CN114376040A - Xinhui mandarin kangpu tea and preparation method thereof - Google Patents

Xinhui mandarin kangpu tea and preparation method thereof Download PDF

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CN114376040A
CN114376040A CN202210152168.9A CN202210152168A CN114376040A CN 114376040 A CN114376040 A CN 114376040A CN 202210152168 A CN202210152168 A CN 202210152168A CN 114376040 A CN114376040 A CN 114376040A
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tea
seed liquid
liquid
secondary seed
xinhui
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廖雪义
李拥军
皮小弟
李佳妮
李馨
杨金霞
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Guangdong Jiangmen Vocational College Of Traditional Chinese Medicine
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Guangdong Jiangmen Vocational College Of Traditional Chinese Medicine
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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23FCOFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
    • A23F3/00Tea; Tea substitutes; Preparations thereof
    • A23F3/06Treating tea before extraction; Preparations produced thereby
    • A23F3/14Tea preparations, e.g. using additives
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23FCOFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
    • A23F3/00Tea; Tea substitutes; Preparations thereof
    • A23F3/16Tea extraction; Tea extracts; Treating tea extract; Making instant tea
    • A23F3/166Addition of, or treatment with, enzymes or microorganisms
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23FCOFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
    • A23F3/00Tea; Tea substitutes; Preparations thereof
    • A23F3/16Tea extraction; Tea extracts; Treating tea extract; Making instant tea
    • A23F3/18Extraction of water soluble tea constituents
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    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14Fungi; Culture media therefor
    • C12N1/16Yeasts; Culture media therefor
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    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14Fungi; Culture media therefor
    • C12N1/16Yeasts; Culture media therefor
    • C12N1/18Baker's yeast; Brewer's yeast
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20Bacteria; Culture media therefor
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00Lactic or propionic acid bacteria
    • A23V2400/11Lactobacillus
    • A23V2400/113Acidophilus
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00Lactic or propionic acid bacteria
    • A23V2400/11Lactobacillus
    • A23V2400/137Delbrueckii
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00Lactic or propionic acid bacteria
    • A23V2400/11Lactobacillus
    • A23V2400/169Plantarum
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00Lactic or propionic acid bacteria
    • A23V2400/11Lactobacillus
    • A23V2400/175Rhamnosus

Abstract

The invention discloses Xinhui mandarin orange kangpu tea, which is prepared by the following steps: s1, preparation of a symbiotic bacteria culture medium: A1. preparing tea liquid; A2. preparing Xinhui citrus juice; A3. compounding; A4. sterilizing; s2, inoculation: B1. primary seed liquid culture; B2. secondary seed liquid culture; B3. compounding strains; B4. inoculating and fermenting; s3, pasteurization. The Xinhui mandarin orange Kangpu tea prepared by the invention can fully improve the resource utilization rate of Xinhui mandarin oranges, combines the unique nutritional values of green tea, probiotics and Xinhui mandarin oranges, and meets the health requirements of modern people on nutritional and healthy beverages on the basis of improving the original poor taste of the Xinhui mandarin orange pulp. The problem that the citrus unshiu pulp is sour, astringent and bitter when being directly eaten is effectively solved, the citrus unshiu pulp is compounded with green tea and mulberry leaves, and the green tea and the mulberry leaves are fermented into the citrus unshiu health and common tea with dual characteristic functions of fermented juice and fermented tea, and the citrus unshiu health and common tea can play roles in regulating intestines and stomach of a human body, removing fat and resisting oxidation.

Description

Xinhui mandarin kangpu tea and preparation method thereof
Technical Field
The invention relates to the field of healthy tea, in particular to Xinhui mandarin orange Kangpu tea and a preparation method thereof.
Background
The Xinhui citrus, also called as tangerine peel, is mainly produced in Xinhui regions in Jiangmen City, is one of ten major medicinal materials in Guangdong province, belongs to a Chinese geographical sign product, has the characteristics of dual purposes of medicine and food, and has the effects of regulating qi, strengthening spleen, harmonizing stomach, preventing vomiting, eliminating dampness and eliminating phlegm and the like. At present, the development and utilization of the citrus reticulata blanco are mostly limited to the fruit peel, the fruit pulp is not ideal for direct eating due to a plurality of sour seeds, limonin can be generated after juicing, and the citrus reticulata blanco is not durable to store after peeling and is randomly discarded, so that not only is great resource waste caused, but also the environment is polluted. The Xinhui citrus pulp is rich in saccharides, organic acids, vitamins, dietary fibers, flavones, polyphenols and other bioactive substances, and still has higher development and utilization values. There are also scholars who produce fruit wine and fermented beverages from citrus pulp of the Xinhui, such as: a Xinhui citrus fruit wine (CN 108753542A) is prepared by removing seeds from pulp of Xinhui citrus, squeezing to obtain juice, adding white sugar, Saccharomyces cerevisiae and pectinase, fermenting, soaking and brewing with oak, filtering, sterilizing, and packaging to obtain final product; a preparation method (CN 109845939A) of Xinhui mandarin orange meat probiotic fermented beverage comprises the steps of removing seeds of Xinhui mandarin oranges, juicing, debitterizing, sterilizing, fermenting with lactic acid bacteria and the like to obtain the Xinhui mandarin orange meat probiotic fermented beverage.
The conpu tea is favored by consumers in many regions all over the world due to the unique flavor, the luscious mouthfeel and the efficacy attribute of benefiting intestines and stomach and natural health, and becomes one of the most representative fine categories of fermented beverages. At present, no related product for preparing Kangpu tea by using Xinhui oranges exists.
Disclosure of Invention
The invention aims to provide Xinhui mandarin orange Kangpu tea and a preparation method thereof.
