC 24434; the acetic acid bacteria is Pasteur acetic acid bacteria (Acetobacter passarianus) G3-2, and the preservation number is CGMCC No. 12930.

Further, in the step S3, the inoculation amount of acetic acid bacteria is 10⁸-10¹⁰Per kg; the inoculation amount of the lactic acid bacteria is 10¹²-10¹⁵Per kg; the inoculation amount of the bacillus is 10⁸-10¹⁰One per kg.

Further, in the step S1, the enzymolysis is to use cellulase and pectinase to respectively carry out enzymolysis on the fruit pulp, and after the enzymolysis, the two kinds of enzymolysis fruit pulp are mixed according to the ratio of 5-8:3-4 to obtain the mixed enzymolysis fruit pulp.

Furthermore, the addition amount of the cellulase and the pectinase is 0.4-0.6 percent of the mass of the fruit pulp.

Further, the enzymolysis temperature is 45-50 ℃, and the enzymolysis time is 2-2.5 h.

Further, in the step S4, the filtering is performed by diatomite, the blending is performed by adding honey, and the sterilizing is performed by 140-160MPa high-pressure instant sterilizing for homogenizing and sterilizing.

The second purpose of the invention is to provide the fruit ferment prepared by the method.

The invention has the beneficial effects that:

the dark plum fruit vinegar is prepared by adopting high-quality dark plum fruit pulp and performing synergistic fermentation on dark plum fruit by utilizing saccharomycetes, lactic acid bacteria, bacillus and acetic acid bacteria. The yeast utilizes an enzyme system of the yeast to hydrolyze sugar in the dark plum into monosaccharide, simultaneously glucose is added to facilitate the growth of the yeast, and lactic acid bacteria and the yeast are further added to perform synergistic action to perform fermentation together, so that the monosaccharide can be more fully utilized, nutrient functional substance lactic acid is generated, lactobacillus elements with a sterilization function are secreted to inhibit the growth of harmful bacteria, and meanwhile, acidic metabolites generated by the lactic acid bacteria maintain the acidity of an alcohol fermentation environment. Further, bacillus is added in the acetic fermentation process, and the bacillus can generate more phenolic substances after fermentation, so that the antioxidant activity of the fermentation liquor is enhanced. Meanwhile, lactic acid bacteria are added, and further fermentation of the dark plum by the lactic acid bacteria provides essential amino acids and various vitamins which can be utilized by growth and reproduction for fermentation microorganisms, promotes the growth and reproduction of the fermentation microorganisms, and can also improve the fermentation capacity of the fermentation enzyme system to the dark plum. Lactic acid generated by fermentation of lactic acid bacteria not only enables the ferment to have antibacterial and anti-inflammatory functions, but also enables the taste and flavor to be better.

Detailed Description

The present invention is further described below in conjunction with specific examples to enable those skilled in the art to better understand the present invention and to practice it, but the examples are not intended to limit the present invention.

Example 1: preparation method of high-quality enzyme with high polyphenol and flavone content

(1) Pretreatment of raw materials: cleaning smoked plums, removing cores, pulping, dividing pulp into two parts, respectively adding cellulase and pectinase which are 0.5 percent of the total mass of raw materials, wherein the enzymolysis temperature is 45 ℃, and the enzymolysis time is 2.5 hours, so as to obtain two kinds of enzymolysis pulp, wherein the two kinds of enzymolysis pulp are 6: 4 mixing for later use;

(2) alcohol fermentation: adding 10 into the fruit pulp obtained in the step (1)¹⁰Fermenting with Saccharomyces cerevisiae CICC1012 at 30 deg.C for 24 hr, supplementing 1% glucose, and adding 10% glucose¹²Fermenting for 6 days in a closed environment, wherein each kg of lactobacillus fermentum CGMCC No.12934 is stirred once every 1 day;

(3) acetic acid fermentation: 10 is inoculated into the fermentation liquor obtained in the step (2)¹²Lactobacillus plantarum CGMCC No.18388 and 10 per kg⁸Heating Bacillus belgii CICC24434 to 35 deg.C, fermenting for 2 days, inoculating 10⁸Mixing and fermenting acetic acid bacteria per kg of the acetic acid bacteria CGMCC No.12930 at 32 ℃ for 4 days to obtain fermentation enzyme, wherein the stirring speed of the whole acetic acid fermentation process is 200 r/min;

(4) and (3) filtering: adding diatomite which is 0.3 percent of the weight of the ferment liquid into the fermented vinegar liquid obtained in the step (3) for filtering treatment to obtain clear ferment liquid;

(5) blending: adding 7% of honey into the clear enzyme liquid obtained in the step (4) for blending;

(6) high-pressure homogenizing and sterilizing: carrying out homogenizing sterilization treatment on the ferment instantly under the high pressure of 160 MPa;

(7) filling: and (5) sterile filling to finally prepare the high-quality ferment with high polyphenol and flavone content.

