CN104543982A - Truffle composition and preparation method thereof - Google Patents
Truffle composition and preparation method thereof Download PDFInfo
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- CN104543982A CN104543982A CN201510058878.5A CN201510058878A CN104543982A CN 104543982 A CN104543982 A CN 104543982A CN 201510058878 A CN201510058878 A CN 201510058878A CN 104543982 A CN104543982 A CN 104543982A
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- truffle
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to the technical field of health foods, and particularly relates to a truffle composition and a preparation method thereof. The composition contains the following raw materials in parts by weight: 0.1-90 parts of truffle, 1-99 parts of coffee and/or cocoa powder, and 1-80 parts of red wine polyphenols. Pharmacological experiments show that the composition disclosed by the invention has the effects of resisting aging and delaying the formation of pigments.
Description
Technical field
The invention belongs to food technology field, be specifically related to a kind of truffle composition and method of making the same.
Background technology
Along with improving constantly of people's living standard; enriching of material life and improving constantly of cultural quality; people pursue self perfect theory and also constantly distil, and people not only pursue healthy health, more focus on beautifying face and moistering lotion; people are not only confined to make to apply some make up; namely outside skin sparing, and more focus on the improvement of meals, the improvement of life style; the improvement of drink, to reach from interior demand of carrying out beautifying face and moistering lotion.
Recent study shows, the old and feeble biomembranous lipid peroxidation caused with free radical causes membrane structure to be damaged and functionally inactive has substantial connection, wherein the attack of methoxy radical pair lipid biomembrane and senile plaque expelling have very large relation, it can make unrighted acid be oxidized to peroxide, this peroxide decomposites a large amount of aldehydes, alcohols more further, wherein MDA (MDA) very easily with phospholipoprotein qualitative response, be the reason forming old color spot.In addition, human body is in order to defend the damage and fracture of free radical in metabolism and other vital movements, containing some polyphenoils comprising superoxide dismutase (SOD) in tissue, they can prevent free radical from causing the degeneration of tissue cell insult and organ to change by effects such as lipid peroxidations in vivo, thus play anti-ageing, to delay pigment formation effect.
Summary of the invention
For these reasons, applicant is on the basis studied for many years, based on modern medical theory, find a kind of new composition, said composition comprises truffle, coffee and/or cocoa power, Red Wine Polyphenols, pharmacology test and clinical testing test show, the present composition have well anti-ageing, delay pigment formed pharmacological action.
The present invention is achieved through the following technical solutions.
A kind of truffle composition, comprises 0.1-90 weight portion truffle, 1-99 weight portion coffee and/or cocoa power, 1-80 weight portion Red Wine Polyphenols.
A kind of truffle composition, comprises 0.1-90 weight portion truffle, 1-99 weight portion coffee and/or cocoa power, 1-80 weight portion Red Wine Polyphenols, 1-80 parts by weight of saccharide.
Described truffle is one or more in truffle alpha-androstanol microcapsule powder, truffle microcapsule powder, truffle polysaccharide, fresh truffle, truffle Ultramicro-powder, fermentation truffle bacterium powder, fermentation truffle extract, truffle ferment, truffle endophyte tunning.
Described coffee is one or more in coffee bean, coffee bean particle, state's aroma ground coffee, instant coffee powder.
Described carbohydrate is one or more in sucrose, fructose, honey, glucose, cellobiose, white sugar, brown sugar, brown sugar, Arabinose, HFCS, D-ribose, date palm syrup, soyabean oligosaccharides, FOS, isomalto-oligosaccharide, galactooligosaccharide, xylo-oligosaccharide, stachyose, raffinose, chitosan oligomer, xylitol, D-sorbite, maltitol, antierythrite.
A symbiotic fermentation preparation method for truffle alpha-androstanol microcapsule powder, comprises the following steps:
Step 1: by truffle aroma former seed liquor according in the volume ratio access Oak Tree endophyte zymotic fluid of 5-30% in temperature be 20-70 DEG C, speed of agitator is 50-600 rev/min of condition bottom fermentation 1-20 days, obtains symbiotic fermentation liquid;
Step 2: by truffle aroma former tunning and concentration be the ethanol of 60%-100% by weight 100: after the ratio of (1-200) mixes, in 20-70 DEG C, ultrasonic wave process 1-120 minute under 20-80KHz; Supercritical carbon dioxide extracting still is placed in by through the truffle aroma former tunning of ultrasonic wave process or symbiotic fermentation liquid, extraction kettle pressure be 20-50MPa, temperature extracts 1-24 hour under being 30-60 DEG C of condition, collect extract and carry out reduced pressure concentration to obtain truffle alpha-androstanol crude extract;
Described truffle aroma former tunning is prepared by following methods:
Symbiotic fermentation liquid is added and is heated to 80-150 DEG C after enzyme preparation carries out enzymolysis 1-15 hour at 20-85 DEG C and is incubated 1-15 hour; Symbiotic fermentation liquid reduced pressure concentration after enzymolysis or nitrogen are blown concentrated after at 10-80 DEG C vacuum drying make water content be 5%-11%, obtain truffle aroma former tunning; Described enzyme preparation is one or more in cellulase, interior raw Penicillium notatum cellulase, flavor protease, bacillus protein enzyme, pectase and beta-glucosidase;
Step 3: be (0.1-5) by weight ratio: 1: the trehalose of (1-5), sucrose, glucose, add in 1-20 water doubly to mix and be heated to 70-98 DEG C and become molten condition, be cooled to add while stirring during 40-60 DEG C the truffle alpha-androstanol crude extract and/or truffle slurries that account for water weight 1-70%, 1-30 minute is stirred under 2000-5000rpm rotating speed, adopt material homogeneous 0.1-5 hour at 20-40 DEG C that high-pressure homogeneous equipment will be stirred, the mixed liquor crossed by homogeneous carries out spraying dry or freeze drying, obtain truffle alpha-androstanol microcapsule powder.
