CN103859122A - Maotai flavored coffee and preparation method thereof - Google Patents
Maotai flavored coffee and preparation method thereof Download PDFInfo
- Publication number
- CN103859122A CN103859122A CN201410128974.8A CN201410128974A CN103859122A CN 103859122 A CN103859122 A CN 103859122A CN 201410128974 A CN201410128974 A CN 201410128974A CN 103859122 A CN103859122 A CN 103859122A
- Authority
- CN
- China
- Prior art keywords
- coffee
- maotai
- aroma
- screening
- ferfas
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Tea And Coffee (AREA)
Abstract
The invention belongs to the technical field of foods, and particularly relates to a Maotai flavored coffee which comprises 1-99.99 parts by weight of coffee and 0.01-30 parts by weight of Maotai wine flavored microorganism or sauce-producing functional bacterium. The invention provides a preparation method of the Maotai flavored coffee. The Maotai flavored coffee has a mellow wine flavor and contains physiological activators such as caffeine in the coffee. The coffee meets the pursue of modern to the health, and is capable of meeting the demand for wine flavor when people drink the Maotai flavored coffee.
Description
Technical field
The invention belongs to food technology field, be specifically related to a kind of state aroma coffee and preparation method thereof.
Background technology
Coffee smell is aromatic, bitter alcohol and, there is arousing brain, raising spirit, effect of stomach invigorating beauty treatment, drink after long times of aftertaste, coffee is gradually popular to people in China, is particularly subject to liking of young man, consumption figure rises increasingly.Wine has long history, has warm taste, promoting blood circulation, flourish effect of supporting skin, and responsible drinking can be excited spiritual, and active atmosphere can damage health but drink beyond one's capacity, especially unfavorable to liver.Coffee perfume (or spice), aroma are melted into a whole, and effect complementation, is a kind of graceful health drink of enjoying.
Summary of the invention
The object of the invention is to utilize fermentation technique that a kind of state aroma coffee and manufacture method thereof are provided.Under process conditions of the present invention, make the state's aroma coffee that not only there is mellow aroma but also comprise the physiological activators such as the caffeine in coffee.This coffee meets modern to healthy pursuit, can in drink coffee, also meet the needs to aroma.
The present invention is achieved through the following technical solutions.
A kind of state aroma coffee, is characterized in that by 1-99.99 weight portion coffee, 0.01-30 weight portion Maotai producing mellow microorganism or produces the fragrant function bacterium of sauce forming.
Described coffee is one or more in coffee bean, coffee bean particle, ground coffee, instant coffee powder.
Described Maotai producing mellow microorganism is to produce one or more compositions in the Aspergillus obtaining with separation and purification Daqu, white mucor, rhizopus, pears head mould, false yeast, endomycopsis, Hansenula yeast, lactic acid bacteria, acetic acid bacteria, bacillus licheniformis, bacillus subtilis, bacillus alcalophilus, cured shape bacillus, bacillus pumilus, bacillus amyloliquefaciens, bafillus natto from Maotai.
Described Maotai producing mellow microorganism is prepared by following methods:
Get the bent powder of 1-10 weight portion Maotai high-temperature daqu and add in the 90-99 weight portion sterilized water or SPSS that bead is housed, 1-10 hour vibrates on 10-70 ℃, 10-200rpm shaking table; Gradient dilution spread plate, cultivates 1-90 hour in 10-70 ℃; From flat board select growth better, the different bacterium colonies of proterties are inoculated in respectively isolation medium inclined-plane, as screening bacterial strain; The screening obtaining is inoculated respectively in screening and culturing base with bacterial strain, cultivated 1-90 hour for 10-70 ℃, the fragrant dominant strain of the product obtaining is Maotai producing mellow microorganism;
The Maotai producing mellow microorganism screening is bacillus, gemma clostridium, pseudomonas, Aspergillus, Penicillium notatum, saccharomycete, and bacterial classification can be bacterium song or bacterium liquid form;
Described isolation medium is coffee 0-300g/L, glucose 1-200g/L, tryptone 1-50g/L, yeast extract 1-50g/L, sodium chloride 0-50g/L, agar 1-50g/L, and pH4-8 removes agar and is screening and culturing base.
The preparation method of a kind of state aroma coffee:
Step 1: get Maotai producing mellow microorganism and be inoculated on ready slant medium, be positioned over and in the constant incubator of 10-70 ℃, cultivate 1-90h and make actication of culture, again the bacterial classification of activation is inoculated in the LB fluid nutrient medium that adds sugared source, in 10-70 ℃ of constant temperature culture 1-90h, by medium centrifugal reject supernatant, precipitation thalline is even with normal saline dilution concussion, is the activated seed liquid obtaining; Described slant medium is beef extract-peptone, PDA culture medium or minimal medium;
Step 2: coffee is pulverized and put into the compound protease of 20-80 ℃ after homogenate and/or the mixed liquor of flavor protease soaks 1-90h, and water in mass ratio 1: 0.1-30 refluxing extraction was filtered after 1-8 hour, at 115 ℃ of sterilizing 100min; In above-mentioned mixed liquor, add and account for the sugared source of above-mentioned mixed liquor weight 0.1-10%, the inorganic salts of 0.1-10%, compound amino acid and/or the vitamin C of 0-10%, obtain coffee fluid nutrient medium;
Step 3: after resuspended to the wet thallus of Maotai producing mellow microorganism, pulvis or freeze-dried powder, according in the coffee fluid nutrient medium of inoculum concentration access step 2 gained of 1-20% at temperature 10-60 ℃ of bottom fermentation 1-20 days, after filtering with microporous membrane degerming, continue fermentation 1-10 days, after fermentation ends, be incubated 0.1-50h at 55-120 ℃; By the coffee zymotic fluid fermenting, sheet frame membrane filtration after bacterial classification deactivation, reduced pressure concentration or freeze drying are condensed into paste again, according to 0: the mass ratio of 1-10 mixes paste mutually with extractant, paste is dissolved fully, after refiltering, concentrate filtrate to except after desolventizing, obtain fermentation of coffee medicinal extract; Fermentation of coffee medicinal extract is cured to 1-50min in 10-200 ℃, pack in airtight oak barrel after cooling and store and within 1-90 days, obtain state's aroma coffee; Described extract, extractant are water or ethanol, or both composite solutions.
