Preparation method of honey kangpu tea based on synergistic fermentation of pure bacteria

Abstract

The invention discloses a preparation method of honey kangpu tea based on pure bacterium synergistic fermentation, and relates to the field of tea preparation. Comprising the following steps: culturing strains, namely culturing koal fotamentosa, zygosaccharomyces bayer and lactobacillus plantarum; black tea is added into boiling water to be soaked, sterilization is performed after filtering, and acacia honey is added after cooling to be evenly mixed; fermenting, namely inoculating the fermentation substrate with the foal foal bacillus, the zygosaccharomyces bayer and the lactobacillus plantarum, sealing with gauze, and carrying out aerobic fermentation; sterilizing and filling: after fermenting and planting, immediately performing sterile filtration or clear liquid, and then filling after pasteurization. Fermentation can be completed in 14 days through synergistic metabolism of three bacteria, the fermentation efficiency is improved, the fermentation period is shortened, unique sweet and sour balance is formed through synergistic effect of acetic acid and lactic acid, meanwhile, pure bacteria are used for fermentation, infectious microbe pollution is avoided in the fermentation process, and harmful metabolites such as aflatoxin in the honey kangpu tea are avoided.

Description

Technical Field

[0001] The present invention relates to the field of tea preparation, and in particular to a method for preparing honey kombucha based on pure bacteria collaborative fermentation. Background Art

[0002] Kombucha has the effects of clearing heat and detoxifying, moistening the lungs and relieving cough, and lowering blood lipids. The honey in it has the effects of supplementing nutrition, moistening the intestines and promoting bowel movements, promoting digestion, enhancing immunity, and helping to improve sleep quality.

[0003] Kombucha is a traditional fermented tea beverage typically fermented with a mixed bacterial community of Kombucha (SCOBY). This can lead to complex bacterial communities, uncontrollable metabolites, and the potential for contamination by other bacteria. Honey, as a natural carbon source, is rich in fructose and glucose, but its high osmotic pressure inhibits the activity of conventional bacterial strains.

[0004] In the prior art, mixed bacteria fermented honey kombucha has the following defects:

[0005] 1. Long fermentation cycle (usually more than 21 days);

[0006] 2. The product contains high levels of residual sugar and limited flavor compounds;

[0007] 3. Competition among mixed bacterial communities leads to unstable metabolic pathways, making large-scale production difficult.

[0008] To this end, the present invention proposes a method for preparing honey kombucha based on pure bacteria collaborative fermentation. Summary of the Invention

[0009] The purpose of the present invention is to solve the shortcomings of the prior art and to propose a method for preparing honey kombucha based on pure bacteria collaborative fermentation.

[0010] In order to achieve the above object, the present invention adopts the following technical solutions:

[0011] A method for preparing honey kombucha based on pure bacterial synergistic fermentation, wherein Bacillus sucroseus, Zygosaccharomyces bailii, and Lactobacillus plantarum are used as fermentation strains, and the preparation method comprises the following steps:

[0012] S1: bacterial culture, culturing Bacillus sucroseus, Bacillus baumannii and Lactobacillus plantarum;

[0013] S2: Preparation of fermentation matrix, black tea extract: soak black tea in boiling water, filter and sterilize, cool and add acacia honey and mix well;

[0014] S3: fermentation, inoculating Bacillus sucroseus, Zygosaccharomyces bailii, and Lactobacillus plantarum into the fermentation medium, sealing it with gauze, and then performing aerobic fermentation;

[0015] S4: Sterilization and filling: After fermentation and cultivation, sterile filtration or clear liquid is immediately carried out, and then filling is carried out after pasteurization.

[0016] Preferably: in the step S1:

[0017] Bacillus sucroseiflorus and Zygosaccharomyces bailii were cultured at 30°C for 2 days, with the final concentrations of 3.0×10 8 CFU / mL, 2.5×10 8 CFU / mL;

[0018] Lactobacillus plantarum: cultured at 37°C for 2 days, final concentration 2.7×10 8 CFU / mL.

[0019] Preferably: in the step S2:

[0020] The ratio of black tea, boiling water and acacia honey is 7-9g:900-1100ml:65-75g, and the soaking time is 15 minutes;

[0021] The sterilization temperature is 120±5 degrees Celsius and the sterilization time is 15 minutes;

[0022] The cooling temperature is 50-60 degrees Celsius.

