CN112825935A - Method for preparing black tea with high gallic acid content by using monascus purpureus - Google Patents
Method for preparing black tea with high gallic acid content by using monascus purpureus Download PDFInfo
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Abstract
The invention discloses a method for preparing black tea with high gallic acid content by using monascus purpureus, belonging to the technical field of tea processing. The black tea with low ester type catechin content and high gallic acid content is prepared by inoculating monascus to raw tea raw materials. The feasibility of inoculating the tea by the monascus is proved, the application range of the monascus is widened, and a foundation is laid for further and deeply researching the monascus to inoculate the tea in the follow-up process; the content of ester catechin in the prepared black tea is reduced, the content of gallic acid is greatly improved, the fact that the Monascus inoculation can reduce the bitter and astringent feeling of tea leaves and add flavor is shown, and the black tea obtained by fermentation has a certain health-care function and can be further developed and utilized as a health-care beverage in industry.
Description
Technical Field
The invention belongs to the technical field of tea processing, and particularly relates to a method for preparing black tea with high gallic acid content by using monascus purpureus.
Background
The black tea is one of six tea types in China and belongs to post-fermented tea. The Fuzhuan tea is an important variety in black tea, has the characteristics of black brown and oily color, luxuriant golden flower, lasting fragrance, obvious aging fragrance, clear red and thick, mellow, sweet and soft and smooth, and is widely popular with consumers.
The process of 'flowering' is a key process for forming the special quality of the Fuzhuan tea. Wherein the eurotium cristatum plays a main role as a dominant strain. The essence of the 'flowering' is that the raw dark green tea pressed into green bricks is put into a drying room, and proper conditions are created to enable eurotium cristatum to grow. The eurotium cristatum is also called as eurotium cristatum, and the surface of the finished Fuzhuan tea and the interior of a tea brick are provided with a plurality of visible golden particles, namely capsule closure shells generated by the eurotium cristatum, which are commonly called as golden flowers. Researches prove that the soup color, mellow taste and special fungus flower fragrance of the Fuzhuan tea are mainly formed under the action of eurotium cristatum, the more the content of the 'golden flower' of the produced closed capsule shell is, the better the conversion of the content of the substances in the tea is, the higher the quality of the prepared Fuzhuan tea is, and the more the number of the 'golden flowers' is also used as the characteristic index of the quality of the Fuzhuan tea.
Monascus has application history of more than one thousand years in China, and is one of the most beneficial fungi applied to food in China. At present, the application of monascus is mainly focused on the fields of coloring, corrosion prevention, wine and vinegar brewing, biological catalysis, traditional Chinese medicine and the like. The monascus secondary metabolites have high utilization value, and include monascin compounds, which can significantly regulate blood lipid and human body immunity, and have the effects of lowering blood pressure and inhibiting cholesterol. The application of monascus is less at present as the main producing country of monascus and the country applying monascus at the earliest in China, mainly the export of monascus pigment, and the application of monascus in other fields has huge development potential and space.
At present, the tea production technology mainly adopts eurotium cristatum, and no report related to tea inoculation by monascus exists, and whether monascus can be used for producing black tea or not and what characteristics of inoculated tea are unknown.
Disclosure of Invention
The invention aims to provide a method for preparing black tea with high gallic acid content by utilizing monascus so as to solve the problems in the prior art, provide a method for inoculating monascus to raw tea raw materials, widen the application range of monascus and provide a new idea for a production process of black tea.
In order to achieve the purpose, the invention provides the following scheme:
the invention provides a preparation method of dark green tea with high gallic acid content, which is characterized in that monascus is used for inoculating a raw tea raw material, and the dark green tea with high gallic acid content is obtained through solid state fermentation.
Preferably, the monascus is FM23 red yeast rice strain.
Preferably, the raw tea raw material is green raw tea.
Preferably, the monascus inoculation comprises the following specific steps: controlling the moisture content of the raw tea material to be 35%, and performing steam sterilization for 30 minutes at the temperature of 115 ℃; adding Monascus liquid, and culturing at constant temperature of 32 deg.C.
Preferably, the concentration of the monascus liquid is 104Per ml-1032One per ml.
Preferably, the concentration of the monascus liquid is 1016One per ml.