The preparation method of a kind of state aroma coffee:
Step 1: get the ascocarpous intermediate structure piece of fresh ferfas, skin, epidermis, epidermis soil, the each 5-10 weight portion of mycelia and ferfas and host's exotrophic mycorrhiza in soil, after sterilization, wear into pulpous state with aseptic mortar, join and in sterile distilled water, shake 10-60min, therefrom getting 100-1000 μ L fluid drips is added on screening and culturing base evenly and is coated with and opens, under aerobic or anaerobic condition, be coated with after flat band method is purified and obtain a collection of microbial strains with plate streaking separation or dilution, then carry out fermenting experiment with liquid screening culture medium and therefrom filter out the rich bacterial strain that produces alpha-androstanol; Screening and culturing base is: glucose or sucrose 5-300g/L, peptone 1-100g/L, yeast extract 0.1-20g/L, potassium dihydrogen phosphate 0.1-10g/L, magnesium sulfate 0.01-10g/L, oak leaf or Oak Tree loosen callus lines 1-200g/L, agar 10-30g/L, pH value 4-9, removes agar and is liquid screening culture medium;
Step 2: will ferment in access liquid screening culture medium after the activation of ferfas aroma former, at 15-40 ℃, speed of agitator is to cultivate 1-30 days under 10-800rpm condition; After fermentation ends, by tunning homogenate, add one or more of cellulase, protease, beta-glucosidase, enzymolysis 1-150min going out after enzyme under 10-85 ℃ of condition, obtains ferfas aroma former zymotic fluid; Described ferfas aroma former is one or more of the bacterial classification, the ferfas spore that separate of ferfas ascocarp tissue, ferfas endophyte, ferfas fungal component, ferfas concomitance bacterium;
Step 3: coffee is pulverized and put into the pepsin of 20-80 ℃ after homogenate and the mixed liquor of hydrochloric acid soaks 1-90h, then after mixing with ferfas aroma former zymotic fluid, the loose resuspended liquid of capsule bacterium of access Bifidobacterium Bifidum and the resuspended mixed liquor of lactobacillus acidophilus or coronoid process is in 20-70 ℃ of fermentation 1-120h, after fermentation at 115 ℃ sterilizing 10-20min; In above-mentioned zymotic fluid, add and account for the sugared source of above-mentioned mixed liquor 0.1%-10%, the inorganic salts of 0.1%-10%, compound amino acid and/or the vitamin C of 0.1%-10%, obtain coffee culture medium;
Step 4: after resuspended to the wet thallus of Maotai producing mellow microorganism, pulvis or freeze-dried powder, according in the coffee culture medium of inoculum concentration access step 3 gained of 1-20% at temperature 10-60 ℃ of bottom fermentation 1-20 days, after filtering with microporous membrane degerming, continue fermentation 1-10 days, after fermentation ends, be incubated 0.1-50h at 55-120 ℃; At the centrifugal 1-20min of 3000-8000rpm, by supernatant, by solvent extraction the concentrated extract A that obtains, filter residue is by solvent extraction the concentrated extract B that obtains; After above-mentioned extract A and B are mixed, obtain Maotai and cause fragrant liquid, add respectively emulsifying agent and embedded material, after homogeneous, the dry or freeze drying of spraying makes Maotai and causes fragrant liquid powder; By fluid bed, Maotai is caused to the water-soluble dressing of fragrant liquid powder spray or sustained release coating, make Maotai and cause fragrant liquid microcapsule powder; Described emulsifying agent is one or more of monoglyceride, sucrose ester, phosphatide, starch ester; Described embedded material is one or more of converted starch, maltodextrin, sucrose, shitosan, Arabic gum, cyclodextrin, carragheen, alginate, gelatin, soybean protein, lactalbumin, protein hydrolysate; Described solvent extraction mode is under room temperature, to leave standstill the one of soaking in extraction, mechanical oscillation extraction, ultrasonic extraction, heating reflux extraction, microwave heating extraction, distillation extraction, while distillation extraction, Soxhlet extraction or supercritical fluid extraction; Solvent is any one or a few in water, ethanol, propane diols, glycerine, ether, ethyl acetate, acetonitrile, acetone, carrene, chloroform, cyclohexane, carbon dioxide; Described concentration process is that under normal pressure, distillation and concentration, nitrogen blow the one in concentrated or reduced pressure concentration;
Step 5: the Maotai of step 4 gained is caused to the granulation or mix to pack while hot in oak barrel balance 1-90 days into after dry and pack afterwards and make the coffee product with state's aroma together with ground coffee of fragrant liquid microcapsule powder.
In described step 1, bacterial strain is carried out ultraviolet monochlor(in)ate lithium complex mutation and is beneficial to the raising of alpha-androstanol content.
Described inorganic salts are KH
2pO
4, (NH4)
2sO
4, MgSO
4, or NaCl.
Described sugared source is one or more the mixture in sucrose, glucose, wood sugar, fructose.
Preparation Example
Embodiment 1
A kind of state aroma coffee, is made up of 0.1g Aspergillus, 10g coffee bean.
The preparation method of Aspergillus:
Get the bent powder of 1g Maotai high-temperature daqu and add in the 99g sterilized water that bead is housed, on 10 ℃, 10rpm shaking table, vibrate 1 hour; Gradient dilution spread plate, cultivates 1 hour in 10 ℃; From flat board select growth better, the different bacterium colonies of proterties are inoculated in respectively isolation medium inclined-plane, as screening bacterial strain; The screening obtaining is inoculated respectively in screening and culturing base with bacterial strain, cultivated 1 hour, obtain Aspergillus and be Maotai producing mellow microorganism for 10 ℃;
Described isolation medium is glucose 1g/L, tryptone 1g/L, yeast extract 1g/L, agar 1g/L, and pH4 removes agar and is screening and culturing base.
The preparation method of state's aroma coffee
Step 1: get Aspergillus and be inoculated on ready beef extract-peptone, be positioned over and in the constant incubator of 10 ℃, cultivate 1h and make actication of culture, again the bacterial classification of activation is inoculated in the LB fluid nutrient medium that adds wood sugar, in 10 ℃ of constant temperature culture 1h, by medium centrifugal reject supernatant, precipitation thalline is even with normal saline dilution concussion, is the activated seed liquid obtaining;
Step 2: coffee is pulverized and is put into the compound protease of 20 ℃ after homogenate and soak 1h, with water in mass ratio 1: 0.1 refluxing extraction after 1 hour, filter, at 115 ℃ of sterilizing 100min; In above-mentioned mixed liquor, add and account for above-mentioned mixed liquor weight 0.1% sucrose, 0.1%KH
2pO
4, obtain coffee fluid nutrient medium;
Step 3: after resuspended Aspergillus wet thallus, according in the coffee fluid nutrient medium of 1% inoculum concentration access step 2 gained 10 ℃ of bottom fermentations of temperature 1 day, after filtering with microporous membrane degerming, continue fermentation 1 day, after fermentation ends at 55 ℃ of insulation 0.1h; By the coffee zymotic fluid fermenting, sheet frame membrane filtration after bacterial classification deactivation, then reduced pressure concentration becomes paste, mass ratio according to 1: 1 is mixed with water by paste, paste is dissolved fully, after refiltering, concentrate filtrate to except after desolventizing, obtain fermentation of coffee medicinal extract; Fermentation of coffee medicinal extract is cured to 1min in 10 ℃, pack in airtight oak barrel after cooling and store and within 1 day, obtain state's aroma coffee.
Embodiment 2
A kind of state aroma coffee, is made up of 0.01g bacillus licheniformis, 10g ground coffee.
Bacillus licheniformis is prepared by following methods:
Get the bent powder of 10g Maotai high-temperature daqu and add in the 90g SPSS that bead is housed, on 70 ℃, 200rpm shaking table, vibrate 10 hours; Gradient dilution spread plate, cultivates 90 hours in 70 ℃; From flat board select growth better, the different bacterium colonies of proterties are inoculated in respectively isolation medium inclined-plane, as screening bacterial strain; The screening obtaining is inoculated respectively in screening and culturing base with bacterial strain, cultivated 90 hours, obtain bacillus licheniformis and be Maotai producing mellow microorganism for 70 ℃;
Described isolation medium is coffee 300g/L, glucose 200g/L, tryptone 50g/L, yeast extract 50g/L, sodium chloride 50g/L, agar 50g/L, and pH8 removes agar and is screening and culturing base.
