CN115349586A - Enzyme for multi-source enzyme beverage and preparation method thereof - Google Patents

Enzyme for multi-source enzyme beverage and preparation method thereof Download PDF

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CN115349586A
CN115349586A CN202211076696.7A CN202211076696A CN115349586A CN 115349586 A CN115349586 A CN 115349586A CN 202211076696 A CN202211076696 A CN 202211076696A CN 115349586 A CN115349586 A CN 115349586A
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parts
weight
concentrated juice
ferment
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魏启山
邓宇初
陈淑萍
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Bazhou Xindeyuan Food Co ltd
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/38—Other non-alcoholic beverages
    • A23L2/382—Other non-alcoholic beverages fermented
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/42—Preservation of non-alcoholic beverages
    • A23L2/44—Preservation of non-alcoholic beverages by adding preservatives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/70—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter
    • A23L2/84—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter using microorganisms or biological material, e.g. enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

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Abstract

The invention discloses a ferment for a multi-source ferment beverage and a preparation method thereof, wherein the ferment for the multi-source ferment beverage is prepared from the following raw materials: sucrose, pineapple concentrated juice, blueberry concentrated juice, dragon fruit concentrated juice, cranberry concentrated juice, mulberry concentrated juice, lemon concentrated juice, honey, apple vinegar, licorice powder, burdock powder, water and bud paste vegetable water extract. Uniformly mixing the multi-source ferment for the ferment beverage through the S1 raw material; s2, sterilizing; s3, fermenting; s4, filtering to obtain the product. The enzyme polyphenol for the multi-source enzyme beverage prepared by the invention has high flavone content, high DPPH free radical clearance rate and low sugar content, and has wide market potential.

Description

Multi-source enzyme for enzyme beverage and preparation method thereof
Technical Field
The invention belongs to the technical field of enzymes, and particularly relates to an enzyme for a multi-source enzyme beverage and a preparation method of the enzyme.
Background
The traditional fruit compound beverage or fruit juice beverage has the defects of high sugar content and high calorie, and is easy to cause obesity, so that excessive intake is not suitable. The ferment is a mixed fermentation broth which is prepared by taking fresh vegetables, fruits, medicinal and edible traditional Chinese medicines and the like as raw materials, juicing or extracting a series of processes, and fermenting by microorganisms and contains abundant nutrient components such as saccharides, organic acids, mineral substances, vitamins, phenols, terpenes and the like and some important bioactive substances such as enzymes and the like. Through fermentation, the raw material tissue is disintegrated under the action of microorganisms, the functional components are fully released and converted into micromolecular nutrient substances which are easily absorbed by human bodies, the efficacy can be further improved through fermentation and conversion, the side effects can be reduced, or bioactive substances with the functions of promoting gastrointestinal motility, reducing blood fat, resisting oxidation and improving the body immunity can be generated.
The invention patent CN 105124581A provides a preparation method of an enzyme stock solution for a multi-source enzyme beverage. The composite material consists of the following raw materials in percentage by weight: 20% of grapefruit, 5% of traditional Chinese medicine extract, 5% of composite esterifying enzyme ecological microbial inoculum, 2% of pomegranate seeds, 2% of lychee seeds, 0.5% of dendrobium stem flowers, 0.5% of luffa flowers, 0.3% of cassia bark, 5% of carrot leaves, 0.3% of hops, 5% of spinach, 1% of water caltrop leaves, 0.5% of citric acid residues, 5% of brown sugar and the balance of distilled water. According to the invention, the composite esterifying enzyme ecological microbial inoculum is adopted to replace the traditional yeast to participate in fermentation, so that the style of the ferment stock solution is more typical, the natural flavor of the ferment is kept, the obvious effect is achieved, the fragrance of the ferment is improved, and the production cost is reduced while the yield of the high-quality ferment stock solution is improved.
Although the taste and flavor of the ferment are improved, the nutrient content of the ferment is not improved.
With the development of the economic society of China, consumers not only have higher and higher requirements on the taste of foods, but also pay more attention to the quality of the foods. Therefore, in order to meet the diversity, balance and easy absorption of the nutritional requirements of modern people, a novel enzyme food containing multiple active enzymes is produced. Most of the existing researches are on how to improve the nutrient content of the ferment. There are few reports on how to reduce the sugar content in ferments. Therefore, it is necessary to improve the nutrient content of the ferment and reduce the sugar content of the ferment.
Disclosure of Invention
Based on the above drawbacks of the prior art, the present invention aims to increase the nutrient content of ferment and reduce the sugar content of ferment.
In order to achieve the purpose, the invention provides an enzyme for multi-source enzyme beverage and a preparation method thereof.
The technical scheme adopted by the invention is as follows:
the multi-source ferment for the ferment beverage is prepared from the following raw materials:
sucrose, pineapple concentrated juice, blueberry concentrated juice, dragon fruit concentrated juice, cranberry concentrated juice, mulberry concentrated juice, lemon concentrated juice, honey, apple vinegar, licorice powder, burdock powder and water.
Preferably, the raw material composition further comprises: water extract of sprout paste vegetable.
Further preferably, the multi-source ferment beverage ferment is prepared from the following raw materials:
50-80 parts of cane sugar, 10-30 parts of pineapple concentrated juice, 10-30 parts of blueberry concentrated juice, 10-30 parts of dragon fruit concentrated juice, 10-30 parts of cranberry concentrated juice, 10-30 parts of mulberry concentrated juice, 10-30 parts of lemon concentrated juice, 5-10 parts of honey, 3-10 parts of apple vinegar, 2-5 parts of licorice powder, 2-5 parts of burdock powder and 80-120 parts of water.