According to one aspect of the invention, the Xinhui mandarin orange Compu tea is provided, and the preparation method comprises the following steps:
s1, preparation of a symbiotic bacteria culture medium:
A1. preparing tea liquid: weighing folium Camelliae sinensis, cooling to 85-90 deg.C with boiling water, adding folium Camelliae sinensis in an amount of 0.6g per 100ml water, maintaining for 20-30min, and filtering to remove tea residue with sieve to obtain tea solution;
A2. preparing Xinhui citrus juice: selecting 9-11 months peeled Xinhui citrus pulp, wherein the Xinhui citrus pulp meets the requirements of bright color, no plant diseases and insect pests and no mechanical damage, and after removing the core of the Xinhui citrus pulp, mixing the Xinhui citrus pulp with purified water according to the ratio of 5: 3, putting the mixture into a juicer to be juiced, and filtering the juiced mixture by using a sieve to obtain citrus unshiu juice for later use;
A3. compounding: mixing the tea liquid and the Xinhui citrus juice according to the volume ratio of 5-7: compounding according to the proportion of 3-5, and uniformly mixing to obtain a compound liquid;
A4. and (3) sterilization: adding white granulated sugar and potassium dihydrogen phosphate into the compound liquid, subpackaging in a conical flask according to 40% liquid loading amount, and sterilizing at 105 deg.C for 10min to obtain sterilized tea fruit juice;
s2, inoculation:
B1. primary seed liquid culture: inoculating lactobacillus glycerol tube into MRS broth culture medium at an inoculum size of 2%, and standing at 37 deg.C for 14 hr to obtain first-class seed solution of lactobacillus; inoculating 2% of yeast glycerol tube into YPD liquid culture medium, placing in a shaking table at 26 deg.C, and culturing at rotation speed of 120r/min for 14 hr to obtain primary seed liquid of yeast; inoculating 2% of acetobacter aceti glycerol tube into an acetobacter aceti liquid culture medium, placing the acetobacter aceti glycerol tube in a shaking table at the temperature of 28 ℃, and culturing for 14h at the rotating speed of 120r/min to prepare a first-stage seed solution of the acetobacter aceti;
B2. secondary seed liquid culture: respectively inoculating the primary seed liquid of the lactic acid bacteria, the primary seed liquid of the saccharomycetes and the primary seed liquid of the acetobacter in a ratio of 5% into corresponding MRS broth culture medium, YPD liquid culture medium or acetobacter liquid culture medium, wherein the culture conditions are the same as those in the step B1, and the culture time is 10 hours, so as to prepare a secondary seed liquid of the lactic acid bacteria, a secondary seed liquid of the saccharomycetes and a secondary seed liquid of the acetobacter;
B3. strain compounding: taking the secondary seed liquid of lactic acid bacteria, the secondary seed liquid of saccharomycetes and the secondary seed liquid of acetobacter respectively, and compounding according to a certain volume ratio to prepare symbiotic bacterial liquid;
B4. inoculation and fermentation: inoculating symbiotic bacteria liquid into the sterilized tea fruit juice according to the inoculation amount of 4-6%, and standing, fermenting and culturing for 3-7 days at the temperature of 28-30 ℃ to obtain fermentation liquid;
s3, pasteurization: centrifuging the fermentation liquor, collecting supernatant, sterilizing the supernatant at 80 deg.C for 30min, and packaging to obtain XINHUIKANGPU tea.
The invention has the beneficial effects that: (1) the fermentation raw material is the Xinhui citrus pulp, so that the problems of resource waste and environmental pollution of the Xinhui citrus pulp are effectively solved, the problems of sourness, astringency and bitterness of the Xinhui citrus pulp which is directly eaten are solved through a fermentation process, the processing form of the Xinhui citrus is enriched, the high-value utilization of the Xinhui citrus pulp is realized, and the Xinhui citrus pulp is compounded with green tea and mulberry leaves, so that the product has the effects of regulating the intestines and stomach of a human body, removing fat and resisting oxidation;
(2) the traditional Kangpu tea is fermented by taking symbiotic flora (SCOBY) composed of bacteria and yeast, which is often called black tea fungus, as a starting strain, and the black tea fungus contains various microorganism varieties, so that the product quality is not easy to control due to lack of understanding and control on the complex microorganism flora of the black tea fungus. The invention takes the strains in the' list of strains for food and the strains with traditional use habits as starting strains, and performs strain compounding according to a certain proportion, thereby not only ensuring the quality and stability of the product, but also leading the product to have the characteristics of the traditional Kangpu tea;
(3) beverages with an ethanol content > 0.5% (vol) are defined as alcoholic beverages. According to the invention, by controlling the fermentation system and carrying out pasteurization heat treatment after fermentation, the ethanol content in Xinhui orange kangpu tea is controlled to be below 0.5%, and the consistent product quality can be ensured to a certain extent.
In some embodiments, the lactic acid bacteria are one or more of lactobacillus plantarum, lactobacillus acidophilus, lactobacillus rhamnosus, or lactobacillus delbrueckii subsp.
In some embodiments, the secondary seed liquid of lactobacillus in step B3 includes a secondary seed liquid of lactobacillus plantarum, a secondary seed liquid of lactobacillus rhamnosus, and a secondary seed liquid of lactobacillus acidophilus, and the volume ratio of the secondary seed liquid of lactobacillus plantarum, the secondary seed liquid of lactobacillus rhamnosus, and the secondary seed liquid of lactobacillus acidophilus is: 2:1:1.
In some embodiments, the secondary seed liquid of lactic acid bacteria in step B3 comprises a secondary seed liquid of lactobacillus plantarum, a secondary seed liquid of lactobacillus rhamnosus, and a secondary seed liquid of lactobacillus delbrueckii subsp. 2:1:1.
In some embodiments, the yeast is one or both of kluyveromyces marxianus, saccharomyces cerevisiae.
In some embodiments, the secondary seed liquid of yeast in step B3 includes kluyveromyces marxianus seed liquid and saccharomyces cerevisiae seed liquid, and the volume ratio of the kluyveromyces marxianus seed liquid to the saccharomyces cerevisiae seed liquid is: 1:1.
In some embodiments, the acetobacter is acetobacter pasteurianus.
In some embodiments, the tea leaves in the step a1 comprise green tea leaves and dried mulberry leaves, and the mass ratio of the green tea leaves to the dried mulberry leaves is 5-7: 3 to 5.
In some embodiments, the mesh size of the screen is 300 mesh.
In some embodiments, the amount of white granulated sugar added in step a4 is 3% to 5%, and the amount of monopotassium phosphate added is 0.1%.
Drawings
FIG. 1 is a graph of the in vitro antioxidant activity of a Xinhui citrus kanpu tea according to an embodiment of the present invention.
Detailed Description
The present invention will be described in further detail with reference to examples.
