CN101491362A - Red tea fungus beverage produced by mixed culture fermentation of pure fungus - Google Patents
Red tea fungus beverage produced by mixed culture fermentation of pure fungus Download PDFInfo
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Abstract
The invention provides a black tea fungus drink which is produced by the mixed fermentation of pure fungi. Three kinds of fungi of saccharomyces cerivisiae, bacillus aceticus and lactobacillus plantarum are compounded, and mixed and fermented to produce the black tea fungus drink. In the drink, the combination of the pure fungi and the mixed fermentation are utilized, and the formula of the culture medium of the black tea fungus and the fermentation process conditions are optimized, thereby obviously shortening the fermentation time, avoiding the pollution of mixed fungi and ensuring the product safety; management methods for culture, rejuvenation and preservation of the fungus are scientific and reasonable, the product quality is stable, and a good basis is laid for the industrialized production of the black tea fungus; and the prepared black tea fungus drink not only tastes good, but also contains various effective components, and is a microbial fermentation drink with high utility value.
Description
Technical field
The invention belongs to microbial technology field, more specifically relate to a kind of pure bacterium mixed culture fermentation production black-tea fungus drink that utilizes.
Background technology
Raising along with science and technology development and people's living standard, many people have more seen clearly the correlation between disease, health and the food, another has proposed the viewpoint of dietotherapy: the composition that improves eating condition and food, the physiological regulation function of performance food itself is to reach the purpose that improves level of human health.Based on this consumption concept, people more and more pay attention to quality healthy and the pursuit beverage.Fermented tea is the probio that is of value to health by three kinds, and promptly the syntaxial system of saccharomycete, acetic acid bacteria, lactic acid bacteria composition by self special role, is converted into extremely rich nutrient substances with raw materials such as tealeaves through the biotechnology zymotechnique.Owing to contain nutriments such as gluconic acid, acetic acid, glucose, fructose, protein, amino acid, vitamin, trace element, Tea Polyphenols, caffeine, ethanol and carbon dioxide in the fermented tea, multiple magical effects such as therefore be considered to a kind of has the health drink of various health-care to human body always, and it has the cleaning stomach, reduces blood sugar, blood pressure, row's calculus, clearing lung-heat, cough-relieving, relievings asthma, anticancer.Though fermented tea has long history in China, its scientific research is started late, still rest at present on the basis that family markets one's own products, the report of relevant successful product is not arranged on the market as yet.The production of fermented tea fermented beverage relates to many-sided problems such as microorganism, medical hygiene, industry, product quality and safety, because at present a lot of fermented teas all are small workshop mode production, adopt mixed bacterium natural fermented, the microbe species that different fermented teas contain is different with vigor, the composition of product and content are also inconsistent in the product, can not fundamentally guarantee the quality and the safety of drink, but also can cause because of misoperation polluting, going bad.The more important thing is that bacterial classification morphs easily, fails and death owing to cultivation, the management of the science of shortage; In addition, owing to adopt natural mixed fungus fermentation, fermentation period is long, is generally about 10 days.Therefore study the science preparation method of black-tea fungus drink, make the production that is confined to the fermented tea realization industrialization on the traditional manufacturing technique to have important economy and society meaning.
Summary of the invention
The purpose of this invention is to provide a kind of pure bacterium mixed culture fermentation production black-tea fungus drink that utilizes, this method is utilized composite, the mixed culture fermentation of pure culture, and fermented tea culture medium prescription, technological condition for fermentation are optimized, and significantly shortens fermentation time, avoid living contaminants, guarantee product safety; Management methods such as bacterial classification cultivation, rejuvenation, preservation are scientific and reasonable, and constant product quality is for the industrialization production of fermented tea is laid a good foundation; The fermented tea drink of preparation is brisk taste not only, and contains the multiple efficacies composition, is a kind of microbial fermentation drink that the value utilized is arranged very much.
