CN106754418A - A kind of method of Rhizopus oryzae and its fermentation fermented Chinese gall herb and tea leaves - Google Patents

Abstract

The invention relates to Rhizopus oryzae and a method for fermenting Baiyao decoction, which can effectively solve the problem of difficult measurement of fermentation process parameters and difficult process control in solid fermentation, resulting in poor product stability, low repeatability, and unstable quality of Baiyao decoction. problem, the technical solution for its solution is to mix the Galla gall powder with the distiller's yeast containing pure BYJ.G-1 bacteria according to the mass ratio of 10:0.4-4, add tea residue and mix with 20-30% tea juice to prepare Soft material; or the spore suspension of BYJ.G-1 bacteria with a concentration of 5% ^-8 % of gallnut powder weight 1×108/mL and gallnut powder, tea residue, 20-30% tea The juice is mixed evenly, and fermented for 60h-66h under the condition of relative humidity of 85%. The present invention utilizes distiller's yeast and spore suspension containing this pure Rhizopus oryzae CCTCC M2016583 to ferment Baiyao decoction. The quality is stable, the repeatability is high, and the tanning process is effectively improved. The efficiency of quality conversion into gallic acid provides technical support for the quality control and standardized production of Baiyaojian.

Description

A kind of rhizopus oryzae and the method for fermenting and decocting a hundred herbs

technical field

The invention relates to the field of microbial fermentation, in particular to a rhizopus strain and its application in the fermentation of the traditional Chinese medicine Galla chinensis to prepare Baiyao decoction.

Background technique

Baiyaojian is a fermented product of traditional Chinese medicine, which was first recorded in "Danxi Xinfa". Nature and flavor are sweet and sour, flat. It has the effects of moistening the lungs and resolving phlegm, promoting body fluid and quenching thirst. Clinically, it is mostly used to treat chronic cough with excessive phlegm, sore throat, blood in the stool, chronic dysentery and prolapse of the anus, aphthous ulcer, dental malnutrition, carbuncle and sore. The main active ingredient of gallnut is tannin, which is easy to combine with the protein in the gastrointestinal tract to form a precipitate, which stimulates the gastrointestinal mucosa and causes adverse reactions such as loss of appetite in a few people. After fermented into Baiyao decoction, the tannin content is reduced, and its astringent effect is enhanced, which may reduce the irritation to the gastrointestinal mucosa, reduce and eliminate adverse reactions such as loss of appetite. At present, Anqi brewing koji is mostly used in production. It is believed that Aspergillus niger is the key strain in the fermentation process of Baiyaojian. The solid fermentation of Gallic gall raw material uses Aspergillus niger strains which are more suitable for the production of tannin enzymes, and solid fermentation has the difficulty of measuring the parameters of the fermentation process. Difficult process control, insufficient bacterial growth, fermentation, and uneven distribution of metabolites in the fermentation system lead to poor product stability and low repeatability, which brings inconvenience to the fermentation process of Baiyaojian and makes the quality of Baiyaojian poor. Stability issue. Therefore, it is an urgent technical problem to study a better fermentation method of Baiyao decoction to control the quality stability of Baiyao decoction, improve the efficiency of converting tannin into gallic acid, and ensure its safe clinical use.

Contents of the invention

In view of the above situation, in order to solve the defects of the prior art, the purpose of the present invention is to provide a kind of rhizopus oryzae and a method for fermenting herbal decoction, which can effectively solve the problem that solid fermentation has fermentation process parameters that are difficult to measure, process control difficulties, and thalline Insufficient growth, fermentation, and uneven distribution of metabolites in the fermentation system lead to poor product stability and low repeatability, which brings inconvenience to the fermentation process of Baiyaojian and unstable quality of Baiyaojian.

The technical scheme that the present invention solves is, its classification name of rhizopus strain described in the present invention is Rhizopus sp. (Rhizopus sp.) BYJ.G-1, has been preserved in China Type Culture Collection Center on October 20, 2016 , the preservation number is: CCTCC NO: M2016583, survived after testing, and the preservation address is: Wuhan University Preservation Center, Wuhan University Campus, No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province.

