CN110771698A - Processing method of controllable Duyun dark tea - Google Patents

Processing method of controllable Duyun dark tea Download PDF

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Publication number
CN110771698A
CN110771698A CN201910372722.2A CN201910372722A CN110771698A CN 110771698 A CN110771698 A CN 110771698A CN 201910372722 A CN201910372722 A CN 201910372722A CN 110771698 A CN110771698 A CN 110771698A
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tea
spore
suspension
leaves
spores
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周才碧
周小露
宋加艳
赵幸运
罗学尹
刘丽明
王佳菜
石悦
周新
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Guizhou Bili Science And Technology Service Co Ltd
Qiannan Normal University for Nationalities
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Guizhou Bili Science And Technology Service Co Ltd
Qiannan Normal University for Nationalities
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23F—COFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
    • A23F3/00—Tea; Tea substitutes; Preparations thereof
    • A23F3/06—Treating tea before extraction; Preparations produced thereby
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23F—COFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
    • A23F3/00—Tea; Tea substitutes; Preparations thereof
    • A23F3/06—Treating tea before extraction; Preparations produced thereby
    • A23F3/08—Oxidation; Fermentation
    • A23F3/10—Fermentation with addition of microorganisms or enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23F—COFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
    • A23F3/00—Tea; Tea substitutes; Preparations thereof
    • A23F3/06—Treating tea before extraction; Preparations produced thereby
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Abstract

The invention discloses a processing method of controllable homogenized black tea, which comprises the steps of picking fresh leaves, spreading, deactivating enzymes, rolling, drying, sterilizing, preparing bacteria, inoculating, piling and drying, and the preparation method is stable, can effectively improve the quality of homogenized black tea leaves, and promotes the aroma and the taste of homogenized black tea leaves, and the prepared homogenized black tea has rich nutrient components, stable tea leaf quality and better color and does not contain T2, HT2 and 4 aflatoxins; the tea provided by the invention is uniform in black brown appearance, mellow and pure in fragrance, red brown in liquor color, mellow in taste and dark brown in leaf bottom.

Description

Processing method of controllable Duyun dark tea
Technical Field
The invention relates to a tea making method, in particular to a processing method of controllable Duyun dark tea.
Background
Dark tea (dark tea) is named because the finished tea is black in appearance. The black tea belongs to one of six major tea types, belongs to post-fermented tea, and has main production areas of Sichuan, Yunnan, Hubei, Hunan, Shaanxi, Anhui and the like. The raw dark green tea adopted by the traditional dark green tea has higher maturity and is a main raw material for pressing compressed tea. The black tea is distributed according to regions and is mainly classified into Hunan black tea (Fuzhuan tea, Qianliang tea, black brick tea, Sanjian tea and the like), Hubei old black tea, Sichuan Tibetan tea (Bian tea), Anhui ancient black tea (Ancha), Yunnan black tea (Pu' er tea), Guangxi Liupao tea and Shaanxi black tea (Fuzhuan tea). The key process of processing the dark tea is pile fermentation, wherein the pile fermentation is a process that the tea leaves generate a series of deep oxidation and decomposition effects on the components in the tea leaves under the combined action of internal moist heat and microorganisms to form the unique quality of the dark tea.
The dark tea is a tea variety which is popular with people, a special process is adopted in the process of making the dark tea, namely 'pile fermentation', and microorganisms play an important role in the process; the dark tea pile fermentation process is suitable for growth of mycotoxin, so that the possibility of mycotoxin pollution exists, and the quality of finished tea is difficult to control due to the fact that the conventional dark tea processing is easily polluted by mixed bacteria during pile fermentation.