Example 1
The preparation method of the Xinhui mandarin orange kangpu tea comprises the following steps:
s1, preparation of a symbiotic bacteria culture medium:
A1. preparing tea liquid: according to the mass ratio of 5: 3 weighing green tea and dry folium Mori, mixing to obtain tea, cooling to 85 deg.C with boiling water, adding tea into water in an amount of 0.6g per 100ml water, maintaining for 20min, and filtering with 300 mesh sieve to remove tea residue to obtain tea solution;
A2. preparing Xinhui citrus juice: selecting 9-11 months peeled Xinhui citrus pulp, wherein the Xinhui citrus pulp meets the requirements of bright color, no plant diseases and insect pests and no mechanical damage, and after removing the core of the Xinhui citrus pulp, mixing the Xinhui citrus pulp with purified water according to the ratio of 5: 3, putting the mixture into a juicer for juicing, and filtering by using a 300-mesh sieve to obtain citrus reticulata fruit juice for later use;
A3. compounding: mixing the tea liquid with the Xinhui citrus juice according to the volume ratio of 5: 5, compounding and uniformly mixing to obtain a compound liquid;
A4. and (3) sterilization: adding 4% of white granulated sugar and 0.1% of potassium dihydrogen phosphate into the compound liquid, stirring uniformly, subpackaging in a conical flask according to 40% of liquid loading amount, and sterilizing at 105 ℃ for 10min to obtain sterilized tea fruit juice;
s2, inoculation:
B1. primary seed liquid culture: inoculating lactobacillus plantarum glycerol tube, lactobacillus rhamnosus glycerol tube and lactobacillus acidophilus glycerol tube into MRS broth culture medium respectively in an inoculum size of 2%, and performing static culture at 37 deg.C for 14h to obtain primary seed liquid of lactobacillus plantarum, primary seed liquid of lactobacillus rhamnosus and primary seed liquid of lactobacillus acidophilus; inoculating Kluyveromyces marxianus glycerol tube and Saccharomyces cerevisiae glycerol tube into YPD liquid culture medium at inoculation amount of 2%, respectively, placing in a shaking table at 26 deg.C, and culturing at rotation speed of 120r/min for 14 hr to obtain first-stage seed liquid of Kluyveromyces marxianus and first-stage seed liquid of Saccharomyces cerevisiae; inoculating 2% of acetobacter pasteurianus glycerol tube into an acetobacter aceti liquid culture medium, placing the acetobacter aceti glycerol tube in a shaking table at the temperature of 28 ℃, and culturing for 14h at the rotating speed of 120r/min to prepare a first-stage seed liquid of acetobacter pasteurianus;
B2. secondary seed liquid culture: respectively inoculating the primary seed liquid of lactobacillus plantarum, the primary seed liquid of lactobacillus rhamnosus, the primary seed liquid of lactobacillus acidophilus, the primary seed liquid of kluyveromyces marxianus, the primary seed liquid of saccharomyces cerevisiae and the primary seed liquid of acetobacter pasteurianus into corresponding MRS broth culture medium, YPD liquid culture medium or acetobacter aceticus culture medium in a proportion of 5%, wherein the culture conditions are the same as those in the step B1, the culture time is 10 hours, and preparing the secondary seed liquid of lactobacillus plantarum, the secondary seed liquid of lactobacillus rhamnosus, the secondary seed liquid of lactobacillus acidophilus, the secondary seed liquid of kluyveromyces marxianus, the secondary seed liquid of saccharomyces cerevisiae and the secondary seed liquid of acetobacter pasteurianus;
B3. strain compounding: mixing the secondary seed liquid of lactobacillus plantarum, the secondary seed liquid of lactobacillus rhamnosus and the secondary seed liquid of lactobacillus acidophilus according to a volume ratio of 2:1:1 to obtain a secondary seed liquid of lactobacillus, and mixing the two seed liquids according to a volume ratio of 1: mixing the second-stage seed liquid of kluyveromyces marxianus and the second-stage seed liquid of saccharomyces cerevisiae to obtain the second-stage seed liquid of saccharomyces cerevisiae, respectively taking the second-stage seed liquid of lactobacillus, the second-stage seed liquid of saccharomyces cerevisiae and the second-stage seed liquid of acetobacter in a volume ratio of 1: 1:1, compounding to prepare symbiotic bacterial liquid;
B4. inoculation and fermentation: inoculating symbiotic bacteria liquid into the sterilized tea fruit juice according to the inoculation amount of 4%, and standing, fermenting and culturing for 3 days at the temperature of 28 ℃ to obtain fermentation liquid;
s3, pasteurization: centrifuging the fermentation liquor, collecting supernatant, sterilizing the supernatant at 80 deg.C for 30min, and packaging to obtain XINHUIKANGPU tea.
Example 2
The preparation method of the Xinhui mandarin orange Kangpu tea comprises the following steps:
s1, preparation of a symbiotic bacteria culture medium:
A1. preparing tea liquid: according to the mass ratio of 6: weighing green tea and dried folium Mori, mixing to obtain tea, cooling to 90 deg.C with boiling water, adding 0.6g of tea into 100ml of water, maintaining for 30min, and filtering with 300 mesh sieve to remove tea residue to obtain tea solution;
A2. preparing Xinhui citrus juice: selecting 9-11 months peeled Xinhui citrus pulp, wherein the Xinhui citrus pulp meets the requirements of bright color, no plant diseases and insect pests and no mechanical damage, and after removing the core of the Xinhui citrus pulp, mixing the Xinhui citrus pulp with purified water according to the ratio of 5: 3, putting the mixture into a juicer for juicing, and filtering by using a 300-mesh sieve to obtain citrus reticulata fruit juice for later use;
A3. compounding: mixing the tea liquid with the Xinhui citrus juice according to the volume ratio of 5: 5, compounding and uniformly mixing to obtain a compound liquid;
A4. and (3) sterilization: adding 5% white sugar and 0.1% potassium dihydrogen phosphate into the compound solution, stirring, subpackaging in a conical flask according to 40% liquid loading amount, and sterilizing at 105 deg.C for 10min to obtain sterilized tea fruit juice;
s2, inoculation:
B1. primary seed liquid culture: inoculating lactobacillus plantarum glycerol tube, lactobacillus rhamnosus glycerol tube and lactobacillus acidophilus glycerol tube into MRS broth culture medium respectively in an inoculum size of 2%, and performing static culture at 37 deg.C for 14h to obtain primary seed liquid of lactobacillus plantarum, primary seed liquid of lactobacillus rhamnosus and primary seed liquid of lactobacillus acidophilus; inoculating Kluyveromyces marxianus glycerol tube and Saccharomyces cerevisiae glycerol tube into YPD liquid culture medium at inoculation amount of 2%, respectively, placing in a shaking table at 26 deg.C, and culturing at rotation speed of 120r/min for 14 hr to obtain first-stage seed liquid of Kluyveromyces marxianus and first-stage seed liquid of Saccharomyces cerevisiae; inoculating 2% of acetobacter pasteurianus glycerol tube into an acetobacter aceti liquid culture medium, placing the acetobacter aceti glycerol tube in a shaking table at the temperature of 28 ℃, and culturing for 14h at the rotating speed of 120r/min to prepare a first-stage seed liquid of acetobacter pasteurianus;