The preparation method is as follows: use distiller's yeast containing Rhizopus oryzae as raw material, apply dilution coating plate method on potato dextrose agar medium, cultivate at 30°C constant temperature, isolate strains with typical characteristics of Rhizopus, and carry out isolation and cultivation , screened out a rhizopus with the highest gallic acid content in Baiyaojian, named BYJ.G-1, and was identified as Rhizopus oryzae; the potato dextrose agar medium was composed of: potato 200g, glucose 20g, Made from 15-20g of agar and 1000ml of tap water.

The method of using Rhizopus oryzae to ferment Baiyao decoction is:

Wash the medicinal material Galla chinensis, dry at 60°C for 10 hours, crush, pass through an 80-mesh sieve, and set aside; take 1-3g of tea leaves for every 10g of gallnut powder, grind the tea leaves into coarse powder, add 8-10 times the amount of distilled water to decoct 10- 15min, put it at 23-27°C, filter, separate the tea residue and tea juice, get 10mL of tea juice per 1g of tea leaves, mix Galla gall powder and distiller's yeast containing pure BYJ.G-1 bacteria according to the mass ratio of 10:0.4-4 Mix evenly, add tea leaves and 20-30% tea juice and mix evenly to make soft material; or make BYJ.G-1 bacteria with a concentration of 5%-8% relative to the weight of Galla gall powder at a concentration of ¹ ×108/mL The spore suspension is mixed with gallnut powder, tea leaves, and 20-30% tea juice to make soft material; the volume water content of the soft material is controlled at 40-60%, at 28-34 ° C, and the relative humidity is 85%. Ferment for 60h-66h, take it out, bake at 60°C for 10h until the volume water content is 10-12%.

The invention provides a strain of Rhizopus oryzae CCTCC M 2016583 and its application in the fermentation of Baiyao decoction, using the distiller's yeast and spore suspension containing the pure Rhizopus oryzae CCTCC M2016583 to ferment Baiyao decoction, with stable quality and high repeatability , and effectively improve the efficiency of converting tannin into gallic acid, providing technical support for the quality control and standardized production of Baiyaojian.

Description of drawings

Fig. 1 is the front and back observation photos of Rhizopus CCTCC M 2016583 plate colony of the present invention.

Fig. 2 is the photomicrograph (10*40) of Rhizopus CCTCC M 2016583 of the present invention.

Fig. 3 is a response surface diagram and a contour diagram of the interaction between fermentation time and fermentation temperature in the present invention.

Fig. 4 is a response surface diagram and a contour diagram of the interaction between the fermentation time of the present invention and the amount of distiller's yeast added.

Fig. 5 is a response surface diagram and a contour diagram of the interaction between the fermentation temperature and the amount of distiller's yeast added in the present invention.

detailed description

The specific implementation of the present invention will be described in further detail below in conjunction with the examples.

Example 1

Isolation of Rhizopus CCTCC M 2016583:

Using koji for Baiyaojian fermentation as raw material, take 10g of koji in 90mL sterile water, shake fully at 180r/min for 30min at 23-27°C, absorb 1mL and add it to 9mL sterile water, and so on, gradually dilute to 10 ^-2 , 10 ^-3 , 10 ^-4 ... 10 ^-9 , respectively take 200 μL of the bacterial suspension and spread it on a PDA (potato dextrose agar medium) plate, place it in a constant temperature incubator at 30°C, and isolate the rhizopus typical The characteristic bacterial strain is isolated and cultured, cultivated for 2-3 days, and the white hyphae cover the entire surface of the medium, white, dense, in the form of cotton wool, and grow many black spores, which is Rhizopus oryzae of the present invention.