The preparation method of the ferment for the multi-source ferment beverage comprises the following steps:
s1, raw material mixing: sequentially adding 50-80 parts by weight of cane sugar, 10-30 parts by weight of pineapple concentrated juice, 10-30 parts by weight of blueberry concentrated juice, 10-30 parts by weight of dragon fruit concentrated juice, 10-30 parts by weight of cranberry concentrated juice, 10-30 parts by weight of mulberry concentrated juice, 10-30 parts by weight of lemon concentrated juice, 5-10 parts by weight of honey, 3-10 parts by weight of apple vinegar, 2-5 parts by weight of licorice powder, 2-5 parts by weight of burdock powder and 80-120 parts by weight of water into a container, and stirring and mixing uniformly to obtain mixed pulp;
s2, sterilization: pasteurizing the mixed slurry at 60-80 ℃ for 30-45 min;
s3, fermentation: after the sterilized mixed slurry is cooled, inoculating activated strain liquid into a sterile operating platform, uniformly mixing, and placing in a constant-temperature incubator at 28-32 ℃ for culturing for 36-48 h;
s4, filtering: and filtering the fermented mixed pulp by using gauze to remove filter residues, thereby obtaining the ferment raw pulp.
More preferably, the multi-source ferment for the ferment beverage is prepared from the following raw materials:
50-80 parts of cane sugar, 10-30 parts of pineapple concentrated juice, 10-30 parts of blueberry concentrated juice, 10-30 parts of dragon fruit concentrated juice, 10-30 parts of cranberry concentrated juice, 10-30 parts of mulberry concentrated juice, 10-30 parts of lemon concentrated juice, 5-10 parts of honey, 3-10 parts of apple vinegar, 2-5 parts of licorice powder, 2-5 parts of burdock powder, 80-120 parts of water and 5-10 parts of bud paste vegetable water extract.
More preferably, the preparation method of the ferment for the multi-source ferment beverage comprises the following steps:
s1, uniformly mixing raw materials: sequentially adding 50-80 parts by weight of cane sugar, 10-30 parts by weight of pineapple concentrated juice, 10-30 parts by weight of blueberry concentrated juice, 10-30 parts by weight of dragon fruit concentrated juice, 10-30 parts by weight of cranberry concentrated juice, 10-30 parts by weight of mulberry concentrated juice, 10-30 parts by weight of lemon concentrated juice, 5-10 parts by weight of honey, 3-10 parts by weight of apple vinegar, 2-5 parts by weight of licorice powder, 2-5 parts by weight of burdock powder and 80-120 parts by weight of water into a container, and stirring and mixing uniformly to obtain mixed pulp;
s2, sterilization: pasteurizing the mixed slurry at 60-80 ℃ for 30-45 min;
s3, fermentation: after the sterilized mixed slurry is cooled, inoculating activated strain liquid into a sterile operating platform, uniformly mixing, and placing in a constant-temperature incubator at 28-32 ℃ for culturing for 36-48 h;
s4, filtering: filtering the fermented mixed slurry by using gauze to remove filter residue, adding 5-10 parts by weight of the water extract of the sprouting vegetable into the filtrate, and uniformly stirring to obtain the ferment protoplasm.
The preparation method of the sundew aqueous extract comprises the following steps: weighing 80-120g of fresh sundew, placing in a round-bottom flask, adding 250-350mL of water, heating to 95-105 ℃, refluxing for 3-5h, condensing, filtering, and concentrating the filtrate to 35-65mL by evaporation to obtain sundew aqueous extract.
The strain comprises one or more of streptococcus thermophilus, lactobacillus acidophilus, lactobacillus paracasei, leuconostoc lactis and vessella.
Preferably, the strain is formed by mixing lactobacillus acidophilus and leuconostoc lactis.
The inoculation amount of the activated strain liquid is 0.2 to 1 weight part.
The strain activation process is as follows:
inoculating 0.1 mass% of bacterial powder into an MRS liquid culture medium, standing at 30 ℃ for 36h, putting 0.5mL of bacterial liquid on an MRS solid culture medium, standing at 30 ℃ for 36h, continuously culturing for three generations, then carrying out streak culture, selecting a single bacterial colony growing vigorously after the single bacterial colony grows out, inoculating the single bacterial colony growing vigorously into the sterilized MRS liquid culture medium for amplification culture, and standing at 30 ℃ for 36h to obtain a strain activating solution, wherein the concentration of the strain activating solution is 10 10 CFU/g。
MRS liquid medium formula: 10g of peptone; 5g of yeast extract; 10g of beef extract; 20g of glucose; 2g of dipotassium phosphate; 1g of triammonium citrate; 5g of sodium acetate; magnesium sulfate 0.8g; 0.25g of manganese sulfate; dissolve in 1000mL of water.
MRS solid medium formula: 10g of peptone; 5g of yeast extract; 10g of beef extract; 20g of glucose; 2g of dipotassium phosphate; 1g of triammonium citrate; 5g of sodium acetate; magnesium sulfate 0.8g; 0.25g of manganese sulfate; and (4) stirring uniformly.
The strain also comprises black tea fungus.
Activation of black tea fungus: adding 10.0g of sucrose into boiling water at 100 ℃, stirring until the sucrose is dissolved, adding 2g of black tea when cooling to 80-85 ℃, keeping for 20min, filtering out tea residues, wrapping with gauze and kraft paper, pasteurizing at 60 ℃ for 30min, cooling to room temperature for later use, inoculating 0.1 mass percent of black tea fungus extract into a prepared culture medium, stirring uniformly, sealing with sterile gauze, and standing and culturing at 30 ℃.