State's aroma coffee preparation method
Step 1: get bacillus licheniformis and be inoculated on ready PDA culture medium, be positioned over and in the constant incubator of 70 ℃, cultivate 90h and make actication of culture, again the bacterial classification of activation is inoculated in the LB fluid nutrient medium that adds sucrose, in 70 ℃ of constant temperature culture 90h, by medium centrifugal reject supernatant, precipitation thalline is even with normal saline dilution concussion, is the activated seed liquid obtaining;
Step 2: coffee is pulverized and is put into the compound protease of 80 ℃ after homogenate and the mixed liquor of flavor protease soaks 90h, with water in mass ratio 1: 30 refluxing extraction after 8 hours, filter, at 115 ℃ of sterilizing 100min; In above-mentioned mixed liquor, add and account for the glucose, 10%NaCl of above-mentioned mixed liquor weight 10%, 10% compound amino acid, obtain coffee fluid nutrient medium;
Step 3: after resuspended bacillus licheniformis pulvis, according in the coffee fluid nutrient medium of 20% inoculum concentration access step 2 gained temperature 60 C bottom fermentation 20 days, after filtering with microporous membrane degerming, continue fermentation 10 days, after fermentation ends at 120 ℃ of insulation 50h; By the coffee zymotic fluid fermenting, sheet frame membrane filtration after bacterial classification deactivation, then freeze drying is condensed into paste, according to the mass ratio of 1: 10, paste is mixed mutually with ethanol, paste is dissolved fully, after refiltering, concentrate filtrate to except after desolventizing, obtain fermentation of coffee medicinal extract; Fermentation of coffee medicinal extract is cured to 50min in 200 ℃, pack in airtight oak barrel after cooling and store and within 90 days, obtain state's aroma coffee.
Embodiment 3
A kind of state aroma coffee, is made up of 1g bacillus subtilis, 20g coffee.
The preparation method of bacillus subtilis:
Get the bent powder of 5g Maotai high-temperature daqu and add in the 30g sterilized water that bead is housed, on 30 ℃, 100rpm shaking table, vibrate 4 hours; Gradient dilution spread plate, cultivates 10 hours in 30 ℃; From flat board select growth better, the different bacterium colonies of proterties are inoculated in respectively isolation medium inclined-plane, as screening bacterial strain; The screening obtaining is inoculated respectively in screening and culturing base with bacterial strain, cultivated 40 hours for 30 ℃, the bacillus subtilis obtaining is Maotai producing mellow microorganism;
Described isolation medium is coffee 100g/L, glucose 20g/L, tryptone 10g/L, yeast extract 20g/L, sodium chloride 10g/L, agar 30g/L, and pH4.8 removes agar and is screening and culturing base.
The preparation method of state's aroma coffee:
Step 1: get bacillus subtilis and be inoculated on ready minimal medium, be positioned over and in the constant incubator of 30 ℃, cultivate 10h and make actication of culture, again the bacterial classification of activation is inoculated in the LB fluid nutrient medium that adds fructose, in 20 ℃ of constant temperature culture 10h, by medium centrifugal reject supernatant, precipitation thalline is even with normal saline dilution concussion, is the activated seed liquid obtaining;
Step 2: coffee is pulverized to the mixed liquor of putting into the flavor protease of 30 ℃ after homogenate and soaks 10h, with water in mass ratio 1: 10 refluxing extraction after 3 hours, filter, at 115 ℃ of sterilizing 100min; In above-mentioned mixed liquor, add and account for the wood sugar of above-mentioned mixed liquor weight 3%, 4% (NH4)
2sO
4, 1% compound amino acid and 2% vitamin C, obtain coffee fluid nutrient medium;
Step 3: after resuspended bacillus subtilis freeze-dried powder, according in the coffee fluid nutrient medium of 2% inoculum concentration access step 2 gained 30 ℃ of bottom fermentations of temperature 5 days, after filtering with microporous membrane degerming, continue fermentation 2 days, after fermentation ends at 70 ℃ of insulation 0.5h; By the coffee zymotic fluid fermenting, sheet frame membrane filtration after bacterial classification deactivation, freeze drying is condensed into paste again, according to the mass ratio of 1: 3, paste is mixed mutually with 70% ethanolic solution, paste is dissolved fully, after refiltering, concentrate filtrate to except after desolventizing, obtain fermentation of coffee medicinal extract; Fermentation of coffee medicinal extract is cured to 10min in 100 ℃, pack in airtight oak barrel after cooling and store and within 10 days, obtain state's aroma coffee.
Embodiment 4
A kind of state aroma coffee, is made up of 4g bacillus alcalophilus, 99g coffee.
Bacillus alcalophilus preparation method:
Get the bent powder of 10g Maotai high-temperature daqu and add in the 90g SPSS that bead is housed, on 20 ℃, 50rpm shaking table, vibrate 3 hours; Gradient dilution spread plate, cultivates 30 hours in 30 ℃; From flat board select growth better, the different bacterium colonies of proterties are inoculated in respectively isolation medium inclined-plane, as screening bacterial strain; The screening obtaining is inoculated respectively in screening and culturing base with bacterial strain, cultivated 20 hours for 20 ℃, the bacillus alcalophilus strain obtaining is Maotai producing mellow microorganism;
Described isolation medium is coffee 50g/L, glucose 20g/L, tryptone 15g/L, yeast extract 10g/L, sodium chloride 5g/L, agar 20g/L, and pH5 removes agar and is screening and culturing base.
State's aroma coffee preparation method
Step 1: get the ascocarpous intermediate structure piece of fresh ferfas, skin, epidermis, epidermis soil, the each 5g of mycelia and ferfas and host's exotrophic mycorrhiza in soil, after sterilization, wear into pulpous state with aseptic mortar, join and in sterile distilled water, shake 10min, therefrom getting 100 μ L fluid drips is added on screening and culturing base evenly and is coated with and opens, under aerobic conditions, separate and obtain a collection of microbial strains with plate streaking, then carry out fermenting experiment with liquid screening culture medium and therefrom filter out the rich bacterial strain that produces alpha-androstanol; Screening and culturing base is: glucose 5g/L, peptone 1g/L, yeast extract 0.1g/L, potassium dihydrogen phosphate 0.1g/L, magnesium sulfate 0.01g/L, oak leaf 1g/L, agar 10g/L, and pH value 4, removes agar and is liquid screening culture medium;
Step 2: will ferment in access step 1 gained liquid screening culture medium after the activation of ferfas concomitance bacterium, at 40 ℃, speed of agitator is to cultivate under 10rpm condition 1 day; After fermentation ends, by tunning homogenate, add cellulase, enzymolysis 1min going out after enzyme under 10 ℃ of conditions, obtains ferfas aroma former zymotic fluid;
Step 3: coffee is pulverized and put into the pepsin of 20 ℃ after homogenate and the mixed liquor of hydrochloric acid soaks 1h, then access Bifidobacterium Bifidum and the resuspended mixed liquor of lactobacillus acidophilus after mixing with step 2 gained ferfas aroma former zymotic fluid, in 20 ℃ of fermentation 1h, after fermentation at 115 ℃ sterilizing 10min, obtain coffee culture medium;
Step 4: after resuspended bacillus alcalophilus wet thallus, according in 1% inoculum concentration access step 3 gained coffee culture medium 10 ℃ of bottom fermentations of temperature 1 day, after filtering with microporous membrane degerming, continue fermentation 1 day, after fermentation ends at 55 ℃ of insulation 0.1h; At the centrifugal 1min of 3000rpm, by supernatant at room temperature water leave standstill to soak extract and carry out under normal pressure distillation and concentration and obtain extract A, filter residue at room temperature water leaves standstill to soak and extracts also reduced pressure concentration and obtain extract B; After above-mentioned extract A and B are mixed, obtain Maotai and cause fragrant liquid, add respectively monoglyceride and Arabic gum, after homogeneous, spraying is dry that Maotai causes fragrant liquid powder; By fluid bed, Maotai is caused to the water-soluble dressing of fragrant liquid powder spray, make Maotai and cause fragrant liquid microcapsule powder;
Step 5: the Maotai of step 4 gained is caused to fragrant liquid microcapsule powder and pack while hot in oak barrel packing after balance 1d after granulating and drying into make the coffee product with state's aroma together with ground coffee.
Embodiment 5
A kind of state aroma coffee, is made up of 25g bacillus amyloliquefaciens, 80g coffee.