The main microorganisms involved in the pile fermentation process of black tea include Aspergillus niger, Penicillium, Saccharomyces and Rhizopus, besides possibly pathogenic bacteria and some fungi producing toxins, such as Aspergillus flavus, Penicillium luteum, Penicillium islandicum, Penicillium citrinum and Aspergillus flavusBacterium B 1(AFB 1) Fungi such as ochratoxin A (OTA), Deoxynivalenol (DON), Patulin (PAT), etc.; mycotoxins are biological toxins produced by some fungi in the growth process, more than 200 types of currently known mycotoxins exist, and the mycotoxins have wide toxicity spectrum, such as suppression of the immune system and the hematopoietic system, neuroendocrine disorder, liver and kidney injury and the like. Sometimes its toxic effects are manifested as endemic, and more seriously, some mycotoxins have been shown to have carcinogenic, teratogenic and mutagenic effects.
In addition, the tea making in Guizhou is too single at present, and mainly comprises green tea and black tea. Therefore, the invention discloses a method for preparing dark green tea by using tea leaves uniformly produced in Guizhou province, can effectively control the growth of microorganisms in the dark green tea pile fermentation process, and has important significance for promoting diversified development of tea industry.
Disclosure of Invention
The invention aims to provide a processing method of controllable Duyun dark tea. The preparation method is stable, the quality of the homogenized black tea can be effectively improved, the aroma and the taste of the homogenized black tea are improved, the homogenized black tea prepared by the method is rich in nutrient content, stable in tea quality and good in color and luster, and does not contain T2, HT2 and 4 aflatoxins; the tea provided by the invention is uniform in black brown appearance, mellow and pure in fragrance, red brown in liquor color, mellow in taste and dark brown in leaf bottom.
The technical scheme of the invention is as follows: a processing method of controllable Duyun dark tea comprises the following steps:
(a) picking fresh leaves: picking fresh leaves of the local variety which are uniformly grown;
(b) spreading: spreading fresh leaves at room temperature of 20-30 deg.C for 1-4 hr, and dehydrating to 60-80%;
(c) de-enzyming: a roller enzyme deactivating machine, the leaf feeding amount is 2-3kg, and the enzyme deactivating is carried out at the temperature of 220 ℃ at 180-; until the green odor disappears and the tea fragrance appears, the leaf color changes from bright green to dark green, and the tea is kneaded and stuck by hands to form a ball;
(d) rolling: rolling for 20-30min while hot, and slightly pressing for 1/4 time, and pressing for 1/2 time and 1/4 time;
(e) and (3) drying: deblocking, sun drying at room temperature of 25 deg.C for 5 hr to form sun-dried flavor, kneading for 5min, and oven drying at 40 deg.C to obtain sun-dried green tea;
(f) and (3) sterilization: taking sun-dried green raw tea, adding water to 35%, sterilizing at 115 deg.C for 30min, and cooling;
(g) preparing bacteria: preparing a filamentous yeast FD spore suspension;
(h) inoculation: according to the weight ratio of sterilized tea leaves to 40 g: performing single inoculation on the FD spore suspension at a ratio of 0.1 mL;
(i) pile fermentation: performing pile fermentation on the inoculated tea leaves at the temperature of 30 ℃ and the humidity of 80% for 25 d;
(j) and (3) drying: taking out the fermented tea leaves, and drying at 45 ℃ to obtain the Duyun black tea.
In the method for processing the controllable Duyun dark tea, the fresh leaves in the step (a) are one bud, three leaves and four leaves.
In the processing method of the controllable homogenized dark tea, the spreading in the step (b) is to spread the fresh leaves indoors for 3 hours at the room temperature of 25 ℃, and the water loss is 70%.
In the method for processing the controllable Duyun dark tea, the water removing in the step (c) is carried out by using a roller water removing machine, the leaf feeding amount is 2.5kg, and the water removing is carried out at the temperature of 200 ℃.
