CN112841356B - Technological method for preparing golden flower fungus fermented tea by liquid fermentation of Xie Washi aspergillus - Google Patents
Technological method for preparing golden flower fungus fermented tea by liquid fermentation of Xie Washi aspergillus Download PDFInfo
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Abstract
A process for preparing a golden flower fungus fermented tea by liquid fermentation of Xie Washi aspergillus comprises the following steps: 1) Activating Xie Washi Aspergillus strain; 2) Preparing shake flask seed suspension; 3) Preparing liquid strains; 4) Inoculating the preparation of the golden flower fungus fermented tea stock solution; 5) Fermenting the preparation of the golden flower fungus fermented tea stock solution; 6) Preparing a golden flower fungus fermented tea stock solution; 7) Preparation of the golden flower fungus fermented tea. The fermentation mode of the golden flower fungus is improved from the fermentation process, the pure liquid secondary fermentation process is used, and the standing-vibration coupling fermentation method is combined, so that the problems of low yield and single quality of golden flower fungus fermentation tea can be solved, the problem of uneven quality of golden flower fungus fermentation tea can be solved, the health care effect of golden flower fungus fermentation tea can be improved, the challenge of safe, high-quality and efficient production of golden flower fungus fermentation tea can be overcome, the quantity (biomass) of golden flower fungus after fermentation can be improved, the quality of fermentation liquid can be improved, and the fermentation period can be shortened.
Description
Technical Field
The invention relates to the technical field of fermentation of golden flower bacteria, in particular to a process method for preparing golden flower bacteria fermented tea by liquid fermentation of Xie Washi aspergillus.
Background
Xie Washi Aspergillus is an advantageous strain in the process of growing flowers of Fuzhuan tea, commonly called as Jin Huajun, is a probiotic, has an indispensable effect on improving the quality of Fuzhuan tea, and can enable Fuzhuan tea to generate unique fungus fragrance and mellow taste under the effect of golden fungus. Therefore, the quantity of the golden fungus is also a standard for evaluating the quality of Fuzhuan tea.
Several researches show that the extracellular polysaccharide produced by the golden flower fungus in the fermentation process has the effects of regulating cholesterol metabolic balance and maintaining blood lipid level and inhibiting tumor; the fermentation of the golden flower fungus can also produce protease, amylase, lipase and the like, and the enzymes are beneficial to the human body to reduce the fat accumulated in the human body; functional components capable of promoting pepsin activity and inhibiting lipase activity exist in the fermentation liquid of the golden flower fungus, so that the golden flower fungus has obvious effects of reducing blood fat and losing weight. In addition, the fermentation liquid of the golden flower fungus also contains a plurality of active substances, and has the effects of resisting oxidation, aging, bacteria, enhancing the immune activity of human bodies and the like.
Tea is known to be beneficial to health, and tea drinks are also known as one of three drinks in the world. The organic and mineral elements in tea beverages contain a number of substances with nutritional and bioactive effects such as: tea polyphenols, tea pigments, amino acids, tea polysaccharide, organic acids, vitamins, etc.
The black tea is the fermentation tea of the golden flower fungus, belongs to the category of traditional solid state fermentation, has complicated manufacturing process flow, consumes long time and has certain requirements on the quality and variety of tea. With the more intensive research of the fermentation products of the golden fungus, the benefits of the golden fungus tea on human beings are continuously explored, and the demands of people on the high-quality golden fungus tea are continuously rising, so that challenges are presented for the safe and high-quality efficient production of the golden fungus tea, and how to solve the problems is a great trend in the future golden fungus fermentation application research field.
Disclosure of Invention
The invention aims to solve the problem that products of the golden fungus fermented tea in the current market are few and poor, improve the health care effect of the golden fungus fermented tea, fill the blank of golden fungus liquid fermented tea drinks in the market, solve the problems of long-term production, long time consumption, less yield and uneven quality limited by a solid state fermentation process with a certain safety risk, provide a process method for preparing the golden fungus fermented tea by Xie Washi aspergillus liquid fermentation, improve the fermentation mode of golden fungus from the fermentation process, apply a pure liquid secondary fermentation process and combine a standing-oscillating coupling fermentation method, not only solve the problems of low yield and single category of the golden fungus fermented tea, but also solve the problem of uneven quality of the golden fungus fermented tea, further improve the health care effect of the golden fungus fermented tea, overcome the challenges of safe, high-quality and efficient production of the golden fungus fermented tea, improve the quantity (biomass) of the golden fungus after fermentation, and further shorten the fermentation period.
In order to achieve the above purpose, the invention adopts the following technical scheme:
the invention takes tea juice, fruit juice or the mixed liquor after the combination of the tea juice and the fruit juice as a culture medium, obtains the golden fungus fermentation tea stock solution through golden fungus fermentation technology process, prepares golden fungus fermentation tea drink through mixing, and the technology process of specific preparation comprises the following steps:
1) Activating Xie Washi Aspergillus strain;
2) Preparing shake flask seed suspension;
3) Preparing liquid strains;
4) Inoculating the preparation of the golden flower fungus fermented tea stock solution;
5) Fermenting the preparation of the golden flower fungus fermented tea stock solution;
6) Preparing a golden flower fungus fermented tea stock solution;
7) Preparation of the golden flower fungus fermented tea.
1) Activating Xie Washi Aspergillus strain;
in the step 1), the Xie Washi aspergillus strain is activated by transferring Xie Washi aspergillus strain onto a plate containing PDA culture medium and placing the plate in an incubator at 28-32 ℃ for 6-8 d.
2) Preparing shake flask seed suspension;
in the step 2), the preparation method of the shake flask seed suspension comprises the following steps: hooking 4-6 blocks of 0.5cm by inoculation 2 Inoculating the size-activated Xie Washi aspergillus (Jin Huajun) strain into a sterilized 250mL triangular flask containing 80-100 mL of seed liquid culture medium, sealing a bottle mouth by using a absorbent cotton plug, setting the rotating speed of a shaking table to 140-180 rpm, and carrying out shaking culture for 5-7 d at 28-32 ℃ to obtain Xie Washi aspergillus (Jin Huajun) seed suspension (primary liquid strain);
Wherein the seed liquid culture medium is PDA liquid culture medium, and the sterilization temperature is 121 ℃ for 20min.
3) Preparing liquid strains;
in the step 3), the preparation method of the liquid strain comprises the following steps: inoculating Xie Washi aspergillus (Jin Huajun) seed suspension into a triangular flask containing a liquid strain culture medium, wherein the inoculum size is 3% -5%, and continuously shaking and culturing for 6-8 d at the rotating speed of 140-180 rpm under the condition of 28-32 ℃ after inoculation to obtain Xie Washi aspergillus secondary liquid strain;
the preparation method of the liquid strain culture medium comprises the steps of weighing 200g of peeled potato, cutting into fragments, putting into a pot, adding 1000mL of tea juice (tea: water is 1:80, tea can be black tea, green tea, oolong tea and the like) into the pot, heating to boil, maintaining for 20-30 min, filtering by two layers of gauze, adding 20g of sucrose into the filtrate, fully dissolving, and supplementing distilled water to 1000mL, wherein the sterilization temperature of the culture medium is 121 ℃ for 20min.