Described truffle aroma former seed liquor is prepared by following methods:
Step 1: get the ascocarpous intermediate structure block of fresh truffle, epidermis, epidermis soil, mycelia in soil, the each 5-10g of exotrophic mycorrhiza of truffle and host, pulpous state is worn into sterile mortar after sterilization, aseptically join in sterile distilled water and shake 10-60min, therefrom getting 100-1000 μ l fluid drips is added on screening and culturing base evenly spreadable, be separated with plate streaking under aerobic or anaerobic condition or dilute to be coated with after flat band method is purified and obtain a collection of microbial strains, then carry out fermenting experiment with liquid screening medium and therefrom filter out the truffle aroma former that the rich bacterial strain producing alpha-androstanol is screening, bacterial classification is bacterium song or bacterium liquid form, the content of alpha-androstanol can be improved by natural variation or artificial induction's variation to bacterial strain, 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 juice or Oak Tree endophyte zymotic fluid 1-900ml/l, agar 1-30g/l, pH value 4-9, remove agar and be liquid screening medium.
Step 2: truffle aroma former slant strains is inoculated in seed culture medium, at 20-40 DEG C, shaken cultivation 1-40 hour under rotating speed 10-150rpm condition, obtained truffle aroma former seed liquor; Described seed culture medium is: glucose 1-50g/l, oak leaf juice or Oak Tree endophyte zymotic fluid 1-900ml/l, peptone 1-10g/l, yeast extract 1-10g/l, magnesium sulfate 0.1-5g/l, potassium dihydrogen phosphate 0.1-5g/l, pH4-10.
Described Oak Tree endophyte zymotic fluid is prepared by following methods:
Step 1: choosing symbiosis has the skin of the Oak Tree of truffle, leaf, stem, flower, fruit and mycorhiza, first use aseptic water washing surface dirt, then aseptically by the alcohol immersion 30 seconds that volume ratio is 75%, sterilized water washes surperficial ethanol off, 30 seconds are soaked again with the mercuric chloride that mass fraction is 0.1%, aseptic water washing 3-5 time, then be inoculated on PDA culture medium flat plate respectively with the fritter that aseptic cutter is cut into 3 millimeters * 3 millimeters afterwards, 20-50 DEG C of constant temperature culture; Treat obviously to grow bacterium colony around sample, the bacterium colony of picking different shape, forward screening and culturing base inclined-plane to, 20-50 DEG C of purifying cultivates the Oak Tree endophyte that the dominant strain obtained is screening; 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, fresh truffle juice 1-900ml/l, agar 1-30g/l, pH value 4-9;
Step 2: the Oak Tree endophyte of separation is inoculated in activation medium inclined-plane, obtains Oak Tree endophyte slant strains for 1-48 hour in 20-50 DEG C of activation, is inoculated in MS fluid nutrient medium and cultivates 1-50 days, obtain Oak Tree endophyte zymotic fluid; Activation medium is: glucose 20-50 grams per liter, peptone 1-20g/l, magnesium sulfate 1-10g/l, potassium dihydrogen phosphate 1-10g/l, agar 10-30g/l, pH4-8.
The technique effect that the present composition is useful:
(1) make the aging of people, pigmentation symptom mentions and alleviate significantly and improve.
(2) composition quality of the present invention is controlled, use safety.
Embodiment
Below by way of the description of detailed description of the invention, the invention will be further described, but this is not limitation of the present invention, those skilled in the art are according to basic thought of the present invention, various amendment or improvement can be made, but only otherwise depart from basic thought of the present invention, all within the scope of the present invention.
Test example 1 alpha-androstanol determination test
Adopt the method for testing of the embodiment 1 (comparative example) of patent 200810000570.5, measure content and the productive rate of alpha-androstanol, the results are shown in Table 1.
Table 1 Isoginkgetin extracting method test result
| Comparative example | Embodiment 1 | Embodiment 3 | Embodiment 4 | Embodiment 5 | |
| Purity | 87.4％ | 97.3％ | 99.0％ | 99.6％ | 99.3％ |
| Recovery rate | 0.76 | 3.72 | 3.87 | 3.86 | 4.02 |
The test of pesticide effectiveness 1: on the impact of anti-oxidative ability of mice
Experimental technique: get mouse 50, male and female half and half, be divided into 5 groups at random, sub-cage rearing.Administration group gives the heavy dose of group (4.54g/kg) of embodiment 1 products obtained therefrom, middle dosage group (3.03g/kg), small dose group (1.52g/kg) bowel lavage 0.5ml respectively; BAIXIAO DAN group (2.42g/kg) bowel lavage 0.5ml; Blank group gives isopyknic physiological saline, once a day, after continuous 15 days, measures the content of liver superoxide dismutase (SOD) and MDA (MDA).Experimental result is in table 2:
Table 2: mouse liver SOD and MDA content
| Group | Dosage | Quantity | SOD(unmg/mg) | MDA(nmol/mg) |
| Blank group | - | 10 | 32.39±0.38 | 4.38±0.13 |
| BAIXIAO DAN group | 2.42 | 10 | 35.45±0.89* | 3.71±0.78** |
| Heavy dose of group | 7.53 | 10 | 47.44±1.11** | 2.13±0.89** |
| Middle dosage group | 6.34 | 10 | 47.17±0.87** | 2.22±0.87** |
| Small dose group | 5.36 | 10 | 45.73±0.56** | 2.33±0.97** |
Note: compare * P < 0.05**P < 0.01 with blank group
Conclusion: composition of the present invention effectively can improve the superoxide dismutase (SOD) in Mice Body and reduce the content of MDA (MDA), and the effect increasing SOD is better than BAIXIAO DAN.