Of the present inventionly utilize pure bacterium mixed culture fermentation to produce black-tea fungus drink: adopt saccharomyces cerevisiae, acetic acid acetobacter and three kinds of bacterium of Lactobacillus plantarum composite, black-tea fungus drink is produced in mixed culture fermentation; The proportioning of described saccharomyces cerevisiae, acetic acid acetobacter and Lactobacillus plantarum, saccharomyces cerevisiae: acetic acid acetobacter: (proportioning of described three kinds of bacterium is: various bacterial cell concentration are controlled at 10 in Lactobacillus plantarum=1: 1: 1～3
8Volume ratio during cfu/mL); Three kinds of total inoculum concentrations of bacterium are: 5%～10% by volume; Described fermentation condition is: fermented tea fermentation culture liquid amount: the 400mL zymotic fluid is packed in the 500mL bottle, rotating speed 100r/min, and 30 ℃ of shaken cultivation, with this understanding, and fermentation 55-60h, the pH value stabilization of fermented tea is between 3.0-3.2.
Remarkable advantage of the present invention is: the present invention is by studying functional black-tea fungus drink: at first will separate obtaining the fermented tea pure culture, and the classification position of understanding bacterial classification, could solve the bacterial classification holding time, inoculum concentration control, anti-pollution in advance, keep many-sided problems such as spawn activity and control product quality, just might be to the various metabolic characteristics of each bacterial classification and their various combinations, condition of culture, zymotic fluid composition and possible functional effect carry out intensive research, by separating voluntarily, screening obtains three kinds of pure cultures: saccharomyces cerevisiae (Saccharomyces cerevisiae Meyen ex Hansen), acetic acid acetobacter (Acetobacter aceti) and Lactobacillus plantarum (Lactobacillus plantarum).Utilize composite, the mixed culture fermentation of pure culture, and fermented tea culture medium prescription, technological condition for fermentation are optimized, make fermentation time shorten to 60 hours from traditional 10 days; Owing to adopt sterile fermentation, avoided living contaminants, guaranteed product safety; Simultaneously because management methods such as the bacterial classification cultivation of formation one cover science, rejuvenation, preservation have been stablized product quality, for the industrialization production of fermented tea is laid a good foundation.The fermented tea drink of the present invention's development not only has sour-sweet suitable, pleasant to the palate local flavor, and contains the multiple efficacies composition, is a kind of microbial fermentation drink that the value utilized is arranged very much.
The specific embodiment
1, bacterial screening is identified
Screening obtains saccharomycete, acetic acid bacteria and lactic acid bacteria from food such as fermented tea, pickles, further separation and purification, identify that through Institute of Microorganism, Academia Sinica three kinds of bacterium are respectively saccharomyces cerevisiae (Saccharomyces cerevisiae Meyen exHansen), acetic acid acetobacter (Acetobacter aceti) and Lactobacillus plantarum (Lactobacillus plantarum).
2, the fermented tea fermentation medium is optimized
With red tea fungus fermented date liquid pH value and local flavor is index, determines that the fermented tea fermentative medium formula is: sucrose 5g, black tea powder 0.2g, water 100mL.Wherein black tea powder is provided by big Fujian, ZhangZhou, Fujian Food Co., Ltd.
3, three kinds of bacterium proportionings and inoculum concentration determines
Saccharomyces cerevisiae 2-3 articulating after the picking activation is gone in the fluid nutrient medium, places under 30 ℃, and 100r/min shaken cultivation 32-36h, the Liquid Culture based formulas of described saccharomyces cerevisiae is: yeast extract 1g, peptone 1g, glucose 2g, water 100mL; Acetic acid acetobacter 2-3 articulating after the picking activation is gone into fluid nutrient medium, places under 30 ℃, and 200r/min shaken cultivation 32-36h, the Liquid Culture based formulas of described acetic acid acetobacter is: yeast extract 1g, glucose 1g, water 100mL, absolute ethyl alcohol 3mL; Lactobacillus plantarum 2-3 articulating after the picking activation is gone into the MRS fluid nutrient medium, place under 30 ℃, leave standstill and cultivate 32-36h, the prescription of described MRS fluid nutrient medium is: peptone 1g, beef extract 1g, yeast extract 0.5g, glucose 2g, sodium acetate 0.5g, Tween 80 0.1mL, citric acid diamines 0.2g, MgSO
47H
2O 0.2g, MnSO
44H
2O 0.005g, K
2HPO
40.2g, water 100mL, pH6.2-6.6.Cultivate and finish back mensuration absorbance OD separately
580(is reference with the blank culture medium of liquid separately).Concentration (10 according to three kinds of bacterium
8Cfu/mL) and OD
580Linear relationship and the actual OD of three kinds of bacterium
580, calculate inoculum concentration, insert in the fermented tea fermentation medium and cultivate.