4) Inoculating the preparation of the golden flower fungus fermented tea stock solution;
in the step 4), the inoculation of the preparation of the stock solution of the fermentation tea of the golden flower fungus is that the second-level liquid strain of the golden flower fungus obtained in the step 3) is inoculated in a triangular flask containing a culture medium of the fermentation tea of the golden flower fungus, and the inoculation amount is 8-15 percent;
The preparation method of the culture medium comprises weighing 10-14 g tea (black tea, green tea, oolong tea, etc.), adding 1000mL of water, heating to boil, maintaining for 5-8 min, filtering with two layers of gauze, adding 40-50 g sucrose and 2-4 g soybean cake soaked powder into the filtrate, fully dissolving, and supplementing distilled water to 1000mL, wherein the sterilization temperature of the culture medium is 105-115 ℃ for 10-15 min (the culture medium can be prepared by proportionally adding fruit juice to prepare the culture medium of the fermentation fruit juice of the fungus or preparing the culture medium of the fermentation fruit juice of the fungus by using fruit juice alone).
5) Controlling fermentation process of the preparation of the golden flower fungus fermented tea stock solution, including standing-vibration coupling control, secondary fermentation material supplementing control and fermentation condition control;
in the step 5), the standing-shaking coupling control is that after the fermentation tea culture medium of the golden flower fungus is inoculated, standing is firstly carried out for 10 to 16 hours, and then shaking culture is started; or firstly shake culturing for 1d, then standing for 10-16 h, and then starting shake culturing; or alternatively culturing by static culture and shaking culture, wherein the alternative period is 34-40 h, and the time ratio of the static culture to the shaking culture is 1:3;
the secondary fermentation feeding is controlled to be carried out for 4-5 d after primary fermentation, feeding operation is carried out, the culture medium used for feeding is a golden flower fungus fermentation tea culture medium, the volume of the feeding culture medium is 30% -70% of the total volume of the primary fermentation, secondary oscillation fermentation is carried out after feeding is finished, and the oscillation fermentation time is 3-5 d;
The fermentation condition is controlled to be that the culture temperature is set at 28-32 ℃, and the shaking culture rotating speed is set at 140-180 rpm.
6) Preparing a golden flower fungus fermented tea stock solution;
in the step 6), the preparation method of the golden flower fungus fermented tea stock solution comprises the steps of standing still and settling for 3-5 h after the golden flower fungus fermented tea stock solution is prepared and fermented, filtering settled impurities, and collecting filtrate to obtain the golden flower fungus fermented tea stock solution.
7) The preparation of the golden flower fungus fermented tea comprises the preparation of golden flower fungus fermented tea with various flavors such as original flavor golden flower fungus fermented tea, fruit juice golden flower fungus fermented tea, milk flavor golden flower fungus fermented tea and the like;
in the step 7), the original flavor golden flower fungus fermented tea comprises the following components in percentage by mass: 70-85% of golden flower fungus fermented tea stock solution, 0.1-0.2% of sodium hexametaphosphate, 0.01-0.015% of vitamin C, 0.007-0.01% of stevioside and the balance of water;
the fruit-flavored golden flower fungus fermented tea comprises the following components in percentage by mass: 65 to 80 percent of golden flower fungus fermented tea stock solution, 2 to 3 percent of sucrose, 0.5 to 1 percent of concentrated juice, 0.01 to 0.015 percent of vitamin C, 0.03 to 0.06 percent of food essence and the balance of water;
The fruit juice golden flower fungus fermented tea comprises the following components in percentage by mass: 60-75% of golden flower fungus fermented tea stock solution, 2-3% of sucrose, 5-10% of fruit juice, 0.01-0.015% of vitamin C, 0.03-0.06% of food essence and the balance of water;
the milk flavor golden flower fungus fermented tea comprises the following components in percentage by mass: 60-75% of golden flower fungus fermented tea stock solution, 1.5-3% of milk powder, 1-2% of sucrose, 0.01-0.015% of vitamin C, 0.03-0.06% of food essence and the balance of water;
in addition to the above-listed formulations, formulations of the golden flower fungus fermented beverages similar thereto are considered as the present invention.
Compared with the prior art, the technical scheme of the invention has the beneficial effects that:
1. according to the invention, the Xie Washi aspergillus liquid fermentation technology is adopted, and fermentation processes such as standing, shaking, secondary feed fermentation and the like are combined, so that the method is applied to the preparation of the golden flower fungus fermented tea stock solution, and compared with the traditional preparation of the golden flower fungus fermented tea stock solution, the quality of the stock solution can be improved, the yield of the stock solution can be increased, and the preparation period of the stock solution can be shortened.
2. The fermentation strain Xie Washi aspergillus liquid fermentation technology can not only overcome the problem of complicated and long-time consumption of the process, but also solve the problem of few tea products of the fermentation of the golden fungus caused by the limitation of the tea leaves in the traditional natural fermentation (the invention can be applied to various tea products of the fermentation of the golden fungus such as green tea, black tea, oolong tea, white tea and the like), and compared with the traditional solid natural fermentation, the safety of the produced golden fungus fermentation tea stock solution is easier to control, the quality is easier to be ensured, and the invention is beneficial to the industrialized standardized production of golden fungus fermentation tea drinks.
Detailed Description
In order to make the technical problems, technical schemes and beneficial effects to be solved more clear and apparent, the invention is further described in detail below with reference to the embodiments. The specific embodiments described herein are to be considered in a descriptive sense only and not for purposes of limitation. The specific technical features of the embodiments described below can be combined with each other as long as they do not constitute mutual conflict.
Example 1
A technical process for preparing original flavor golden flower fungus fermented tea by pure aspergillus Xie Washi (Jin Huajun) liquid state standing-oscillation coupling secondary submerged fermentation specifically comprises the following steps:
1) Activation of Xie Washi aspergillus strain: the stored Xie Washi Aspergillus slant strain was transferred to a plate containing PDA medium and placed in an incubator at 30℃for 7d of activation.
2) Preparation of shake flask seed suspension: hooking 5 blocks of 0.5cm by inoculation 2 The activated strain of Aspergillus Xie Washi (Jin Huajun) was inoculated into a sterilized 250mL Erlenmeyer flask containing 100mL of seed liquid medium, the flask mouth was then sealed with a cotton wool stopper, the shaking table was set at 150rpm, and shaking culture was performed at 30℃for 6d to obtain the strain of Aspergillus Xie Washi (Jin Huajun) as a seed suspension (primary liquid strain).
Wherein the seed liquid culture medium is PDA liquid culture medium, and the sterilization temperature is 121 ℃ for 20min.
3) Preparing liquid strains: inoculating Xie Washi Aspergillus (Jin Huajun) seed suspension (primary liquid strain) into triangular flask containing liquid strain culture medium with an inoculum size of 5%, and continuously culturing at 30deg.C and rotation speed of 150rpm for 7d to obtain secondary liquid strain.
The preparation method of the liquid strain culture medium comprises weighing 200g of peeled potato, cutting into pieces, placing into a pot, adding 1000mL of tea juice (black tea: water is 1:80, obtained by hot extraction), heating to boil, maintaining for 20min, filtering with two layers of gauze, adding 20g of sucrose into the filtrate, fully dissolving, and supplementing distilled water to 1000mL, wherein the sterilization temperature of the culture medium is 121 ℃ for 20min.
4) Inoculating preparation of the golden flower fungus fermented tea stock solution: inoculating the second-level liquid strain of the golden flower fungus obtained in the step 3) into a triangular flask containing a golden flower fungus fermentation tea culture medium, wherein the inoculation amount is 8%;
the preparation method of the culture medium comprises weighing black tea leaf with weight of 12g, adding 1000mL of water, heating to boil, maintaining for 6min, filtering with two layers of gauze, adding sucrose with weight of 40g and soybean cake soaked powder with weight of 4g into filtrate, sufficiently dissolving, and adding distilled water to 1000mL, wherein the sterilization temperature of the culture medium is 110deg.C for 10min.