The invention researches a multi-source ferment beverage enzyme with high nutrient content and low sugar content. The invention adopts a probiotic fermentation method to prepare the ferment, firstly, the fermented probiotics are screened, and lactobacillus acidophilus is preferably selected, and the main basis is that the concentrated juice is acidic, the growth capacity (acid resistance) of the lactobacillus acidophilus in a meta-acid environment and the capacity of lactic acid bacteria to metabolize and produce ethanol. Further, the lactobacillus acidophilus and the leuconostoc lactis mutually benefit and coexist in the fermentation process, so that the balance is good, the contents of flavone and polyphenol in the ferment are obviously improved after fermentation, and the DPPH free radical clearance rate is further improved. In addition, black tea fungus is additionally introduced into the fermentation liquor, which is prepared by taking tea sugar water as a raw material and fermenting a plurality of microorganisms such as acetic acid bacteria, saccharomycetes, lactic acid bacteria and the like together. The fermentation process of the black tea fungus comprises the following steps: the saccharomycetes firstly decompose sucrose into glucose and fructose, then preferentially utilize the fructose and the glucose to generate ethanol, and the acetic acid bacteria convert the glucose into gluconic acid and then convert the ethanol into acetic acid through metabolism. Therefore, the sugar content of the fermented ferment is further reduced.
The invention also improves the formula of the multi-source enzyme raw material, and adds the water extract of the raw material bud paste vegetable, namely Drosera peltata (J.E. Smith), which is a plant of Drosera, perennial herb and widely distributed in southern provinces of China. The herbs are usually dried and added into the medicine, warm in nature and pungent in flavor. Has effects in expelling pathogenic wind, removing dampness, promoting blood circulation, relieving pain, treating rheumatic osteodynia, and resisting bacteria and inflammation. Researches show that the sundew aqueous extract has a good antibacterial effect, and the sundew aqueous extract added into the fermented ferment can inhibit the growth of bacteria and prolong the shelf life of the ferment.
Compared with the prior art, the invention has the beneficial effects that:
the invention realizes the catabolism of carbohydrate in the fermentation source by a fermentation method, and converts the fermentation source into active substances with flavor and beneficial to human health by probiotics.
According to the invention, the purely natural bacteriostatic sundew aqueous extract is added into the enzyme, so that the addition of a chemical preservative is avoided, and the enzyme is ensured to have a longer shelf life.
Detailed Description
In the examples, the sources of the raw materials are as follows:
the concentrated juice used in the examples is prepared by the method disclosed in example 1 in a preparation method of concentrated juice according to patent CN 111955628A.
Lactobacillus acidophilus powder: the name of Latin is: lactobacillus acidophilus, strain accession number: CICC 20244, purchased from China center for culture Collection of Industrial microorganisms.
Lactobacillus paracasei powder: name of Latin: lactobacillus paracasei, strain accession number: CICC 20241, purchased from China center for culture Collection of Industrial microorganisms.
Leuconostoc lactis: the name of Latin is: leuconostoc lactis, strain accession number: CICC 20715, purchased from China center for culture Collection of Industrial microorganisms.
Black tea fungus extract: purchased from western national hao biotechnology limited.
Fresh sundew: is obtained by picking from western part of Fujian province.
Example 1
A preparation method of ferment for multi-source ferment beverage comprises the following steps:
s1, uniformly mixing raw materials: sequentially adding 60 parts by weight of cane sugar, 20 parts by weight of pineapple concentrated juice, 20 parts by weight of blueberry concentrated juice, 20 parts by weight of dragon fruit concentrated juice, 20 parts by weight of cranberry concentrated juice, 20 parts by weight of mulberry concentrated juice, 20 parts by weight of lemon concentrated juice, 6 parts by weight of honey, 4 parts by weight of apple vinegar, 3 parts by weight of licorice powder, 2 parts by weight of burdock powder and 100 parts by weight of distilled water into a container, and uniformly stirring and mixing to obtain mixed pulp;
s2, sterilization: pasteurizing the mixed slurry at 70 deg.C for 30min;
s3, fermentation: cooling the sterilized mixed slurry, inoculating 0.2 weight part of lactobacillus acidophilus activating solution into a sterile operating platform, uniformly mixing, and culturing in a constant temperature incubator at 30 ℃ for 48 hours;
s4, filtering: and filtering the fermented mixed pulp by using gauze to remove filter residues to obtain the enzyme raw pulp.
The strain activation process is as follows:
inoculating Lactobacillus acidophilus powder 0.1 wt% into MRS liquid culture medium, standing at 30 deg.C for 36 hr, adding 0.5mL of bacteria solution onto MRS solid culture medium, standing at 30 deg.C for 36 hr, continuously culturing for three generations, performing streak culture, and selecting the strain with vigorous growth after single strain grows outInoculating the single colony growing vigorously to sterilized MRS liquid culture medium, performing amplification culture, and standing at 30 deg.C for 36 hr to obtain Lactobacillus acidophilus activating solution with concentration of 10 10 CFU/g。
MRS liquid medium formula: 10g of peptone; 5g of yeast extract; 10g of beef extract; 20g of glucose; 2g of dipotassium phosphate; 1g of triammonium citrate; 5g of sodium acetate; magnesium sulfate 0.8g; 0.25g of manganese sulfate; dissolved in 1000mL of distilled water.
MRS solid medium formula: 10g of peptone; 5g of yeast extract; 10g of beef extract; 20g of glucose; 2g of dipotassium phosphate; 1g of triammonium citrate; 5g of sodium acetate; magnesium sulfate 0.8g; 0.25g of manganese sulfate; and (4) stirring uniformly.