Example 2

The method of using Rhizopus oryzae CCTCC M 2016583 to ferment Baiyaojian: activate the preserved strain BYJ.G-1 on PDA (potato dextrose agar medium) slant medium for 4 days, take 50mL distilled water and sterile glass balls in 200mL Sterilize in a Erlenmeyer flask at 121°C for 20 minutes, let it cool, scrape 2-4 cyclospores into sterilized distilled water, shake and mix well, and make a spore suspension of 1× ¹⁰⁸ /mL; Wash, dry at 60°C for 10 hours, pulverize, pass through a 80-mesh sieve, and set aside; take 1g of tea leaves for every 10g gallnut powder, grind the tea leaves into coarse powder, add 10 times the amount of distilled water to decoct for 15 minutes, place at 25°C, filter, and put the tea leaves The juice and the tea leaves are separated, and 10 mL of tea juice is obtained for every 1 g of tea leaves. Gallnut powder is mixed with tea leaves, 25% tea juice, and 5% BYJ. ℃, fermentation time 66h under the condition of relative humidity 85%, take it out, and dry at 60 ℃, after testing, gallic acid content reaches 33.32%; The described potato dextrose agar medium is made up of: potato 200g, glucose 20g, agar 15- 20g and 1000ml of tap water.

Example 3

The method of fermenting Baiyao decoction with distiller's yeast of pure rhizopus oryzae CCTCC M 2016583:

Wash the Galla gall medicinal material, dry at 60°C for 10 hours, pulverize, pass through an 80-mesh sieve, and set aside; take 1g of tea leaves and 1.2g of distiller’s yeast containing pure BYJ. Decoct 10 times the amount of distilled water for 15 minutes, place at 23°C, filter, separate the tea juice and tea residues, mix gallnut powder with koji containing BYJ.G-1 bacteria, add tea residues, 30% tea juice, and mix to make soft The materials were fermented for 66 hours under the conditions of 31.8°C and 85% relative humidity, taken out, and dried at 60°C. After testing, the content of gallic acid reached 34.22%.

The present invention classifies and identifies Rhizopus, and studies the process optimization of distiller's koji fermentation Baiyao decoction using pure Rhizopus oryzae CCTCC M 2016583, and relevant experimental data are as follows:

1. Identification of Rhizopus CCTCC M 2016583

1 Traditional colony morphology observation Use an inoculation needle to take a small amount of spores, inoculate them on PDA medium, and then culture them in a constant temperature incubator at 30°C. Results After the strains were cultured on the PDA plate at 30°C for 48 hours, white transparent hyphae grew on the surface of the medium. After 60 hours, the white mycelium covered the entire surface of the medium, white, dense, cotton-like, and many black spores ( As shown in Figure 1).

2. Observation of microstructure. The fungus to be identified was inoculated at the junction of the cover glass and the culture medium by insert culture method, so that the hyphae could attach to the cover glass during the growth process, and cultured in a constant temperature incubator at 30°C. The coverslips were taken out with sterile tweezers every day from the 1st day, stained with lactic acid-Medan stain, and placed under a microscope to observe the fungal structure. Under the microscope, the hyphae have no septum, have branches and rhizoids, and have sporangia at the top. The sporangia are approximately spherical, and the spores are spherical or approximately spherical (as shown in Figure 2).

3 Modern Molecular Biological Identification The cryopreserved Rhizopus CCTCC M 2016583 was activated on PDA medium and cultured at 30°C for 2-3 days. After contamination, the DNA was extracted according to the instructions of the Sangon kit, and the quality of the DNA extraction was detected by 2% agarose gel electrophoresis. The PCR amplification primers used general primers ITS1 (TCCG TAGG TGAA CCTG CGG) and ITS4 (TCCT CCGCTTAT TGAT ATGC). Amplify the 18SrRNA gene fragment in the fungal genomic DNA. The qualified PCR products were sequenced. The determination result of the 18S rRNA gene sequence of Rhizopus CCTCC M 2016583 is:

TCTAGTATAAATAACTTTATATTGTGAAACTGCGAATGGCTCATTAAATCAGTTATGATCTACGTGACAAATTCTTTACTACTTGGATAACCGTGGTAATTCTAGAGCTAATACATGCAAAAAAGCCCTGACTTACGAAGGGGTGCACTTATTAGATAAAACCAACGCGGGGTAAAACCTGTTTCTTGGTGAATCATAATAATTAAGCGGATCGCATGGCCTTGTGCCGGCGACGGTCCACTCGATTTTCTGCCCTATCATGGTTGAGATTGTAAGATAGAGGCTTACAATGCCTACAACGGGTAACGGGGAATTAGGGTTCGATTCCGGAGAGGGAGCCTGAGAAACGGCTACCACATCCAAGGAAGGCAGCAGGCGCGCAAATTACCCAATCCCGACACGGGGAGGTAGTGACAATACATAACAATGCAGGGCCTTTAAGGTCTTGCAATTGGAATGAGTACAATTTAAATCCCTTAACGAGGATCAATTGGAGGGCAAGTCTGGTGCCAGCAGCCGCGGTAATTCCAGCTCCAATAGCGTATATTAAAGTTGTTGCAGTTAAAACGTCCGTAGTCAAACTTTAGTCTTACCGGCGTAGTGGCCTGGTCTTCATTGACCAAGCTCATTGCTGCCGGAGACTCCACGTCCATTGACTCCTAGTCCTCGTGGCTAGGGTTTTCTGGACAATTACCATGAGCAAATCAGAGTGTTTAAAGCAGGCTTTTAAGCTTGAATGTGTTAGCATGGAATAATGAAATATGACTTTAGTCCTATTTTCGTTGGTTTAGGTACTTCAGTAATGATGAATAGAAACGGTTAGGGGCATTTGTATTTGGTCGCTAGAGGTGAAATTCTTGGATTGACCGAAGACAAACTACTGCGAAAGCATTTGACCCGGGACGTTTTCATTGATCAAGGTCTAAAGTTAAGGGATCGAAGACGATTAGATACCGTCGTAGTCTTTAACCACAAACTATGCCGACTAGAGATTGGGCGT GTTTATTATGACTCGCTCAGCATCTTAGCGAAAGTAAAGTTTTTGGGTTCTGGGGGGAGTATGGGACGCAAGGCTGAAACTTAAAGGAATTGACGGAAGGGCACCACCAGGAGTGGAGCCTGCGGCTTAATTTGACTCAACACGGGGAAACTCACCAGGTCCAGACATAGTAAGGATTGACAGATTGAAAGCTCTTTCTAGATTCTATGGGTGGTGGTGCATGGCCGTTCTTAGTTCGTGGAGTGATTTGTCTGGTTAATTCCGATAACGAACGAGACCTTATTCTGCTAATTAGACAGGCTAACTCTTTCGGGTTGGTTTATATTTAATATTTAACTGGCTTCTTAGAGAGACTATCGGCTTCAAGCCGAAGGAAGTTTTAGGCAATAACAGGTCTGTGATGCCCTTAGATGTTCTGGGCCGCACGCGCGCTACACTGATGAAGTCAGCGAGTTTATAACCTTGGCCGGAAGGTCTGGGTAAACTTTTGAAACTTCATCGTGCTGGGGATAGAGCATTGTAATTATTGCTCTTCAACGAGGAATTCCTAGTAAGCGCAAGTCATCAGCTTGCGTTGATTACGTCCCTGCCCTTTGTACACACCGCCCGTCGCTACTACCGATTGAATGGTTATAGTGAGCATATGGGATCAGTAGGATTTGACTGGCAACAGTCATTTCCTGCAGAGAACTATGGCAAACTAGGCTATTAGAGAAAGTAAAAA

The sequence of the sequencing results was compared with the effectively published strain sequences in the GenBank database by BLAST, and combined with traditional morphological identification and molecular biological identification, the final identification result was Rhizopus oryzae (Rhizopus oryzae).

2. Process optimization of fermenting Baiyaojian with distiller's yeast containing pure Rhizopus oryzae CCTCC M 2016583

1 Single-factor investigation of the effect of fermentation time on the content of gallic acid in the fermentation of Baiyaojian

The Rhizopus fermented Baiyao decoction of the present invention, under the same other conditions, take out quantitatively every 12 hours, dry, pulverize, measure the content of gallic acid, it can be concluded that the gallic acid content reaches the highest when fermented for 66 hours, the results are shown in the table 1.