5) The fermentation process control of the preparation of the golden flower fungus fermented tea stock solution: placing the culture solution inoculated with the strain of the golden flower fungus into a shaking incubator with the culture temperature set at 30 ℃ for culture, firstly standing for 12 hours, and then starting shaking culture (the shaking culture rotating speed is set at 150 rpm), wherein the culture time is 5d, and the primary fermentation is carried out; after the first fermentation culture is finished, feeding operation is carried out, the culture medium used for feeding is a golden flower fungus fermentation tea culture medium, the volume of the feeding culture medium is 50% of the total volume of the first fermentation, secondary shaking fermentation is carried out after the feeding is finished, the shaking fermentation time is 4d, and the shaking culture rotating speed is set at 150rpm.
6) Preparing a golden flower fungus fermented tea stock solution: after the preparation and fermentation of the golden flower fungus fermented tea stock solution are finished, standing still and settling for 4 hours, filtering settled impurities, and collecting filtrate to obtain the golden flower fungus fermented tea stock solution.
7) Preparation of original-taste golden flower fungus fermented tea drink:
the original flavor golden flower fungus fermented tea comprises the following components in percentage by mass: 80% of golden flower fungus fermented tea stock solution, 0.15% of sodium hexametaphosphate, 0.012% of vitamin C, 0.008% of stevioside and the balance of water;
example 2
A technical process for preparing fruit flavored golden flower fungus fermented tea by pure aspergillus Xie Washi (Jin Huajun) liquid state standing-oscillation coupling secondary submerged fermentation specifically comprises the following steps:
1) Activation of Xie Washi aspergillus strain: the stored Xie Washi Aspergillus slant strain was transferred to a plate containing PDA medium and placed in an incubator at 32℃for 8d of activation.
2) Preparation of shake flask seed suspension: 6 pieces of 0.5cm are hooked by inoculation 2 The activated strain of Aspergillus Xie Washi (Jin Huajun) was inoculated into a sterilized 250mL Erlenmeyer flask containing 80mL of seed liquid medium, the flask mouth was then sealed with a cotton wool stopper, the shaking table was set at 180rpm, and the culture was performed under shaking at 32℃for 7d to obtain the seed suspension (first-order liquid strain) of Aspergillus Xie Washi (Jin Huajun).
Wherein the seed liquid culture medium is PDA liquid culture medium, and the sterilization temperature is 121 ℃ for 20min.
3) Preparing liquid strains: inoculating Xie Washi Aspergillus (Jin Huajun) seed suspension (primary liquid strain) into a triangular flask containing liquid strain culture medium, wherein the inoculation amount is 4%, and continuously culturing at 32deg.C and rotation speed of 180rpm for 8d to obtain secondary liquid strain.
The preparation method of the liquid strain culture medium comprises weighing 200g of peeled potato, cutting into pieces, placing into a pot, adding 1000mL of tea juice (oolong tea: water is 1:80, obtained by hot extraction), heating to boil, maintaining for 30min, filtering with two layers of gauze, adding 20g of sucrose into the filtrate, fully dissolving, adding distilled water to 1000mL, and sterilizing at 121deg.C for 20min.
4) Inoculating preparation of the golden flower fungus fermented tea stock solution: inoculating the second-level liquid strain of the golden flower fungus obtained in the step 3) into a triangular flask containing golden flower fungus fermentation tea culture medium, wherein the inoculation amount is 15%;
the preparation method of the culture medium comprises weighing oolong tea leaf with weight of 14g, adding 1000mL of water, heating to boil, maintaining for 8min, filtering with two layers of gauze, adding sucrose with weight of 50g and soybean cake powder with weight of 2g into filtrate, dissolving thoroughly, adding distilled water to 1000mL, and sterilizing at 115deg.C for 13min.
5) The fermentation process control of the preparation of the golden flower fungus fermented tea stock solution: placing the culture solution inoculated with the strain of the golden flower fungus into a shaking incubator with the culture temperature set at 32 ℃ for culture, firstly shaking the culture (the shaking culture rotating speed is set at 180 rpm) for 1d, then standing for 16h, and then starting shaking the culture for 6d, wherein the culture time is one-time fermentation; after the first fermentation culture is finished, feeding operation is carried out, the culture medium used for feeding is a golden flower fungus fermentation tea culture medium, the volume of the feeding culture medium is 70% of the total volume of the first fermentation, secondary shaking fermentation is carried out after the feeding is finished, the shaking fermentation time is 4d, and the shaking culture rotating speed is set at 180rpm.
6) Preparing a golden flower fungus fermented tea stock solution: after the preparation and fermentation of the golden flower fungus fermented tea stock solution are finished, standing still and settling for 5 hours, filtering settled impurities, and collecting filtrate to obtain the golden flower fungus fermented tea stock solution.
7) Preparation of fruit-flavored golden flower fungus fermented tea beverage:
the fruit-flavored golden flower fungus fermented tea comprises the following components in percentage by mass: 70% of the golden flower fungus fermented tea stock solution, 3% of sucrose, 0.5% of concentrated juice, 0.015% of vitamin C, 0.06% of food essence and the balance of water;
example 3
A technology for preparing fruit juice golden flower fungus fermented tea by Xie Washi aspergillus (Jin Huajun) pure liquid state standing-oscillation coupling secondary submerged fermentation specifically comprises the following steps:
1) Activation of Xie Washi aspergillus strain: the stored Xie Washi Aspergillus slant strain was transferred to a plate containing PDA medium and placed in an incubator at 28℃for activation for 6d.
2) Preparation of shake flask seed suspension: hooking 4 blocks of 0.5cm by inoculation 2 The activated strain of Aspergillus Xie Washi (Jin Huajun) was inoculated into a sterilized 250mL Erlenmeyer flask containing 90mL of seed liquid medium, the flask mouth was then sealed with a cotton wool stopper, the shaking table rotation speed was set at 140rpm, and shaking culture was performed at 28℃for 5 days to obtain the strain of Aspergillus Xie Washi (Jin Huajun) as a seed suspension (primary liquid strain).
Wherein the seed liquid culture medium is PDA liquid culture medium, and the sterilization temperature is 121 ℃ for 20min.
3) Preparing liquid strains: inoculating Xie Washi Aspergillus (Jin Huajun) seed suspension (primary liquid strain) into a triangular flask containing liquid strain culture medium, wherein the inoculation amount is 5%, and continuously culturing at 28deg.C and 140rpm for 6d to obtain secondary liquid strain.
The preparation method of the liquid strain culture medium comprises weighing 200g of peeled potato, cutting into pieces, placing into a pot, adding 1000mL of tea juice (green tea 1: water is 1:80, obtained by hot extraction), heating to boil, maintaining for 25min, filtering with two layers of gauze, adding 20g of sucrose into the filtrate, fully dissolving, and supplementing distilled water to 1000mL, wherein the sterilization temperature of the culture medium is 121deg.C for 20min.