The test of pesticide effectiveness 2: the effect of anti-wrinkle and color spot
Description of test: the characteristic feature of skin senescence is wrinkle and old color spot, modern medicine study shows, old color spot is relevant with the deposition of lipofuscin, the formation of wrinkle is then relevant with the hydratability of skin, therefore, this research by illustrating the anti-wrinkle of combination of the present invention and the effect of pigmented spots to the research of rat liver lipofuscin content, skin histology hydratability, thus reaches whitening function.
Experimental technique: get Wistar rat, female, without conceived, 20 ~ 22 monthly ages, body weight 250 ~ 350g.Old rats is divided into aged controls group, high dose group, middle dosage group, small dose group, BAIXIAO DAN group at random by body weight, and in addition, remainder rat in about 5 week age is as young control group.Embodiment 2 resulting composition three dosage component do not press 4g/kg, 2g/kg, 1g/kg administration, aged group and Younger group give the water (2ml/100g) of same volume, daily 1 time, after continuous 2 months, eye socket is got blood and is put to death animal, gets liver and skin histology carries out index determining.The results are shown in Table 3,4:
Table 3: old rats liver lipofuscin content
| Group | Dosage | Quantity | Liver L f (ug/g tissue) |
| Aged controls group | - | 10 | 69.37±3.45 |
| Young control group | - | 10 | 37.33±0.32** |
| BAIXIAO DAN group | 2.0 | 10 | 62.64±0.97 |
| Heavy dose of group | 4.0 | 10 | 43.12±0.36** |
| Middle dosage group | 2.0 | 10 | 44.78±0.53** |
| Small dose group | 1.0 | 10 | 46.31±0.63** |
Note: compare * P < 0.05**P < 0.01 with aged controls group
Table 4: the impact of old rats skin histology water content
| Group | Dosage | Quantity | Water content (%) |
| Aged controls group | - | 10 | 57.77±0.32 |
| Young control group | - | 10 | 66.43±6.64** |
| BAIXIAO DAN group | 2.0 | 10 | 62.64±0.97 |
| Heavy dose of group | 4.0 | 10 | 68.58±0.34** |
| Middle dosage group | 2.0 | 10 | 66.57±0.83** |
| Small dose group | 1.0 | 10 | 60.01±0.47 |
Note: with aged controls group comparatively * P < 0.05**P < 0.01
Conclusion: composition of the present invention effectively can reduce the content of lipofuscin in liver organization, the formation of check melanin.Meanwhile, increase skin histology water content, delay and prevent the generation of wrinkle, therefore, there is the beautification function of dewrinkling spot-eliminating.
The raw materials such as truffle of the present invention, Red Wine Polyphenols are all purchased from Mao Wei bio tech ltd, Guizhou;
Embodiment 1
A kind of truffle composition, comprises 1g truffle alpha-androstanol microcapsule powder, 1g coffee, 10g Red Wine Polyphenols.
Preparation method: 1g truffle alpha-androstanol microcapsule powder, 1g coffee, 10g Red Wine Polyphenols are ground rear mistake 100 mesh sieve respectively, mixes rear filling capsule and obtain capsule.
The preparation method of Oak Tree endophyte zymotic fluid:
Step 1: choosing symbiosis has the skin of the Oak Tree of truffle, leaf, stem, flower, fruit and mycorhiza, first use aseptic water washing surface dirt, then aseptically by the alcohol immersion 30 seconds that volume ratio is 75%, sterilized water washes surperficial ethanol off, 30 seconds are soaked again with the mercuric chloride that mass fraction is 0.1%, aseptic water washing 3-5 time, then be inoculated on PDA culture medium flat plate respectively with the fritter that aseptic cutter is cut into 3 millimeters * 3 millimeters afterwards, 20 DEG C of constant temperature culture; Treat obviously to grow bacterium colony around sample, the bacterium colony of picking different shape, forward screening and culturing base inclined-plane to, 20 DEG C of purifying cultivate the Oak Tree endophyte that the dominant strain obtained is screening; Screening and culturing base is: glucose 50g/l, peptone 10g/l, yeast extract 0.1g/l, potassium dihydrogen phosphate 0.1g/l, magnesium sulfate 0.01g/l, fresh truffle juice 100ml/l, agar 10g/l, pH value 4;
Step 2: the Oak Tree endophyte of separation is inoculated in activation medium inclined-plane, obtains Oak Tree endophyte slant strains for 18 hours in 25 DEG C of activation, is inoculated in MS fluid nutrient medium and cultivates 10 days, obtain Oak Tree endophyte zymotic fluid; Activation medium is: glucose 25g/l, peptone 1-20g/l, magnesium sulfate 1-10g/l, potassium dihydrogen phosphate 1-10g/l, agar 10-30g/l, pH4-8.