In the processing method of the controllable homogenized black tea, the filamentous fungus FD spore suspension in the step (g) is prepared by taking the filamentous fungus FD, inoculating the filamentous fungus FD on a PDA slant culture medium, culturing at 20-40 ℃ for 3d-9d, after the strain is mature, washing off the spores by using 0.3-1.5% of normal saline, transferring the spores into a triangular conical flask filled with glass beads, fully oscillating to uniformly distribute the spores in the suspension, and filtering by absorbent cotton to obtain a single spore suspension; on a dry clean blood cell counting plate, cover the glass slide, drop 1 drop spore suspension on the edge of the glass slide, make it penetrate into the counting chamber by capillary action, when the spore sinks to the bottom of the counting plate, count and adjust the spore concentration 10 7cfu/mL。
In the method for processing the controllable Duyun dark tea, the filamentous yeast FD spore suspension in the step (g) is prepared by inoculating the filamentous yeast FD on a PDA slant culture medium, culturing at 25-31 ℃ for 4d-8d, and after the strain is mature, using 0.7-1.1% physiological saline to mix the spores with the PDA slant culture mediumWashing, transferring into triangular conical bottle filled with glass beads, oscillating sufficiently to uniformly distribute spores in the suspension, and filtering with absorbent cotton to obtain monospore suspension; on a dry clean blood cell counting plate, cover the glass slide, drop 1 drop spore suspension on the edge of the glass slide, make it penetrate into the counting chamber by capillary action, when the spore sinks to the bottom of the counting plate, count and adjust the spore concentration 10 7cfu/mL。
In the processing method of the controllable homogenized black tea, the filamentous fungus FD spore suspension in the step (g) is prepared by taking the filamentous fungus FD, inoculating the filamentous fungus FD on a PDA slant culture medium, culturing at 28 ℃ for 5d-7d, after the strain is mature, washing off the spores by using 0.9% physiological saline, transferring the spores into a triangular conical flask filled with glass beads, fully oscillating to uniformly distribute the spores in the suspension, and filtering by absorbent cotton to obtain a single spore suspension; on a dry clean blood cell counting plate, cover the glass slide, drop 1 drop spore suspension on the edge of the glass slide, make it penetrate into the counting chamber by capillary action, when the spore sinks to the bottom of the counting plate, count and adjust the spore concentration 10 7cfu/mL。
In the processing method of the controllable Duyun dark tea, the Trichosporon sp FD is obtained by taking aged Pu' er tea as a raw material, performing streak separation by utilizing gradient concentration and analyzing an ITS sequence.
Compared with the prior art, the pile-fermentation type green tea after-fermentation tea is prepared by pile-fermentation of sunned green tea made of all-uniform native varieties, and the humidifying pile fermentation is a key process for forming the quality of the black tea; the microbial fermentation plays a decisive role in the quality formation of the homogenized black tea and is a necessary condition for the quality formation of the homogenized black tea. The invention is characterized in that fresh leaves of one bud, three leaves and four leaves of native local varieties in a yun-yun area are picked to prepare the sun-dried green raw tea, the green raw tea is singly inoculated with the hyphomycete FD after being sterilized, and the yun-yun raw tea is prepared into the yun-yun black tea through pile fermentation.
To verify the effect of the present invention, the inventors conducted the following experiments:
experimental example:
firstly, preparing sun-dried green tea prepared from Duyun native varieties:
(1) fresh leaves: one bud, three leaves and four leaves; the fresh leaves are all blended into native local variety.
(2) Spreading: spreading indoors for 3h at room temperature of 25 ℃, and dehydrating to 70%;
(3) de-enzyming: a roller enzyme deactivating machine, the leaf feeding amount is 2.5kg, and the enzyme deactivating is carried out at 200 ℃; until the green odor disappears and the tea fragrance appears, the leaf color changes from bright green to dark green, and the tea is kneaded and sticky by hand to form a ball.
(4) Rolling: rolling for 25min while hot, and slightly pressing for 1/4 time, and moderately pressing for 1/2 time and 1/4 time;
(5) and (3) drying: deblocking, sun drying at room temperature of 25 deg.C for 5 hr (forming sun-drying smell), kneading for 5min, and oven drying at 40 deg.C.