4) Inoculating preparation of the golden flower fungus fermented tea stock solution: inoculating the second-level liquid strain of the golden flower fungus obtained in the step 3) into a triangular flask containing a golden flower fungus fermentation tea culture medium, wherein the inoculation amount is 10%;
the preparation method of the culture medium comprises weighing 10g of green tea leaves, adding 1000mL of water, heating to boil, maintaining for 5min, filtering with two layers of gauze, adding 45g of sucrose and 3g of soybean cake soaked powder into the filtrate, sufficiently dissolving, and supplementing distilled water to 1000mL, wherein the sterilization temperature of the culture medium is 105 ℃ for 15min (the culture medium can be prepared by proportionally adding fruit juice or the culture medium can be prepared by singly using fruit juice).
5) The fermentation process control of the preparation of the golden flower fungus fermented tea stock solution: placing the culture solution inoculated with the strain of the golden flower fungus into a shaking incubator with the culture temperature set at 28 ℃ for culture, and carrying out stationary culture and shaking culture (the shaking culture rotating speed is set at 140 rpm) alternate culture, wherein the alternate period is 40 hours, the time ratio of the stationary culture to the shaking culture alternate culture is 1:3, and the total culture time is 6d, which is one-time fermentation; after the first fermentation culture is finished, feeding operation is carried out, the culture medium used for feeding is a golden flower fungus fermentation tea culture medium, the volume of the feeding culture medium is 60% of the total volume of the first fermentation, secondary shaking fermentation is carried out after the feeding is finished, the shaking fermentation time is 4d, and the shaking culture rotating speed is set at 140rpm.
6) Preparing a golden flower fungus fermented tea stock solution: after the preparation and fermentation of the golden flower fungus fermented tea stock solution are finished, standing still and settling for 3 hours, filtering settled impurities, and collecting filtrate to obtain the golden flower fungus fermented tea stock solution.
7) Preparation of fruit juice golden flower fungus fermented tea beverage:
the fruit juice golden flower fungus fermented tea comprises the following components in percentage by mass: 65% of the golden flower fungus fermented tea stock solution, 2% of sucrose, 7% of fruit juice, 0.013% of vitamin C, 0.04% of food essence and the balance of water;
Example 4
A technical process for preparing milk flavor golden flower fungus fermented tea by pure aspergillus Xie Washi (Jin Huajun) liquid state standing-oscillation coupling secondary submerged fermentation specifically comprises the following steps:
1) Activation of Xie Washi aspergillus strain: the stored Xie Washi Aspergillus slant strain was transferred to a plate containing PDA medium and placed in an incubator at 29℃for activation for 8 days.
2) Preparation of shake flask seed suspension: by inoculation with6 pieces of 0.5cm are hooked 2 The activated strain of Aspergillus Xie Washi (Jin Huajun) was inoculated into a sterilized 250mL Erlenmeyer flask containing 85mL of seed liquid medium, the flask mouth was then sealed with a cotton wool stopper, the rotation speed of the shaker was set at 160rpm, and shaking culture was performed at 29℃for 7d to obtain the suspension of Aspergillus Xie Washi (Jin Huajun) seed (primary liquid strain).
Wherein the seed liquid culture medium is PDA liquid culture medium, and the sterilization temperature is 121 ℃ for 20min.
3) Preparing liquid strains: inoculating Xie Washi Aspergillus (Jin Huajun) seed suspension (primary liquid strain) into a triangular flask containing liquid strain culture medium, wherein the inoculation amount is 3.5%, and continuously culturing at 29 deg.C and 160rpm for 8d to obtain secondary liquid strain.
The preparation method of the liquid strain culture medium comprises weighing 200g of peeled potato, cutting into pieces, placing into a pot, adding 1000mL of tea juice (black tea: water is 1:80, obtained by hot extraction), heating to boil, maintaining for 26min, filtering with two layers of gauze, adding 20g of sucrose into the filtrate, fully dissolving, and supplementing distilled water to 1000mL, wherein the sterilization temperature of the culture medium is 121 ℃ for 20min.
4) Inoculating preparation of the golden flower fungus fermented tea stock solution: inoculating the second-level liquid strain of the golden flower fungus obtained in the step 3) into a triangular flask containing golden flower fungus fermentation tea culture medium, wherein the inoculation amount is 11%;
the preparation method of the culture medium comprises weighing 11g black tea leaves, adding 1000mL of water, heating to boil, maintaining for 7min, filtering with two layers of gauze, adding 47g of sucrose and 3.2g of soybean cake powder into the filtrate, sufficiently dissolving, adding distilled water to 1000mL, and sterilizing at 107 ℃ for 13min.
5) The fermentation process control of the preparation of the golden flower fungus fermented tea stock solution: placing the culture solution inoculated with the strain of the golden flower fungus into a shaking incubator with the culture temperature set at 29 ℃ for culture, and carrying out standing culture and shaking culture (the shaking culture rotating speed is set at 160 rpm) alternate culture, wherein the alternate period is 36 hours, the time ratio of the standing culture to the shaking culture alternate culture is 1:3, the primary fermentation is carried out, and the fermentation time is 5d; after the first fermentation culture is finished, feeding operation is carried out, the culture medium used for feeding is a golden flower fungus fermentation tea culture medium, the volume of the feeding culture medium is 55% of the total volume of the first fermentation, secondary shaking fermentation is carried out after the feeding is finished, the shaking fermentation time is 4d, and the shaking culture rotating speed is set at 160rpm.
6) Preparing a golden flower fungus fermented tea stock solution: after the preparation and fermentation of the golden flower fungus fermented tea stock solution are finished, standing still and settling for 3 hours, filtering settled impurities, and collecting filtrate to obtain the golden flower fungus fermented tea stock solution.
7) Preparation of milk-flavored golden flower fungus fermented tea beverage:
the milk-flavored golden flower fungus fermented tea comprises the following components in percentage by mass: 65% of golden flower fungus fermented tea stock solution, 2% of milk powder, 1.5% of sucrose, 0.012% of vitamin C, 0.05% of food essence and the balance of water.
Example 5
A method for improving the biomass of a pure-breed liquid fermentation of aspergillus Xie Washi (Jin Huajun), which comprises the following steps:
1) Activation of Xie Washi aspergillus strain: the stored Xie Washi Aspergillus slant strain was transferred to a plate containing PDA medium and placed in an incubator at 30℃for 7d of activation.
2) Preparation of shake flask seed suspension: hooking 5 blocks of 0.5cm by inoculation 2 The activated strain of Aspergillus Xie Washi (Jin Huajun) was inoculated into a sterilized 250mL Erlenmeyer flask containing 100mL of seed liquid medium, the flask mouth was then sealed with a cotton wool stopper, the shaking table was set at 150rpm, and shaking culture was performed at 30℃for 6d to obtain the strain of Aspergillus Xie Washi (Jin Huajun) as a seed suspension (primary liquid strain).
Wherein the seed liquid culture medium is PDA liquid culture medium, and the sterilization temperature is 121 ℃ for 20min.
3) Preparing liquid strains: inoculating Xie Washi Aspergillus (Jin Huajun) seed suspension (primary liquid strain) into triangular flask containing liquid strain culture medium with an inoculum size of 5%, and continuously culturing at 30deg.C and rotation speed of 150rpm for 7d to obtain secondary liquid strain.
The preparation method of the liquid strain culture medium comprises weighing 200g of peeled potato, cutting into pieces, placing into a pot, adding 1000mL of tea juice (oolong tea: water is 1:80, obtained by hot extraction), heating to boil, maintaining for 20min, filtering with two layers of gauze, adding 20g of sucrose into the filtrate, fully dissolving, adding distilled water to 1000mL, and sterilizing at 121deg.C for 20min.