Table 1 Effect of different fermentation time on gallic acid content in Baiyao decoction

Fermentation time/h 0 12 twenty four 36 48 60 66 72 84 Gallic acid content mg/g 13.38 13.59 14.33 62.89 161.34 337.09 409.27 387.77 392.77

2 Single factor investigation of the effect of different distiller's yeast additions on the gallic acid content in the fermentation of Baiyaojian

Under the same other conditions, when the amount of distiller’s yeast added is 0.4, 1, 2, 3, 4g/10g gallnut powder, ferment for 66 hours at 30°C and a relative humidity of 85%, take it out, dry it, crush it, and measure it For the content of gallic acid, it can be concluded that every 10g gallnut powder added with 1g distiller's yeast fermented Baiyao decoction has the highest gallic acid content. The results are shown in Table 2.

Table 2 Determination results of gallic acid content in fermented Baiyao decoction with different koji additions

3 Single factor investigation on the effect of different fermentation temperatures on the content of gallic acid in the fermentation of Baiyaojian

Under the same other conditions, when the fermentation temperature is 28°C, 30°C, 32°C, and 34°C, and the relative humidity is 85%, it is fermented for 66 hours. Finally, the optimal fermentation temperature is 30°C, and the results are shown in Table 3.

Table 3 Determination results of gallic acid content in Baiyao decoction fermented at different fermentation temperatures

Fermentation temperature/℃ Gallic acid content mg/g 28 381.26 30 409.97 32 326.23 34 351.19

4 Response surface design results and analysis

On the basis of the single factor experiment, the Box-Behnken experiment design was used to optimize the fermentation process. Three factors and three levels were adopted. The content of gallic acid is the response value, and the factor levels and codes are shown in Table 1; the response surface experiment groups and results are shown in Table 2.

Table 4 Experimental level and coding of Box-Behnken response surface test design factors

Table 5 Response surface experiment groups and results

Table 6 Response Surface Analysis of Variance

source of variance sum of square degrees of freedom mean square F value P value significant Model 2.015E+005 9 22385.45 9.13 0.0041 significantly A 38418.32 1 38418.32 15.66 0.0055 B 54.67 1 54.67 0.022 0.8855 C 6574.39 1 6574.39 2.68 0.1456 AB 2395.33 1 2395.33 0.98 0.3560 AC 128.86 1 128.86 0.053 0.8253 BC 643.54 1 643.54 0.26 0.6243 A2 25771.88 1 25771.88 10.51 0.0142 B2 6817.04 1 6817.04 2.78 0.1394 C2 12666.40 1 12666.40 5.16 0.0573 residual 17171.67 7 2453.10 Lack of fit 6824.86 3 2274.95 0.88 0.5230 Not obvious pure error 10346.81 4 2586.70 total deviation 2.186E+005 16

4.1 Response surface methodology to optimize the fermentation process results of Baiyaojian

Using Design-Expert software for regression model analysis and significance test, the results obtained multiple regression equations of fermentation time, fermentation temperature, added amount of koji and gallic acid content. The regression equation is: Y=-402.61+1617.56A-15.82B -308.43C+104.00AB-35.63AC+42.23BC-998.69A ² -90.69B ² -301.01C ² , variance analysis and system significance test are shown in Table 6. It can be seen from Table 6 that all items are significant, the lack of fit item P=0.5230 is not significant, and R ² =0.9215 indicates that the model fully fits the test data.

By solving the regression equation, the optimal fermentation conditions were obtained as fermentation time 65.99h, fermentation temperature 31.79℃, adding 1.22g distiller's yeast per 10g gallnut powder.

Through Design-Expert software software processing, the response surface and contour line of the interaction of each factor are shown in Figure 3, Figure 4, and Figure 5. From Figure 3, Figure 4, and Figure 5, it can be known that the fermentation time has an effect on the gallic acid in the fermented Baiyao decoction. The content of koji has the greatest impact, followed by the amount of koji added and the fermentation temperature. It can be seen from Figure 3 that when the fermentation time is short and the temperature is low, the surface is relatively flat. As the fermentation time increases and the temperature increases, the surface becomes steeper, and the contour line It shows that the interaction between the two is better; Figure 4 is the response surface diagram and contour diagram of the interaction between fermentation time and koji addition. With the increase of fermentation time and koji addition, the surface is smooth at first, then steep and finally tends It is flat, indicating that the content of gallic acid is higher when the fermentation time and the amount of koji added are moderate. Figure 5 shows the effects of fermentation temperature and koji addition on the gallic acid content in fermented Baiyao decoction. The contour lines are elliptical, indicating that the interaction is better.