Example 2
A preparation method of ferment for multi-source ferment beverage comprises the following steps:
s1, uniformly mixing raw materials: sequentially adding 60 parts by weight of cane sugar, 20 parts by weight of pineapple concentrated juice, 20 parts by weight of blueberry concentrated juice, 20 parts by weight of dragon fruit concentrated juice, 20 parts by weight of cranberry concentrated juice, 20 parts by weight of mulberry concentrated juice, 20 parts by weight of lemon concentrated juice, 6 parts by weight of honey, 4 parts by weight of apple vinegar, 3 parts by weight of licorice powder, 2 parts by weight of burdock powder and 100 parts by weight of distilled water into a container, and uniformly stirring and mixing to obtain mixed slurry;
s2, sterilization: pasteurizing the mixed slurry at 70 deg.C for 30min;
s3, fermentation: after the sterilized mixed slurry is cooled, 0.2 part by weight of lactobacillus paracasei activation solution is inoculated into a sterile operation table, uniformly mixed and placed into a constant temperature incubator at 30 ℃ for culturing for 48 hours;
s4, filtering: and filtering the fermented mixed pulp by using gauze to remove filter residues to obtain the enzyme raw pulp.
The strain activation process is as follows:
inoculating 0.1% by mass of Lactobacillus casei into MRS liquid culture medium, standing at 30 deg.C for 36 hr, placing 0.5mL of bacterial liquid on MRS solid culture medium, standing at 30 deg.C for 36 hr, continuously culturing for three generations, performing streak culture, and selecting the strongly growing single strain after single strain grows outInoculating single colony growing vigorously to sterilized MRS liquid culture medium, performing amplification culture, and standing at 30 deg.C for 36 hr to obtain Lactobacillus paracasei activating solution with concentration of 10 10 CFU/g。
MRS liquid medium formula: 10g of peptone; 5g of yeast extract; 10g of beef extract; 20g of glucose; 2g of dipotassium phosphate; 1g of triammonium citrate; 5g of sodium acetate; magnesium sulfate 0.8g; 0.25g of manganese sulfate; dissolved in 1000mL of distilled water.
MRS solid medium formula: 10g of peptone; 5g of yeast extract; 10g of beef extract; 20g of glucose; 2g of dipotassium phosphate; 1g of triammonium citrate; 5g of sodium acetate; magnesium sulfate 0.8g; 0.25g of manganese sulfate; and (5) stirring uniformly.
Example 3
A preparation method of ferment for multi-source ferment beverage comprises the following steps:
s1, uniformly mixing raw materials: sequentially adding 60 parts by weight of cane sugar, 20 parts by weight of pineapple concentrated juice, 20 parts by weight of blueberry concentrated juice, 20 parts by weight of dragon fruit concentrated juice, 20 parts by weight of cranberry concentrated juice, 20 parts by weight of mulberry concentrated juice, 20 parts by weight of lemon concentrated juice, 6 parts by weight of honey, 4 parts by weight of apple vinegar, 3 parts by weight of licorice powder, 2 parts by weight of burdock powder and 100 parts by weight of distilled water into a container, and uniformly stirring and mixing to obtain mixed pulp;
s2, sterilization: pasteurizing the mixed slurry at 70 deg.C for 30min;
s3, fermentation: after the sterilized mixed slurry is cooled, inoculating 0.2 weight part of Leuconostoc lactis activation solution into a sterile operation table, uniformly mixing, and culturing in a constant-temperature incubator at 30 ℃ for 48 hours;
s4, filtering: and filtering the fermented mixed pulp by using gauze to remove filter residues to obtain the enzyme raw pulp.
The strain activation process is as follows:
inoculating 0.1% mass of Leuconostoc lactis into MRS liquid culture medium, standing at 30 deg.C for 36 hr, placing 0.5mL of bacterial liquid on MRS solid culture medium, standing at 30 deg.C for 36 hr, continuously culturing for three generations, performing streak culture, and selecting the vigorously grown single bacterium after single bacterium grows outInoculating the single colony growing vigorously into sterilized MRS liquid culture medium, performing amplification culture, and standing at 30 deg.C for 36 hr to obtain Leuconostoc lactis activating solution with concentration of 10 10 CFU/g。
MRS liquid medium formula: 10g of peptone; 5g of yeast extract; 10g of beef extract; 20g of glucose; 2g of dipotassium phosphate; 1g of triammonium citrate; 5g of sodium acetate; magnesium sulfate 0.8g; 0.25g of manganese sulfate; dissolved in 1000mL of distilled water.
MRS solid medium formula: 10g of peptone; 5g of yeast extract; 10g of beef extract; 20g of glucose; 2g of dipotassium phosphate; 1g of triammonium citrate; 5g of sodium acetate; magnesium sulfate 0.8g; 0.25g of manganese sulfate; and (5) stirring uniformly.
Example 4
A preparation method of ferment for multi-source ferment beverage comprises the following steps:
s1, raw material mixing: sequentially adding 60 parts by weight of cane sugar, 20 parts by weight of pineapple concentrated juice, 20 parts by weight of blueberry concentrated juice, 20 parts by weight of dragon fruit concentrated juice, 20 parts by weight of cranberry concentrated juice, 20 parts by weight of mulberry concentrated juice, 20 parts by weight of lemon concentrated juice, 6 parts by weight of honey, 4 parts by weight of apple vinegar, 3 parts by weight of licorice powder, 2 parts by weight of burdock powder and 100 parts by weight of distilled water into a container, and uniformly stirring and mixing to obtain mixed pulp;
s2, sterilization: pasteurizing the mixed slurry at 70 deg.C for 30min;
s3, fermentation: after the sterilized mixed slurry is cooled, 0.1 part by weight of lactobacillus acidophilus activating solution and 0.1 part by weight of leuconostoc lactis activating solution are added into a sterile operation table, uniformly mixed and placed into a constant-temperature incubator at 30 ℃ for culturing for 48 hours;
s4, filtering: and filtering the fermented mixed pulp by using gauze to remove filter residues to obtain the enzyme raw pulp.