4) Inoculating preparation of the golden flower fungus fermented tea stock solution: inoculating the second-level liquid strain of the golden flower fungus obtained in the step 3) into a triangular flask containing a golden flower fungus fermentation tea culture medium, wherein the inoculation amount is 10%;
the preparation method of the culture medium comprises weighing black tea leaf with weight of 12g, adding 1000mL of water, heating to boil, maintaining for 6min, filtering with two layers of gauze, adding sucrose with weight of 40g and soybean cake soaked powder with weight of 4g into filtrate, sufficiently dissolving, and adding distilled water to 1000mL, wherein the sterilization temperature of the culture medium is 110deg.C for 10min.
5) Controlling the fermentation process of the golden flower fungus fermented tea: the culture solution inoculated with the strain of the golden flower fungus is placed in a shaking incubator with the culture temperature set at 30 ℃ for culture, and is firstly kept stand for 12 hours and then shaking culture is started (the shaking culture rotating speed is set at 150 rpm), and the culture time is 6d.
6) Determination of the biological amount of the golden flower fungus: vacuum suction filtration is carried out by using a suction filter, the culture solution of the fermentation tea of the golden flower fungus is filtered on filter paper to obtain mycelium, the mycelium is baked at 100-105 ℃ until the quality is constant, and the measurement results are shown in table 1.
Example 6
A method for improving the biomass of a pure-breed liquid fermentation of aspergillus Xie Washi (Jin Huajun), which comprises the following steps:
1) Activation of Xie Washi aspergillus strain: the stored Xie Washi Aspergillus slant strain was transferred to a plate containing PDA medium and placed in an incubator at 30℃for 7d of activation.
2) Preparation of shake flask seed suspension: hooking 5 blocks of 0.5cm by inoculation 2 The size of the activated A. Xie Washi (Jin Huajun) strain was inoculated into sterilized 250mL Erlenmeyer flasks containing 100mL of seed liquid medium and then tampons were usedSealing the bottle mouth, setting the rotation speed of the shaking table to 150rpm, and culturing for 6d under the condition of 30 ℃ to obtain the Xie Washi aspergillus (Jin Huajun) seed suspension (primary liquid strain).
Wherein the seed liquid culture medium is PDA liquid culture medium, and the sterilization temperature is 121 ℃ for 20min.
3) Preparing liquid strains: inoculating Xie Washi Aspergillus (Jin Huajun) seed suspension (primary liquid strain) into triangular flask containing liquid strain culture medium with an inoculum size of 5%, and continuously culturing at 30deg.C and rotation speed of 150rpm for 7d to obtain secondary liquid strain.
The preparation method of the liquid strain culture medium comprises weighing 200g of peeled potato, cutting into pieces, placing into a pot, adding 1000mL of tea juice (oolong tea: water is 1:80, obtained by hot extraction), heating to boil, maintaining for 20min, filtering with two layers of gauze, adding 20g of sucrose into the filtrate, fully dissolving, adding distilled water to 1000mL, and sterilizing at 121deg.C for 20min.
4) Inoculating preparation of the golden flower fungus fermented tea stock solution: inoculating the second-level liquid strain of the golden flower fungus obtained in the step 3) into a triangular flask containing a golden flower fungus fermentation tea culture medium, wherein the inoculation amount is 10%;
the preparation method of the culture medium comprises weighing black tea leaf with weight of 12g, adding 1000mL of water, heating to boil, maintaining for 6min, filtering with two layers of gauze, adding sucrose with weight of 40g and soybean cake soaked powder with weight of 4g into filtrate, sufficiently dissolving, and adding distilled water to 1000mL, wherein the sterilization temperature of the culture medium is 110deg.C for 10min.
5) Controlling the fermentation process of the golden flower fungus fermented tea: placing the culture solution inoculated with the strain of the golden flower fungus into a shaking incubator with the culture temperature set at 30 ℃ for culture, and carrying out stationary culture and shaking culture (the shaking culture rotating speed is set at 150 rpm) alternate culture, wherein the alternate period is 40 hours, the time ratio of the stationary culture to the shaking culture alternate culture is 1:3, and the total culture time is 6 days.
6) Determination of the biological amount of the golden flower fungus: vacuum suction filtration is carried out by using a suction filter, the culture solution of the fermentation tea of the golden flower fungus is filtered on filter paper to obtain mycelium, the mycelium is baked at 100-105 ℃ until the quality is constant, and the measurement results are shown in table 1.
Example 7
A method for improving the biomass of a pure-breed liquid fermentation of aspergillus Xie Washi (Jin Huajun), which comprises the following steps:
1) Activation of Xie Washi aspergillus strain: the stored Xie Washi Aspergillus slant strain was transferred to a plate containing PDA medium and placed in an incubator at 30℃for 7d of activation.
2) Preparation of shake flask seed suspension: hooking 5 blocks of 0.5cm by inoculation 2 The activated strain of Aspergillus Xie Washi (Jin Huajun) was inoculated into a sterilized 250mL Erlenmeyer flask containing 100mL of seed liquid medium, the flask mouth was then sealed with a cotton wool stopper, the shaking table was set at 150rpm, and shaking culture was performed at 30℃for 6d to obtain the strain of Aspergillus Xie Washi (Jin Huajun) as a seed suspension (primary liquid strain).
Wherein the seed liquid culture medium is PDA liquid culture medium, and the sterilization temperature is 121 ℃ for 20min.
3) Preparing liquid strains: inoculating Xie Washi Aspergillus (Jin Huajun) seed suspension (primary liquid strain) into triangular flask containing liquid strain culture medium with an inoculum size of 5%, and continuously culturing at 30deg.C and rotation speed of 150rpm for 7d to obtain secondary liquid strain.
The preparation method of the liquid strain culture medium comprises weighing 200g of peeled potato, cutting into pieces, placing into a pot, adding 1000mL of tea juice (oolong tea: water is 1:80, obtained by hot extraction), heating to boil, maintaining for 20min, filtering with two layers of gauze, adding 20g of sucrose into the filtrate, fully dissolving, adding distilled water to 1000mL, and sterilizing at 121deg.C for 20min.
4) Inoculating preparation of the golden flower fungus fermented tea stock solution: inoculating the second-level liquid strain of the golden flower fungus obtained in the step 3) into a triangular flask containing a golden flower fungus fermentation tea culture medium, wherein the inoculation amount is 10%;
the preparation method of the culture medium comprises weighing black tea leaf with weight of 12g, adding 1000mL of water, heating to boil, maintaining for 6min, filtering with two layers of gauze, adding sucrose with weight of 40g and soybean cake soaked powder with weight of 4g into filtrate, sufficiently dissolving, and adding distilled water to 1000mL, wherein the sterilization temperature of the culture medium is 110deg.C for 10min.
5) Controlling the fermentation process of the golden flower fungus fermented tea: the culture solution inoculated with the strain of the golden flower fungus is placed in a shaking incubator with the culture temperature set at 30 ℃ for culture, shaking culture is carried out firstly (the shaking culture rotating speed is set at 150 rpm) for 1d, then standing for 16h, and shaking culture is started again, wherein the culture time is 6d.
6) Determination of the biological amount of the golden flower fungus: vacuum suction filtration is carried out by using a suction filter, the culture solution of the fermentation tea of the golden flower fungus is filtered on filter paper to obtain mycelium, the mycelium is baked at 100-105 ℃ until the quality is constant, and the measurement results are shown in table 1.
Example 8
A method for improving the biomass of a pure-breed liquid fermentation of aspergillus Xie Washi (Jin Huajun), which comprises the following steps:
1) Activation of Xie Washi aspergillus strain: the stored Xie Washi Aspergillus slant strain was transferred to a plate containing PDA medium and placed in an incubator at 30℃for 7d of activation.