Preferably, the volume-mass ratio of the monascus purpureus bacterial liquid to the raw tea material is 1ml to 40 g.
The invention also provides the tea prepared by the preparation method.
The invention discloses the following technical effects:
the invention proves the feasibility of inoculating the tea by the monascus, widens the application range of the monascus, and lays a foundation for further and deeply researching the tea inoculated by the monascus in the follow-up process.
In the black tea prepared by the method for inoculating the monascus to the raw tea, the content of ester catechin is reduced, and the monascus inoculation can reduce the bitter and astringent feeling of tea and add flavor; the content of gallic acid with the effects of sterilization, inflammation diminishing and the like is greatly improved, and the results prove that the black tea obtained by inoculating and fermenting monascus has a certain health-care function and can be further developed and utilized in industry as a health-care beverage.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.
FIG. 1 shows the results of five samplings after the tea leaves were inoculated with Monascus purpureus strain in example 2; samples at 0d, 5d, 15d, 20d and 25d from left to right in the figure;
FIG. 2 is a bar graph of catechin content in tea leaves at different inoculation times; five columns corresponding to GA in the figure are CK, 5d, 10d, 15d and 20d from left to right respectively; five columns corresponding to the GC are CK, 5d, 10d, 15d and 20d from left to right respectively; five columns corresponding to the EGC are CK, 5d, 10d, 15d and 20d from left to right respectively; the content of C is 0; when four columns corresponding to the EGCG are CK, 5d, 10d and 15d from left to right respectively, and the detection content is 0 when 20 d; five columns corresponding to EC are CK, 5d, 10d, 15d and 20d from left to right respectively; when four columns corresponding to GCG are CK, 5d, 10d, 15d from left to right and 20d respectively, the detection content is 0; when the three columns corresponding to the ECG are CK, 5d and 15d from left to right and 10d and 20d respectively, the detection content is 0; a column corresponding to CG is CK, and the detection content of the rest samples is 0.
Detailed Description
Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Further, for numerical ranges in this disclosure, it is understood that each intervening value, between the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The specification and examples are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.
The materials and reagents used in the present invention are commercially available, unless otherwise specified.
Example 1
Determination of method for inoculating monascus to green tea
1. Materials and methods
1.1 Experimental materials
Green tea is purchased from Guangxi Zhaoping county, and FM23 red yeast rice strain is purchased from the Chinese bacterial center.
1.2 Experimental reagents
Trifluoroacetic acid (analytically pure), methanol (chromatographically pure), and catechin monomers (chromatographically pure), wherein the catechin monomers include Gallocatechin (GC), Epigallocatechin (EGC), catechin (C), epigallocatechin gallate (EGCG), Epicatechin (EC), gallocatechin gallate (GCG), epicatechin gallate (ECG), and Catechin Gallate (CG).
1.3 Experimental instruments
AB204-N type electronic balance (Shanghai Mei Teller-Tolliduo instruments Co., Ltd.), vertical pressure steam sterilizer (Shanghai Boxun industries Co., Ltd.), SW-CJ-2F type double-side purification work bench (Suzhou purification equipments Co., Ltd.), DHG-101 oven (New Yogyang oven manufacturing plant of Wujiang, Jiangsu), GXZ-250A constant temperature incubator (Ningbo science park New Yongnan instruments Co., Ltd.), DK-8D electric heating constant temperature water bath (Yongming medical instruments Co., Ltd., Beijing) and high performance liquid chromatograph (Agilent 1200).
2. Experimental methods
2.1 Strain culture
Storing the FM23 red yeast rice strain test tube in an environment of 4 ℃, taking out the test tube in advance when in use, and placing the test tube in a constant temperature incubator (the temperature is set to be 32 ℃) for 24 hours for activation. The culture medium is Potato glucose agar (PDA) culture medium, after the strain is activated, the hyphae are picked up by using inoculating loop, streaking culture is carried out on the culture medium, and then the culture medium is continuously placed in a constant temperature incubator at 32 ℃ to be cultured until monascus colonies are obtained.