Second, strain isolation and identification
2.1 Strain isolation and morphological characterization
Screening high-quality 2010 Hongrunlai Pu-Er ripe tea, and making into 10 -2-10 -7Diluting, separating and culturing the strain, purifying and preserving. According to morphological characteristics such as shape, size, color and surface characteristics of the primary colony, the obtained strains are subjected to primary morphological identification by combining Chinese journal.
2.2 molecular characterization
Firstly, extracting strain DNA by adopting an improved CTAB method; secondly, performing PCR amplification on a target fragment by using KOD high-fidelity DNA polymerase, performing 3730XL conventional sequencing reaction, and completing a full-sequence gene according to ABI standard program operation; finally, comparing in an NCBI database, determining the classification status of the strains, and constructing an evolutionary tree; the result of the ITS sequence analysis was Trichosporon sp FD.
2.3 preparation of fermentation samples
(S3) group: preparation 10 7cfu/mL spore suspension of Trichosporon sp FD; taking 40g of the prepared sun-dried green raw tea, filling the sun-dried green raw tea with the moisture content of 35% into a 500mL triangular flask, sterilizing for 30min at 115 ℃, cooling, inoculating 0.1mL of spore suspension of the hyphomycete FD, culturing for 25d in an artificial climate box at the temperature of 30 ℃ and the humidity of 80%, shaking the flask once every 5d, taking out the green raw tea after fermentation is completed, and drying at 45 ℃;
(S1) group: taking 40g of the prepared sun-dried green raw tea, and taking natural fermentation (namely the moisture content is 35%, no bacteria are added in a sterile manner, and by means of the natural bacterial strains in the sun-dried green raw tea of the uniform native variety and air, culturing for 25d at the temperature of 30 ℃ and the humidity of 80%, shaking the bottle once every 5d, taking out the bottle after the fermentation is finished, and drying at the temperature of 45 ℃) as positive control (S1);
40g of the prepared sun-dried green tea is sterilized at 115 ℃ for 30min, and no strain is added as a negative control (S2).
Three treatments, each 3 replicates, produced 9 pile ferments.
2.4 fermentation sample detection
Water extract GB/T8305-.
Third, results and analysis
3.1 sensory analysis of pile fermented samples
The three groups of pile fermentation samples S1, S2 and S3 are shown in FIG. 1, and Table 1 is a sensory analysis table of the three groups of pile fermentation samples S1, S2 and S3; table 2 shows sensory scores, physicochemical indexes and safety of the fermented products of the three fermentation groups S1, S2 and S3
Table 1 sensory tables of pile fermentation at S1, S2, S3;
Figure RE-GDA0002329898300000071
TABLE 2 sensory evaluation, physicochemical index and safety of the fermented product of S1, S2 and S3
S3 has the best treatment effect, the Pu 'er tea dominant bacteria FD is obtained through separation and purification, and then the Pu' er tea dominant bacteria FD is inoculated to the sunned green raw tea prepared from the uniform native varieties and is obtained through pile fermentation, at the moment, 51.43% of water extract, 4.21% of soluble sugar, 2.98% of tea polysaccharide, 4.01% of caffeine, 7.39% of catechin, 0.13% of theaflavin, 0.12% of thearubigin, 10.01% of theabrownin, 1.37% of flavonoids and 9.81% of tea polyphenol.
3.2 toxicological analysis of pile fermentation samples
3.2.1 Standard Curve preparation
(1) Optimization of instrumental analysis conditions
Under the optimized instrument conditions, the extracted ion chromatograms of T2, HT2 and 4 standard solutions (10 μ g/L) of aflatoxin compounds are shown in FIG. 2;
(2) linear relationship, detection limit and quantitation limit of detection methods
The instrument limit of quantitation (ILOQ) for 6 targets was 0.1-0.5 μ g/L and the limit of quantitation of the detection Method (MLOQ) was between 0.5-2.5 pg/L (see table 3) calculated as S/N-10, indicating that the method had higher sensitivity.