2) Preparation of shake flask seed suspension: hooking 5 blocks of 0.5cm by inoculation 2 The activated strain of Aspergillus Xie Washi (Jin Huajun) was inoculated into a sterilized 250mL Erlenmeyer flask containing 100mL of seed liquid medium, the flask mouth was then sealed with a cotton wool stopper, the shaking table was set at 150rpm, and shaking culture was performed at 30℃for 6d to obtain the strain of Aspergillus Xie Washi (Jin Huajun) as a seed suspension (primary liquid strain).
Wherein the seed liquid culture medium is PDA liquid culture medium, and the sterilization temperature is 121 ℃ for 20min.
3) Preparing liquid strains: inoculating Xie Washi Aspergillus (Jin Huajun) seed suspension (primary liquid strain) into triangular flask containing liquid strain culture medium with an inoculum size of 5%, and continuously culturing at 30deg.C and rotation speed of 150rpm for 7d to obtain secondary liquid strain.
The preparation method of the liquid strain culture medium comprises weighing 200g of peeled potato, cutting into pieces, placing into a pot, adding 1000mL of tea juice (oolong tea: water is 1:80, obtained by hot extraction), heating to boil, maintaining for 20min, filtering with two layers of gauze, adding 20g of sucrose into the filtrate, fully dissolving, adding distilled water to 1000mL, and sterilizing at 121deg.C for 20min.
4) Inoculating preparation of the golden flower fungus fermented tea stock solution: inoculating the second-level liquid strain of the golden flower fungus obtained in the step 3) into a triangular flask containing a golden flower fungus fermentation tea culture medium, wherein the inoculation amount is 10%;
the preparation method of the culture medium comprises weighing black tea leaf with weight of 12g, adding 1000mL of water, heating to boil, maintaining for 6min, filtering with two layers of gauze, adding sucrose with weight of 40g and soybean cake soaked powder with weight of 4g into filtrate, sufficiently dissolving, and adding distilled water to 1000mL, wherein the sterilization temperature of the culture medium is 110deg.C for 10min.
5) Controlling the fermentation process of the golden flower fungus fermented tea: placing the culture solution inoculated with the golden flower fungus strain into a shaking incubator with the culture temperature set at 30 ℃ for culture, firstly standing for 12 hours, then starting shaking culture (the shaking culture rotating speed is set at 150 rpm), wherein the culture time is 6d, after the primary fermentation culture is finished, performing feeding operation, wherein the volume of the feeding culture medium is 50% of the total primary fermentation volume, performing secondary shaking fermentation after the feeding is finished, the shaking fermentation rotating speed is set at 150rpm, and the shaking culture rotating speed is set at 4 d.
6) Determination of the biological amount of the golden flower fungus: vacuum suction filtration is carried out by using a suction filter, the culture solution of the fermentation tea of the golden flower fungus is filtered on filter paper to obtain mycelium, the mycelium is baked at 100-105 ℃ until the quality is constant, and the measurement results are shown in table 1.
Example 9
A method for improving the biomass of a pure-breed liquid fermentation of aspergillus Xie Washi (Jin Huajun), which comprises the following steps:
1) Activation of Xie Washi aspergillus strain: the stored Xie Washi Aspergillus slant strain was transferred to a plate containing PDA medium and placed in an incubator at 30℃for 7d of activation.
2) Preparation of shake flask seed suspension: hooking 5 blocks of 0.5cm by inoculation 2 The size of the activated A. Xie Washi strain (Jin Huajun) was inoculated into sterilizedIn a 250mL triangular flask containing 100mL of seed liquid culture medium, then sealing the bottle mouth with a absorbent cotton plug, setting the rotation speed of a shaking table to 150rpm, and carrying out shaking culture for 6d at 30 ℃ to obtain the Xie Washi aspergillus (Jin Huajun) seed suspension (primary liquid strain).
Wherein the seed liquid culture medium is PDA liquid culture medium, and the sterilization temperature is 121 ℃ for 20min.
3) Preparing liquid strains: inoculating Xie Washi Aspergillus (Jin Huajun) seed suspension (primary liquid strain) into triangular flask containing liquid strain culture medium with an inoculum size of 5%, and continuously culturing at 30deg.C and rotation speed of 150rpm for 7d to obtain secondary liquid strain.
The preparation method of the liquid strain culture medium comprises weighing 200g of peeled potato, cutting into pieces, placing into a pot, adding 1000mL of tea juice (oolong tea: water is 1:80, obtained by hot extraction), heating to boil, maintaining for 20min, filtering with two layers of gauze, adding 20g of sucrose into the filtrate, fully dissolving, adding distilled water to 1000mL, and sterilizing at 121deg.C for 20min.
4) Inoculating preparation of the golden flower fungus fermented tea stock solution: inoculating the second-level liquid strain of the golden flower fungus obtained in the step 3) into a triangular flask containing a golden flower fungus fermentation tea culture medium, wherein the inoculation amount is 10%;
The preparation method of the culture medium comprises weighing black tea leaf with weight of 12g, adding 1000mL of water, heating to boil, maintaining for 6min, filtering with two layers of gauze, adding sucrose with weight of 40g and soybean cake soaked powder with weight of 4g into filtrate, sufficiently dissolving, and adding distilled water to 1000mL, wherein the sterilization temperature of the culture medium is 110deg.C for 10min.
5) Controlling the fermentation process of the golden flower fungus fermented tea: placing the culture solution inoculated with the golden flower fungus strain into a shaking incubator with the culture temperature set at 30 ℃ for culture, carrying out standing culture and shaking culture (the shaking culture rotating speed is set at 150 rpm) for alternate culture, wherein the alternate period is 40h, the time ratio of the standing culture to the shaking culture alternate culture is 1:3, the total culture time is 6d, after the primary fermentation culture is finished, carrying out feeding operation, wherein the culture medium used for feeding is golden flower fungus fermentation tea culture medium, the volume of the feeding culture medium is 60% of the total volume of the primary fermentation, carrying out secondary shaking fermentation after the feeding is finished, the shaking fermentation rotating speed is set at 150rpm, and the shaking fermentation rotating speed is set at 4 d.
6) Determination of the biological amount of the golden flower fungus: vacuum suction filtration is carried out by using a suction filter, the culture solution of the fermentation tea of the golden flower fungus is filtered on filter paper to obtain mycelium, the mycelium is baked at 100-105 ℃ until the quality is constant, and the measurement results are shown in table 1.
Comparative example 1
A method for improving the biomass of a pure-breed liquid fermentation of aspergillus Xie Washi (Jin Huajun), which comprises the following steps:
1) Activation of Xie Washi aspergillus strain: the stored Xie Washi Aspergillus slant strain was transferred to a plate containing PDA medium and placed in an incubator at 30℃for 7d of activation.
2) Preparation of shake flask seed suspension: hooking 5 blocks of 0.5cm by inoculation 2 The activated strain of Aspergillus Xie Washi (Jin Huajun) was inoculated into a sterilized 250mL Erlenmeyer flask containing 100mL of seed liquid medium, the flask mouth was then sealed with a cotton wool stopper, the shaking table was set at 150rpm, and shaking culture was performed at 30℃for 6d to obtain the strain of Aspergillus Xie Washi (Jin Huajun) as a seed suspension (primary liquid strain).
Wherein the seed liquid culture medium is PDA liquid culture medium, and the sterilization temperature is 121 ℃ for 20min.