The strain activation process is as follows:
inoculating 0.1% Leuconostoc lactis powder or 0.1% Lactobacillus acidophilus powder into MRS liquid culture medium, standing at 30 deg.C for 36 hr, adding 0.5mL of bacterial liquid into MRS solid culture medium, standing at 30 deg.C for 36 hr, continuously culturing for three generations, and adding into MRS liquid culture mediumPerforming streak culture, after single colony growth, selecting the single colony with vigorous growth, inoculating the single colony with vigorous growth into sterilized MRS liquid culture medium, performing amplification culture, and performing static culture at 30 deg.C for 36 hr to obtain Leuconostoc lactis activation solution with concentration of 10 10 CFU/g。
MRS liquid medium formula: 10g of peptone; 5g of yeast extract; 10g of beef extract; 20g of glucose; 2g of dipotassium phosphate; 1g of triammonium citrate; 5g of sodium acetate; magnesium sulfate 0.8g; 0.25g of manganese sulfate; dissolved in 1000mL of distilled water.
MRS solid medium formula: 10g of peptone; 5g of yeast extract; 10g of beef extract; 20g of glucose; 2g of dipotassium phosphate; 1g of triammonium citrate; 5g of sodium acetate; magnesium sulfate 0.8g; 0.25g of manganese sulfate; and (4) stirring uniformly.
Example 5
A preparation method of ferment for multi-source ferment beverage comprises the following steps:
s1, uniformly mixing raw materials: sequentially adding 60 parts by weight of cane sugar, 20 parts by weight of pineapple concentrated juice, 20 parts by weight of blueberry concentrated juice, 20 parts by weight of dragon fruit concentrated juice, 20 parts by weight of cranberry concentrated juice, 20 parts by weight of mulberry concentrated juice, 20 parts by weight of lemon concentrated juice, 6 parts by weight of honey, 4 parts by weight of apple vinegar, 3 parts by weight of licorice powder, 2 parts by weight of burdock powder and 100 parts by weight of distilled water into a container, and uniformly stirring and mixing to obtain mixed slurry;
s2, sterilization: pasteurizing the mixed slurry at 70 deg.C for 30min;
s3, fermentation: after the sterilized mixed slurry is cooled, 0.1 part by weight of lactobacillus casei-like activation liquid and 0.1 part by weight of leuconostoc lactis activation liquid are inoculated into a sterile operation table, uniformly mixed and placed into a constant-temperature incubator at 30 ℃ for culturing for 48 hours;
s4, filtering: and filtering the fermented mixed pulp by using gauze to remove filter residues to obtain the enzyme raw pulp.
The strain activation process is as follows:
inoculating 0.1% wt. Leuconostoc lactis powder or 0.1% wt. Lactobacillus casei powder into MRS liquid culture medium, standing at 30 deg.C for culturing for 36 hr, and adding 0.5mL of bacterial liquid into the culture mediumPerforming static culture for 36h at 30 ℃ on an MRS solid culture medium, performing streak culture after continuous culture for three generations, selecting a single colony growing vigorously after the single colony grows out, inoculating the single colony growing vigorously into a sterilized MRS liquid culture medium for amplification culture, and performing static culture for 36h at 30 ℃ to obtain the leuconostoc lactis activation solution, wherein the concentration of the leuconostoc lactis activation solution is 10 10 CFU/g。
MRS liquid medium formula: 10g of peptone; 5g of yeast extract; 10g of beef extract; 20g of glucose; 2g of dipotassium phosphate; 1g of triammonium citrate; 5g of sodium acetate; magnesium sulfate 0.8g; 0.25g of manganese sulfate; dissolved in 1000mL of distilled water.
MRS solid medium formula: 10g of peptone; 5g of yeast extract; 10g of beef extract; 20g of glucose; dipotassium phosphate 2g; 1g of triammonium citrate; 5g of sodium acetate; magnesium sulfate 0.8g; 0.25g of manganese sulfate; and (4) stirring uniformly.
Example 6
A preparation method of ferment for multi-source ferment beverage comprises the following steps:
s1, raw material mixing: sequentially adding 60 parts by weight of cane sugar, 20 parts by weight of pineapple concentrated juice, 20 parts by weight of blueberry concentrated juice, 20 parts by weight of dragon fruit concentrated juice, 20 parts by weight of cranberry concentrated juice, 20 parts by weight of mulberry concentrated juice, 20 parts by weight of lemon concentrated juice, 6 parts by weight of honey, 4 parts by weight of apple vinegar, 3 parts by weight of licorice powder, 2 parts by weight of burdock powder and 100 parts by weight of distilled water into a container, and uniformly stirring and mixing to obtain mixed slurry;
s2, sterilization: pasteurizing the mixed slurry at 70 deg.C for 30min;
s3, fermentation: after the sterilized mixed slurry is cooled, inoculating 0.1 part by weight of Lactobacillus acidophilus activating solution, 0.1 part by weight of Leuconostoc lactis activating solution and 0.1 part by weight of black tea fungus activating solution into a sterile operating platform, uniformly mixing, and culturing in a constant-temperature incubator at 30 ℃ for 48 hours;
s4, filtering: and filtering the fermented mixed pulp by using gauze to remove filter residues to obtain the enzyme raw pulp.
The strain activation process is as follows:
inoculating 0.1% by mass of Ruming kebab into MRS liquid culture mediumCulturing 0.5mL of bacterial liquid on an MRS solid culture medium for 36h at 30 ℃ in a standing way or 0.1 mass percent of acidophilic lactobacillus powder for three generations in a standing way, carrying out streak culture after continuous culture for three generations, selecting a single colony which grows vigorously after the single colony grows out, inoculating the single colony which grows vigorously into a sterilized MRS liquid culture medium for amplification culture, and carrying out standing culture at 30 ℃ for 36h to obtain the leuconostoc lactis activating solution, wherein the concentrations of the leuconostoc lactis activating solution and the acidophilic lactobacillus acidophilic activating solution are both 10 10 CFU/g。
MRS liquid medium formula: 10g of peptone; 5g of yeast extract; 10g of beef extract; 20g of glucose; 2g of dipotassium phosphate; 1g of triammonium citrate; 5g of sodium acetate; magnesium sulfate 0.8g; 0.25g of manganese sulfate; dissolved in 1000mL of distilled water.