Verification test: Combined with the actual production under the optimal optimal fermentation conditions mentioned above, 3 experiments were carried out under the condition of 66 hours of fermentation time, 31.8°C of fermentation temperature, and 1.2g of distiller’s yeast per 10g of Chinese gallnut powder, and the average value of the experimental results was 337.2mg/g , the relative error with the theoretical prediction value of 353mg/g is less than 5%. It shows that the equation fits well with the actual results.

3. The effect of different inoculum amounts of CCTCC M 2016583 on the content of gallic acid in the fermentation of Baiyaojian

Activate the preserved strains on PDA solid medium for 3-4 days, take 50mL distilled water and several glass balls and sterilize them in a 200mL Erlenmeyer flask at 121°C for 20min, let cool, scrape 2-4 ring spores into the sterilized Mix well in distilled water to make 1× ¹⁰⁸ /mL spore suspension.

Wash the Galla gall medicinal material, dry at 60°C for 10 hours, pulverize, pass through an 80-mesh sieve, and set aside; take 1g of tea leaves for every 10g of Gallnut powder, grind the tea leaves into coarse powder, add 10 times the amount of distilled water to decoct for 15 minutes, place at 25°C, and filter , separate the tea juice from the tea leaves, get 10mL of tea juice per 1g of tea leaves, inoculate the above-mentioned BYJ.G-1 spore suspension 1%, 5%, 10%, respectively 15%, made of soft wood, placed at 31.8°C and 85% relative humidity for 66 hours of fermentation, taken out, dried at 60°C, and measured for gallic acid content. The results showed that the content of gallic acid in Baiyaojian fermented with 5% inoculum was the highest. Several tests were carried out after repeatedly adjusting the parameters according to the foregoing conditions of the present invention. The test results showed that the gallic acid content of the fermented Baiyao decoction could be continuously maintained at a relatively high level when the inoculum amount was 5-8%.

The invention provides a strain of Rhizopus oryzae and its application in the fermentation of Baiyao decoction. The distiller's yeast and spore suspension containing the pure Rhizopus oryzae CCTCC M2016583 are used to ferment Baiyao decoction, with stable quality, high repeatability and effective Improve the efficiency of converting tannin into gallic acid, provide technical support for the quality control and standardized production of Baiyaojian, and are suitable for large-scale application and promotion.

SEQUENCE LISTING

<110> Henan University of Traditional Chinese Medicine

<120> A kind of Rhizopus oryzae and method for fermenting Baiyao decoction thereof

<130> 1

<160> 1

<170> PatentIn version 3.3

<210> 1

<211> 1724

<212>DNA

<213> Rhizopus oryzae

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ctgcagagaa ctatggcaaa ctaggctatt agagaaagta aaaa 1724

Claims (6)