The preparation method of truffle aroma former seed liquor:
Step 1: get the ascocarpous intermediate structure block of fresh truffle, epidermis, epidermis soil, mycelia in soil, the each 5g of exotrophic mycorrhiza of truffle and host, pulpous state is worn into sterile mortar after sterilization, aseptically join in sterile distilled water and shake 10min, therefrom getting 100 μ l fluid drips is added on screening and culturing base evenly spreadable, be separated with plate streaking under aerobic conditions or dilute to be coated with after flat band method is purified and obtain a collection of microbial strains, then carry out fermenting experiment with liquid screening medium and therefrom filter out the truffle aroma former that the rich bacterial strain producing alpha-androstanol is screening, bacterial classification is the curved formula of bacterium, the content of alpha-androstanol can be improved by natural variation to bacterial strain, screening and culturing base is: glucose 50g/l, peptone 10g/l, yeast extract 0.1g/l, potassium dihydrogen phosphate 0.1g/l, magnesium sulfate 0.01g/l, oak leaf juice or Oak Tree endophyte zymotic fluid 10ml/l, agar 10g/l, pH value 4, remove agar and be liquid screening medium.
Step 2: truffle aroma former slant strains is inoculated in seed culture medium, 20 DEG C, shaken cultivation 10 hours under rotating speed 100rpm condition, obtained truffle aroma former seed liquor; Described seed culture medium is: glucose 5g/l, oak leaf juice 900ml/l, peptone 1g/l, yeast extract 1g/l, magnesium sulfate 0.1g/l, potassium dihydrogen phosphate 0.1g/l, pH4.
The symbiotic fermentation preparation method of truffle alpha-androstanol microcapsule powder:
Step 1: by truffle aroma former seed liquor according in the volume ratio access Oak Tree endophyte zymotic fluid of 5% in temperature be 20 DEG C, speed of agitator is 50 revs/min of condition bottom fermentations 1 day, obtains symbiotic fermentation liquid;
Step 2: symbiotic fermentation liquid is added enzyme preparation and carry out enzymolysis be heated to 80 DEG C after 1 hour and be incubated 1 hour at 20 DEG C; Symbiotic fermentation liquid reduced pressure concentration after enzymolysis or nitrogen are blown concentrated after at 10 DEG C vacuum drying make water content be 5%, obtain truffle aroma former tunning; Described enzyme preparation is cellulase, interior raw Penicillium notatum cellulase, flavor protease;
Step 3: by truffle aroma former tunning and concentration be 100% the ratio of ethanol by weight 100: 10 mix after, in 20 DEG C, ultrasonic wave process 10 minutes under 20KHz; Truffle aroma former tunning through ultrasonic wave process is obtained truffle alpha-androstanol crude extract;
Step 4: be trehalose, sucrose, the glucose of 0.1: 1: 1 by weight ratio, add in the water of 1 times to mix and be heated to 70 DEG C and become molten condition, be cooled to when 40 DEG C, add the truffle alpha-androstanol crude extract accounting for water weight 1% while stirring, 1-30 minute is stirred under 2000rpm rotating speed, adopt high-pressure homogeneous equipment by the material that is stirred homogeneous 0.1 hour at 20 DEG C, the mixed liquor crossed by homogeneous carries out spraying dry, obtains truffle alpha-androstanol microcapsule powder.
Embodiment 2
A kind of truffle composition, comprises 10g truffle Ultramicro-powder, 5g cocoa power, 30g Red Wine Polyphenols, 40g lactose.
Preparation method: 10g truffle Ultramicro-powder, 5g cocoa power, 30g Red Wine Polyphenols are ground rear mistake 100 mesh sieve respectively, adds lactose 40g, sodium carboxymethyl starch 6g after mixing, talcum powder 3.5g mixes; Ready press sheet mixture is mixed compressing tablet after granulating with PVP, to obtain final product.
Embodiment 3
A kind of truffle composition, comprises 3g truffle alpha-androstanol microcapsule powder, 15g cocoa power, 45g Red Wine Polyphenols, 45g Icing Sugar.
Preparation method: 3g truffle alpha-androstanol microcapsule powder, 15g cocoa power, 45g Red Wine Polyphenols are ground rear mistake 100 mesh sieve respectively, adds Icing Sugar 45g after mixing, low-substituted hydroxypropyl cellulose 3.2g mixes; Mixture is mixed rear granulation with PVP, mixes rear filling capsule with superfine silica gel powder 5g and obtain capsule.