MRS solid medium formula: 10g of peptone; 5g of yeast extract; 10g of beef extract; 20g of glucose; dipotassium phosphate 2g; 1g of triammonium citrate; 5g of sodium acetate; magnesium sulfate 0.8g; 0.25g of manganese sulfate; and (4) stirring uniformly.
Activation of black tea fungus: adding 10.0g sucrose into 500mL distilled water at 100 deg.C, stirring to dissolve, cooling to 80 deg.C, adding 2g black tea, maintaining for 20min, filtering to remove tea residue, wrapping with gauze and kraft paper, pasteurizing at 60 deg.C for 30min, cooling to room temperature, adding 0.1% black tea fungus extract into the prepared culture medium, stirring, sealing with sterile gauze, standing at 30 deg.C for 36h to obtain a black tea fungus activation solution with a concentration of 10 10 CFU/g。
Example 7
A preparation method of ferment for multi-source ferment beverage comprises the following steps:
s1, uniformly mixing raw materials: sequentially adding 60 parts by weight of cane sugar, 20 parts by weight of pineapple concentrated juice, 20 parts by weight of blueberry concentrated juice, 20 parts by weight of dragon fruit concentrated juice, 20 parts by weight of cranberry concentrated juice, 20 parts by weight of mulberry concentrated juice, 20 parts by weight of lemon concentrated juice, 6 parts by weight of honey, 4 parts by weight of apple vinegar, 3 parts by weight of licorice powder, 2 parts by weight of burdock powder and 100 parts by weight of distilled water into a container, and uniformly stirring and mixing to obtain mixed pulp;
s2, sterilization: pasteurizing the mixed slurry at 70 deg.C for 30min;
s3, fermentation: after the sterilized mixed serous fluid is cooled, inoculating 0.1 weight part of lactobacillus acidophilus activating liquid, 0.1 weight part of leuconostoc lactis activating liquid and 0.1 weight part of black tea fungus activating liquid into a sterile operation table, uniformly mixing, and culturing in a constant-temperature incubator at 30 ℃ for 48 hours;
s4, filtering: and filtering the fermented mixed slurry by using gauze to remove filter residues, adding 5 parts by weight of the water extract of the sprout ointment into the filtrate, and uniformly stirring to obtain the multi-source ferment for the ferment beverage.
The strain activation process is as follows:
inoculating 0.1 mass percent of leuconostoc lactis powder or 0.1 mass percent of acidophilic lactobacillus-like powder into an MRS liquid culture medium, standing for 36 hours at 30 ℃, taking 0.5mL of bacterial liquid on the MRS solid culture medium, standing for 36 hours at 30 ℃, carrying out streak culture after three continuous cultures, selecting a single bacterial colony which grows vigorously after the single bacterial colony grows out, inoculating the single bacterial colony which grows vigorously into the sterilized MRS liquid culture medium for expanding culture, and standing for 36 hours at 30 ℃ to obtain the leuconostoc lactis activation liquid or acidophilic lactobacillus-like activation liquid, wherein the concentrations of the leuconostoc lactis activation liquid and the acidophilic lactobacillus-like activation liquid are both 10 10 CFU/g。
MRS liquid medium formula: 10g of peptone; 5g of yeast extract; 10g of beef extract; 20g of glucose; 2g of dipotassium phosphate; 1g of triammonium citrate; 5g of sodium acetate; magnesium sulfate 0.8g; 0.25g of manganese sulfate; dissolved in 1000mL of distilled water.
MRS solid medium formula: 10g of peptone; 5g of yeast extract; 10g of beef extract; 20g of glucose; 2g of dipotassium phosphate; 1g of triammonium citrate; 5g of sodium acetate; magnesium sulfate 0.8g; 0.25g of manganese sulfate; and (4) stirring uniformly.
Activation of black tea fungus: adding 10.0g sucrose into 500mL distilled water at 100 deg.C, stirring to dissolve, cooling to 80 deg.C, adding 2g black tea, maintaining for 20min, filtering to remove tea residue, wrapping with gauze and kraft paper, pasteurizing at 60 deg.C for 30min, cooling to room temperature to obtain black tea fungus culture medium, adding 0.1% black tea fungus extract into the prepared black tea fungus culture medium, stirring, sealing with sterile gauze, standing at 30 deg.C, standing, and standingCulturing for 36h to obtain black tea fungus activating solution with concentration of 10 10 CFU/g。
The preparation method of the sundew aqueous extract comprises the following steps: weighing 100g of fresh sundew, placing in a round-bottom flask, adding 300mL of distilled water, raising the temperature to 100 ℃, refluxing for 4h, condensing, filtering, and evaporating and concentrating the filtrate to 50mL to obtain the sundew aqueous extract.
Test example:
1. measurement of flavone content
The method for measuring flavone comprises the following steps: preparing rutin standard substance into standard solution with concentration of 0, 0.04mg/mL, 0.08mg/mL, 0.12mg/mL, 0.16mg/mL and 0.20mg/mL, adding 1.0mL of standard solution into a test tube, adding 0.75mL of sodium nitrite solution with mass fraction of 5%, mixing uniformly, standing for 6min, adding 0.75mL of aluminum nitrate solution with mass fraction of 10%, mixing uniformly, standing for 6min, adding 4.0mL of 4% sodium hydroxide solution, standing for 10min, standing at 510nm for measuring light absorption value, and drawing a standard curve with rutin concentration as abscissa and light absorption as ordinate.