Endomycopsis preparation method:
Get the bent powder of 4g Maotai high-temperature daqu and add in the 96g SPSS that bead is housed, on 60 ℃, 80rpm shaking table, vibrate 5 hours; Gradient dilution spread plate, cultivates 80 hours in 30 ℃; From flat board select growth better, the different bacterium colonies of proterties are inoculated in respectively isolation medium inclined-plane, as screening bacterial strain; The screening obtaining is inoculated respectively in screening and culturing base with bacterial strain, cultivated 55 hours for 35 ℃, the endomycopsis obtaining is Maotai producing mellow microorganism;
Described isolation medium is coffee 50g/L, glucose 180g/L, tryptone 25g/L, yeast extract 6g/L, sodium chloride 18g/L, agar 45g/L, and pH7 removes agar and is screening and culturing base.
State's aroma coffee preparation method
Step 1: get the ascocarpous intermediate structure piece of fresh ferfas, skin, epidermis, epidermis soil, the each 10g of mycelia and ferfas and host's exotrophic mycorrhiza in soil, after sterilization, wear into pulpous state with aseptic mortar, join and in sterile distilled water, shake 60min, therefrom getting 1000 μ L fluid drips is added on screening and culturing base evenly and is coated with and opens, under anaerobic be coated with after flat band method is purified and obtain a collection of microbial strains with dilution, then carry out fermenting experiment with liquid screening culture medium and therefrom filter out the rich bacterial strain that produces alpha-androstanol; Bacterial strain is carried out ultraviolet monochlor(in)ate lithium complex mutation and is beneficial to the raising of alpha-androstanol content; Screening and culturing base is: sucrose 300g/L, peptone 100g/L, yeast extract 20g/L, potassium dihydrogen phosphate 10g/L, magnesium sulfate 10g/L, Oak Tree loose callus lines 200g/L, agar 30g/L, and pH value 9, removing agar had been both liquid screening culture medium;
Step 2: will ferment in access liquid screening culture medium after the activation of ferfas fungal component, at 40 ℃, speed of agitator is to cultivate under 800rpm condition 30 days; After fermentation ends, by tunning homogenate, add protease, enzymolysis 150min going out after enzyme under 85 ℃ of conditions, obtains ferfas aroma former zymotic fluid;
Step 3: coffee is pulverized and is put into the pepsin of 80 ℃ after homogenate and the mixed liquor of hydrochloric acid soaks 90h, after then mixing with ferfas aroma former zymotic fluid the loose resuspended liquid of capsule bacterium of access coronoid process in 70 ℃ of fermentation 120h, after fermentation at 115 ℃ sterilizing 20min; In above-mentioned zymotic fluid, add the glucose that accounts for above-mentioned mixed liquor 10%, obtain coffee culture medium;
Step 4: after resuspended endomycopsis pulvis, according in 20% inoculum concentration access coffee culture medium temperature 60 C bottom fermentation 20 days, after filtering with microporous membrane degerming, continue fermentation 10 days, after fermentation ends at 120 ℃ of insulation 50h; At the centrifugal 20min of 8000rpm, supernatant is carried out to supercritical fluid extraction nitrogen by carbon dioxide and blow the concentrated extract A that obtains, filter residue extracts by ethyl acetate Soxhlet and distillation and concentration obtains extract B; After above-mentioned extract A and B are mixed, obtain Maotai and cause fragrant liquid, add respectively starch ester and soybean protein, homogeneous postlyophilization makes Maotai and causes fragrant liquid powder; By fluid bed, Maotai is caused to fragrant liquid powder spray sustained release coating, make Maotai and cause fragrant liquid microcapsule powder;
Step 5: by Maotai cause fragrant liquid microcapsule powder and ground coffee mix pack into while hot after dry in oak barrel balance 90 days afterwards packing make the coffee product with state's aroma.
Embodiment 6
A kind of state aroma coffee, is made up of 2g bafillus natto, 1g pears head mould, 42g coffee.
Bafillus natto preparation method:
Get the bent powder of 7g Maotai high-temperature daqu and add in the 93g sterilized water that bead is housed, on 50 ℃, 180rpm shaking table, vibrate 3 hours; Gradient dilution spread plate, cultivates 48 hours in 25 ℃; From flat board select growth better, the different bacterium colonies of proterties are inoculated in respectively isolation medium inclined-plane, as screening bacterial strain; The screening obtaining is inoculated respectively in screening and culturing base with bacterial strain, cultivated 72 hours for 25 ℃, the bafillus natto obtaining is Maotai producing mellow microorganism;
Described isolation medium is coffee 30g/L, glucose 100g/L, tryptone 25g/L, yeast extract 18g/L, agar 50g/L, and pH5 removes agar and is screening and culturing base.
Pears head mould preparation method:
Get the bent powder of 7g Maotai high-temperature daqu and add in the 93g sterilized water that bead is housed, on 50 ℃, 180rpm shaking table, vibrate 3 hours; Gradient dilution spread plate, cultivates 48 hours in 25 ℃; From flat board select growth better, the different bacterium colonies of proterties are inoculated in respectively isolation medium inclined-plane, as screening bacterial strain; The screening obtaining is inoculated respectively in screening and culturing base with bacterial strain, cultivated 72 hours for 25 ℃, the pears head mould obtaining is Maotai producing mellow microorganism;
Described isolation medium is glucose 10g/L, tryptone 45g/L, yeast extract 20g/L, sodium chloride 10g/L, agar 20g/L, and pH6 removes agar and is screening and culturing base.
State's aroma coffee preparation method:
Step 1: get the ascocarpous intermediate structure piece of fresh ferfas, skin, epidermis, epidermis soil, the each 8g of mycelia and ferfas and host's exotrophic mycorrhiza in soil, after sterilization, wear into pulpous state with aseptic mortar, join and in sterile distilled water, shake 30min, therefrom getting 500 μ L fluid drips is added on screening and culturing base evenly and is coated with and opens, under aerobic conditions, be coated with after flat band method is purified and obtain a collection of microbial strains with dilution, then carry out fermenting experiment with liquid screening culture medium and therefrom filter out the rich bacterial strain that produces alpha-androstanol; Screening and culturing base is: glucose 30g/L, peptone 10g/L, yeast extract 1g/L, potassium dihydrogen phosphate 1g/L, magnesium sulfate 0.1g/L, oak leaf 100g/L, agar 13g/L, and pH value 4.9, removes agar and is liquid screening culture medium;
Step 2: will ferment in access liquid screening culture medium after the activation of ferfas spore, at 20 ℃, speed of agitator is to cultivate under 80rpm condition 13 days; After fermentation ends, by tunning homogenate, add cellulase, beta-glucosidase, enzymolysis 15min going out after enzyme under 15 ℃ of conditions, obtains ferfas aroma former zymotic fluid;
Step 3: coffee is pulverized and put into the pepsin of 30 ℃ after homogenate and the mixed liquor of hydrochloric acid soaks 10h, then after mixing with ferfas aroma former zymotic fluid, access Bifidobacterium Bifidum and the resuspended mixed liquor of lactobacillus acidophilus be in 30 ℃ of fermentation 10h, after fermentation at 115 ℃ sterilizing 12min; In above-mentioned zymotic fluid, add and account for above-mentioned mixed liquor 1% sucrose, 10%KH
2pO
4, obtain coffee culture medium;
Step 4: after resuspended the wet thallus of bafillus natto, pears head mould, according in the coffee culture medium of 5% inoculum concentration access step 3 gained 20 ℃ of bottom fermentations of temperature 10 days, after filtering with microporous membrane degerming, continue fermentation 3 days, after fermentation ends, be incubated 5h at 70 ℃; At the centrifugal 5min of 5000rpm, supernatant is carried out to microwave heating extraction reduced pressure concentration acquisition extract A by propane diols, filter residue carries out ultrasonic extraction by acetonitrile and reduced pressure concentration obtains extract B; After above-mentioned extract A and B are mixed, obtain Maotai and cause fragrant liquid, add respectively starch ester and Arabic gum, homogeneous postlyophilization makes Maotai and causes fragrant liquid powder; By fluid bed, Maotai is caused to the water-soluble dressing of fragrant liquid powder spray, make Maotai and cause fragrant liquid microcapsule powder;
Step 5: by the Maotai of step 4 gained cause fragrant liquid microcapsule powder and ground coffee mix pack into while hot after dry in oak barrel balance 10 days afterwards packing make the coffee product with state's aroma.