Conventional oese is adopted in inoculation, and ring diameter is at 2-3mm; Conventional method activation is all adopted in the activation of three kinds of bacterium.
The proportioning of saccharomyces cerevisiae, acetic acid acetobacter and Lactobacillus plantarum, saccharomyces cerevisiae: acetic acid acetobacter: (proportioning of described three kinds of bacterium is: various bacterial cell concentration are controlled at 10 in Lactobacillus plantarum=1: 1: 1～3
8Volume ratio during cfu/mL); Three kinds of total inoculum concentrations of bacterium are: 5%～10% by volume.
And adopt orthogonal experiment to study the proportioning and the inoculum concentration of three kinds of bacterium the bests.
Adopt orthogonal experiment research inoculum concentration (5%, 7.5%, 10%) and three kinds of bacterium proportionings (saccharomycete: acetic acid bacteria: lactic acid bacteria=1: 1: 1,1: 1: 2,1: 1: 3) influence to the fermented tea ferment effect, comprehensive zymotic fluid pH value and local flavor are considered, determine the best proportioning of three kinds of bacterium, saccharomyces cerevisiae: acetic acid acetobacter: (proportioning of described three kinds of bacterium is: various bacterial cell concentration are controlled at 10 to Lactobacillus plantarum=1: 1: 1
8Volume ratio during cfu/mL); Best total inoculum concentration is: 5% by volume.
4, optimization of fermentation condition
On above-mentioned research basis, adopt orthogonal experiment L
9(4
3) determine that further the optimum technological condition for fermentation of fermented tea is: fermented tea fermentation culture liquid amount: the 400mL zymotic fluid is packed in the 500mL bottle, rotating speed 100r/min, 30 ℃ of shaken cultivation, with this understanding, fermentation 55-60h, the pH value stabilization of fermented tea has sour-sweet suitable, pleasant to the palate local flavor simultaneously between 3.0-3.2.
Most preferred embodiment
1, seed culture
Saccharomyces cerevisiae 2-3 articulating after the picking activation is gone in the fluid nutrient medium (yeast extract 1g, peptone 1g, glucose 2g, water 100mL), places under 30 ℃, and 100r/min shaken cultivation 36h measures OD
580Acetic acid acetobacter 2-3 articulating after the picking activation is gone into fluid nutrient medium (yeast extract 1g, glucose 1g, water 100mL, absolute ethyl alcohol 3mL), places under 30 ℃, and 200r/min shaken cultivation 32h measures OD
580Lactobacillus plantarum 2-3 articulating after the picking activation is gone into MRS fluid nutrient medium (peptone 1g, beef extract 1g, yeast extract 0.5g, glucose 2g, sodium acetate 0.5g, Tween 80 0.1mL, citric acid diamines 0.2g, MgSO
47H
2O 0.2g, MnSO
44H
2O 0.005g, K
2HPO
40.2g, water 100mL, pH6.2-6.6), place under 30 ℃, leave standstill and cultivate 32h, measure OD
580
2, the fermentation of fermented tea
Measure three kinds of bacterium seed liquor OD respectively
580, and according to saccharomyces cerevisiae: the acetic acid acetobacter: (proportioning of described three kinds of bacterium is: various bacterial cell concentration are controlled at 10 to Lactobacillus plantarum=1: 1: 1
8Volume ratio during cfu/mL), total inoculum concentration (volume ratio) 5%, calculate the inoculum concentration of various bacterium, insert fermented tea culture medium (sucrose 5g, black tea powder 0.2g, water 100mL), fermented tea fermentation culture liquid amount: the 400mL zymotic fluid is packed in the 500mL bottle, place under 30 ℃, 100r/min shaken cultivation 60h, the zymotic fluid pH value that obtains is 3.14.