Taking an enzyme sample, uniformly mixing, taking out a proper amount of the enzyme sample, centrifuging (5000r, 5min), taking supernatant, adding a proper amount of methanol for diluting, adding 0.75mL of sodium nitrite solution with the mass fraction of 5%, uniformly mixing, standing for 6min, adding 0.75mL of aluminum nitrate solution with the mass fraction of 10%, uniformly mixing, standing for 6min, adding 4.0mL of 4% sodium hydroxide solution, standing for 10min, centrifuging under the same condition to remove floccules, and standing at 510nm for measuring the light absorption value. And calculating the content of the flavone in the fermentation liquor according to the standard curve and the dilution times of the rutin standard solution.
2. Total phenol determination method
Preparing 1.0g/L of gallic acid standard solution, then respectively transferring 0.1mL, 0.2mL, 0.3mL, 0.4mL and 0.5mL of gallic acid, adding deionized water, fixing the volume to 1.0mL, placing the mixture in a 15mL test tube, adding 5mL of a 10% forskolin phenol reagent, uniformly mixing, standing for 5min, adding 4mL of 7.5% sodium carbonate solution, standing for 60min, measuring the light absorption value at 765nm, and drawing a standard curve by taking the concentration of the gallic acid as the abscissa and the light absorption as the ordinate.
Taking an enzyme sample, uniformly mixing, taking out a proper amount, centrifuging (5000r, 5min), taking supernatant for determination, adding 1.0mL of sample to be determined which is diluted by a proper time into a test tube, adding 5mL of 10% Fulin phenol reagent, uniformly mixing, standing for 5min, adding 4mL of 7.5% sodium carbonate solution, standing for 60min, determining the light absorption value at 765nm, and calculating the total phenol content according to a standard curve.
3. Total sugar determination
The glucose control was oven dried at 105 ℃ to constant weight and two portions, 100mg each, were weighed. Adding distilled water according to the concentration of 1mg/mL to a constant volume, and respectively drawing a total sugar standard curve and a reducing sugar standard curve; 5.00g of analytically pure phenol is accurately weighed, redistilled to prepare crystalline phenol, and the crystalline phenol is put into a 100mL brown volumetric flask for constant volume and sealed preservation for later use. Weighing 0.650g of multisource enzyme sample, dissolving with 50mL of distilled water, putting into a 100mL brown measuring flask, adding 32.5mL of 2mol/L NaOH solution and 4.5g of glycerol, and fixing the volume with distilled water for later use.
Drawing a total sugar standard curve of the sample
Precisely measuring a prepared part of 1mg/mL glucose reference substance solution 0mL, 0.1mL, 0.3mL, 0.5mL, 0.7mL, 0.9mL, 1.1mL and 1.3mL in 8 20mL graduated test tubes, adding distilled water to 2mL, and mixing uniformly; adding 1mL of phenol aqueous solution (5%), shaking, rapidly adding 5mL of concentrated sulfuric acid, standing for 10min, and boiling in a water bath for 20min; after the water bath is finished, the test tube is rapidly cooled to room temperature by flowing cold water, a blank tube is used as a control, and the absorbance is detected at 487 nm. And drawing a standard curve by taking the mass (mg) of the glucose as an abscissa and the absorbance value as an ordinate.
Determination of total sugar in a sample
And (3) under the condition of a wavelength of 487nm, adjusting a zero point by using a sample blank, measuring an absorbance value (A) of the enzyme sample, and calculating the total sugar content.
DPPH radical clearance assay
The sample pretreatment method comprises the following steps: and (3) uniformly mixing 1.0mL of centrifuged sample with 9.0mL of deionized water to be detected. 0.4mL of the diluted sample was added to 3.6mL10 -4 DPPH (1,1 diphenyl-2-trinitrophenylhydrazine) free radical solution, reacting for 30min at normal temperature in the dark, and measuring the light absorption value at the wavelength of 517 nm. 0.4mL of absolute ethanol and 3.6mL of LDPPH solution were combined as a blank. Each sample was assayed in 3 replicates.
TABLE 1 measurement of flavone, total phenol, total sugar content and DPPH radical scavenging ratio
Figure BDA0003831757090000151
5. Bacteriostasis test
And performing an in vitro bacteriostasis test by using a tube-disc method. Preparing a culture plate and a proper amount of agar powder. And (5) placing the sterilized test article on a clean operation table, and irradiating for 30min by using an ultraviolet lamp. And when the agar is cooled to 55-60 ℃, pouring the agar into the sterilized plates (15-20 mL of agar needs to be poured into each plate), gently shaking until the agar is uniformly spread, putting the plates on a horizontal operating platform, and waiting for the agar to solidify. After the agar is completely solidified, the cultured bacterial suspension is dipped by a cotton swab and is evenly smeared on the surface of the agar. Each sample was tested for 3 bacteria and then the oxford cup was gently placed on the agar surface (3 oxford cups per plate) with clean tweezers and marked with a marker. The test samples were added separately to the oxford cups of the plate and each bacteria was repeated 3 times for each different concentration of drug. The plates were then incubated in a 37 ℃ incubator for 24h. The size of the inhibition zone is measured, and the average value is calculated.