[0023] Preferably, in step S3, the inoculation volume ratio is: Bacillus sucroseus liquid: Zygosaccharomyces bailii liquid: Lactobacillus plantarum liquid = (1.5-2.5)%: (1.5-2.5)%: (0.5-1.5)%.

[0024] Preferably, in step S3, the inoculation volume ratio is: Bacillus sucroseus liquid: Bayer's combined yeast liquid: Lactobacillus plantarum liquid = 2%: 2%: 1%.

[0025] Preferably: in the step S3, the fermentation temperature is 30 degrees Celsius, and sampling and testing are carried out during the fermentation process.

[0026] Preferably, in step S3, the sampling time is: sampling and testing every 0, 2, 4, 7, 9, 11, and 14 days.

[0027] Preferably, in step S4, the fermentation endpoint determination conditions are: pH ≤ 3.0, lactic acid ≥ 0.08 g / L, acetic acid ≥ 9.4 g / L; sucrose residue ≤ 1.2 g / L, fructose residue ≤ 1.7 g / L, glucose residue ≤ 4.7 g / L.

[0028] Preferably: in the step S4, the sterilization temperature is 70 degrees Celsius, the sterilization time is 15 minutes, and a 0.22 μm microporous filter membrane is used for filtration.

[0029] Preferably: use of the prepared honey kombucha in functional beverages.

[0030] The beneficial effects of the present invention are:

[0031] 1. The present invention completes fermentation in 14 days through the synergistic metabolism of three bacteria, thereby improving fermentation efficiency and shortening the fermentation cycle. Acetic acid and lactic acid work synergistically to form a unique sweet and sour balance. At the same time, pure bacteria are used for fermentation to avoid contamination by other bacteria during fermentation and the presence of harmful metabolites such as aflatoxin in honey kombucha.

[0032] 2. The kombucha prepared in the present invention has an improved sugar conversion rate, which reduces the greasy sweetness of honey kombucha, while increasing the lactic acid content and antioxidant substances.

[0033] 3. The process of the present invention has high repeatability and is suitable for industrial scale-up. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 This is a flow chart of the method for preparing honey kombucha based on pure bacteria collaborative fermentation proposed by the present invention. DETAILED DESCRIPTION

[0035] The technical solution of the present invention will be further described in detail below in conjunction with specific implementation methods.

[0036] In the description of the present invention, it should be noted that, unless otherwise expressly specified or limited, the terms "installed," "connected," "connected," and "disposed" should be understood in a broad sense. For example, they may refer to fixed connection or disposition, detachable connection or disposition, or integral connection or disposition. Those skilled in the art will understand the specific meanings of the above terms in the present invention based on specific circumstances.

[0037] Example 1: Preparation and testing of honey kombucha

[0038] A method for preparing honey kombucha based on pure bacterial synergistic fermentation, wherein Bacillus sucroseus, Zygosaccharomyces bailii, and Lactobacillus plantarum are used as fermentation strains, and the preparation method comprises the following steps:

[0039] S1: bacterial culture, culture Bacillus sucroseus, Zygosaccharomyces bailii and Lactobacillus plantarum;

[0040] S2: Preparation of fermentation matrix, black tea extract: soak black tea in boiling water, filter and sterilize, cool and add acacia honey and mix well;

[0041] S3: fermentation, inoculating Bacillus sucroseus, Zygosaccharomyces bailii, and Lactobacillus plantarum into the fermentation medium, sealing it with gauze, and then performing aerobic fermentation;

[0042] S4: Sterilization and filling: After fermentation and cultivation, sterile filtration or clear liquid is immediately carried out, and then filling is carried out after pasteurization.

[0043] In the S1 step:

[0044] Bacillus sucroseiflorus and Zygosaccharomyces bailii were cultured at 30°C for 2 days, with the final concentrations of 3.0×10 8 CFU / mL, 2.5×10 8 CFU / mL;

[0045] Lactobacillus plantarum: cultured at 37°C for 2 days, final concentration 2.7×10 8 CFU / mL.