Embodiment 7
A kind of state aroma coffee, is made up of 4g endomycopsis, 48g coffee.
Endomycopsis preparation method:
Get the bent powder of 10g Maotai high-temperature daqu and add in the 90g sterilized water that bead is housed, on 60 ℃, 180rpm shaking table, vibrate 9 hours; Gradient dilution spread plate, cultivates 85 hours in 65 ℃; From flat board select growth better, the different bacterium colonies of proterties are inoculated in respectively isolation medium inclined-plane, as screening bacterial strain; The screening obtaining is inoculated respectively in screening and culturing base with bacterial strain, cultivated 36 hours for 25 ℃, the endomycopsis obtaining is Maotai producing mellow microorganism;
Described isolation medium is coffee 80g/L, glucose 130g/L, tryptone 42g/L, yeast extract 36g/L, sodium chloride 8g/L, agar 43g/L, and pH7.3 removes agar and is screening and culturing base.
State's aroma coffee preparation method:
Step 1: the each 9g of exotrophic mycorrhiza that gets the ascocarpous intermediate structure piece of fresh ferfas, skin, epidermis and host, after sterilization, wear into pulpous state with aseptic mortar, join and in sterile distilled water, shake 50min, therefrom getting 800 μ L fluid drips is added on screening and culturing base evenly and is coated with and opens, under anaerobic be coated with after flat band method is purified and obtain a collection of microbial strains with dilution, then carry out fermenting experiment with liquid screening culture medium and therefrom filter out the rich bacterial strain that produces alpha-androstanol; Screening and culturing base is: sucrose 140g/L, peptone 78g/L, yeast extract 2g/L, potassium dihydrogen phosphate 3g/L, magnesium sulfate 0.8g/L, Oak Tree loose callus lines 70g/L, agar 16g/L, and pH value 5.2, removes agar and is liquid screening culture medium;
Step 2: will ferment in access liquid screening culture medium after the activation of ferfas aroma former, at 27 ℃, speed of agitator is to cultivate under 530rpm condition 27 days; After fermentation ends, by tunning homogenate, add beta-glucosidase, enzymolysis 58min going out after enzyme under 64 ℃ of conditions, obtains ferfas aroma former zymotic fluid; Described ferfas aroma former is the bacterial classification of ferfas ascocarp tissue separation, the mixture of ferfas spore;
Step 3: coffee is pulverized and is put into the pepsin of 74 ℃ after homogenate and the mixed liquor of hydrochloric acid soaks 72h, after then mixing with ferfas aroma former zymotic fluid the loose resuspended liquid of capsule bacterium of access coronoid process in 48 ℃ of fermentation 12h, after fermentation at 115 ℃ sterilizing 18min; In above-mentioned zymotic fluid, add the compound amino acid and 10% vitamin C that account for above-mentioned mixed liquor 10%, obtain coffee culture medium;
Step 4: after resuspended endomycopsis freeze-dried powder, according in the coffee culture medium of 8% inoculum concentration access step 3 gained 48 ℃ of bottom fermentations of temperature 7 days, after filtering with microporous membrane degerming, continue fermentation 3 days, after fermentation ends at 98 ℃ of insulation 36h; At the centrifugal 8min of 6500rpm, supernatant is obtained to extract A by cyclohexane with mechanical oscillation extraction reduced pressure concentration, filter residue uses distillation extraction extraction simultaneously nitrogen to blow the concentrated extract B that obtains by acetone; After above-mentioned extract A and B are mixed, obtain Maotai and cause fragrant liquid, add respectively monoglyceride and cyclodextrin, homogeneous postlyophilization makes Maotai and causes fragrant liquid powder; By fluid bed, Maotai is caused to fragrant liquid powder spray sustained release coating, make Maotai and cause fragrant liquid microcapsule powder;
Step 5: by the Maotai of step 4 gained cause fragrant liquid microcapsule powder after granulation, pack into while hot together with ground coffee in oak barrel balance 80 days afterwards packing make the coffee product with state's aroma.
Embodiment 8
A kind of state aroma coffee, is made up of the cured shape bacillus of 7g, 30g coffee.
Cured shape bacillus preparation method:
Get the bent powder of 5g Maotai high-temperature daqu and add in the 95g SPSS that bead is housed, on 55 ℃, 108rpm shaking table, vibrate 5 hours; Gradient dilution spread plate, cultivates 65 hours in 40 ℃; From flat board select growth better, the different bacterium colonies of proterties are inoculated in respectively isolation medium inclined-plane, as screening bacterial strain; The screening obtaining is inoculated respectively in screening and culturing base with bacterial strain, cultivated 58 hours for 35 ℃, the cured shape bacillus obtaining is Maotai producing mellow microorganism;
Described isolation medium is coffee 180g/L, glucose 150g/L, tryptone 35g/L, yeast extract 42g/L, sodium chloride 8g/L, agar 36g/L, and pH7.4 removes agar and is screening and culturing base.
State's aroma coffee preparation method:
Step 1: get the ascocarpous intermediate structure piece of fresh ferfas, skin, epidermis, epidermis soil, the each 6g of mycelia and ferfas and host's exotrophic mycorrhiza in soil, after sterilization, wear into pulpous state with aseptic mortar, join and in sterile distilled water, shake 35min, therefrom getting 800 μ L fluid drips is added on screening and culturing base evenly and is coated with and opens, after under anaerobic separating with plate streaking, obtain a collection of microbial strains, then carry out fermenting experiment with liquid screening culture medium and therefrom filter out the rich bacterial strain that produces alpha-androstanol, bacterial strain is carried out ultraviolet monochlor(in)ate lithium complex mutation and is beneficial to the raising of alpha-androstanol content, screening and culturing base is: sugarcane 350g/L, peptone 70g/L, yeast extract 0.8g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.7g/L, Oak Tree loose callus lines 180g/L, agar 27g/L, and pH value 8.2, removes agar and is liquid screening culture medium,
Step 2: will ferment in access liquid screening culture medium after the activation of ferfas aroma former, at 30 ℃, speed of agitator is to cultivate under 600rpm condition 10 days; After fermentation ends, by tunning homogenate, add cellulase, protease, beta-glucosidase, enzymolysis 106min going out after enzyme under 65 ℃ of conditions, obtains ferfas aroma former zymotic fluid; Described ferfas aroma former is the mixture of ferfas spore, ferfas endophyte, ferfas concomitance bacterium;
Step 3: coffee is pulverized and put into the pepsin of 70 ℃ after homogenate and the mixed liquor of hydrochloric acid soaks 65h, then after mixing with ferfas aroma former zymotic fluid, access Bifidobacterium Bifidum and the resuspended mixed liquor of lactobacillus acidophilus be in 60 ℃ of fermentation 105h, after fermentation at 115 ℃ sterilizing 20min; In above-mentioned zymotic fluid, add and account for above-mentioned mixed liquor 8% wood sugar, 0.5% fructose, 3%MgSO
4, 4%NaCl, 7% compound amino acid and 1% vitamin C, obtain coffee culture medium;
Step 4: after resuspended the wet thallus of cured shape bacillus, according in the coffee culture medium of 7% inoculum concentration access step 3 gained temperature 50 C bottom fermentation 17 days, after filtering with microporous membrane degerming, continue fermentation 6 days, after fermentation ends at 94 ℃ of insulation 5h; At the centrifugal 12min of 7800rpm, supernatant is extracted by glycerol distillation and reduced pressure concentration acquisition extract A, filter residue extracts by carrene Soxhlet and distillation and concentration obtains extract B; After above-mentioned extract A and B are mixed, obtain Maotai and cause fragrant liquid, add respectively sucrose ester, phosphatide and maltodextrin, shitosan, Arabic gum, homogeneous postlyophilization makes Maotai and causes fragrant liquid powder; By fluid bed, Maotai is caused to the water-soluble dressing of fragrant liquid powder spray, make Maotai and cause fragrant liquid microcapsule powder;
Step 5: by the Maotai of step 4 gained cause fragrant liquid microcapsule powder and ground coffee mix pack into while hot after dry in oak barrel balance 80 days afterwards packing make the coffee product with state's aroma.