3, the mensuration of fermented tea functional component
By 3 batches of red tea fungus fermented date liquids of above-mentioned process conditions preparation, measure main functional component content wherein.The average content of glucose, total acid, polysaccharide, glucuronic acid, Tea Polyphenols and ethanol is respectively 34.95g/L, 22.14g/L, 0.74g/L, 5.35g/L, 0.047g/L, 8.79g/L in the red tea fungus fermented date liquid.
Embodiment 2
(1) seed culture: the saccharomyces cerevisiae 2-3 articulating after the picking activation is gone in the fluid nutrient medium, places under 30 ℃, and 100r/min shaken cultivation 32h, the Liquid Culture based formulas of described saccharomyces cerevisiae is: yeast extract 1g, peptone 1g, glucose 2g, water 100mL; Acetic acid acetobacter 2-3 articulating after the picking activation is gone into fluid nutrient medium, places under 30 ℃, and 200r/min shaken cultivation 32h, the Liquid Culture based formulas of described acetic acid acetobacter is: yeast extract 1g, glucose 1g, water 100mL, absolute ethyl alcohol 3mL; Lactobacillus plantarum 2-3 articulating after the picking activation is gone into the MRS fluid nutrient medium, place under 30 ℃, leave standstill and cultivate 32h, the prescription of described MRS fluid nutrient medium is: peptone 1g, beef extract 1g, yeast extract 0.5g, glucose 2g, sodium acetate 0.5g, Tween 80 0.1mL, citric acid diamines 0.2g, MgSO
47H
2O 0.2g, MnSO
44H
2O 0.005g, K
2HPO
40.2g, water 100mL, pH6.2-6.6;
(2) fermentation of fermented tea: according to the proportioning of described saccharomyces cerevisiae, acetic acid acetobacter and Lactobacillus plantarum is that (proportioning of described three kinds of bacterium is: various bacterial cell concentration were controlled at 10 in 1: 1: 2
8Volume ratio during cfu/mL) and total inoculum concentration (volume ratio) be 7.5%, calculate the inoculum concentration of various bacterium, inserting fermented tea fermentation culture (sucrose 5g, black tea powder 0.2g, water 100mL) ferments, fermentation condition is: fermented tea fermentation culture liquid amount: the 400mL zymotic fluid is packed in the 500mL bottle, rotating speed 100r/min, 30 ℃ of shaken cultivation, with this understanding, fermentation 55-60h, the pH value of fermented tea is 3.12.
Embodiment 3
(1) seed culture: the saccharomyces cerevisiae 2-3 articulating after the picking activation is gone in the fluid nutrient medium, places under 30 ℃, and 100r/min shaken cultivation 34h, the Liquid Culture based formulas of described saccharomyces cerevisiae is: yeast extract 1g, peptone 1g, glucose 2g, water 100mL; Acetic acid acetobacter 2-3 articulating after the picking activation is gone into fluid nutrient medium, places under 30 ℃, and 200r/min shaken cultivation 35h, the Liquid Culture based formulas of described acetic acid acetobacter is: yeast extract 1g, glucose 1g, water 100mL, absolute ethyl alcohol 3mL; Lactobacillus plantarum 2-3 articulating after the picking activation is gone into the MRS fluid nutrient medium, place under 30 ℃, leave standstill and cultivate 35h, the prescription of described MRS fluid nutrient medium is: peptone 1g, beef extract 1g, yeast extract 0.5g, glucose 2g, sodium acetate 0.5g, Tween 80 0.1mL, citric acid diamines 0.2g, MgSO
47H
2O 0.2g, MnSO
44H
2O 0.005g, K
2HPO
40.2g, water 100mL, pH6.2-6.6;
(2) fermentation of fermented tea: according to the weight proportion of described saccharomyces cerevisiae, acetic acid acetobacter and Lactobacillus plantarum is that (proportioning of described three kinds of bacterium is: various bacterial cell concentration were controlled at 10 in 1: 1: 3
8Volume ratio during cfu/mL) and total inoculum concentration (volume ratio) be 10%, inserting the fermented tea fermentation culture ferments, fermentation condition is: fermented tea fermentation culture liquid amount: the 400mL zymotic fluid is packed in the 500mL bottle, rotating speed 100r/min, 30 ℃ of shaken cultivation, with this understanding, fermentation 55-60h, the pH value of fermented tea is 3.17.