3) Preparing liquid strains: inoculating Xie Washi Aspergillus (Jin Huajun) seed suspension (primary liquid strain) into triangular flask containing liquid strain culture medium with an inoculum size of 5%, and continuously culturing at 30deg.C and rotation speed of 150rpm for 7d to obtain secondary liquid strain.
The preparation method of the liquid strain culture medium comprises weighing 200g of peeled potato, cutting into pieces, placing into a pot, adding 1000mL of tea juice (oolong tea: water is 1:80, obtained by hot extraction), heating to boil, maintaining for 20min, filtering with two layers of gauze, adding 20g of sucrose into the filtrate, fully dissolving, adding distilled water to 1000mL, and sterilizing at 121deg.C for 20min.
4) Inoculating preparation of the golden flower fungus fermented tea stock solution: inoculating the second-level liquid strain of the golden flower fungus obtained in the step 3) into a triangular flask containing a golden flower fungus fermentation tea culture medium, wherein the inoculation amount is 10%;
the preparation method of the culture medium comprises weighing black tea leaf with weight of 12g, adding 1000mL of water, heating to boil, maintaining for 6min, filtering with two layers of gauze, adding sucrose with weight of 40g and soybean cake soaked powder with weight of 4g into filtrate, sufficiently dissolving, and adding distilled water to 1000mL, wherein the sterilization temperature of the culture medium is 110deg.C for 10min.
5) Controlling the fermentation process of the golden flower fungus fermented tea: the culture solution inoculated with the strain of the golden flower fungus is placed in a shaking incubator with the culture temperature set at 30 ℃ for shaking culture, the shaking culture rotating speed is set at 150rpm, and the culture time is 6d.
6) Determination of the biological amount of the golden flower fungus: vacuum suction filtration is carried out by using a suction filter, the culture solution of the fermentation tea of the golden flower fungus is filtered on filter paper to obtain mycelium, the mycelium is baked at 100-105 ℃ until the quality is constant, and the measurement results are shown in table 1.
Comparative example 2
A method for improving the biomass of a pure-breed liquid fermentation of aspergillus Xie Washi (Jin Huajun), which comprises the following steps:
1) Activation of Xie Washi aspergillus strain: the stored Xie Washi Aspergillus slant strain was transferred to a plate containing PDA medium and placed in an incubator at 30℃for 7d of activation.
2) Preparation of shake flask seed suspension: hooking 5 blocks of 0.5cm by inoculation 2 The activated strain of Aspergillus Xie Washi (Jin Huajun) was inoculated into a sterilized 250mL Erlenmeyer flask containing 100mL of seed liquid medium, the flask mouth was then sealed with a cotton wool stopper, the shaking table was set at 150rpm, and shaking culture was performed at 30℃for 6d to obtain the strain of Aspergillus Xie Washi (Jin Huajun) as a seed suspension (primary liquid strain).
Wherein the seed liquid culture medium is PDA liquid culture medium, and the sterilization temperature is 121 ℃ for 20min.
3) Preparing liquid strains: inoculating Xie Washi Aspergillus (Jin Huajun) seed suspension (primary liquid strain) into triangular flask containing liquid strain culture medium with an inoculum size of 5%, and continuously culturing at 30deg.C and rotation speed of 150rpm for 7d to obtain secondary liquid strain.
The preparation method of the liquid strain culture medium comprises weighing 200g of peeled potato, cutting into pieces, placing into a pot, adding 1000mL of tea juice (oolong tea: water is 1:80, obtained by hot extraction), heating to boil, maintaining for 20min, filtering with two layers of gauze, adding 20g of sucrose into the filtrate, fully dissolving, adding distilled water to 1000mL, and sterilizing at 121deg.C for 20min.
4) Inoculating preparation of the golden flower fungus fermented tea stock solution: inoculating the second-level liquid strain of the golden flower fungus obtained in the step 3) into a triangular flask containing a golden flower fungus fermentation tea culture medium, wherein the inoculation amount is 10%;
the preparation method of the culture medium comprises weighing black tea leaf with weight of 12g, adding 1000mL of water, heating to boil, maintaining for 6min, filtering with two layers of gauze, adding sucrose with weight of 40g and soybean cake soaked powder with weight of 4g into filtrate, sufficiently dissolving, and adding distilled water to 1000mL, wherein the sterilization temperature of the culture medium is 110deg.C for 10min.
5) Controlling the fermentation process of the golden flower fungus fermented tea: placing the culture solution inoculated with the golden flower fungus strain into a shaking incubator with the culture temperature set at 30 ℃ for shaking culture, wherein the shaking culture rotating speed is set at 150rpm, the culture time is 6d, the operation of supplementing sterile water is carried out after the primary fermentation culture is finished, the volume of the supplemented sterile water is 50% of the total volume of the primary fermentation, the secondary shaking fermentation is carried out after the water supplementing is finished, the shaking fermentation rotating speed is set at 150rpm, and the shaking culture rotating speed is set at 4 d.
6) Determination of the biological amount of the golden flower fungus: vacuum suction filtration is carried out by using a suction filter, the culture solution of the fermentation tea of the golden flower fungus is filtered on filter paper to obtain mycelium, the mycelium is baked at 100-105 ℃ until the quality is constant, and the measurement results are shown in table 1.
Comparative example 3
A method for improving the biomass of a pure-breed liquid fermentation of aspergillus Xie Washi (Jin Huajun), which comprises the following steps:
1) Activation of Xie Washi aspergillus strain: the stored Xie Washi Aspergillus slant strain was transferred to a plate containing PDA medium and placed in an incubator at 30℃for 7d of activation.
2) Preparation of shake flask seed suspension: hooking 5 blocks of 0.5cm by inoculation 2 The activated strain of Aspergillus Xie Washi (Jin Huajun) was inoculated into a sterilized 250mL Erlenmeyer flask containing 100mL of seed liquid medium, the flask mouth was then sealed with a cotton wool stopper, the shaking table was set at 150rpm, and shaking culture was performed at 30℃for 6d to obtain the strain of Aspergillus Xie Washi (Jin Huajun) as a seed suspension (primary liquid strain).
Wherein the seed liquid culture medium is PDA liquid culture medium, and the sterilization temperature is 121 ℃ for 20min.
3) Preparing liquid strains: inoculating Xie Washi Aspergillus (Jin Huajun) seed suspension (primary liquid strain) into triangular flask containing liquid strain culture medium with an inoculum size of 5%, and continuously culturing at 30deg.C and rotation speed of 150rpm for 7d to obtain secondary liquid strain.
The preparation method of the liquid strain culture medium comprises weighing 200g of peeled potato, cutting into pieces, placing into a pot, adding 1000mL of tea juice (oolong tea: water is 1:80, obtained by hot extraction), heating to boil, maintaining for 20min, filtering with two layers of gauze, adding 20g of sucrose into the filtrate, fully dissolving, adding distilled water to 1000mL, and sterilizing at 121deg.C for 20min.
4) Inoculating preparation of the golden flower fungus fermented tea stock solution: inoculating the second-level liquid strain of the golden flower fungus obtained in the step 3) into a triangular flask containing a golden flower fungus fermentation tea culture medium, wherein the inoculation amount is 10%;
the preparation method of the culture medium comprises weighing black tea leaf with weight of 12g, adding 1000mL of water, heating to boil, maintaining for 6min, filtering with two layers of gauze, adding sucrose with weight of 40g and soybean cake soaked powder with weight of 4g into filtrate, sufficiently dissolving, and adding distilled water to 1000mL, wherein the sterilization temperature of the culture medium is 110deg.C for 10min.