Embodiment 9
A kind of state aroma coffee, is made up of 10g bafillus natto, 60g coffee.
Bafillus natto preparation method:
Get the bent powder of 30g Maotai high-temperature daqu and add in the 970g SPSS that bead is housed, on 55 ℃, 80rpm shaking table, vibrate 3 hours; Gradient dilution spread plate, cultivates 84 hours for 50 ℃; From flat board select growth better, the different bacterium colonies of proterties are inoculated in respectively isolation medium inclined-plane, as screening bacterial strain; The screening obtaining is inoculated respectively in screening and culturing base with bacterial strain, cultivated 30 hours for 60 ℃, the bafillus natto obtaining is Maotai producing mellow microorganism;
Described isolation medium is coffee 190g/L, glucose 70g/L, tryptone 32g/L, yeast extract 24g/L, sodium chloride 4.8g/L, agar 36g/L, and pH5.7 removes agar and is screening and culturing base.
State's aroma coffee preparation method:
Step 1: get Maotai producing mellow microorganism and be inoculated on ready PDA culture medium, be positioned over and in the constant incubator of 45 ℃, cultivate 78h and make actication of culture, again the bacterial classification of activation is inoculated in the LB fluid nutrient medium that adds glucose, in 40 ℃ of constant temperature culture 50h, by medium centrifugal reject supernatant, precipitation thalline is even with normal saline dilution concussion, is the activated seed liquid obtaining;
Step 2: coffee is pulverized and is put into the compound protease of 50 ℃ after homogenate and soak 70h, with water in mass ratio 1: 18 refluxing extraction after 6 hours, filter, at 115 ℃ of sterilizing 100min; In above-mentioned mixed liquor, add and account for above-mentioned mixed liquor weight 7% fructose, 0.80%MgSO
4, 5% compound amino acid and 3% vitamin C, obtain coffee fluid nutrient medium;
Step 3: after resuspended bafillus natto wet thallus, according in the coffee fluid nutrient medium of 14% inoculum concentration access step 2 gained temperature 45 C bottom fermentation 18 days, after filtering with microporous membrane degerming, continue fermentation 6 days, after fermentation ends, be incubated 40h at 100 ℃; By the coffee zymotic fluid fermenting, sheet frame membrane filtration after bacterial classification deactivation, freeze drying is condensed into paste again, according to the mass ratio of 1: 8, paste is mixed mutually with 98% ethanol water, paste is dissolved fully, after refiltering, concentrate filtrate to except after desolventizing, obtain fermentation of coffee medicinal extract; Fermentation of coffee medicinal extract is cured to 30min in 150 ℃, pack in airtight oak barrel after cooling and store and within 70 days, obtain state's aroma coffee.
Test example
Caffeine detection method:
1.1 materials and instrument
Caffeine standard items are all analyzed pure; Methyl alcohol is chromatographically pure, and water is secondary deionized water;
Instrument: Waters2695 high performance liquid chromatograph is joined PDAD.
1.2 chromatographic condition
Mobile phase: A methyl alcohol, B water, gradient elution (in table 1); Sample size: 10L; Column temperature: 25 ℃.
Chromatographic column: Zorbax C18 (4.6 × 250mm); Detect wavelength: 271nm; Flow velocity: 1.0mL/min;
1.3 sample pretreatment
Accurately measure each embodiment sample and the each 200g of nest's normal coffee is dissolved in 1000ml water, get 5.0mL sample, be settled to 50mL with secondary deionized water dilution, 40 ℃ of ultrasonic extractions, through 0.45 μ m filtering with microporous membrane, through high-performance liquid chromatogram determination.
The preparation of 1.4 standard liquids
Accurately take respectively 0.0100g caffeine in 10.0mL volumetric flask, respectively by methanol constant volume to scale, obtain the each 1000g/mL of standard reserving solution, pipette respectively 1.0mL standard reserving solution in 10.0mL volumetric flask, methanol constant volume is to scale, and the standard that obtains is mixed intermediate liquid 100g/mL.
Table 1 mobile phase ratio
| min | A | B |
| 0.0 | 25 | 75 |
| 3.0 | 25 | 75 |
| 5.0 | 40 | 60 |
| 10.0 | 40 | 60 |
| 10.5 | 25 | 75 |
| 12.0 | 25 | 75 |
2.2 calibration curves drafting
With methyl alcohol be mixed with coffee because 0.1,0.2,0.5,1.0,5.0,10.0, the hybrid standard series of 20.0g/mL, take concentration as abscissa, peak area is that ordinate is made calibration curve, and external standard method is quantitative, obtains linear equation and is: caffeine: y=22859x+31.25; Coefficient correlation: r
2=0.9995
2.3 detection limit
With the standard liquid sample introduction of 0.1g/mL, obtain minimum detectability with 3 times of signal to noise ratios, measure with 1.0g/mL standard liquid sample introduction, obtain the method precision (n=6), detect and be limited to 0.0g/mL, precision is 1.8458%.
2.4 the rate of recovery
Commercially available sample Jilibao is added to caffeine standard reference material, according to 1.3 sample pretreating methods, through liquid chromatographic detection, obtain the rate of recovery scope that caffeine obtains, the results are shown in Table 2.
Table 2 rate of recovery experimental result (n=6)
2.5 sample determination
Detect the caffeine in embodiment 1-9 sample, result is as table 3
2.6 total acid and ester detection methods are with reference to GB/T10345-2007.
Caffeine and total acid and ester content in table 3 embodiment 1-9 sample
As seen from the above table, state's aroma coffee of gained of the present invention has not only been preserved the caffeine in former coffee, has preserved caffeine, also contains the fragrant acid and the ester that possess of requiring of white wine, and therefore it has possessed coffee perfume (or spice) and aroma simultaneously.
Claims (9)
1. state's aroma coffee, is characterized in that by 1-99.99 weight portion coffee, 0.01-30 weight portion Maotai producing mellow microorganism or produces the fragrant function bacterium of sauce forming.
2. a kind of state according to claim 1 aroma coffee, is characterized in that described coffee is one or more in coffee bean, coffee bean particle, ground coffee, instant coffee powder.
3. a kind of state according to claim 1 aroma coffee, is characterized in that described Maotai producing mellow microorganism is to produce one or more compositions in the Aspergillus obtaining with separation and purification Daqu, white mucor, rhizopus, pears head mould, false yeast, endomycopsis, Hansenula yeast, lactic acid bacteria, acetic acid bacteria, bacillus licheniformis, bacillus subtilis, bacillus alcalophilus, cured shape bacillus, bacillus pumilus, bacillus amyloliquefaciens, bafillus natto from Maotai.