1. A kind of Rhizopus oryzae, characterized in that, its classification name is Rhizopus sp. (Rhizopus sp.) BYJ.G-1, has been preserved in China Center for Type Culture Collection, and the preservation number is: CCTCC NO: M2016583. 2. the preparation method of the described rhizopus oryzae of claim 1 is characterized in that, with the distiller's yeast containing rhizopus oryzae as raw material, adopts dilution coating plate method, is coated on the potato dextrose agar medium, in 30 ℃ constant temperature culture , isolate the bacterial strain with the typical characteristics of Rhizopus, carry out isolation culture, screen out a strain of Rhizopus, name BYJ.G-1, be identified as Rhizopus oryzae; Described potato dextrose agar medium is made of: potato 200g , glucose 20g, agar 15-20g and tap water 1000ml. 3. The preparation method of Rhizopus oryzae according to claim 2, is characterized in that, take the koji for fermentation of Baiyao decoction as raw material, get koji 10g in 90mL sterile water, under 23-27 ℃, fully oscillate at 180r/min 30min, draw 1mL and add to 9mL sterile water, and so on, gradually dilute to 10 ^-2 , 10 ^-3 , 10 ^-4 ... 10 ^-9 , respectively take 200μL of bacterial suspension and spread on potato dextrose agar medium plate Put it in a constant temperature incubator at 30°C, isolate the strain with the typical characteristics of Rhizopus, and carry out isolation culture. After 2-3 days of culture, the white hyphae cover the entire surface of the medium, white, dense, cotton-like, and Many black spores grow, which is Rhizopus oryzae. 4. utilize the method for the rhizopus oryzae described in claim 1 or the preparation method described in any one of claim 2-3 to prepare the rhizopus oryzae fermentation hundred medicine decoction, it is characterized in that, Galla gall medicinal material is cleaned, 60 ℃ Dry for 10 hours, crush, pass through a 80-mesh sieve, and set aside; take 1-3g of tea leaves for every 10g of gallnut powder, grind the tea leaves into coarse powder, add 8-10 times the amount of distilled water to decoct for 10-15min, and put it at 23-27℃ , filter, separate the tea leaves and tea juice, get 10mL of tea juice per 1g of tea leaves, mix Galla chinensis powder and distiller’s yeast containing pure BYJ.G-1 bacteria according to the mass ratio of 10:0.4-4, add tea leaves and 20 Mix -30% tea juice to make a soft material; or mix the spore suspension of ^BYJ . , tea dregs, and 20-30% tea juice are mixed evenly to make a soft material; the volume water content of the soft material is controlled at 40-60%, and it is fermented at 28-34°C and a relative humidity of 85% for 60h-66h, taken out, Bake at 60°C for 10 hours until the volume moisture content is 10-12%. 5. the method for Rhizopus oryzae fermented Baiyao decoction according to claim 4, is characterized in that, the preservation bacterial classification BYJ.G-1 is activated on the potato dextrose agar slant medium for 4 days, gets 50 mL distilled water and without Sterilize the sterile glass ball in a 200mL Erlenmeyer flask at 121 ℃ for 20 minutes, let it cool, scrape 2-4 ring spores into sterilized distilled water, shake and mix well, and make 1×10 ⁸ spores/mL Suspension: Wash galla chinensis medicinal material, dry at 60°C for 10 hours, pulverize, pass through an 80-mesh sieve, and set aside; take 1g of tea leaves for every 10g of gallnut powder, grind the tea leaves into coarse powder, add 10 times the amount of distilled water to decoct for 15 minutes, and place for 25 minutes ℃, filter, separate the tea juice and tea residue, get 10mL of tea juice per 1g of tea leaves, mix Galla gall powder with tea residue, 25% tea juice, and BYJ.G-1 spore suspension with 5% weight of gall powder to prepare The soft material is fermented for 66 hours under the conditions of 31.8°C and 85% relative humidity, taken out, and dried at 60°C. After testing, the gallic acid content reaches 33.32%; the potato dextrose agar medium is composed of: 200g of potatoes, Glucose 20g, agar 15-20g and tap water 1000ml are made. 6. the method for rhizopus oryzae fermentation hundred medicine decoction according to claim 4, is characterized in that, galla gall medicinal material is washed, 60 ℃ of drying 10h, pulverizes, crosses 80 mesh sieves, for subsequent use; Take tealeaves according to every 10g gallnut powder 1g, 1.2g distiller's yeast containing pure BYJ.G-1 bacteria, the tea leaves are ground into coarse powder, add 10 times the amount of distilled water to decoct for 15 minutes, place at 23°C, filter, separate the tea juice and tea dregs, gallnut powder and containing BYJ .Mix the koji of G-1 bacteria, add tea leaves and 30% tea juice, mix to make soft material, put it at 31.8°C and 85% relative humidity for 66 hours of fermentation, take it out, dry it at 60°C, and test it , the content of gallic acid reaches 34.22%.