TABLE 3 correlation coefficients, instrument detection limits and method quantitation limits for various standard curves
Figure RE-GDA0002329898300000081
3.2.2 detection of pile-fermented samples
According to the method, 9 fermentation samples are detected, no positive sample is found, and the total ion flow chart of the 9 fermentation samples is shown in figure 3.
Fourth, discuss and conclude
4.1 safety analysis of pile-fermented samples of Duyun dark tea
The pile-fermented type homogenized dark tea is a result of microbial enzyme promotion and damp-heat action, so that substances in the tea are subjected to a series of reactions such as oxidation, degradation and condensation, and the basic characteristics of reddish brown color, brownish red soup color, mellow taste and obvious old aroma are formed. The safety of the dark tea is a concern during pile fermentation. The results of the studies by Mogensen j.m. et al were similar and mycotoxins ochratoxin a, fumonisin B2 and B4 were not detected in the yushui black tea, thus confirming the safety of the yushui black tea.
9 fermentation samples are detected by using an ultra performance liquid chromatography-mass spectrometry/mass spectrometry detection method, and no positive sample is found. The results show that: the dominant strain of the filamentous yeast, a single strain is fermented under the conditions of temperature and humidity control (30 ℃ and 80 percent), and does not produce T2, HT2 and 4 aflatoxins; and the temperature and humidity control (30 ℃, 80%) of the fermentation of the natural strain is not detected.
4.2 conclusion
The pile-fermentation type after-fermentation tea of the uniformly-mixed dark tea is prepared by pile-fermentation of the sunned green tea made of uniformly-mixed native varieties, and the humidifying pile fermentation is a key process for forming the quality of the dark tea; the microbial fermentation plays a decisive role in the quality formation of the homogenized black tea and is a necessary condition for the quality formation of the homogenized black tea. According to the research, aged Pu 'er tea is used as a raw material, dominant bacteria FD of the Pu' er tea is obtained by gradient concentration streaking separation, and the dominant bacteria FD is analyzed as filamentous yeast through an ITS sequence.
And (3) taking the uniformly-mixed native variety of the sunned raw tea as a raw material, separating to obtain FD, and performing pile fermentation by controlling temperature, humidity, moisture and fermentation time to obtain a pile fermentation sample. And detecting T2, HT2 and 4 aflatoxins in the pile fermentation sample by using an ultra performance liquid chromatography-mass spectrometry/mass spectrometry detection method. Researches show that the dark tea is uniformly fermented by pile fermentation at 30 ℃ and 80%, and the safety of the dominant strain, namely the filamentous yeast, is higher.
The S3 treatment effect is the best, and the best quality is obtained by inoculating the yun black tea dominant bacteria FD to the sunned green tea made of yun native varieties and performing pile fermentation after the yun black tea dominant bacteria FD is obtained through separation and purification.
In conclusion, the preparation method is stable, the quality of the homogenized black tea can be effectively improved, the aroma and the taste of the homogenized black tea are improved, the homogenized black tea prepared by the method is rich in nutrient content, stable in tea quality and good in color, and does not contain T2, HT2 and 4 aflatoxins; the tea provided by the invention is uniform in black brown appearance, mellow and pure in fragrance, red brown in liquor color, mellow in taste and dark brown in leaf bottom.
Drawings
FIG. 1 is a photograph of the appearance, leaf bottom and soup color of three sets of pile fermentation samples in the experimental example;
FIG. 2 is a MRM chromatogram of standard solutions of T2, HT2 and 4 aflatoxin compounds in experimental examples;
FIG. 3 is a total ion flow graph of T2, HT2 and 4 aflatoxins.
Detailed Description
The present invention is further illustrated by the following examples, which are not to be construed as limiting the invention.