Example 2: preparation method of high-quality enzyme with high polyphenol and flavone content

(1) Pretreatment of raw materials: cleaning smoked plums, removing cores, pulping, dividing pulp into two parts, respectively adding cellulase and pectinase which are 0.5 percent of the total mass of raw materials, wherein the enzymolysis temperature is 45 ℃, and the enzymolysis time is 2.5 hours, so as to obtain two kinds of enzymolysis pulp, and the two kinds of enzymolysis pulp are 7: 3 mixing for later use;

(2) alcohol fermentation: adding 10 into the fruit pulp obtained in the step (1)¹²Brewing wine of one/kgFermenting with yeast CICC1012 at 30 deg.C for 24 hr, supplementing 2% glucose, and adding 10%¹³Fermenting for 6 days in a closed environment, wherein each kg of lactobacillus fermentum CGMCC No.12934 is stirred once every 1 day;

(3) acetic acid fermentation: 10 is inoculated into the fermentation liquor obtained in the step (2)¹⁴Lactobacillus plantarum CGMCC No.18388 and 10 per kg¹⁰Heating Bacillus belgii CICC24434 to 35 deg.C, fermenting for 3 days, inoculating 10⁹Mixing and fermenting acetic acid bacteria per kg of the acetic acid bacteria of Pasteur CGMCC No.12930 at 32 ℃ for 5 days to obtain fermentation enzyme, wherein the stirring speed of the whole acetic acid fermentation process is 200 r/min;

(4) and (3) filtering: adding diatomite which is 0.3 percent of the weight of the ferment liquid into the fermented vinegar liquid obtained in the step (3) for filtering treatment to obtain clear ferment liquid;

(5) blending: adding 7% of honey into the clear enzyme liquid obtained in the step (4) for blending;

(6) high-pressure homogenizing and sterilizing: carrying out homogenizing sterilization treatment on the ferment instantly under the high pressure of 160 MPa;

(7) filling: and (5) sterile filling to finally prepare the high-quality ferment with high polyphenol and flavone content.

Comparative example 1:

(1) pretreatment of raw materials: cleaning smoked plums, removing cores, pulping, dividing pulp into two parts, respectively adding cellulase and pectinase which are 0.5 percent of the total mass of raw materials, wherein the enzymolysis temperature is 45 ℃, and the enzymolysis time is 2.5 hours, so as to obtain two kinds of enzymolysis pulp, wherein the two kinds of enzymolysis pulp are 6: 4 mixing for later use;

(2) alcohol fermentation: adding 10 into the fruit pulp obtained in the step (1)¹⁰Fermenting the saccharomyces cerevisiae CICC1012 of each kg for 6 days in a closed environment at the temperature of 30 ℃, and stirring once every 1 day;

(3) acetic acid fermentation: 10 is inoculated into the fermentation liquor obtained in the step (2)¹²Lactobacillus plantarum CGMCC No.18388 and 10 per kg⁸Heating Bacillus belgii CICC24434 to 35 deg.C, fermenting for 2 days, inoculating 10⁸Mixing and fermenting the raw materials at 32 deg.C for 4 days to obtain fermentation liquorStirring the acetic acid fermentation at the stirring speed of 200r/min in the whole process;

(4) and (3) filtering: adding diatomite which is 0.3 percent of the weight of the ferment liquid into the fermented vinegar liquid obtained in the step (3) for filtering treatment to obtain clear ferment liquid;

(5) blending: adding 7% of honey into the clear enzyme liquid obtained in the step (4) for blending;

(6) high-pressure homogenizing and sterilizing: carrying out homogenizing sterilization treatment on the ferment instantly under the high pressure of 160 MPa;

(7) filling: and (5) sterile filling to finally obtain the ferment.