[0046] In the S2 step:

[0047] The ratio of black tea, boiling water and acacia honey is 8g:1000ml:70g, and the soaking time is 15 minutes;

[0048] The sterilization temperature is 121 degrees Celsius and the sterilization time is 15 minutes;

[0049] The cooling temperature is 50-60 degrees Celsius.

[0050] In the step S3, the inoculation volume ratio is: Bacillus sucroseus liquid: Zygosaccharomyces bailii liquid: Lactobacillus plantarum liquid = 2%: 2%: 1%.

[0051] In step S3, the fermentation temperature is 30 degrees Celsius, and samples are taken for testing during the fermentation process.

[0052] In the step S3, the sampling time is: sampling and testing every 0, 2, 4, 7, 9, 11, and 14 days.

[0053] Detection:

[0054] pH value: dropped from the initial 5.8 to 3.0 (measured by pH meter);

[0055] Sugar content: HILIC chromatography detection showed that sucrose, fructose, and glucose were reduced to 1.12 g / L, 1.63 g / L, and 4.64 g / L, respectively;

[0056] Organic acids: T3 column detected lactic acid at 0.09 g / L, HPX-87C column detected acetic acid at 9.47 g / L;

[0057] Volatile components: GC-MS identified 50 substances, including 9 carboxylic acids, 8 esters and 4 characteristic terpenes.

[0058] In the step S4, the fermentation endpoint determination conditions are: pH ≤ 3.0, lactic acid ≥ 0.08 g / L, acetic acid ≥ 9.4 g / L; sucrose residue ≤ 1.2 g / L, fructose residue ≤ 1.7 g / L, and glucose residue ≤ 4.7 g / L.

[0059] In the step S4, the sterilization temperature is 70 degrees Celsius, the sterilization time is 15 minutes, and the filtration is performed using a 0.22 μm microporous filter membrane.

[0060] Example 2:

[0061] A method for preparing honey kombucha based on pure bacterial synergistic fermentation, wherein Bacillus sucroseus, Zygosaccharomyces bailii, and Lactobacillus plantarum are used as fermentation strains, and the preparation method comprises the following steps:

[0062] S1: bacterial culture, culture Bacillus sucroseus, Zygosaccharomyces bailii and Lactobacillus plantarum;

[0063] S2: Preparation of fermentation matrix, black tea extract: soak black tea in boiling water, filter and sterilize, cool and add acacia honey and mix well;

[0064] S3: fermentation, inoculating Bacillus sucroseus, Zygosaccharomyces bailii, and Lactobacillus plantarum into the fermentation medium, sealing it with gauze, and then performing aerobic fermentation;

[0065] S4: Sterilization and filling: After fermentation and cultivation, sterile filtration or clear liquid is immediately carried out, and then filling is carried out after pasteurization.

[0066] In the S1 step:

[0067] Bacillus sucroseiflorus and Zygosaccharomyces bailii were cultured at 30°C for 2 days, with the final concentrations of 3.0×10 8 CFU / mL, 2.5×10 8 CFU / mL;

[0068] Lactobacillus plantarum: cultured at 37°C for 2 days, final concentration 2.7×10 8 CFU / mL.

[0069] In the S2 step:

[0070] The ratio of black tea, boiling water and acacia honey is 7-9g:900-1100ml:65-75g, and the soaking time is 15 minutes;

[0071] The sterilization temperature is 120±5 degrees Celsius and the sterilization time is 15 minutes;

[0072] The cooling temperature is 50-60 degrees Celsius.

[0073] In the step S3, the inoculation volume ratio is: Bacillus sucroseus liquid: Zygosaccharomyces bailii liquid: Lactobacillus plantarum liquid = 1.5%: 2%: 0.5%.

[0074] In step S3, the fermentation temperature is 30 degrees Celsius, and samples are taken for testing during the fermentation process.

[0075] In the step S3, the sampling time is: sampling and testing every 0, 2, 4, 7, 9, 11, and 14 days.

[0076] In the step S4, the fermentation endpoint determination conditions are: pH ≤ 3.0, lactic acid ≥ 0.08 g / L, acetic acid ≥ 9.4 g / L; sucrose residue ≤ 1.2 g / L, fructose residue ≤ 1.7 g / L, and glucose residue ≤ 4.7 g / L.