The preparation method of Oak Tree endophyte zymotic fluid:
Step 1: choosing symbiosis has the skin of the Oak Tree of truffle, leaf, stem, flower, fruit and mycorhiza, first use aseptic water washing surface dirt, then aseptically by the alcohol immersion 30 seconds that volume ratio is 75%, sterilized water washes surperficial ethanol off, 30 seconds are soaked again with the mercuric chloride that mass fraction is 0.1%, aseptic water washing 3-5 time, then be inoculated on PDA culture medium flat plate respectively with the fritter that aseptic cutter is cut into 3 millimeters * 3 millimeters afterwards, 50 DEG C of constant temperature culture; Treat obviously to grow bacterium colony around sample, the bacterium colony of picking different shape, forward screening and culturing base inclined-plane to, 25 DEG C of purifying cultivate the Oak Tree endophyte that the dominant strain obtained is screening; Screening and culturing base is: yeast extract 2g/l, potassium dihydrogen phosphate 1g/l, magnesium sulfate 0.1g/l, fresh truffle juice 500ml/l, agar 30g/l, pH value 9;
Step 2: the Oak Tree endophyte of separation is inoculated in activation medium inclined-plane, obtains Oak Tree endophyte slant strains for 48 hours in 25 DEG C of activation, is inoculated in MS fluid nutrient medium and cultivates 15 days, obtain Oak Tree endophyte zymotic fluid; Activation medium is: glucose 50 grams per liter, peptone 12g/l, magnesium sulfate 3g/l, potassium dihydrogen phosphate 3.6g/l, agar 20g/l, pH4.8.
Described truffle aroma former seed liquor is prepared by following methods:
Step 1: get the ascocarpous intermediate structure block of fresh truffle, epidermis, epidermis soil, mycelia in soil, the each 10g of exotrophic mycorrhiza of truffle and host, pulpous state is worn into sterile mortar after sterilization, aseptically join in sterile distilled water and shake 60min, therefrom getting 1000 μ l fluid drips is added on screening and culturing base evenly spreadable, under anaerobic be separated with plate streaking or dilute to be coated with after flat band method is purified and obtain a collection of microbial strains, then carry out fermenting experiment with liquid screening medium and therefrom filter out the truffle aroma former that the rich bacterial strain producing alpha-androstanol is screening, bacterial classification is bacterium song or bacterium liquid form, can be made a variation by artificial induction to bacterial strain and improve the content of alpha-androstanol, screening and culturing base is: sucrose 300g/l, peptone 10g/l, yeast extract 0.2g/l, potassium dihydrogen phosphate 1g/l, magnesium sulfate 0.1g/l, Oak Tree endophyte zymotic fluid 10ml/l, agar 30g/l, pH value 4.9, remove agar and be liquid screening medium.
Step 2: truffle aroma former slant strains is inoculated in seed culture medium, 24 DEG C, shaken cultivation 4 hours under rotating speed 10rpm condition, obtained truffle aroma former seed liquor; Described seed culture medium is: glucose 15g/l, Oak Tree endophyte zymotic fluid 190ml/l, peptone 3g/l, yeast extract 5g/l, magnesium sulfate 0.5g/l, potassium dihydrogen phosphate 0.5g/l, pH6.
A kind of symbiotic fermentation preparation method of truffle alpha-androstanol microcapsule powder:
Step 1: by truffle aroma former seed liquor according in the volume ratio access Oak Tree endophyte zymotic fluid of 10% in temperature be 27 DEG C, speed of agitator is 100 revs/min of condition bottom fermentations 12 days, obtains symbiotic fermentation liquid;
Step 2: symbiotic fermentation liquid is placed in supercritical carbon dioxide extracting still, extraction kettle pressure be 30MPa, temperature be 36 DEG C of conditions under extraction 12 hours, collect extract and carry out reduced pressure concentration and obtain truffle alpha-androstanol crude extract;
Step 3: be trehalose, sucrose, the glucose of 5: 1: 5 by weight ratio, add in the water of 10 times to mix and be heated to 98 DEG C and become molten condition, be cooled to when 60 DEG C, add the truffle alpha-androstanol crude extract accounting for water weight 10% while stirring, stir 10 minutes under 3000rpm rotating speed, adopt high-pressure homogeneous equipment by the material that is stirred homogeneous 0.5 hour at 40 DEG C, the mixed liquor crossed by homogeneous carries out freeze drying, obtains truffle alpha-androstanol microcapsule powder.
Embodiment 4
A kind of truffle composition, comprises 13g truffle alpha-androstanol microcapsule powder, 75g cocoa power, 85g Red Wine Polyphenols, 54g sweet mellow wine.
Preparation method: mix after 13g truffle alpha-androstanol microcapsule powder, 75g cocoa power, the grinding of 85g Red Wine Polyphenols; In mixture, add pure water, granulate; In mixture, add sweet mellow wine 54g, PVPP 15g, dolomol 2.5g mixes; Ready press sheet mixture is mixed rear direct tablet compressing with PVP.
The preparation method of Oak Tree endophyte zymotic fluid:
Step 1: choosing symbiosis has the skin of the Oak Tree of truffle, leaf, stem, flower, fruit and mycorhiza, first use aseptic water washing surface dirt, then aseptically by the alcohol immersion 30 seconds that volume ratio is 75%, sterilized water washes surperficial ethanol off, 30 seconds are soaked again with the mercuric chloride that mass fraction is 0.1%, aseptic water washing 3-5 time, then be inoculated on PDA culture medium flat plate respectively with the fritter that aseptic cutter is cut into 3 millimeters * 3 millimeters afterwards, 35 DEG C of constant temperature culture; Treat obviously to grow bacterium colony around sample, the bacterium colony of picking different shape, forward screening and culturing base inclined-plane to, 40 DEG C of purifying cultivate the Oak Tree endophyte that the dominant strain obtained is screening; Screening and culturing base is: sucrose 150g/l, peptone 70g/l, yeast extract 2g/l, potassium dihydrogen phosphate 0.5g/l, magnesium sulfate 2g/l, fresh truffle juice 600ml/l, agar 20g/l, pH value 8;
Step 2: the Oak Tree endophyte of separation is inoculated in activation medium inclined-plane, obtains Oak Tree endophyte slant strains for 36 hours in 45 DEG C of activation, is inoculated in MS fluid nutrient medium and cultivates 30 days, obtain Oak Tree endophyte zymotic fluid; Activation medium is: glucose 40g/l, peptone 18g/l, magnesium sulfate 9g/l, potassium dihydrogen phosphate 3g/l, agar 30g/l, pH7.5.