B2. secondary seed liquid culture: respectively inoculating the primary seed liquid of lactobacillus plantarum, the primary seed liquid of lactobacillus rhamnosus, the primary seed liquid of lactobacillus acidophilus, the primary seed liquid of kluyveromyces marxianus, the primary seed liquid of saccharomyces cerevisiae and the primary seed liquid of acetobacter pasteurianus into corresponding MRS broth culture medium, YPD liquid culture medium or acetobacter aceticus culture medium in a proportion of 5%, wherein the culture conditions are the same as those in the step B1, the culture time is 10 hours, and preparing the secondary seed liquid of lactobacillus plantarum, the secondary seed liquid of lactobacillus rhamnosus, the secondary seed liquid of lactobacillus acidophilus, the secondary seed liquid of kluyveromyces marxianus, the secondary seed liquid of saccharomyces cerevisiae and the secondary seed liquid of acetobacter pasteurianus;
B3. strain compounding: according to the volume ratio of 2:1:1 mixing the secondary seed liquid of lactobacillus plantarum, the secondary seed liquid of lactobacillus rhamnosus and the secondary seed liquid of lactobacillus acidophilus to obtain the secondary seed liquid of lactobacillus according to the volume ratio of 1: mixing the second-stage seed liquid of kluyveromyces marxianus and the second-stage seed liquid of saccharomyces cerevisiae to obtain the second-stage seed liquid of saccharomyces cerevisiae, respectively taking the second-stage seed liquid of lactobacillus, the second-stage seed liquid of saccharomyces cerevisiae and the second-stage seed liquid of acetobacter in a volume ratio of 2:1: 2, compounding to prepare symbiotic bacterial liquid;
B4. inoculation and fermentation: inoculating the symbiotic bacterial liquid into the sterilized tea fruit juice according to the inoculation amount of 5%, and standing, fermenting and culturing for 5 days at the temperature of 30 ℃ to obtain fermentation liquid;
s3, pasteurization: centrifuging the fermentation liquor, collecting supernatant, sterilizing the supernatant at 80 deg.C for 30min, and packaging to obtain XINHUIKANGPU tea.
Example 3
The preparation method of the Xinhui mandarin orange Kangpu tea comprises the following steps:
s1, preparation of a symbiotic bacteria culture medium:
A1. preparing tea liquid: according to the mass ratio of 7: 3 weighing green tea and dry folium Mori, mixing to obtain tea, cooling to 87.5 deg.C with boiling water, adding tea into water at an amount of 0.6g per 100ml water, maintaining for 30min, and filtering with 300 mesh sieve to remove tea residue to obtain tea solution;
A2. preparing Xinhui citrus juice: selecting 9-11 months peeled Xinhui citrus pulp, wherein the Xinhui citrus pulp meets the requirements of bright color, no plant diseases and insect pests and no mechanical damage, and after removing the core of the Xinhui citrus pulp, mixing the Xinhui citrus pulp with purified water according to the ratio of 5: 3, putting the mixture into a juicer for juicing, and filtering by using a 300-mesh sieve to obtain citrus reticulata fruit juice for later use;
A3. compounding: mixing the tea liquid with the Xinhui citrus juice according to the volume ratio of 5: 5, compounding and uniformly mixing to obtain a compound liquid;
A4. and (3) sterilization: adding 4.5% white sugar and 0.1% potassium dihydrogen phosphate into the compound solution, stirring, packaging into conical flask according to 40% liquid loading amount, and sterilizing at 105 deg.C for 10min to obtain sterilized tea fruit juice;
s2, inoculation:
B1. primary seed liquid culture: inoculating lactobacillus plantarum glycerol tube, lactobacillus rhamnosus glycerol tube and lactobacillus delbrueckii subspecies bulgaricus glycerol tube into MRS broth culture medium respectively in an inoculum size of 2%, and standing and culturing at 37 deg.C for 14h to obtain primary seed liquid of lactobacillus plantarum, primary seed liquid of lactobacillus rhamnosus and primary seed liquid of lactobacillus delbrueckii subspecies bulgaricus; inoculating Kluyveromyces marxianus glycerol tube and Saccharomyces cerevisiae glycerol tube into YPD liquid culture medium at inoculation amount of 2%, respectively, placing in a shaking table at 26 deg.C, and culturing at rotation speed of 120r/min for 14 hr to obtain first-stage seed liquid of Kluyveromyces marxianus and first-stage seed liquid of Saccharomyces cerevisiae; inoculating 2% of acetobacter pasteurianus glycerol tube into an acetobacter aceti liquid culture medium, placing the acetobacter aceti glycerol tube in a shaking table at the temperature of 28 ℃, and culturing for 14h at the rotating speed of 120r/min to prepare a first-stage seed liquid of acetobacter pasteurianus;
B2. secondary seed liquid culture: respectively inoculating the primary seed liquid of lactobacillus plantarum, the primary seed liquid of lactobacillus rhamnosus, the primary seed liquid of lactobacillus delbrueckii subspecies bulgaricus, the primary seed liquid of kluyveromyces marxianus, the primary seed liquid of saccharomyces cerevisiae and the primary seed liquid of acetobacter pasteurianus into corresponding MRS broth culture medium, YPD liquid culture medium or acetobacter aceticus liquid culture medium in a proportion of 5%, wherein the culture conditions are the same as those in step B1, the culture time is 10 hours, and the secondary seed liquid of lactobacillus plantarum, the secondary seed liquid of lactobacillus rhamnosus, the secondary seed liquid of lactobacillus delbrueckii subspecies bulgaricus, the secondary seed liquid of kluyveromyces marxianus, the secondary seed liquid of saccharomyces cerevisiae and the secondary seed liquid of acetobacter pasteurianus are prepared;
B3. strain compounding: according to the volume ratio of 2:1:1, mixing the secondary seed liquid of lactobacillus plantarum, the secondary seed liquid of lactobacillus rhamnosus and the secondary seed liquid of lactobacillus delbrueckii subsp bulgaricus to obtain the secondary seed liquid of lactobacillus according to the volume ratio of 1: mixing the second-stage seed liquid of kluyveromyces marxianus and the second-stage seed liquid of saccharomyces cerevisiae to obtain the second-stage seed liquid of saccharomyces cerevisiae, respectively taking the second-stage seed liquid of lactobacillus, the second-stage seed liquid of saccharomyces cerevisiae and the second-stage seed liquid of acetobacter in a volume ratio of 1: 1:1, compounding to prepare symbiotic bacterial liquid;
B4. inoculation and fermentation: inoculating the symbiotic bacterial liquid into the sterilized tea fruit juice according to the inoculation amount of 6%, and standing, fermenting and culturing for 6 days at 29 ℃ to obtain fermentation liquid;
s3, pasteurization: centrifuging the fermentation liquor, collecting supernatant, sterilizing the supernatant at 80 deg.C for 30min, and packaging to obtain XINHUIKANGPU tea.