Comparative example 2:

(1) pretreatment of raw materials: cleaning smoked plums, removing cores, pulping, dividing pulp into two parts, respectively adding cellulase and pectinase which are 0.5 percent of the total mass of raw materials, wherein the enzymolysis temperature is 45 ℃, and the enzymolysis time is 2.5 hours, so as to obtain two kinds of enzymolysis pulp, wherein the two kinds of enzymolysis pulp are 6: 4 mixing for later use;

(2) alcohol fermentation: adding 10 into the fruit pulp obtained in the step (1)¹⁰Fermenting with Saccharomyces cerevisiae CICC1012 at 30 deg.C for 24 hr, supplementing 1% glucose, and adding 10% glucose¹²Fermenting for 6 days in a closed environment, wherein each kg of lactobacillus fermentum CGMCC No.12934 is stirred once every 1 day;

(3) acetic acid fermentation: 10 is inoculated into the fermentation liquor obtained in the step (2)¹²Lactobacillus plantarum CGMCC No. 18388/kg, heating to 35 deg.C, fermenting for 2 days, inoculating 10⁸Mixing and fermenting acetic acid bacteria per kg of the acetic acid bacteria CGMCC No.12930 at 32 ℃ for 4 days to obtain fermentation enzyme, wherein the stirring speed of the whole acetic acid fermentation process is 200 r/min;

(4) and (3) filtering: adding diatomite which is 0.3 percent of the weight of the ferment liquid into the fermented vinegar liquid obtained in the step (3) for filtering treatment to obtain clear ferment liquid;

(5) blending: adding 7% of honey into the clear enzyme liquid obtained in the step (4) for blending;

(6) high-pressure homogenizing and sterilizing: carrying out homogenizing sterilization treatment on the ferment instantly under the high pressure of 160 MPa;

(7) filling: and (5) sterile filling to finally obtain the ferment.

Comparative example 3:

(1) pretreatment of raw materials: cleaning smoked plums, removing cores, pulping, dividing pulp into two parts, respectively adding cellulase and pectinase which are 0.5 percent of the total mass of raw materials, wherein the enzymolysis temperature is 45 ℃, and the enzymolysis time is 2.5 hours, so as to obtain two kinds of enzymolysis pulp, wherein the two kinds of enzymolysis pulp are 6: 4 mixing for later use;

(2) alcohol fermentation: adding 10 into the fruit pulp obtained in the step (1)¹⁰Fermenting with Saccharomyces cerevisiae CICC1012 at 30 deg.C for 24 hr, supplementing 1% glucose, and adding 10% glucose¹²Fermenting for 6 days in a closed environment, wherein each kg of lactobacillus fermentum CGMCC No.12934 is stirred once every 1 day;

(3) acetic acid fermentation: 10 is inoculated into the fermentation liquor obtained in the step (2)⁸Heating Bacillus belgii CICC24434 to 35 deg.C, fermenting for 2 days, inoculating 10⁸Mixing and fermenting acetic acid bacteria per kg of the acetic acid bacteria CGMCC No.12930 at 32 ℃ for 4 days to obtain fermentation enzyme, wherein the stirring speed of the whole acetic acid fermentation process is 200 r/min;

(4) and (3) filtering: adding diatomite which is 0.3 percent of the weight of the ferment liquid into the fermented vinegar liquid obtained in the step (3) for filtering treatment to obtain clear ferment liquid;

(5) blending: adding 7% of honey into the clear enzyme liquid obtained in the step (4) for blending;

(6) high-pressure homogenizing and sterilizing: carrying out homogenizing sterilization treatment on the ferment instantly under the high pressure of 160 MPa;

(7) filling: and (5) sterile filling to finally obtain the ferment.

Comparative example 4:

(1) pretreatment of raw materials: cleaning mume fructus, removing core, pulping, adding cellulase and pectinase 0.5% of the total weight of the raw materials, performing enzymolysis at 45 deg.C for 2.5 hr to obtain enzymolysis pulp;

(2) alcohol fermentation: adding 10 into the fruit pulp obtained in the step (1)¹⁰Fermenting with Saccharomyces cerevisiae CICC1012 at 30 deg.C for 24 hr, and supplementing 1% of glucoseAdding glucose, and adding 10¹²Fermenting for 6 days in a closed environment, wherein each kg of lactobacillus fermentum CGMCC No.12934 is stirred once every 1 day;

(3) acetic acid fermentation: 10 is inoculated into the fermentation liquor obtained in the step (2)¹²Lactobacillus plantarum CGMCC No.18388 and 10 per kg⁸Heating Bacillus belgii CICC24434 to 35 deg.C, fermenting for 2 days, inoculating 10⁸Mixing and fermenting acetic acid bacteria per kg of the acetic acid bacteria CGMCC No.12930 at 32 ℃ for 4 days to obtain fermentation enzyme, wherein the stirring speed of the whole acetic acid fermentation process is 200 r/min;

(4) and (3) filtering: adding diatomite which is 0.3 percent of the weight of the ferment liquid into the fermented vinegar liquid obtained in the step (3) for filtering treatment to obtain clear ferment liquid;

(5) blending: adding 7% of honey into the clear enzyme liquid obtained in the step (4) for blending;

(6) high-pressure homogenizing and sterilizing: carrying out homogenizing sterilization treatment on the ferment instantly under the high pressure of 160 MPa;

(7) filling: and (5) sterile filling to finally prepare the high-quality ferment with high polyphenol and flavone content.