Described truffle aroma former seed liquor is prepared by following methods:
Step 1: get the ascocarpous intermediate structure block of fresh truffle, epidermis, epidermis soil, mycelia in soil, the each 8g of exotrophic mycorrhiza of truffle and host, pulpous state is worn into sterile mortar after sterilization, aseptically join in sterile distilled water and shake 50min, therefrom getting 900 μ l fluid drips is added on screening and culturing base evenly spreadable, under anaerobic be coated with after flat band method is purified with dilution and obtain a collection of microbial strains, then carry out fermenting experiment with liquid screening medium and therefrom filter out the truffle aroma former that the rich bacterial strain producing alpha-androstanol is screening, bacterial classification is bacterium liquid form, the content of alpha-androstanol can be improved by natural variation or artificial induction's variation to bacterial strain, screening and culturing base is: sucrose 50g/l, peptone 60g/l, yeast extract 0.8g/l, potassium dihydrogen phosphate 0.7g/l, magnesium sulfate 1g/l, Oak Tree endophyte zymotic fluid 10ml/l, agar 20g/l, pH value 6, remove agar and be liquid screening medium.
Step 2: truffle aroma former slant strains is inoculated in seed culture medium, 35 DEG C, shaken cultivation 30 hours under rotating speed 120rpm condition, obtained truffle aroma former seed liquor; Described seed culture medium is: glucose 30g/l, Oak Tree endophyte zymotic fluid 20ml/l, peptone 7g/l, yeast extract 4g/l, magnesium sulfate 0.8g/l, potassium dihydrogen phosphate 1.5g/l, pH7.4.
A kind of symbiotic fermentation preparation method of truffle alpha-androstanol microcapsule powder:
Step 1: by truffle aroma former seed liquor according in the volume ratio access Oak Tree endophyte zymotic fluid of 10% in temperature be 60 DEG C, speed of agitator is 550 revs/min of condition bottom fermentations 16 days, obtains symbiotic fermentation liquid;
Step 2: symbiotic fermentation liquid is placed in supercritical carbon dioxide extracting still, extraction kettle pressure be 20-50MPa, temperature extracts 1-24 hour under being 30-60 DEG C of condition, collect extract and carry out reduced pressure concentration to obtain truffle alpha-androstanol crude extract;
Step 3: be trehalose, sucrose, the glucose of 3: 1: 2 by weight ratio, add in the water of 15 times to mix and be heated to 98 DEG C and become molten condition, be cooled to when 45 DEG C, add the truffle alpha-androstanol crude extract accounting for water weight 10% while stirring, stir 13 minutes under 2500rpm rotating speed, adopt high-pressure homogeneous equipment by the material that is stirred homogeneous 0.5 hour at 30 DEG C, the mixed liquor crossed by homogeneous carries out freeze drying, obtains truffle alpha-androstanol microcapsule powder.
Embodiment 5
A kind of truffle composition, comprises 54g truffle alpha-androstanol microcapsule powder, 35g ground coffee, 65g Red Wine Polyphenols.
Preparation method: mix after 54g truffle alpha-androstanol microcapsule powder, 35g ground coffee, the grinding of 65g Red Wine Polyphenols; In mixture, add pure water, granulate; In mixture, add superfine silica gel powder 1.2g mix, divide to be filled in capsule and make capsule.
The preparation method of Oak Tree endophyte zymotic fluid:
Step 1: choosing symbiosis has the skin of the Oak Tree of truffle, leaf, stem, flower, fruit and mycorhiza, first use aseptic water washing surface dirt, then aseptically by the alcohol immersion 30 seconds that volume ratio is 75%, sterilized water washes surperficial ethanol off, 30 seconds are soaked again with the mercuric chloride that mass fraction is 0.1%, aseptic water washing 3-5 time, then be inoculated on PDA culture medium flat plate respectively with the fritter that aseptic cutter is cut into 3 millimeters * 3 millimeters afterwards, 40 DEG C of constant temperature culture; Treat obviously to grow bacterium colony around sample, the bacterium colony of picking different shape, forward screening and culturing base inclined-plane to, 30 DEG C of purifying cultivate the Oak Tree endophyte that the dominant strain obtained is screening; Screening and culturing base is: glucose 80g/l, peptone 13g/l, yeast extract 5g/l, potassium dihydrogen phosphate 2g/l, magnesium sulfate 0.5g/l, fresh truffle juice 700ml/l, agar 10g/l, pH value 7;
Step 2: the Oak Tree endophyte of separation is inoculated in activation medium inclined-plane, obtains Oak Tree endophyte slant strains for 24 hours in 30 DEG C of activation, is inoculated in MS fluid nutrient medium and cultivates 25 days, obtain Oak Tree endophyte zymotic fluid; Activation medium is: glucose 40g/l, peptone 18g/l, magnesium sulfate 7g/l, potassium dihydrogen phosphate 2g/l, agar 30g/l, pH8.