[0077] In the step S4, the sterilization temperature is 70 degrees Celsius, the sterilization time is 15 minutes, and the filtration is performed using a 0.22 μm microporous filter membrane.

[0078] Example 3:

[0079] A method for preparing honey kombucha based on pure bacterial synergistic fermentation, wherein Bacillus sucroseus, Zygosaccharomyces bailii, and Lactobacillus plantarum are used as fermentation strains, and the preparation method comprises the following steps:

[0080] S1: bacterial culture, culture Bacillus sucroseus, Zygosaccharomyces bailii and Lactobacillus plantarum;

[0081] S2: Preparation of fermentation matrix, black tea extract: soak black tea in boiling water, filter and sterilize, cool and add acacia honey and mix well;

[0082] S3: fermentation, inoculating Bacillus sucroseus, Zygosaccharomyces bailii, and Lactobacillus plantarum into the fermentation medium, sealing it with gauze, and then performing aerobic fermentation;

[0083] S4: Sterilization and filling: After fermentation and cultivation, sterile filtration or clear liquid is immediately carried out, and then filling is carried out after pasteurization.

[0084] In the S1 step:

[0085] Bacillus sucroseiflorus and Zygosaccharomyces bailii were cultured at 30°C for 2 days, with the final concentrations of 3.0×10 8 CFU / mL, 2.5×10 8 CFU / mL;

[0086] Lactobacillus plantarum: cultured at 37°C for 2 days, final concentration 2.7×10 8 CFU / mL.

[0087] In the S2 step:

[0088] The ratio of black tea, boiling water and acacia honey is 7-9g:900-1100ml:65-75g, and the soaking time is 15 minutes;

[0089] The sterilization temperature is 120±5 degrees Celsius and the sterilization time is 15 minutes;

[0090] The cooling temperature is 50-60 degrees Celsius.

[0091] In the step S3, the inoculation volume ratio is: Bacillus sucroseus liquid: Zygosaccharomyces bailii liquid: Lactobacillus plantarum liquid = 2.5%: 1.5%: 1%.

[0092] In step S3, the fermentation temperature is 30 degrees Celsius, and samples are taken for testing during the fermentation process.

[0093] In the step S3, the sampling time is: sampling and testing every 0, 2, 4, 7, 9, 11, and 14 days.

[0094] In the step S4, the fermentation endpoint determination conditions are: pH ≤ 3.0, lactic acid ≥ 0.08 g / L, acetic acid ≥ 9.4 g / L; sucrose residue ≤ 1.2 g / L, fructose residue ≤ 1.7 g / L, and glucose residue ≤ 4.7 g / L.

[0095] In the step S4, the sterilization temperature is 70 degrees Celsius, the sterilization time is 15 minutes, and the filtration is performed using a 0.22 μm microporous filter membrane.

[0096] Example 4:

[0097] A method for preparing honey kombucha based on pure bacterial synergistic fermentation, wherein Bacillus sucroseus, Zygosaccharomyces bailii, and Lactobacillus plantarum are used as fermentation strains, and the preparation method comprises the following steps:

[0098] S1: bacterial culture, culture Bacillus sucroseus, Zygosaccharomyces bailii and Lactobacillus plantarum;

[0099] S2: Preparation of fermentation matrix, black tea extract: soak black tea in boiling water, filter and sterilize, cool and add acacia honey and mix well;

[0100] S3: fermentation, inoculating Bacillus sucroseus, Zygosaccharomyces bailii, and Lactobacillus plantarum into the fermentation medium, sealing it with gauze, and then performing aerobic fermentation;

[0101] S4: Sterilization and filling: After fermentation and cultivation, sterile filtration or clear liquid is immediately carried out, and then filling is carried out after pasteurization.

[0102] In the S1 step:

[0103] Bacillus sucroseiflorus and Zygosaccharomyces bailii were cultured at 30°C for 2 days, with the final concentrations of 3.0×10 8 CFU / mL, 2.5×10 8 CFU / mL;

[0104] Lactobacillus plantarum: cultured at 37°C for 2 days, final concentration 2.7×10 8 CFU / mL.

[0105] In the S2 step:

[0106] The ratio of black tea, boiling water and acacia honey is 7g:1000ml:65g, and the soaking time is 15 minutes;

[0107] The sterilization temperature is 120±5 degrees Celsius and the sterilization time is 15 minutes;

[0108] The cooling temperature is 50-60 degrees Celsius.