Example 4
The preparation method of the Xinhui mandarin orange Kangpu tea comprises the following steps:
s1, preparation of a symbiotic bacteria culture medium:
A1. preparing tea liquid: according to the mass ratio of 6: weighing green tea and dried folium Mori, mixing to obtain tea, cooling to 89 deg.C with boiling water, adding 0.6g of tea into 100ml of water, maintaining for 25min, and filtering with 300 mesh sieve to remove tea residue to obtain tea solution;
A2. preparing Xinhui citrus juice: selecting 9-11 months peeled Xinhui citrus pulp, wherein the Xinhui citrus pulp meets the requirements of bright color, no plant diseases and insect pests and no mechanical damage, and after removing the core of the Xinhui citrus pulp, mixing the Xinhui citrus pulp with purified water according to the ratio of 5: 3, putting the mixture into a juicer for juicing, and filtering by using a 300-mesh sieve to obtain citrus reticulata fruit juice for later use;
A3. compounding: mixing the tea liquid with the Xinhui citrus juice according to the volume ratio of 6: 4, compounding and uniformly mixing to obtain a compound liquid;
A4. and (3) sterilization: adding 5% white sugar and 0.1% potassium dihydrogen phosphate into the compound solution, stirring, subpackaging in a conical flask according to 40% liquid loading amount, and sterilizing at 105 deg.C for 10min to obtain sterilized tea fruit juice;
s2, inoculation:
B1. primary seed liquid culture: inoculating lactobacillus plantarum glycerol tube, lactobacillus rhamnosus glycerol tube and lactobacillus delbrueckii subspecies bulgaricus glycerol tube into MRS broth culture medium respectively in an inoculum size of 2%, and standing and culturing at 37 deg.C for 14h to obtain primary seed liquid of lactobacillus plantarum, primary seed liquid of lactobacillus rhamnosus and primary seed liquid of lactobacillus delbrueckii subspecies bulgaricus; inoculating Kluyveromyces marxianus glycerol tube and Saccharomyces cerevisiae glycerol tube into YPD liquid culture medium at inoculation amount of 2%, respectively, placing in a shaking table at 26 deg.C, and culturing at rotation speed of 120r/min for 14 hr to obtain first-stage seed liquid of Kluyveromyces marxianus and first-stage seed liquid of Saccharomyces cerevisiae; inoculating 2% of acetobacter pasteurianus glycerol tube into an acetobacter aceti liquid culture medium, placing the acetobacter aceti glycerol tube in a shaking table at the temperature of 28 ℃, and culturing for 14h at the rotating speed of 120r/min to prepare a first-stage seed liquid of acetobacter pasteurianus;
B2. secondary seed liquid culture: respectively inoculating the primary seed liquid of lactobacillus plantarum, the primary seed liquid of lactobacillus rhamnosus, the primary seed liquid of lactobacillus delbrueckii subspecies bulgaricus, the primary seed liquid of kluyveromyces marxianus, the primary seed liquid of saccharomyces cerevisiae and the primary seed liquid of acetobacter pasteurianus into corresponding MRS broth culture medium, YPD liquid culture medium or acetobacter aceticus liquid culture medium in a proportion of 5%, wherein the culture conditions are the same as those in step B1, the culture time is 10 hours, and the secondary seed liquid of lactobacillus plantarum, the secondary seed liquid of lactobacillus rhamnosus, the secondary seed liquid of lactobacillus delbrueckii subspecies bulgaricus, the secondary seed liquid of kluyveromyces marxianus, the secondary seed liquid of saccharomyces cerevisiae and the secondary seed liquid of acetobacter pasteurianus are prepared;
B3. strain compounding: according to the volume ratio of 2:1:1, mixing the secondary seed liquid of lactobacillus plantarum, the secondary seed liquid of lactobacillus rhamnosus and the secondary seed liquid of lactobacillus delbrueckii subsp bulgaricus to obtain the secondary seed liquid of lactobacillus according to the volume ratio of 1: mixing the second-stage seed liquid of kluyveromyces marxianus and the second-stage seed liquid of saccharomyces cerevisiae to obtain the second-stage seed liquid of saccharomyces cerevisiae, respectively taking the second-stage seed liquid of lactobacillus, the second-stage seed liquid of saccharomyces cerevisiae and the second-stage seed liquid of acetobacter in a volume ratio of 2:1: 2, compounding to prepare symbiotic bacterial liquid;
B4. inoculation and fermentation: inoculating the symbiotic bacterial liquid into the sterilized tea fruit juice according to the inoculation amount of 6%, and standing, fermenting and culturing for 5 days at the temperature of 28 ℃ to obtain fermentation liquid;
s3, pasteurization: centrifuging the fermentation liquor, collecting supernatant, sterilizing the supernatant at 80 deg.C for 30min, and packaging to obtain XINHUIKANGPU tea.