Described truffle aroma former seed liquor is prepared by following methods:
Step 1: get the ascocarpous intermediate structure block of fresh truffle, epidermis, epidermis soil, mycelia in soil, the each 8g of exotrophic mycorrhiza of truffle and host, pulpous state is worn into sterile mortar after sterilization, aseptically join in sterile distilled water and shake 40min, therefrom getting 800 μ l fluid drips is added on screening and culturing base evenly spreadable, under anaerobic be separated with plate streaking or dilute to be coated with after flat band method is purified and obtain a collection of microbial strains, then carry out fermenting experiment with liquid screening medium and therefrom filter out the truffle aroma former that the rich bacterial strain producing alpha-androstanol is screening, bacterial classification is bacterium song or bacterium liquid form, can be made a variation by artificial induction to bacterial strain and improve the content of alpha-androstanol, screening and culturing base is: sucrose 250g/l, peptone 90g/l, yeast extract 4g/l, potassium dihydrogen phosphate 2g/l, magnesium sulfate 6g/l, oak leaf juice 800ml/l, agar 30g/l, pH value 5.7, remove agar and be liquid screening medium.
Step 2: truffle aroma former slant strains is inoculated in seed culture medium, 30 DEG C, shaken cultivation 36 hours under rotating speed 110rpm condition, obtained truffle aroma former seed liquor; Described seed culture medium is: glucose 40g/l, Oak Tree endophyte zymotic fluid 20ml/l, peptone 6g/l, yeast extract 4g/l, magnesium sulfate 3.8g/l, potassium dihydrogen phosphate 2.1g/l, pH9.
A symbiotic fermentation preparation method for truffle alpha-androstanol microcapsule powder, is characterized in that, comprise the following steps:
Step 1: by truffle aroma former seed liquor according in the volume ratio access Oak Tree endophyte zymotic fluid of 20% in temperature be 60 DEG C, speed of agitator is 400 revs/min of condition bottom fermentations 18 days, obtains symbiotic fermentation liquid;
Step 2: symbiotic fermentation liquid is added enzyme preparation and carry out enzymolysis be heated to 130 DEG C after 8 hours and be incubated 8 hours at 55 DEG C; Symbiotic fermentation liquid nitrogen after enzymolysis is blown concentrated after at 70 DEG C vacuum drying make water content be 9%, obtain truffle aroma former tunning; Described enzyme preparation is interior raw Penicillium notatum cellulase, flavor protease, bacillus protein enzyme, beta-glucosidase;
Step 3: by truffle aroma former tunning and concentration be 80% the ratio of ethanol by weight 100: 150 mix after, in 40 DEG C, ultrasonic wave process 100 minutes under 60KHz; Truffle aroma former tunning through ultrasonic wave process is carried out reduced pressure concentration and obtains truffle alpha-androstanol crude extract;
Step 4: be trehalose, sucrose, the glucose of 3: 1: 4 by weight ratio, add in the water of 16 times to mix and be heated to 88 DEG C and become molten condition, be cooled to when 55 DEG C, add the truffle alpha-androstanol crude extract accounting for water weight 60% while stirring, stir 18 minutes under 3000rpm rotating speed, adopt high-pressure homogeneous equipment by the material that is stirred homogeneous 3 hours at 35 DEG C, the mixed liquor crossed by homogeneous carries out freeze drying, obtains truffle alpha-androstanol microcapsule powder.
Embodiment 6:
A kind of truffle composition, comprises 90g truffle polysaccharide, 19g state aroma ground coffee, 10g Red Wine Polyphenols, cellobiose 67g.
Preparation method: 90g truffle polysaccharide, 19g state aroma ground coffee, 10g Red Wine Polyphenols are ground rear mistake 80 mesh sieve respectively, adds cellobiose 67g, sodium carboxymethyl starch 5g after mixing, talcum powder 3g mixes; Ready press sheet mixture is mixed rear direct tablet compressing with PVP, to obtain final product.
Embodiment 7:
A kind of truffle composition, comprises 70g truffle polysaccharide, 43g coffee bean particle, 32g Red Wine Polyphenols, FOS 54g.
Preparation method: 70g truffle polysaccharide, 43g coffee bean particle, 32g Red Wine Polyphenols are ground rear mistake 100 mesh sieve respectively, adds FOS 54g, sodium carboxymethyl starch 6g after mixing, talcum powder 3.5g mixes; Ready press sheet mixture is mixed compressing tablet after granulating with PVP, to obtain final product.
Embodiment 8:
A kind of truffle composition, comprises 30g fermentation truffle bacterium powder, 63g instant coffee powder, 72g Red Wine Polyphenols, FOS 38g.
Preparation method: truffle of being fermented by 30g bacterium powder, 63g instant coffee powder, 72g Red Wine Polyphenols grind rear mistake 100 mesh sieve respectively, add FOS 38g, low-substituted hydroxypropyl cellulose 3.2g mixes after mixing; Mixture is mixed rear granulation with PVP, mixes rear filling capsule with superfine silica gel powder 5g and obtain capsule.
Embodiment 9:
A kind of truffle composition, comprises 18g fermentation truffle extract, 95g cocoa power, 36g Red Wine Polyphenols, xylitol 42g.