[0109] In the step S3, the inoculation volume ratio is: Bacillus sucroseus liquid: Zygosaccharomyces bailii liquid: Lactobacillus plantarum liquid = 2%: 2%: 1%.

[0110] In step S3, the fermentation temperature is 30 degrees Celsius, and samples are taken for testing during the fermentation process.

[0111] In the step S3, the sampling time is: sampling and testing every 0, 2, 4, 7, 9, 11, and 14 days.

[0112] In the step S4, the fermentation endpoint determination conditions are: pH ≤ 3.0, lactic acid ≥ 0.08 g / L, acetic acid ≥ 9.4 g / L; sucrose residue ≤ 1.2 g / L, fructose residue ≤ 1.7 g / L, and glucose residue ≤ 4.7 g / L.

[0113] In the step S4, the sterilization temperature is 70 degrees Celsius, the sterilization time is 15 minutes, and the filtration is performed using a 0.22 μm microporous filter membrane.

[0114] Example 5:

[0115] A method for preparing honey kombucha based on pure bacterial synergistic fermentation, wherein Bacillus sucroseus, Zygosaccharomyces bailii, and Lactobacillus plantarum are used as fermentation strains, and the preparation method comprises the following steps:

[0116] S1: bacterial culture, culture Bacillus sucroseus, Zygosaccharomyces bailii and Lactobacillus plantarum;

[0117] S2: Preparation of fermentation matrix, black tea extract: soak black tea in boiling water, filter and sterilize, cool and add acacia honey and mix well;

[0118] S3: fermentation, inoculating Bacillus sucroseus, Zygosaccharomyces bailii, and Lactobacillus plantarum into the fermentation medium, sealing it with gauze, and then performing aerobic fermentation;

[0119] S4: Sterilization and filling: After fermentation and cultivation, sterile filtration or clear liquid is immediately carried out, and then filling is carried out after pasteurization.

[0120] In the S1 step:

[0121] Bacillus sucroseiflorus and Zygosaccharomyces bailii were cultured at 30°C for 2 days, with the final concentrations of 3.0×10 8 CFU / mL, 2.5×10 8 CFU / mL;

[0122] Lactobacillus plantarum: cultured at 37°C for 2 days, final concentration 2.7×10 8 CFU / mL.

[0123] In the S2 step:

[0124] The ratio of black tea, boiling water and acacia honey is 9g:1100ml:70g, and the soaking time is 15 minutes;

[0125] The sterilization temperature is 120±5 degrees Celsius and the sterilization time is 15 minutes;

[0126] The cooling temperature is 50-60 degrees Celsius.

[0127] In the step S3, the inoculation volume ratio is: Bacillus sucroseus liquid: Zygosaccharomyces bailii liquid: Lactobacillus plantarum liquid = 2%: 2%: 1%.

[0128] In step S3, the fermentation temperature is 30 degrees Celsius, and samples are taken for testing during the fermentation process.

[0129] In the step S3, the sampling time is: sampling and testing every 0, 2, 4, 7, 9, 11, and 14 days.

[0130] In the step S4, the fermentation endpoint determination conditions are: pH ≤ 3.0, lactic acid ≥ 0.08 g / L, acetic acid ≥ 9.4 g / L; sucrose residue ≤ 1.2 g / L, fructose residue ≤ 1.7 g / L, and glucose residue ≤ 4.7 g / L.

[0131] In the step S4, the sterilization temperature is 70 degrees Celsius, the sterilization time is 15 minutes, and the filtration is performed using a 0.22 μm microporous filter membrane.

[0132] Example 6: Scaled production verification

[0133] Based on Example 1, a 500 L fermenter was used to scale up the production according to the same process. The indicators of the finished product were consistent with those of the laboratory batch (p>0.05), confirming that the process can be applied on a large scale.

[0134] The present invention completes fermentation in 14 days through the synergistic metabolism of three bacteria, thereby improving fermentation efficiency and shortening the fermentation cycle. Acetic acid and lactic acid work synergistically to form a unique sweet and sour balance. At the same time, pure bacteria are used for fermentation to avoid contamination by miscellaneous bacteria during fermentation and to prevent the presence of harmful metabolites such as aflatoxin in honey kombucha.