Example 5
The preparation method of the Xinhui mandarin orange Kangpu tea comprises the following steps:
s1, preparation of a symbiotic bacteria culture medium:
A1. preparing tea liquid: according to the mass ratio of 6: weighing green tea and dried folium Mori, mixing to obtain tea, cooling to 89 deg.C with boiling water, adding 0.6g of tea into 100ml of water, maintaining for 25min, and filtering with 300 mesh sieve to remove tea residue to obtain tea solution;
A2. preparing Xinhui citrus juice: selecting 9-11 months peeled Xinhui citrus pulp, wherein the Xinhui citrus pulp meets the requirements of bright color, no plant diseases and insect pests and no mechanical damage, and after removing the core of the Xinhui citrus pulp, mixing the Xinhui citrus pulp with purified water according to the ratio of 5: 3, putting the mixture into a juicer for juicing, and filtering by using a 300-mesh sieve to obtain citrus reticulata fruit juice for later use;
A3. compounding: mixing the tea liquid with the Xinhui citrus juice according to the volume ratio of 6: 4, compounding and uniformly mixing to obtain a compound liquid;
A4. and (3) sterilization: adding 5% white sugar and 0.1% potassium dihydrogen phosphate into the compound solution, stirring, subpackaging in a conical flask according to 40% liquid loading amount, and sterilizing at 105 deg.C for 10min to obtain sterilized tea fruit juice;
s2, inoculation:
B1. primary seed liquid culture: inoculating lactobacillus plantarum glycerol tube, lactobacillus rhamnosus glycerol tube and lactobacillus delbrueckii subspecies bulgaricus glycerol tube into MRS broth culture medium respectively in an inoculum size of 2%, and standing and culturing at 37 deg.C for 14h to obtain primary seed liquid of lactobacillus plantarum, primary seed liquid of lactobacillus rhamnosus and primary seed liquid of lactobacillus delbrueckii subspecies bulgaricus; inoculating Kluyveromyces marxianus glycerol tube and Saccharomyces cerevisiae glycerol tube into YPD liquid culture medium at inoculation amount of 2%, respectively, placing in a shaking table at 26 deg.C, and culturing at rotation speed of 120r/min for 14 hr to obtain first-stage seed liquid of Kluyveromyces marxianus and first-stage seed liquid of Saccharomyces cerevisiae; inoculating 2% of acetobacter pasteurianus glycerol tube into an acetobacter aceti liquid culture medium, placing the acetobacter aceti glycerol tube in a shaking table at the temperature of 28 ℃, and culturing for 14h at the rotating speed of 120r/min to prepare a first-stage seed liquid of acetobacter pasteurianus;
B2. secondary seed liquid culture: respectively inoculating the primary seed liquid of lactobacillus plantarum, the primary seed liquid of lactobacillus rhamnosus, the primary seed liquid of lactobacillus delbrueckii subspecies bulgaricus, the primary seed liquid of kluyveromyces marxianus, the primary seed liquid of saccharomyces cerevisiae and the primary seed liquid of acetobacter pasteurianus into corresponding MRS broth culture medium, YPD liquid culture medium or acetobacter aceticus liquid culture medium in a proportion of 5%, wherein the culture conditions are the same as those in step B1, the culture time is 10 hours, and the secondary seed liquid of lactobacillus plantarum, the secondary seed liquid of lactobacillus rhamnosus, the secondary seed liquid of lactobacillus delbrueckii subspecies bulgaricus, the secondary seed liquid of kluyveromyces marxianus, the secondary seed liquid of saccharomyces cerevisiae and the secondary seed liquid of acetobacter pasteurianus are prepared;
B3. strain compounding: according to the volume ratio of 2:1:1, mixing the secondary seed liquid of lactobacillus plantarum, the secondary seed liquid of lactobacillus rhamnosus and the secondary seed liquid of lactobacillus delbrueckii subsp bulgaricus to obtain the secondary seed liquid of lactobacillus according to the volume ratio of 1: mixing the second-stage seed liquid of kluyveromyces marxianus and the second-stage seed liquid of saccharomyces cerevisiae to obtain the second-stage seed liquid of saccharomyces cerevisiae, respectively taking the second-stage seed liquid of lactobacillus, the second-stage seed liquid of saccharomyces cerevisiae and the second-stage seed liquid of acetobacter in a volume ratio of 1: 1: 3, compounding to prepare symbiotic bacterial liquid;
B4. inoculation and fermentation: inoculating the symbiotic bacterial liquid into the sterilized tea fruit juice according to the inoculation amount of 6%, and standing, fermenting and culturing for 5 days at the temperature of 28 ℃ to obtain fermentation liquid;
s3, pasteurization: centrifuging the fermentation liquor, collecting supernatant, sterilizing the supernatant at 80 deg.C for 30min, and packaging to obtain XINHUIKANGPU tea.
Example 6
The preparation method of the Xinhui mandarin orange Kangpu tea comprises the following steps:
s1, preparation of a symbiotic bacteria culture medium:
A1. preparing tea liquid: according to the mass ratio of 6: weighing green tea and dried folium Mori, mixing to obtain tea, cooling to 89 deg.C with boiling water, adding 0.6g of tea into 100ml of water, maintaining for 25min, and filtering with 300 mesh sieve to remove tea residue to obtain tea solution;
A2. preparing Xinhui citrus juice: selecting 9-11 months peeled Xinhui citrus pulp, wherein the Xinhui citrus pulp meets the requirements of bright color, no plant diseases and insect pests and no mechanical damage, and after removing the core of the Xinhui citrus pulp, mixing the Xinhui citrus pulp with purified water according to the ratio of 5: 3, putting the mixture into a juicer for juicing, and filtering by using a 300-mesh sieve to obtain citrus reticulata fruit juice for later use;
A3. compounding: mixing the tea liquid with the Xinhui citrus juice according to the volume ratio of 6: 4, compounding and uniformly mixing to obtain a compound liquid;
A4. and (3) sterilization: adding 5% white sugar and 0.1% potassium dihydrogen phosphate into the compound solution, stirring, subpackaging in a conical flask according to 40% liquid loading amount, and sterilizing at 105 deg.C for 10min to obtain sterilized tea fruit juice;
s2, inoculation:
B1. primary seed liquid culture: inoculating lactobacillus plantarum glycerol tube, lactobacillus rhamnosus glycerol tube and lactobacillus delbrueckii subspecies bulgaricus glycerol tube into MRS broth culture medium respectively in an inoculum size of 2%, and standing and culturing at 37 deg.C for 14h to obtain primary seed liquid of lactobacillus plantarum, primary seed liquid of lactobacillus rhamnosus and primary seed liquid of lactobacillus delbrueckii subspecies bulgaricus; inoculating Kluyveromyces marxianus glycerol tube and Saccharomyces cerevisiae glycerol tube into YPD liquid culture medium at inoculation amount of 2%, respectively, placing in a shaking table at 26 deg.C, and culturing at rotation speed of 120r/min for 14 hr to obtain first-stage seed liquid of Kluyveromyces marxianus and first-stage seed liquid of Saccharomyces cerevisiae; inoculating 2% of acetobacter pasteurianus glycerol tube into an acetobacter aceti liquid culture medium, placing the acetobacter aceti glycerol tube in a shaking table at the temperature of 28 ℃, and culturing for 14h at the rotating speed of 120r/min to prepare a first-stage seed liquid of acetobacter pasteurianus;
B2. secondary seed liquid culture: respectively inoculating the primary seed liquid of lactobacillus plantarum, the primary seed liquid of lactobacillus rhamnosus, the primary seed liquid of lactobacillus delbrueckii subspecies bulgaricus, the primary seed liquid of kluyveromyces marxianus, the primary seed liquid of saccharomyces cerevisiae and the primary seed liquid of acetobacter pasteurianus into corresponding MRS broth culture medium, YPD liquid culture medium or acetobacter aceticus liquid culture medium in a proportion of 5%, wherein the culture conditions are the same as those in step B1, the culture time is 10 hours, and the secondary seed liquid of lactobacillus plantarum, the secondary seed liquid of lactobacillus rhamnosus, the secondary seed liquid of lactobacillus delbrueckii subspecies bulgaricus, the secondary seed liquid of kluyveromyces marxianus, the secondary seed liquid of saccharomyces cerevisiae and the secondary seed liquid of acetobacter pasteurianus are prepared;
B3. strain compounding: according to the volume ratio of 2:1:1, mixing the secondary seed liquid of lactobacillus plantarum, the secondary seed liquid of lactobacillus rhamnosus and the secondary seed liquid of lactobacillus delbrueckii subsp bulgaricus to obtain the secondary seed liquid of lactobacillus according to the volume ratio of 1: mixing the second-stage seed liquid of kluyveromyces marxianus and the second-stage seed liquid of saccharomyces cerevisiae to obtain the second-stage seed liquid of saccharomyces cerevisiae, respectively taking the second-stage seed liquid of lactobacillus, the second-stage seed liquid of saccharomyces cerevisiae and the second-stage seed liquid of acetobacter in a volume ratio of 2:1: 3, compounding to prepare symbiotic bacterial liquid;
B4. inoculation and fermentation: inoculating the symbiotic bacterial liquid into the sterilized tea fruit juice according to the inoculation amount of 6%, and standing, fermenting and culturing for 5 days at the temperature of 28 ℃ to obtain fermentation liquid;
s3, pasteurization: centrifuging the fermentation liquor, collecting supernatant, sterilizing the supernatant at 80 deg.C for 30min, and packaging to obtain XINHUIKANGPU tea.