Claims (4)
1. a method of utilizing pure bacterium mixed culture fermentation to produce black-tea fungus drink is characterized in that: adopt saccharomyces cerevisiae, acetic acid acetobacter and three kinds of bacterium of Lactobacillus plantarum composite, mixed culture fermentation production black-tea fungus drink; The proportioning of described saccharomyces cerevisiae, acetic acid acetobacter and Lactobacillus plantarum: saccharomyces cerevisiae: acetic acid acetobacter: Lactobacillus plantarum=1: 1: 1～3, the proportioning of described three kinds of bacterium is: various bacterial cell concentration are controlled at 10
8Volume ratio during cfu/mL; Three kinds of total inoculum concentrations of bacterium are: 5%～10% by volume; Described fermentation condition is: fermented tea fermentation medium liquid amount: the 400mL zymotic fluid is packed in the 500mL bottle, rotating speed 100r/min, and 30 ℃ of shaken cultivation, with this understanding, and fermentation 55～60h, the pH value stabilization of fermented tea is between 3.0-3.2.
2. the method for utilizing pure bacterium mixed culture fermentation to produce black-tea fungus drink according to claim 1, it is characterized in that: the proportioning of described saccharomyces cerevisiae, acetic acid acetobacter and Lactobacillus plantarum, saccharomyces cerevisiae: acetic acid acetobacter: Lactobacillus plantarum=1: 1: 1, the proportioning of described three kinds of bacterium is: various bacterial cell concentration are controlled at 10
8Volume ratio during cfu/mL; Three kinds of total inoculum concentrations of bacterium are: 5% by volume.
3. the method for utilizing pure bacterium mixed culture fermentation to produce black-tea fungus drink according to claim 1, it is characterized in that: described fermented tea fermentative medium formula is: sucrose 5g, black tea powder 0.2g, water 100mL.
4. according to claim 1, the 2 or 3 described methods of utilizing pure bacterium mixed culture fermentation to produce black-tea fungus drink, it is characterized in that: concrete steps are:
(1) seed culture: the saccharomyces cerevisiae 2-3 articulating after the picking activation is gone in the fluid nutrient medium, place under 30 ℃, 100r/min shaken cultivation 32-36h, the Liquid Culture based formulas of described saccharomyces cerevisiae is: yeast extract 1g, peptone 1g, glucose 2g, water 100mL; Acetic acid acetobacter 2-3 articulating after the picking activation is gone into fluid nutrient medium, places under 30 ℃, and 200r/min shaken cultivation 32-36h, the Liquid Culture based formulas of described acetic acid acetobacter is: yeast extract 1g, glucose 1g, water 100mL, absolute ethyl alcohol 3mL; Lactobacillus plantarum 2-3 articulating after the picking activation is gone into the MRS fluid nutrient medium, place under 30 ℃, leave standstill and cultivate 32-36h, the prescription of described MRS fluid nutrient medium is: peptone 1g, beef extract 1g, yeast extract 0.5g, glucose 2g, sodium acetate 0.5g, Tween 80 0.1mL, citric acid diamines 0.2g, MgSO
47H
2O 0.2g, MnSO
44H
2O 0.005g, K
2HPO
40.2g, water 100mL, pH6.2-6.6;
(2) fermentation of fermented tea: according to proportioning and total inoculum concentration of described saccharomyces cerevisiae, acetic acid acetobacter and Lactobacillus plantarum, calculate the inoculum concentration of various bacterium, inserting the fermented tea fermentation culture ferments, fermentation condition is: fermented tea fermentation culture liquid amount: the 400mL zymotic fluid is packed in the 500mL bottle, rotating speed 100r/min, 30 ℃ of shaken cultivation, with this understanding, fermentation 55-60h, the pH value stabilization of fermented tea is between 3.0-3.2; The concentration of three kinds of bacterium seed liquor is 10
8Cfu/mL.
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| CN115141774B (en) * | 2022-07-11 | 2024-01-30 | 澄江老仓醋食品科技有限公司 | Acetobacter aceti, acquisition method, preparation method of composite freeze-dried starter and application of composite freeze-dried starter in vinegar preparation |
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