Control group:

(1) pretreatment of raw materials: cleaning smoked plums, removing cores, pulping, dividing pulp into two parts, respectively adding cellulase and pectinase which are 0.5 percent of the total mass of raw materials, wherein the enzymolysis temperature is 45 ℃, and the enzymolysis time is 2.5 hours, so as to obtain two kinds of enzymolysis pulp, wherein the two kinds of enzymolysis pulp are 6: 4 mixing for later use;

(2) alcohol fermentation: adding 10 into the fruit pulp obtained in the step (1)¹⁰Fermenting per kg of Saccharomyces cerevisiae CICC1012 at 30 deg.C for 6 days in a closed environment, and stirring once every 1 day;

(3) acetic acid fermentation: 10 is inoculated into the fermentation liquor obtained in the step (2)⁸The acetic acid bacteria per kg of the acetic acid bacteria are CGMCC No.12930, and the fermentation enzyme is obtained by fermenting for 4 days at the temperature of 32 ℃, wherein the stirring speed is 200r/min in the whole acetic acid fermentation process;

(4) and (3) filtering: adding diatomite which is 0.3 percent of the weight of the ferment liquid into the fermented vinegar liquid obtained in the step (3) for filtering treatment to obtain clear ferment liquid;

(5) blending: adding 7% of honey into the clear enzyme liquid obtained in the step (4) for blending;

(6) high-pressure homogenizing and sterilizing: carrying out homogenizing sterilization treatment on the ferment instantly under the high pressure of 160 MPa;

(7) filling: and (5) sterile filling to finally obtain the ferment.

Test example:

in order to determine the content of organic acids and total flavonoids in the enzyme, the experiment was divided into 7 groups, which were respectively a control group, an experiment group 1, an experiment group 2, an experiment group 3, an experiment group 4, an experiment group 5, and an experiment group 6. Wherein experimental group 1, experimental group 2, experimental group 3, experimental group 4, experimental group 5, and experimental group 6 correspond to the ferments prepared in example 1, example 2, comparative example 1, comparative example 2, comparative example 3, and comparative example 4.

Method for measuring organic acid: the method for detecting the organic acid in the enzyme is modified by using the method in the national standard GB 5009.157-2016 as a reference. The method is used for measuring the content of various organic acids in a control group and an experimental group, including acetic acid, lactic acid, citric acid, oxalic acid, malic acid, succinic acid, tartaric acid, pyruvic acid, alpha-ketoglutaric acid and fumaric acid. The organic acid content results are shown in table 1.

TABLE 1 enzyme organic acid content

Significant differences were assessed using the duncan multiple comparison test. The difference between different letters within a column is statistically significant (p <0.05), and the same letter indicates no significant difference (p > 0.05).

The method for measuring the total flavone comprises the following steps: the content of the total flavonoids in the control group and the experimental group is determined according to the determination method of the total flavonoids in the national standard GB/T19777-2013.

TABLE 2 enzyme Total Polyphenol flavone content

Significant differences were assessed using the duncan multiple comparison test. The difference between different letters within a column is statistically significant (p <0.05), and the same letter indicates no significant difference (p > 0.05).

TABLE 3 enzyme antioxidant capacity

Significant differences were assessed using the duncan multiple comparison test. The difference between different letters within a column is statistically significant (p <0.05), and the same letter indicates no significant difference (p > 0.05).

From the above data, the types and contents of the organic acids in the high-quality ferment prepared by the invention are all higher than those of the comparative example, wherein the content of the lactic acid reaches 40% of the total organic acid content. In the embodiment, the content of the total flavone serving as the functional component is also obviously higher than that of the comparative example, and the content can reach 6.2 mg/mL. In addition, the antioxidant activity of the examples was better than that of the comparative examples. Therefore, the high-polyphenol-flavone-content high-quality enzyme obtained by fermenting dark plum in a synergistic manner by using multiple strains is sour and refreshing in taste, rich in nutritional functional components and mellow in flavor, and is an enzyme product with good taste and nutritional and health-care effects.

The above-mentioned embodiments are merely preferred embodiments for fully illustrating the present invention, and the scope of the present invention is not limited thereto. The equivalent substitution or change made by the technical personnel in the technical field on the basis of the invention is all within the protection scope of the invention. The protection scope of the invention is subject to the claims.