TABLE 2 sample diameter (mm) of inhibition zone for different bacteria
Figure BDA0003831757090000152
Figure BDA0003831757090000161
Through comparison of examples 1 to 3, lactobacillus acidophilus is screened out as the most suitable fermentation strain, and the nutrient content of the multi-source enzyme can be remarkably improved. The reason for this is that; the main concentrated juice is acidic, and the growth capacity (acid resistance) of lactobacillus acidophilus in a meta-acid environment and the capacity of lactobacillus to metabolize to produce ethanol. Further, the lactobacillus acidophilus and the leuconostoc lactis are adopted in the embodiment 4 for compound fermentation, and the lactobacillus acidophilus and the leuconostoc lactis are found to be in mutual beneficial symbiosis in the fermentation process unexpectedly, so that the balance is good, the content of nutrient substances in the ferment after fermentation is obviously improved, and the DPPH free radical scavenging rate is further improved. Example 6 black tea fungus was additionally introduced into the fermentation broth, which was prepared by co-fermenting tea sugar water as a raw material with various microorganisms such as acetic acid bacteria, yeast and lactic acid bacteria. The fermentation process of the black tea fungus comprises the following steps: the saccharomycetes firstly decompose sucrose into glucose and fructose, then preferentially utilize the fructose and the glucose to generate ethanol, and the acetic acid bacteria convert the glucose into gluconic acid and then convert the ethanol into acetic acid through metabolism. Therefore, the sugar content of the ferment after fermentation is further reduced.
Example 7 adds the water extract of the raw material of the sundew, which shows that the sundew water extract has good antibacterial effect, and the addition of the sundew water extract in the fermented ferment can inhibit the growth of bacteria and prolong the shelf life of the ferment.
The foregoing detailed description of the preferred embodiments of the invention has been presented. It should be understood that numerous modifications and variations can be devised by those skilled in the art in light of the above teachings. Therefore, the technical solutions available to those skilled in the art through logic analysis, reasoning and limited experiments based on the prior art according to the concept of the present invention should be within the scope of protection defined by the claims.

Claims (8)

1. The multi-source ferment for the ferment beverage is characterized by being prepared from the following raw materials:
sucrose, pineapple concentrated juice, blueberry concentrated juice, dragon fruit concentrated juice, cranberry concentrated juice, mulberry concentrated juice, lemon concentrated juice, honey, apple vinegar, licorice powder, burdock powder and water.
2. The multi-source ferment beverage of claim 1, wherein the raw material composition further comprises:
water extract of sprout paste vegetable.
3. The multi-source ferment for the ferment beverage is characterized by being prepared from the following raw materials:
50-80 parts of cane sugar, 10-30 parts of pineapple concentrated juice, 10-30 parts of blueberry concentrated juice, 10-30 parts of dragon fruit concentrated juice, 10-30 parts of cranberry concentrated juice, 10-30 parts of mulberry concentrated juice, 10-30 parts of lemon concentrated juice, 5-10 parts of honey, 3-10 parts of apple vinegar, 2-5 parts of licorice powder, 2-5 parts of burdock powder and 100 parts of water.
4. The multi-source ferment beverage of claim 3, wherein the raw material composition further comprises:
5 to 10 parts of water extract of the bud paste vegetable.
5. The multi-source ferment beverage of claim 3, wherein the preparation method of the multi-source ferment beverage comprises the following steps:
s1, raw material mixing: sequentially adding 50-80 parts by weight of cane sugar, 10-30 parts by weight of pineapple concentrated juice, 10-30 parts by weight of blueberry concentrated juice, 10-30 parts by weight of dragon fruit concentrated juice, 10-30 parts by weight of cranberry concentrated juice, 10-30 parts by weight of mulberry concentrated juice, 10-30 parts by weight of lemon concentrated juice, 5-10 parts by weight of honey, 3-10 parts by weight of apple vinegar, 2-5 parts by weight of licorice powder, 2-5 parts by weight of burdock powder and 80-120 parts by weight of water into a container, and stirring and mixing uniformly to obtain mixed slurry;
s2, sterilization: pasteurizing the mixed slurry at 60-80 ℃ for 30-45 min;
s3, fermentation: after the sterilized mixed slurry is cooled, inoculating activated strain liquid into a sterile operating platform, uniformly mixing, and placing in a constant-temperature incubator at 28-32 ℃ for culturing for 36-48 h;
s4, filtering: and filtering the fermented mixed pulp by using gauze to remove filter residues, thereby obtaining the ferment raw pulp.
6. The multi-source ferment beverage of claim 4, wherein the preparation method of the multi-source ferment beverage comprises the following steps:
s1, raw material mixing: adding 50-80 parts by weight of cane sugar, 10-30 parts by weight of pineapple concentrated juice, 10-30 parts by weight of blueberry concentrated juice, 10-30 parts by weight of dragon fruit concentrated juice, 10-30 parts by weight of cranberry concentrated juice, 10-30 parts by weight of mulberry concentrated juice, 10-30 parts by weight of lemon concentrated juice, 5-10 parts by weight of honey, 3-10 parts by weight of apple vinegar, 2-5 parts by weight of licorice powder, 2-5 parts by weight of burdock powder and 80-120 parts by weight of water into a container in sequence, and stirring and mixing uniformly to obtain mixed pulp;
s2, sterilization: pasteurizing the mixed slurry at 60-80 ℃ for 30-45 min;
s3, fermentation: after the sterilized mixed slurry is cooled, inoculating activated strain liquid into a sterile operating platform, uniformly mixing, and placing in a constant-temperature incubator at 28-32 ℃ for culturing for 36-48 h;
s4, filtering: filtering the fermented mixed slurry by using gauze to remove filter residue, adding 5-10 parts by weight of the water extract of the sprouting vegetable into the filtrate, and uniformly stirring to obtain the ferment protoplasm.
7. The multi-source ferment beverage of claim 5 or 6, wherein the strain comprises one or more of Streptococcus thermophilus, lactobacillus acidophilus, lactobacillus paracasei, leuconostoc lactis, lactobacillus, and Weissella.
8. The multi-source ferment for ferment beverages according to claim 5 or 6, wherein the amount of the activated fungal solution added is 0.2 to 1 part by weight.
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