2.2 spore suspension preparation
After a certain number of monascus spores grow on the surface of the culture medium, preparing a conical flask with glass beads filled with 50mL of sterile water, scraping a proper amount of spores by a blade, fully oscillating in the flask, measuring the concentration of the bacterial liquid by using a blood counting chamber after oscillation is finished, properly adjusting, and recording data. In addition, 4 flasks containing 9mL of sterile water were prepared, and 1mL of the suspension was sequentially transferred to the flasks, and five concentration gradients were set.
2.3 determination of optimal inoculation conditions
Weighing 20g of green tea sample in an erlenmeyer flask, weighing 6 parts in total, wherein one part is used as a control. Controlling the moisture content to be 35%, sealing, and steam sterilizing in an autoclave at 115 deg.C for 30 min. 0.5mL of the bacterial solution was taken from each of the tea samples corresponding to each concentration, and 0.5mL of sterile water was taken from the control group. The tea leaves are sealed and put into a constant temperature incubator at 32 ℃, and the inoculation condition is observed every day. Four groups of gradients are made in the experiment, and the original bacteria liquid concentration is as follows in sequence: 108one/mL, 1016one/mL, 1024one/mL, 1032one/mL.
3. Results of the experiment
First group (original bacteria liquid concentration 10)8pieces/mL): the growth gradient is positively correlated with the concentration of the bacteria liquid, and the highest concentration sample generates less bacteria in 7 daysThe hyphae are more dense at 9 days, and spores grow only at 14 days, so that the overall growth speed is slow and the optimal concentration does not appear.
Second group (original bacteria liquid concentration 10)16pieces/mL): the growth gradient is in positive correlation with the concentration of the bacterial liquid, a small amount of hyphae grow at the highest concentration sample within 3 days, the hyphae are dense within 5 days, and spores can grow at 7 days. The overall growth rate is obviously improved, but the optimal concentration still does not appear.
Third and fourth groups (original bacterial liquid concentration 10)24one/mL, 1032pieces/mL): the growth rate was slightly increased compared to the second group, but the overall difference was not large.
The experimental result shows that the tea inoculated with the monascus can produce hypha to form spores, and the feasibility of inoculating the tea with the monascus is proved. In addition, in a certain concentration range, the growth rate of the monascus inoculated with the tea leaves is positively correlated with the concentration of the bacterial liquid, and the correlation tends to be slow or even unobvious after the monascus is inoculated with the tea leaves. Comprehensive production cost consideration, choose 1016The concentration per mL is the optimal concentration for inoculating the green tea by the monascus.
Example 2
Influence of Monascus inoculation on catechin content in tea
1. Experimental methods
1.1 sample treatment
The amount of the tea leaf sample inoculated with monascus was increased to 250g, and the bacterial solution was inoculated in the ratio according to the method of example 1 (concentration of the bacterial solution used: 2.98X 10)16one/mL), and the original tea sample without any treatment was used as a control sample and incubated at a constant temperature of 32 ℃. Sampling is carried out once at intervals of 5d according to the growth condition of thalli until spores grow on the tea sample. And taking out the sample every time, drying the sample to be dry enough by using an oven, grinding the sample into tea powder, and storing the tea powder in a dryer. All samples were assayed for catechin composition and 3 replicates were set.
1.2 measurement of Catechin content
High Performance Liquid Chromatography (HPLC) assay. The chromatographic column is Agilent Technologies C18(4.6mm × 250mm, 5 μm), and the catechin detection conditions are as follows: the mobile phase A is 0.5 per mill trifluoroacetic acid aqueous solution,b is methanol, gradient elution (change in mobile phase B): 8% for 0-8 min; 8-13% for 8-20 min; 20-25 min, 13-20%; 20-25% in 25-30 min; 25-30% in 30-35 min; 35-40 min, 30-35%; keeping for 35% for 40-45 min; 45-50 min, 35-8%. Column temperature 38 deg.C, flow rate 0.8 mL/min-1The detection wavelength is 278nm, and the sample injection amount is 10 mu L.
2. Results of the experiment
The results of five sampling show that the monascus still grows well after the amount of tea leaves required to be inoculated is enlarged. As can be seen in figure 1, after the monascus is inoculated to the tea, the growth rate of hyphae in the early stage (0-10 d) is relatively slow, the hyphae grow rapidly in the middle stage (10-15d), the hyphae are densely distributed on the surface of a tea sample, the spore production is relatively rapid in the later stage (15-20d), and the inoculation can be completed within 25 days under the experimental concentration. However, the sample was contaminated with other bacterial species at the time of sampling at 25d, and the number of spores was large at 20d, and only a small amount of hyphae was present, so that the 25d sampling result was not included in the data in the measurement of the content of catechin components below.