Example 1. A processing method of controllable Duyun dark tea comprises the following steps:
the method comprises the following steps:
(a) picking fresh leaves: picking fresh leaves of one bud, three leaves and four leaves of a uniform native variety;
(b) spreading: spreading fresh leaves at room temperature of 25 deg.C for 3 hr, and dehydrating to 70%;
(c) de-enzyming: a roller enzyme deactivating machine, the leaf feeding amount is 2.5kg, and the enzyme deactivating is carried out at 200 ℃; until the green odor disappears and the tea fragrance appears, the leaf color changes from bright green to dark green, and the tea is kneaded and stuck by hands to form a ball;
(d) rolling: rolling for 25min, pressing lightly for 5min, pressing at middle pressure for 10min, and pressing lightly for 5 min;
(e) and (3) drying: deblocking, sun drying at room temperature of 25 deg.C for 5 hr to form sun-dried flavor, kneading for 5min, and oven drying at 40 deg.C to obtain sun-dried green tea;
(f) and (3) sterilization: taking sun-dried green raw tea, adding water to 35%, sterilizing at 115 deg.C for 30min, and cooling;
(g) preparing bacteria: taking a hyphomycete FD, inoculating the hyphomycete FD on a PDA slant culture medium, culturing at 28 ℃ for 6 days, after a strain is mature, washing off spores by using a proper amount of 0.9% physiological saline, transferring the spores into a triangular conical flask filled with glass beads, fully oscillating to uniformly distribute the spores in a suspension, and filtering by absorbent cotton to obtain a monospore suspension; on a dry clean blood cell counting plate, cover the glass slide, drop 1 drop spore suspension on the edge of the glass slide, make it penetrate into the counting chamber by capillary action, when the spore sinks to the bottom of the counting plate, count and adjust the spore concentration 10 7cfu/mL；
(h) Inoculation: according to the weight ratio of sterilized tea leaves to 40 g: performing single inoculation on the FD spore suspension at a ratio of 0.1 mL;
(i) pile fermentation: performing pile fermentation on the inoculated tea leaves at the temperature of 30 ℃ and the humidity of 80% for 25 d;
(j) and (3) drying: taking out the fermented tea leaves, and drying at 45 ℃ to obtain the Duyun black tea.
Example 2. A processing method of controllable Duyun dark tea comprises the following steps:
(a) picking fresh leaves: picking fresh leaves of one bud, three leaves and four leaves of a uniform native variety;
(b) spreading: spreading fresh leaves at room temperature of 20 deg.C for 1.5h, and dehydrating to 64%;
(c) de-enzyming: a roller enzyme deactivating machine, the leaf feeding amount is 2kg, and the enzyme deactivating is carried out at 180 ℃; until the green odor disappears and the tea fragrance appears, the leaf color changes from bright green to dark green, and the tea is kneaded and stuck by hands to form a ball;
(d) rolling: rolling for 20min while hot, and slightly pressing for 1/4 time, and moderately pressing for 1/2 time and 1/4 time;
(e) and (3) drying: deblocking, sun drying at room temperature of 25 deg.C for 5 hr to form sun-dried flavor, kneading for 5min, and oven drying at 40 deg.C to obtain sun-dried green tea;
(f) and (3) sterilization: taking sun-dried green raw tea, adding water to 35%, sterilizing at 115 deg.C for 30min, and cooling;
(g) preparing bacteria: taking a hyphomycete FD, inoculating the hyphomycete FD on a PDA slant culture medium, culturing at 28 ℃ for 5 days, after a strain is mature, washing off spores by using a proper amount of 0.9% physiological saline, transferring the spores into a triangular conical flask filled with glass beads, fully oscillating to uniformly distribute the spores in a suspension, and filtering by absorbent cotton to obtain a monospore suspension; on a dry clean blood cell counting plate, cover the glass slide, drop 1 drop spore suspension on the edge of the glass slide, make it penetrate into the counting chamber by capillary action, when the spore sinks to the bottom of the counting plate, count and adjust the spore concentration 10 7cfu/mL；
(h) Inoculation: according to the weight ratio of sterilized tea leaves to 40 g: performing single inoculation on the FD spore suspension at a ratio of 0.1 mL;
(i) pile fermentation: performing pile fermentation on the inoculated tea leaves at the temperature of 30 ℃ and the humidity of 80% for 25 d;
(j) and (3) drying: taking out the fermented tea leaves, and drying at 45 ℃ to obtain the Duyun black tea.