[0135] The kombucha prepared by the present invention has an improved sugar conversion rate, reduces the greasy sweetness of honey kombucha, and at the same time increases the lactic acid content and antioxidant substances.

[0136] The process of the present invention has high repeatability and is suitable for industrial scale-up.

[0137] The above description is only a preferred specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any technician familiar with the technical field, within the technical scope disclosed by the present invention, who makes equivalent replacements or changes based on the technical solution and inventive concept of the present invention, should be covered by the scope of protection of the present invention.

Claims

1. A method for preparing honey kombucha based on pure bacterial cooperative fermentation, which uses Bacillus sucrose, Zygosaccharomyces bailii, and Lactobacillus plantarum as fermentation strains, characterized in that: The preparation method comprises the following steps: S1: bacterial culture, culturing Bacillus sucroseus, Bacillus baumannii and Lactobacillus plantarum; S2: Preparation of fermentation matrix, black tea extract: soak black tea in boiling water, filter and sterilize, cool and add acacia honey and mix well; S3: fermentation, inoculating Bacillus sucroseus, Zygosaccharomyces bailii, and Lactobacillus plantarum into the fermentation medium, sealing it with gauze, and then performing aerobic fermentation; S4: Sterilization and filling: After fermentation and cultivation, sterile filtration or clear liquid is immediately carried out, and then filling is carried out after pasteurization.

2. The method for preparing honey kombucha based on pure bacteria cooperative fermentation according to claim 1, characterized in that: In the S1 step: Bacillus sucroseiflorus and Zygosaccharomyces bailii were cultured at 30°C for 2 days, with the final concentrations of 3.0×10 8 CFU / mL, 2.5×10 8 CFU / mL; Lactobacillus plantarum: cultured at 37°C for 2 days, final concentration 2.7×10 8 CFU / mL.

3. The method for preparing honey kombucha based on pure bacteria cooperative fermentation according to claim 1, characterized in that: In the S2 step: The ratio of black tea, boiling water and acacia honey is 7-9g:900-1100ml:65-75g, and the soaking time is 15 minutes; The sterilization temperature is 120±5 degrees Celsius and the sterilization time is 15 minutes; The cooling temperature is 50-60 degrees Celsius.

4. The method for preparing honey kombucha based on pure bacteria cooperative fermentation according to claim 1, characterized in that: In the step S3, the inoculation volume ratio is: Bacillus sucroseus liquid: Zygosaccharomyces bailii liquid: Lactobacillus plantarum liquid = (1.5-2.5)%: (1.5-2.5)%: (0.5-1.5)%.

5. The method for preparing honey kombucha based on pure bacteria cooperative fermentation according to claim 4, characterized in that: In the step S3, the inoculation volume ratio is: Bacillus sucroseus liquid: Zygosaccharomyces bailii liquid: Lactobacillus plantarum liquid = 2%: 2%: 1%.

6. The method for preparing honey kombucha based on pure bacteria cooperative fermentation according to claim 1, characterized in that: In step S3, the fermentation temperature is 30 degrees Celsius, and samples are taken for testing during the fermentation process.

7. The method for preparing honey kombucha based on pure bacteria cooperative fermentation according to claim 6, characterized in that: In the step S3, the sampling time is: sampling and testing every 0, 2, 4, 7, 9, 11, and 14 days.

8. The method for preparing honey kombucha based on pure bacteria cooperative fermentation according to claim 1, characterized in that: In the step S4, the fermentation endpoint determination conditions are: pH ≤ 3.0, lactic acid ≥ 0.08 g / L, acetic acid ≥ 9.4 g / L; sucrose residue ≤ 1.2 g / L, fructose residue ≤ 1.7 g / L, and glucose residue ≤ 4.7 g / L.

9. The method for preparing honey kombucha based on pure bacteria cooperative fermentation according to claim 1, characterized in that: In the step S4, the sterilization temperature is 70 degrees Celsius, the sterilization time is 15 minutes, and the filtration is performed using a 0.22 μm microporous filter membrane.

10. The method for preparing honey kombucha based on pure bacteria cooperative fermentation according to any one of claims 1 to 9, characterized in that: Application of the prepared honey kombucha in functional beverages.

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