4. a kind of state according to claim 1 aroma coffee, is characterized in that described Maotai producing mellow microorganism prepared by following methods:
Get the bent powder of 1-10 weight portion Maotai high-temperature daqu and add in the 90-99 weight portion sterilized water or SPSS that bead is housed, 1-10 hour vibrates on 10-70 ℃, 10-200rpm shaking table; Gradient dilution spread plate, cultivates 1-90 hour in 10-70 ℃; From flat board select growth better, the different bacterium colonies of proterties are inoculated in respectively isolation medium inclined-plane, as screening bacterial strain; The screening obtaining is inoculated respectively in screening and culturing base with bacterial strain, cultivated 1-90 hour for 10-70 ℃, the fragrant dominant strain of the product obtaining is Maotai producing mellow microorganism;
The Maotai producing mellow microorganism screening is bacillus, gemma clostridium, pseudomonas, Aspergillus, Penicillium notatum, saccharomycete, and bacterial classification can be bacterium song or bacterium liquid form;
Described isolation medium is coffee 0-300g/L, glucose 1-200g/L, tryptone 1-50g/L, yeast extract 1-50g/L, sodium chloride 0-50g/L, agar 1-50g/L, and pH4-8 removes agar and is screening and culturing base.
5. a kind of state according to claim 1 aroma coffee, is characterized in that being prepared by following methods:
Step 1: get Maotai producing mellow microorganism and be inoculated on ready slant medium, be positioned over and in the constant incubator of 10-70 ℃, cultivate 1-90h and make actication of culture, again the bacterial classification of activation is inoculated in the LB fluid nutrient medium that adds sugared source, in 10-70 ℃ of constant temperature culture 1-90h, by medium centrifugal reject supernatant, precipitation thalline is even with normal saline dilution concussion, is the activated seed liquid obtaining; Described slant medium is beef extract-peptone, PDA culture medium or minimal medium;
Step 2: coffee is pulverized and put into the compound protease of 20-80 ℃ after homogenate and/or the mixed liquor of flavor protease soaks 1-90h, and water in mass ratio 1: 0.1-30 refluxing extraction was filtered after 1-8 hour, at 115 ℃ of sterilizing 100min; In above-mentioned mixed liquor, add and account for the sugared source of above-mentioned mixed liquor weight 0.1-10%, the inorganic salts of 0.1-10%, compound amino acid and/or the vitamin C of 0-10%, obtain coffee fluid nutrient medium;
Step 3: after resuspended to the wet thallus of Maotai producing mellow microorganism, pulvis or freeze-dried powder, according in the coffee fluid nutrient medium of inoculum concentration access step 2 gained of 1-20% at temperature 10-60 ℃ of bottom fermentation 1-20 days, after filtering with microporous membrane degerming, continue fermentation 1-10 days, after fermentation ends, be incubated 0.1-50h at 55-120 ℃; By the coffee zymotic fluid fermenting, sheet frame membrane filtration after bacterial classification deactivation, reduced pressure concentration or freeze drying are condensed into paste again, according to 1: the mass ratio of 1-10 mixes paste mutually with extractant, paste is dissolved fully, after refiltering, concentrate filtrate to except after desolventizing, obtain fermentation of coffee medicinal extract; Fermentation of coffee medicinal extract is cured to 1-50min in 10-200 ℃, pack in airtight oak barrel after cooling and store and within 1-90 days, obtain state's aroma coffee; Described extract, extractant are water or ethanol, or both composite solutions.
6. a kind of state according to claim 1 aroma coffee, is characterized in that being prepared by following methods:
Step 1: get the ascocarpous intermediate structure piece of fresh ferfas, skin, epidermis, epidermis soil, the each 5-10 weight portion of mycelia and ferfas and host's exotrophic mycorrhiza in soil, after sterilization, wear into pulpous state with aseptic mortar, join and in sterile distilled water, shake 10-60min, therefrom getting 100-1000 μ L fluid drips is added on screening and culturing base evenly and is coated with and opens, under aerobic or anaerobic condition, be coated with after flat band method is purified and obtain a collection of microbial strains with plate streaking separation or dilution, then carry out fermenting experiment with liquid screening culture medium and therefrom filter out the rich bacterial strain that produces alpha-androstanol; Screening and culturing base is: glucose or sucrose 5-300g/L, peptone 1-100g/L, yeast extract 0.1-20g/L, potassium dihydrogen phosphate 0.1-10g/L, magnesium sulfate 0.01-10g/L, oak leaf or Oak Tree loosen callus lines 1-200g/L, agar 10-30g/L, pH value 4-9, removes agar and is liquid screening culture medium;
Step 2: will ferment in access liquid screening culture medium after the activation of ferfas aroma former, at 15-40 ℃, speed of agitator is to cultivate 1-30 days under 10-800rpm condition; After fermentation ends, by tunning homogenate, add one or more of cellulase, protease, beta-glucosidase, enzymolysis 1-150min going out after enzyme under 10-85 ℃ of condition, obtains ferfas aroma former zymotic fluid; Described ferfas aroma former is one or more of the bacterial classification, the ferfas spore that separate of ferfas ascocarp tissue, ferfas endophyte, ferfas fungal component, ferfas concomitance bacterium;
Step 3: coffee is pulverized and put into the pepsin of 20-80 ℃ after homogenate and the mixed liquor of hydrochloric acid soaks 1-90h, then after mixing with ferfas aroma former zymotic fluid, the loose resuspended liquid of capsule bacterium of access Bifidobacterium Bifidum and the resuspended mixed liquor of lactobacillus acidophilus or coronoid process is in 20-70 ℃ of fermentation 1-120h, after fermentation at 115 ℃ sterilizing 10-20min; In above-mentioned zymotic fluid, add and account for the sugared source of above-mentioned mixed liquor 0.1%-10%, the inorganic salts of 0.1%-10%, compound amino acid and/or the vitamin C of 0.1%-10%, obtain coffee culture medium;
Step 4: after resuspended to the wet thallus of Maotai producing mellow microorganism, pulvis or freeze-dried powder, according in the coffee culture medium of inoculum concentration access step 3 gained of 1-20% at temperature 10-60 ℃ of bottom fermentation 1-20 days, after filtering with microporous membrane degerming, continue fermentation 1-10 days, after fermentation ends, be incubated 0.1-50h at 55-120 ℃; At the centrifugal 1-20min of 3000-8000rpm, by supernatant, by solvent extraction the concentrated extract A that obtains, filter residue is by solvent extraction the concentrated extract B that obtains; After above-mentioned extract A and B are mixed, obtain Maotai and cause fragrant liquid, add respectively emulsifying agent and embedded material, after homogeneous, the dry or freeze drying of spraying makes Maotai and causes fragrant liquid powder; By fluid bed, Maotai is caused to the water-soluble dressing of fragrant liquid powder spray or sustained release coating, make Maotai and cause fragrant liquid microcapsule powder; Described emulsifying agent is one or more of monoglyceride, sucrose ester, phosphatide, starch ester; Described embedded material is one or more of converted starch, maltodextrin, sucrose, shitosan, Arabic gum, cyclodextrin, carragheen, alginate, gelatin, soybean protein, lactalbumin, protein hydrolysate; Described solvent extraction mode is under room temperature, to leave standstill the one of soaking in extraction, mechanical oscillation extraction, ultrasonic extraction, heating reflux extraction, microwave heating extraction, distillation extraction, while distillation extraction, Soxhlet extraction or supercritical fluid extraction; Solvent is any one or a few in water, ethanol, propane diols, glycerine, ether, ethyl acetate, acetonitrile, acetone, carrene, chloroform, cyclohexane, carbon dioxide; Described concentration process is that under normal pressure, distillation and concentration, nitrogen blow the one in concentrated or reduced pressure concentration;
Step 5: the Maotai of step 4 gained is caused to the granulation or mix to pack while hot in oak barrel balance 1-90 days into after dry and pack afterwards and make the coffee product with state's aroma together with ground coffee of fragrant liquid microcapsule powder.
7. the preparation method of a kind of state according to claim 6 aroma coffee, is characterized in that in described step 1 bacterial strain to carry out ultraviolet monochlor(in)ate lithium complex mutation and be beneficial to the raising of alpha-androstanol content.