Example 3. A processing method of controllable Duyun dark tea comprises the following steps:
(a) picking fresh leaves: picking fresh leaves of one bud, three leaves and four leaves of a uniform native variety;
(b) spreading: spreading fresh leaves at room temperature of 30 deg.C for 4 hr, and dehydrating to 78%;
(c) de-enzyming: a roller enzyme deactivating machine, wherein the leaf feeding amount is 3kg, and the enzyme deactivating is carried out at 220 ℃; until the green odor disappears and the tea fragrance appears, the leaf color changes from bright green to dark green, and the tea is kneaded and stuck by hands to form a ball;
(d) rolling: rolling for 30min while hot, and slightly pressing for 1/4 time, and moderately pressing for 1/2 time and 1/4 time;
(e) and (3) drying: deblocking, sun drying at room temperature of 25 deg.C for 5 hr to form sun-dried flavor, kneading for 5min, and oven drying at 40 deg.C to obtain sun-dried green tea;
(f) and (3) sterilization: taking sun-dried green raw tea, adding water to 35%, sterilizing at 115 deg.C for 30min, and cooling;
(g) preparing bacteria: taking a hyphomycete FD, inoculating the hyphomycete FD on a PDA slant culture medium, culturing at 28 ℃ for 7d, after a strain is mature, washing off spores by using a proper amount of 0.9% physiological saline, transferring the spores into a triangular conical flask filled with glass beads, fully oscillating to uniformly distribute the spores in a suspension, and filtering by absorbent cotton to obtain a monospore suspension; on a dry clean blood cell counting plate, cover the glass slide, drop 1 drop spore suspension on the edge of the glass slide, make it penetrate into the counting chamber by capillary action, when the spore sinks to the bottom of the counting plate, count and adjust the spore concentration 10 7cfu/mL；
(h) Inoculation: according to the weight ratio of sterilized tea leaves to 40 g: performing single inoculation on the FD spore suspension at a ratio of 0.1 mL;
(i) pile fermentation: performing pile fermentation on the inoculated tea leaves at the temperature of 30 ℃ and the humidity of 80% for 25 d;
(j) and (3) drying: taking out the fermented tea leaves, and drying at 45 ℃ to obtain the Duyun black tea.

Claims (8)

1. A processing method of controllable Duyun dark tea is characterized in that: the method comprises the following steps:
(a) picking fresh leaves: picking fresh leaves of the local variety which are uniformly grown;
(b) spreading: spreading fresh leaves at room temperature of 20-30 deg.C for 1-4 hr, and dehydrating to 60-80%;
(c) de-enzyming: a roller enzyme deactivating machine, the leaf feeding amount is 2-3kg, and the enzyme deactivating is carried out at the temperature of 220 ℃ at 180-; until the green odor disappears and the tea fragrance appears, the leaf color changes from bright green to dark green, and the tea is kneaded and stuck by hands to form a ball;
(d) rolling: rolling for 20-30min while hot, and slightly pressing for 1/4 time, and pressing for 1/2 time and 1/4 time;
(e) and (3) drying: deblocking, sun drying at room temperature of 25 deg.C for 5 hr to form sun-dried flavor, kneading for 5min, and oven drying at 40 deg.C to obtain sun-dried green tea;
(f) and (3) sterilization: taking sun-dried green raw tea, adding water to 35%, sterilizing at 115 deg.C for 30min, and cooling;
(g) preparing bacteria: preparing a filamentous yeast FD spore suspension;
(h) inoculation: according to the weight ratio of sterilized tea leaves to 40 g: performing single inoculation on the FD spore suspension at a ratio of 0.1 mL;
(i) pile fermentation: performing pile fermentation on the inoculated tea leaves at the temperature of 30 ℃ and the humidity of 80% for 25 d;
(j) and (3) drying: taking out the fermented tea leaves, and drying at 45 ℃ to obtain the Duyun black tea.