Example 7 active ingredient assay
Taking the Xinhui Gankoupu tea prepared in the examples 1 to 6 as a detection object, measuring the content and pH value of total flavone, total phenol, lactic acid and Ethanol, measuring the content of total flavone by spectrophotometry in the content of total flavone according to appendix G of GB/T12143-2008 "general analysis method for beverage", measuring the content of total phenol according to T/AHFIA 005-2018 "spectrophotometry for measuring the content of total polyphenol in plant extract and products thereof", measuring the content of lactic acid according to GB 5009.157-2016 (determination of organic acid in food) ", measuring the content of Ethanol by using an Ethanol Colorimetric Kit (Ethanol Colorimetric Assay Kit E-BC-K891-M), measuring the pH value by using a pH meter, obtaining the content and pH value of total flavone, total phenol, lactic acid and Ethanol in the Xinhui Gankoupu tea prepared in the examples 1 to 6, see table 1 below.
TABLE 1 table of active ingredient assay in Xinhui mandarin orange Kangpu tea
Figure DEST_PATH_IMAGE002
Through the data in the table 1, the contents of total flavone and total phenol in the Xinhui citrus kangpu tea prepared by the invention are higher, the content of the total flavone is higher than 0.856g/kg, the content of the total phenol is higher than 0.16mg/mL, the content of antioxidant active ingredients is higher, the content of ethanol is lower than 0.5%, and the Xinhui citrus kangpu tea does not belong to alcoholic beverages.
Example 8 evaluation of antioxidant Effect of Xinhui Ganku Compu tea
In vitro antioxidant activity of chaga neoforma was evaluated by an Oxygen Radical Absorption Capacity (ORAC) test using the chaga neoforma prepared in example 4 as an evaluation target. Fluorescein sodium, when excited by 485 nm light, emits 527 nm fluorescence, which can be oxidized by free radicals released by AAPH to cause fluorescence quenching. And the antioxidant in the reaction system competes with sodium fluorescein for the oxidation of free radicals to slow down the fluorescence decay rate. Thus, the ORAC assay can be used to determine the free radical scavenging activity of a sample.
The experiments were divided into free radical control (AAPH +) and blank control without AAPH addition (AAPH-), Trolox and Xinhui mandarin orange conpu tea groups. The specific determination method is as follows: the Xinhui kangpu tea prepared in example 4 was added to a 96-well plate, and then potassium phosphate buffer and fluorescein sodium were added, and finally AAPH was added. And (3) placing the 96-well plate in a fluorescent microplate reader at 37 ℃ for starting measurement, recording the change of a fluorescent signal within 120 min, detecting once every 2 min, drawing a fluorescence decay curve and calculating the area under the curve. The ORAC value of the oxygen free radical scavenging capacity of the antioxidant is obtained by comparing the protection area of a fluorescence decay curve of a sample to be detected with the protection area of Trolox (1 mu mol. L-1). The formula is given as ORAC = K [ (AUCSample-AUCAAPH +)/(AUCTRlox-AUCAAPH +) ] and K is the sample dilution factor.
The in vitro antioxidant activity of the Xinhui mandarin orange Compu tea prepared in example 4 is shown in FIG. 1, and as can be seen from FIG. 1, the Xinhui mandarin orange Compu tea has a good in vitro free radical scavenging ability, and the ORAC value of the Xinhui mandarin orange Compu tea is calculated to be 1039.38.
Example 9 sensory evaluation of Xinhui Ganku Compu tea
The Xinhui mandarin orange kongpo tea prepared in the examples 1 to 6 is used as an evaluation object, and sensory evaluation is performed on the Xinhui mandarin orange kongpo tea from the aspects of aroma, taste, clarity and color, and evaluation results are shown in the following table 2.
TABLE 2 sensory evaluation results of Xinhui Gankangpu tea
Figure DEST_PATH_IMAGE004
The Xinhui citrus kangpu tea prepared by the method has good aroma and taste, and has good clarity and color. The Xinhui mandarin orange pulp Kangpu tea prepared by the invention can fully improve the resource utilization rate of Xinhui mandarin oranges, combines the unique nutritional values of green tea, probiotics and Xinhui mandarin oranges, and meets the health requirements of modern people on nutritional and healthy beverages on the basis of improving the original poor taste of the Xinhui mandarin orange pulp. The problem that the citrus unshiu pulp is sour, astringent and bitter when being directly eaten is effectively solved, the citrus unshiu pulp is compounded with green tea and mulberry leaves, and the green tea and the mulberry leaves are fermented into the citrus unshiu health and common tea with dual characteristic functions of fermented juice and fermented tea, and the citrus unshiu health and common tea can play roles in regulating intestines and stomach of a human body, removing fat and resisting oxidation.