The catechin content in the tea leaves at different inoculation times is shown in fig. 2, and as can be seen from fig. 2, the total amount of catechin is in a decreasing trend on the whole, the content of ester-type catechin EGCG and ECG is significantly reduced, and after a large amount of spores are generated, the content is kept at a lower level, and the monascus inoculation is judged to have a certain effect on reducing the bitter taste of the tea leaves. Different from other monomers, the GA content is obviously increased after inoculation, so that the health-care function of the tea can be improved after the monascus is inoculated.
The above-described embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solutions of the present invention can be made by those skilled in the art without departing from the spirit of the present invention, and the technical solutions of the present invention are within the scope of the present invention defined by the claims.
Claims (8)
1. A preparation method of dark tea with high gallic acid content is characterized in that monascus is used for inoculating a raw tea raw material, and dark tea with high gallic acid content is obtained through solid state fermentation.
2. The method for preparing the dark tea with high gallic acid content according to claim 1, wherein the monascus is FM23 red yeast rice strain.
3. The method for preparing the dark tea with high gallic acid content according to claim 1, wherein the raw tea material is green raw tea.
4. The preparation method of the dark tea with high gallic acid content according to claim 1, wherein the specific steps of monascus inoculation comprise: controlling the moisture content of the raw tea material to be 35%, and performing steam sterilization for 30 minutes at the temperature of 115 ℃; adding Monascus liquid, and culturing at constant temperature of 32 deg.C.
5. The method for preparing dark green tea with high gallic acid content according to claim 4, wherein the concentration of the monascus purpureus bacterial liquid is 104Per ml-1032One per ml.
6. The method for preparing dark green tea with high gallic acid content according to claim 4, wherein the concentration of the monascus purpureus bacterial liquid is 1016One per ml.
7. The preparation method of dark green tea with high gallic acid content according to claim 4, wherein the volume to mass ratio of the monascus purpureus bacterial liquid to the raw tea material is 1 ml: 40 g.
8. A dark tea with high gallic acid content prepared by the method of any one of claims 1-7.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107980945A (en) * | 2016-10-26 | 2018-05-04 | 成都体九生物科技有限公司 | A kind of processing method of black tea |
| CN108112715A (en) * | 2016-11-28 | 2018-06-05 | 勐海茶业有限责任公司 | The method for preparing fermented tea, the fermented tea prepared with this method and its application |
| CN109055235A (en) * | 2018-08-20 | 2018-12-21 | 云南农业大学 | A kind of preparation of the inoculating microbes strain for Pu'er tea pile fermentation and its application method |
| CN110393222A (en) * | 2019-08-15 | 2019-11-01 | 湖南省湘茶高科技有限公司 | A method of golden flower dark green tea is produced using Dark Green Tea and green gross tea as raw material |
| CN111011551A (en) * | 2020-01-08 | 2020-04-17 | 云南农业大学 | Pu' er tea cooked tea and preparation method thereof |
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- 2021-02-08 CN CN202110170853.XA patent/CN112825935A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107980945A (en) * | 2016-10-26 | 2018-05-04 | 成都体九生物科技有限公司 | A kind of processing method of black tea |
| CN108112715A (en) * | 2016-11-28 | 2018-06-05 | 勐海茶业有限责任公司 | The method for preparing fermented tea, the fermented tea prepared with this method and its application |
| CN109055235A (en) * | 2018-08-20 | 2018-12-21 | 云南农业大学 | A kind of preparation of the inoculating microbes strain for Pu'er tea pile fermentation and its application method |
| CN110393222A (en) * | 2019-08-15 | 2019-11-01 | 湖南省湘茶高科技有限公司 | A method of golden flower dark green tea is produced using Dark Green Tea and green gross tea as raw material |
| CN111011551A (en) * | 2020-01-08 | 2020-04-17 | 云南农业大学 | Pu' er tea cooked tea and preparation method thereof |
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