5) Controlling the fermentation process of the golden flower fungus fermented tea: the culture solution inoculated with the strain of the golden flower fungus is placed in a shaking incubator with the culture temperature set at 30 ℃ for shaking culture, the shaking culture rotating speed is set at 150rpm, and the culture time is 10d.
6) Determination of the biological amount of the golden flower fungus: vacuum suction filtration is carried out by using a suction filter, the culture solution of the fermentation tea of the golden flower fungus is filtered on filter paper to obtain mycelium, the mycelium is baked at 100-105 ℃ until the quality is constant, and the measurement results are shown in table 1.
TABLE 1
| Examples | Golden flower fungus mycelium biomass (g/L) | Percent biomass improvement (%) |
| Example 5 | 3.612 | 11.10 |
| Example 6 | 3.569 | 9.78 |
| Example 7 | 3.545 | 9.04 |
| Example 8 | 5.248 | 61.43 |
| Example 9 | 5.503 | 69.27 |
| Comparative example 1 | 3.251 | ---- |
| Comparative example 2 | 3.406 | 4.77 |
| Comparative example 3 | 3.384 | 4.09 |
The invention adopts Xie Washi aspergillus liquid fermentation technology, which not only can overcome the problems of complicated flow and long time consumption, but also has no requirements on tea products and varieties, and can realize that different tea products and varieties such as green tea, black tea, oolong tea and the like are fermented by using the golden fungus to obtain different types of golden fungus fermented tea, thus having certain practical significance for the product expansion of golden fungus tea and the popularization of golden fungus tea.
According to the invention, the mycelium of the aspergillus Xie Washi (Jin Huajun) is in a smaller sphere shape, compared with edible fungus spheres, the mycelium is difficult to culture and large, if long-term shaking culture is unfavorable for the growth of the mycelium and the propagation of microorganisms, a standing-shaking coupled fermentation technology is adopted, after shaking for a period of time, a standing environment is provided for the mycelium, the growth and elongation of the mycelium are facilitated, shaking is continued again, the mycelium falls off from the fungus spheres, the fermentation culture is continued, the fallen mycelium can grow into the fungus spheres again, the contact surface between the golden flower fungus and the culture medium is increased, and the utilization of nutrients in the culture medium by the golden flower fungus is facilitated.
The invention adopts the secondary fermentation technology in the liquid fermentation, the secondary fermentation can lead the fermentation material to ferment more fully, the strain activity is stronger, the bacterial quantity is more, the obtained fermentation products and active substances are more, the fermentation time can be shortened, the invention has the advantage that the primary fermentation is difficult to compare, and the biomass and the metabolite quantity are improved by the secondary fermentation technology, thereby improving the industrial production efficiency, shortening the production period and saving the production cost.
Claims (3)
1. A process for preparing a golden flower fungus fermented tea by liquid fermentation of Xie Washi aspergillus is characterized by comprising the following steps:
1) Activation of Xie Washi aspergillus strain: transferring Xie Washi aspergillus strain to a plate containing PDA culture medium, and placing the plate in an incubator at 28-32 ℃ for activation for 6-8 d;
2) Preparation of shake flask seed suspension: inoculating the Xie Washi aspergillus strain after the activation into a sterilized seed liquid culture medium, and carrying out shaking culture for 5-7 d at 28-32 ℃ to obtain Xie Washi aspergillus seed suspension; the seed liquid culture medium is PDA liquid culture medium;
3) Preparing liquid strains: inoculating the Xie Washi aspergillus seed suspension obtained in the step 2) into a liquid strain culture medium, wherein the inoculum size is 3% -5%, and carrying out shaking culture for 6-8 d at 28-32 ℃ after inoculation to obtain Xie Washi aspergillus secondary liquid strain; the preparation method of the liquid strain culture medium comprises cutting peeled potato into pieces, placing into a pot, adding tea juice, boiling, filtering to obtain filtrate, adding sucrose into the filtrate for dissolving, and adding distilled water to obtain the liquid strain culture medium;
4) Inoculating preparation of the golden flower fungus fermented tea stock solution: inoculating Xie Washi aspergillus secondary liquid strain obtained in the step 3) into a fermentation culture medium, wherein the inoculum size is 8% -15%; the fermentation culture medium takes tea juice, fruit juice or a mixed solution of the tea juice and the fruit juice as the culture medium;
5) Fermenting the preparation of the golden flower fungus fermented tea stock solution: comprises standing-oscillating coupling control, secondary fermentation feed supplement control and fermentation condition control; the standing-shaking coupling control is that after the fermentation culture medium is inoculated, standing is firstly carried out, and then shaking culture is started; or firstly shake culturing, standing and then starting shake culturing; or alternatively culturing by standing culture and shaking culture; the secondary fermentation feeding is controlled to be carried out for 4-5 d after primary fermentation, feeding operation is carried out, the culture medium used for feeding is a fermentation culture medium, the volume of the feeding culture medium is 30% -70% of the total volume of the primary fermentation, secondary oscillation fermentation is carried out after feeding is completed, and the oscillation fermentation time is 3-4 d; the fermentation condition is controlled to be that the culture temperature is set to be 28-32 ℃;
6) Preparing a golden flower fungus fermented tea stock solution: after the fermentation of the step 5), standing and settling are started, then impurities after settling are filtered, and filtrate is collected to obtain the golden flower fungus fermented tea stock solution;
7) Preparation of the golden flower fungus fermented tea.
2. The process for preparing the golden flower fungus fermented tea by liquid fermentation of Xie Washi aspergillus as claimed in claim 1, wherein the process comprises the following steps: in the step 7), the original flavor golden flower fungus fermented tea comprises the following components in percentage by mass: 70-85% of golden flower fungus fermented tea stock solution, 0.1-0.2% of sodium hexametaphosphate, 0.01-0.015% of vitamin C, 0.007-0.01% of stevioside and the balance of water; the fruit-flavored golden flower fungus fermented tea comprises the following components in percentage by mass: 65 to 80 percent of golden flower fungus fermented tea stock solution, 2 to 3 percent of sucrose, 0.5 to 1 percent of concentrated juice, 0.01 to 0.015 percent of vitamin C, 0.03 to 0.06 percent of food essence and the balance of water; the fruit juice golden flower fungus fermented tea comprises the following components in percentage by mass: 60-75% of golden flower fungus fermented tea stock solution, 2-3% of sucrose, 5-10% of fruit juice, 0.01-0.015% of vitamin C, 0.03-0.06% of food essence and the balance of water; the milk-flavored golden flower fungus fermented tea comprises the following components in percentage by mass: 60-75% of golden flower fungus fermented tea stock solution, 1.5-3% of milk powder, 1-2% of sucrose, 0.01-0.015% of vitamin C, 0.03-0.06% of food essence and the balance of water.
3. The process for preparing the golden flower fungus fermented tea by liquid fermentation of Xie Washi aspergillus as claimed in any one of claims 1, wherein the process comprises the following steps: the rotation speed of the oscillating shaking table is 140-180 rpm.
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| JP2012080812A (en) * | 2010-10-11 | 2012-04-26 | Riverson:Kk | Method for producing microorganism-fermented tea rich in flagrance of flower |
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| JP2012080812A (en) * | 2010-10-11 | 2012-04-26 | Riverson:Kk | Method for producing microorganism-fermented tea rich in flagrance of flower |
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