The above description is only for the embodiments of the present invention, and it is obvious to those skilled in the art that various changes and modifications can be made without departing from the inventive concept of the present invention, and these changes and modifications are all within the scope of the present invention.

Claims (10)

1. A Xinhui mandarin orange kangpu tea is characterized in that: the preparation method comprises the following steps:
s1, preparation of a symbiotic bacteria culture medium:
A1. preparing tea liquid: weighing tea, cooling to 85-90 deg.C with boiling water, adding the tea in an amount of 0.6g per 100ml water, maintaining for 20-30min, and filtering to remove tea residue with sieve to obtain tea solution;
A2. preparing Xinhui citrus juice: selecting citrus reticulata pulp peeled for 9-11 months, wherein the citrus reticulata pulp meets the requirements of bright color, no plant diseases and insect pests and no mechanical damage, and after the citrus reticulata pulp is denucleated, mixing the denucleated citrus reticulata pulp with purified water according to the ratio of 5: 3, putting the mixture into a juicer to be juiced, and filtering the juiced mixture by using a sieve to obtain citrus unshiu juice for later use;
A3. compounding: and (3) mixing the tea liquid with the citrus reticulata juice according to the volume ratio of 5-7: compounding according to the proportion of 3-5, and uniformly mixing to obtain a compound liquid;
A4. and (3) sterilization: adding white granulated sugar and potassium dihydrogen phosphate into the compound liquid, subpackaging in a conical flask according to 40% liquid loading amount, and sterilizing at 105 deg.C for 10min to obtain sterilized tea fruit juice;
s2, inoculation:
B1. primary seed liquid culture: inoculating lactobacillus glycerol tube into MRS broth culture medium at an inoculum size of 2%, and standing at 37 deg.C for 14 hr to obtain first-class seed solution of lactobacillus; inoculating 2% of yeast glycerol tube into YPD liquid culture medium, placing in a shaking table at 26 deg.C, and culturing at rotation speed of 120r/min for 14 hr to obtain primary seed liquid of yeast; inoculating 2% of acetobacter aceti glycerol tube into an acetobacter aceti liquid culture medium, placing the acetobacter aceti glycerol tube in a shaking table at the temperature of 28 ℃, and culturing for 14h at the rotating speed of 120r/min to prepare a first-stage seed solution of the acetobacter aceti;
B2. secondary seed liquid culture: respectively inoculating the primary seed liquid of the lactic acid bacteria, the primary seed liquid of the saccharomycetes and the primary seed liquid of the acetobacter into corresponding MRS broth culture medium, YPD liquid culture medium or acetobacter liquid culture medium in a proportion of 5%, wherein the culture conditions are the same as those in the step B1, the culture time is 10 hours, and the secondary seed liquid of the lactic acid bacteria, the secondary seed liquid of the saccharomycetes and the secondary seed liquid of the acetobacter are prepared;
B3. strain compounding: taking the secondary seed liquid of the lactic acid bacteria, the secondary seed liquid of the saccharomycetes and the secondary seed liquid of the acetobacter respectively, and compounding according to a certain volume ratio to prepare symbiotic bacteria liquid;
B4. inoculation and fermentation: inoculating the symbiotic bacterial liquid into the sterilized tea fruit juice according to the inoculation amount of 4-6%, and standing, fermenting and culturing for 3-7 days at the temperature of 28-30 ℃ to obtain fermentation liquid;
s3, pasteurization: and centrifuging the fermentation liquor, collecting supernatant, sterilizing the supernatant at 80 ℃ for 30min, and subpackaging to obtain the Xinhui mandarin orange Kangpu tea.
2. The Xinhui mandarin orange Compu tea of claim 1, wherein the lactic acid bacteria is one or more of Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus rhamnosus, or Lactobacillus delbrueckii subsp.
3. The Xinhui mandarin orange Compu tea as claimed in claim 2, wherein in step B3, the secondary seed liquid of lactobacillus comprises the secondary seed liquid of lactobacillus plantarum, the secondary seed liquid of lactobacillus rhamnosus and the secondary seed liquid of lactobacillus acidophilus, wherein the volume ratio of the secondary seed liquid of lactobacillus plantarum, the secondary seed liquid of lactobacillus rhamnosus and the secondary seed liquid of lactobacillus acidophilus is: 2:1:1.
4. The xinhui mandarin orange compl tea as claimed in claim 2, wherein in step B3, the secondary seed liquid of lactic acid bacteria comprises secondary seed liquid of lactobacillus plantarum, secondary seed liquid of lactobacillus rhamnosus and secondary seed liquid of lactobacillus delbrueckii subsp. 2:1:1.
5. The Xinhui citrus kangpu tea as claimed in claim 1, wherein the yeast is one or two of Kluyveromyces marxianus and Saccharomyces cerevisiae.
6. The Xinhui citrus kanpu tea as claimed in claim 5, wherein in step B3, the secondary seed liquid of yeast comprises a secondary seed liquid of Kluyveromyces marxianus and a secondary seed liquid of Saccharomyces cerevisiae, and the volume ratio of the secondary seed liquid of Kluyveromyces marxianus to the secondary seed liquid of Saccharomyces cerevisiae is: 1:1.
7. The Xinhui citrus compta tea according to claim 1, wherein the acetobacter is acetobacter pasteurianus.
8. The Xinhui mandarin orange Kangpu tea as claimed in claim 1, wherein the tea leaves in the step A1 comprise green tea leaves and dry mulberry leaves, and the mass ratio of the green tea leaves to the dry mulberry leaves is 5-7: 3 to 5.
9. The Xinhui citrus compta tea as claimed in claim 1, wherein the size of the sieve is 300 meshes.
10. The Xinhui mandarin orange Compu tea as claimed in claim 1, wherein the white granulated sugar is added in an amount of 3% -5% in the step A4, and the potassium dihydrogen phosphate is added in an amount of 0.1%.
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CN114831198A (en) * 2022-05-25 2022-08-02 江南大学 Method for increasing organic acid in Kangpu tea by adding milk permeate powder
CN114933987A (en) * 2022-05-09 2022-08-23 仙婷(广州)科技研发有限公司 Symbiotic bacteria fermentation process and application thereof

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CN109845939A (en) * 2019-03-18 2019-06-07 天津科技大学 A kind of preparation method of Xinhui citrus reticulata meat probiotic fermentation beverage
CN111500643A (en) * 2020-04-20 2020-08-07 中国农业科学院麻类研究所 Xinhui citrus fermentation liquor and preparation method and application thereof
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