8. according to the preparation method of a kind of state of any one in claim 5,6,7 aroma coffee, wherein said inorganic salts are KH
2pO
4, (NH4)
2sO
4, MgSO
4, or NaCl.
9. according to the preparation method of a kind of state of any one in claim 5,6,7 aroma coffee, wherein said sugared source is one or more the mixture in sucrose, glucose, wood sugar, fructose.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410128974.8A CN103859122A (en) | 2014-04-02 | 2014-04-02 | Maotai flavored coffee and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410128974.8A CN103859122A (en) | 2014-04-02 | 2014-04-02 | Maotai flavored coffee and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103859122A true CN103859122A (en) | 2014-06-18 |
Family
ID=50898583
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410128974.8A Pending CN103859122A (en) | 2014-04-02 | 2014-04-02 | Maotai flavored coffee and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103859122A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105105255A (en) * | 2015-07-16 | 2015-12-02 | 郭景龙 | Beverage and preparation method thereof |
| CN105918576A (en) * | 2016-04-21 | 2016-09-07 | 广州市佰沃生物科技有限公司 | New fermentation type coffee beans and preparation method thereof |
| CN106358765A (en) * | 2016-11-16 | 2017-02-01 | 陕西北极宫茶叶有限公司 | Method of cultivating eurotium cristatum on solid-state medium |
| CN108913425A (en) * | 2018-07-11 | 2018-11-30 | 贵州神奇药业有限公司 | A kind of coffee tea wine and preparation method thereof |
| CN111589315A (en) * | 2020-05-27 | 2020-08-28 | 华南理工大学 | Coffee fragrant wine preparation system and process |
| CN114009562A (en) * | 2021-11-29 | 2022-02-08 | 海南一哟食品有限公司 | Coffee containing natron and preparation method of coffee |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101040653A (en) * | 2005-03-24 | 2007-09-26 | 三得利株式会社 | Method of processing fresh coffee beans by using surface-treated coffee fruits |
| CN100425687C (en) * | 2006-07-07 | 2008-10-15 | 贵州神奇集团控股有限公司 | Method for making caffee milk wine |
| KR20100020121A (en) * | 2008-08-12 | 2010-02-22 | 두두원발효(주) | Coffee fermented with kimchi lactic acid bacteria and production method thereof |
| CN102224873A (en) * | 2011-05-06 | 2011-10-26 | 郭景龙 | Method for preparing food from raw materials comprising coffee beans |
| RU125432U1 (en) * | 2012-07-03 | 2013-03-10 | Федеральное государственное бюджетное образовательное учреждение высшего профессионального образования "Кубанский государственный технологический университет" (ФГБОУ ВПО "КубГТУ") | TECHNOLOGICAL LINE FOR PRODUCING NATURAL AROMATED GROUND COFFEE |
-
2014
- 2014-04-02 CN CN201410128974.8A patent/CN103859122A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101040653A (en) * | 2005-03-24 | 2007-09-26 | 三得利株式会社 | Method of processing fresh coffee beans by using surface-treated coffee fruits |
| CN100425687C (en) * | 2006-07-07 | 2008-10-15 | 贵州神奇集团控股有限公司 | Method for making caffee milk wine |
| KR20100020121A (en) * | 2008-08-12 | 2010-02-22 | 두두원발효(주) | Coffee fermented with kimchi lactic acid bacteria and production method thereof |
| CN102224873A (en) * | 2011-05-06 | 2011-10-26 | 郭景龙 | Method for preparing food from raw materials comprising coffee beans |
| RU125432U1 (en) * | 2012-07-03 | 2013-03-10 | Федеральное государственное бюджетное образовательное учреждение высшего профессионального образования "Кубанский государственный технологический университет" (ФГБОУ ВПО "КубГТУ") | TECHNOLOGICAL LINE FOR PRODUCING NATURAL AROMATED GROUND COFFEE |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105105255A (en) * | 2015-07-16 | 2015-12-02 | 郭景龙 | Beverage and preparation method thereof |
| CN105918576A (en) * | 2016-04-21 | 2016-09-07 | 广州市佰沃生物科技有限公司 | New fermentation type coffee beans and preparation method thereof |
| CN106358765A (en) * | 2016-11-16 | 2017-02-01 | 陕西北极宫茶叶有限公司 | Method of cultivating eurotium cristatum on solid-state medium |
| CN108913425A (en) * | 2018-07-11 | 2018-11-30 | 贵州神奇药业有限公司 | A kind of coffee tea wine and preparation method thereof |
| CN111589315A (en) * | 2020-05-27 | 2020-08-28 | 华南理工大学 | Coffee fragrant wine preparation system and process |
| CN111589315B (en) * | 2020-05-27 | 2023-12-19 | 华南理工大学 | Coffee flavored wine preparation system and process |
| CN114009562A (en) * | 2021-11-29 | 2022-02-08 | 海南一哟食品有限公司 | Coffee containing natron and preparation method of coffee |
| CN114009562B (en) * | 2021-11-29 | 2023-10-27 | 海南一哟食品有限公司 | Latte coffee and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103859122A (en) | Maotai flavored coffee and preparation method thereof | |
| CN101897429B (en) | Complex microbial agent for producing soybean paste in Pixian County and preparation method thereof | |
| CN103783226A (en) | Soy sauce flavor type tea | |
| Lao et al. | A novel combination of enzymatic hydrolysis and fermentation: Effects on the flavor and nutritional quality of fermented Cordyceps militaris beverage | |
| CN104366318A (en) | Dendrobium enzyme with Chinese wine fragrance and preparation method thereof | |
| CN103110180B (en) | Early-flue-cured tobacco quality improving method with eurotium cristatum | |
| US11344043B2 (en) | Method for producing fermented coffee using enteric bacteria of Kopi Luwak | |
| CN101333508B (en) | Lactobacillus brevis for highly producing gamma-aminobutyric acid | |
| CN109370927A (en) | Candidiasis FW922-1 and its application | |
| CN110157623A (en) | A kind of method of reaping hook bacteria strain and its fermenting and producing D-pantoyl lactone hydrolase | |
| CN103271951A (en) | Method for preparing cordyceps preparations with high adenosine contents | |
| CN105077484A (en) | Phellinus igniarius flavored beverage and preparation method thereof | |
| CN104982571A (en) | Endophyte fermented tea and preparing method thereof | |
| CN104277985A (en) | Monascus purpureus M-24 bacterial strain and application thereof for preparation of Monacolin K | |
| CN107475149A (en) | A kind of composite bacteria agent for improving fermentation fermented soya bean antioxidation activity and its preparation method and application | |
| CN103087893A (en) | Preparation method of composite coarse cereals monascus | |
| CN106754507A (en) | A kind of composite flavor microbial inoculum and preparation method thereof and the direct putting type application in soy sauce flavouring | |
| CN110236060A (en) | A kind of mushroom instant solid beverage and preparation method thereof containing biodiasmin | |
| CN104543982A (en) | Truffle composition and preparation method thereof | |
| CN103224894A (en) | Enterobacter mori, and biotransformation method for fermented wheat bran extract used for cigarette and application | |
| JP4325834B2 (en) | Alcohol-containing beverage and method for producing the same | |
| CN106434399A (en) | Mixed distiller's yeast for glutinous rice wine and preparation method of mixed distiller's yeast | |
| CN107988090B (en) | Functional microorganism combined microbial inoculum and application thereof | |
| KR101721836B1 (en) | Method of preparing composition containing active ingredient of culture medium of ceriporia lacerata for anticancer, prevention and improvement of cancer disease and the composition made therefrom | |
| US8673324B2 (en) | Method for producing gamma-aminobutyric acid and food produced thereby |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20140618 |