2. The method for processing the controllable Duyun dark tea as claimed in claim 1, wherein the method comprises the following steps: the fresh leaves in the step (a) are one bud, three leaves and four leaves.
3. The method for processing the controllable Duyun dark tea as claimed in claim 1, wherein the method comprises the following steps: the spreading in the step (b) is to spread the fresh leaves indoors for 3 hours at the room temperature of 25 ℃, and the water loss is 70%.
4. The method for processing the controllable Duyun dark tea as claimed in claim 1, wherein the method comprises the following steps: the water removing in the step (c) is carried out by using a roller water removing machine, the leaf feeding amount is 2.5kg, and the water removing is carried out at the temperature of 200 ℃.
5. The method for processing the controllable Duyun dark tea as claimed in claim 1, wherein the method comprises the following steps: the filamentous spore yeast FD spore suspension in the step (g) is prepared by taking the filamentous yeast FD, inoculating the filamentous yeast FD on a PDA slant culture medium, culturing at 20-40 ℃ for 3d-9d, washing off spores by using 0.3-1.5% of normal saline after strains are mature, transferring the spores into a triangular conical flask filled with glass beads, fully oscillating to uniformly distribute the spores in the suspension, and filtering by absorbent cotton to obtain a monospore suspension; on a dry clean blood cell counting plate, cover the glass slide, drop 1 drop spore suspension on the edge of the glass slide, make it penetrate into the counting chamber by capillary action, when the spore sinks to the bottom of the counting plate, count and adjust the spore concentration 10 7cfu/mL。
6. The method for processing the controlled homogenized dark tea as claimed in claim 5, wherein: the filamentous spore yeast FD spore suspension in the step (g) is prepared by taking the filamentous yeast FD, inoculating the filamentous yeast FD on a PDA slant culture medium, culturing for 4d-8d at the temperature of 25-31 ℃, washing down spores by using 0.7-1.1% of normal saline after strains are mature, transferring the spores into a triangular conical flask filled with glass beads, fully oscillating to uniformly distribute the spores in the suspension, and filtering by absorbent cotton to obtain a monospore suspension; on a dry clean blood cell counting plate, cover the glass slide, drop 1 drop spore suspension on the edge of the glass slide, make it penetrate into the counting chamber by capillary action, when the spore sinks to the bottom of the counting plate, count and adjust the spore concentration 10 7cfu/mL。
7. The method for processing the controllable Duyun dark tea as claimed in claim 6, wherein the method comprises the following steps: the filamentous spore yeast FD spore suspension in the step (g) is prepared by taking the filamentous yeast FD, inoculating the filamentous yeast FD on a PDA slant culture medium, culturing for 5d-7d at 28 ℃, washing off spores by using 0.9% of normal saline after strains are mature, transferring the spores into a triangular conical flask filled with glass beads, fully oscillating to uniformly distribute the spores in the suspension, and filtering by absorbent cotton to obtain a monospore suspension; on a dry clean blood cell counting plate, cover the glass slide, drop 1 drop spore suspension on the edge of the glass slide, make it penetrate into the counting chamber by capillary action, wait for the spore to sink to the bottom of the counting plateSection, count and adjust spore concentration 10 7cfu/mL。
8. A process for the controlled processing of a yun cha as claimed in any one of claims 5 to 7 wherein: the filamentous spore yeast FD is prepared from aged Pu' er tea by gradient concentration streaking separation and ITS sequence analysis.
CN201910372722.2A 2019-05-06 2019-05-06 Processing method of controllable Duyun dark tea Pending CN110771698A (en)

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CN112314726A (en) * 2020-11-20 2021-02-05 安化县亦神芙蓉茶业有限公司 Golden flower raw leaf black tea and processing technology thereof
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Application publication date: 20200211