Preparation method: truffle of being fermented by 18g extract, 95g cocoa power, 36g Red Wine Polyphenols grind rear mistake 100 mesh sieve respectively, add xylitol 42g, low-substituted hydroxypropyl cellulose 7.6g mixes after mixing; Mixture is mixed rear granulation with PVP, mixes rear filling capsule with superfine silica gel powder 3.8g and obtain capsule.
Claims (8)
1. a truffle composition, is characterized in that, comprises 0.1-90 weight portion truffle, 1-99 weight portion coffee and/or cocoa power, 1-80 weight portion Red Wine Polyphenols.
2. a kind of truffle composition according to claim 1, is characterized in that, comprises 0.1-90 weight portion truffle, 1-99 weight portion coffee and/or cocoa power, 1-80 weight portion Red Wine Polyphenols, 1-80 parts by weight of saccharide.
3. a kind of truffle composition according to claim 1, is characterized in that described truffle is one or more in truffle alpha-androstanol microcapsule powder, truffle microcapsule powder, truffle polysaccharide, fresh truffle, truffle Ultramicro-powder, fermentation truffle bacterium powder, fermentation truffle extract, truffle ferment, truffle endophyte tunning.
4. a kind of truffle composition according to claim 1, is characterized in that, described coffee is one or more in coffee bean, coffee bean particle, state's aroma ground coffee, instant coffee powder.
5. a kind of truffle composition according to claim 2, it is characterized in that, described carbohydrate is one or more in sucrose, fructose, honey, glucose, cellobiose, white sugar, brown sugar, brown sugar, Arabinose, HFCS, D-ribose, date palm syrup, soyabean oligosaccharides, FOS, isomalto-oligosaccharide, galactooligosaccharide, xylo-oligosaccharide, stachyose, raffinose, chitosan oligomer, xylitol, D-sorbite, maltitol, antierythrite.
6. a symbiotic fermentation preparation method for truffle alpha-androstanol microcapsule powder, is characterized in that, comprise the following steps:
Step 1: by truffle aroma former seed liquor according in the volume ratio access Oak Tree endophyte zymotic fluid of 5-30% in temperature be 20-70 DEG C, speed of agitator is 50-600 rev/min of condition bottom fermentation 1-20 days, obtains symbiotic fermentation liquid;
Step 2: by truffle aroma former tunning and concentration be the ethanol of 60%-100% by weight 100: after the ratio of (1-200) mixes, in 20-70 DEG C, ultrasonic wave process 1-120 minute under 20-80KHz; Supercritical carbon dioxide extracting still is placed in by through the truffle aroma former tunning of ultrasonic wave process or symbiotic fermentation liquid, extraction kettle pressure be 20-50MPa, temperature extracts 1-24 hour under being 30-60 DEG C of condition, collect extract and carry out reduced pressure concentration to obtain truffle alpha-androstanol crude extract;
Described truffle aroma former tunning is prepared by following methods:
Symbiotic fermentation liquid is added and is heated to 80-150 DEG C after enzyme preparation carries out enzymolysis 1-15 hour at 20-85 DEG C and is incubated 1-15 hour; Symbiotic fermentation liquid reduced pressure concentration after enzymolysis or nitrogen are blown concentrated after at 10-80 DEG C vacuum drying make water content be 5%-11%, obtain truffle aroma former tunning; Described enzyme preparation is one or more in cellulase, interior raw Penicillium notatum cellulase, flavor protease, bacillus protein enzyme, pectase and beta-glucosidase;
Step 3: be (0.1-5) by weight ratio: 1: the trehalose of (1-5), sucrose, glucose, add in 1-20 water doubly to mix and be heated to 70-98 DEG C and become molten condition, be cooled to add while stirring during 40-60 DEG C the truffle alpha-androstanol crude extract and/or truffle slurries that account for water weight 1-70%, 1-30 minute is stirred under 2000-5000rpm rotating speed, adopt material homogeneous 0.1-5 hour at 20-40 DEG C that high-pressure homogeneous equipment will be stirred, the mixed liquor crossed by homogeneous carries out spraying dry or freeze drying, obtain truffle alpha-androstanol microcapsule powder.
7. the symbiotic fermentation preparation method of truffle alpha-androstanol microcapsule powder according to claim 6, is characterized in that, described truffle aroma former seed liquor is prepared by following methods:
Step 1: get the ascocarpous intermediate structure block of fresh truffle, epidermis, epidermis soil, mycelia in soil, the each 5-10g of exotrophic mycorrhiza of truffle and host, pulpous state is worn into sterile mortar after sterilization, aseptically join in sterile distilled water and shake 10-60min, therefrom getting 100-1000 μ l fluid drips is added on screening and culturing base evenly spreadable, be separated with plate streaking under aerobic or anaerobic condition or dilute to be coated with after flat band method is purified and obtain a collection of microbial strains, then carry out fermenting experiment with liquid screening medium and therefrom filter out the truffle aroma former that the rich bacterial strain producing alpha-androstanol is screening, bacterial classification is bacterium song or bacterium liquid form, the content of alpha-androstanol can be improved by natural variation or artificial induction's variation to bacterial strain, 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 juice or Oak Tree endophyte zymotic fluid 1-900ml/l, agar 1-30g/l, pH value 4-9, remove agar and be liquid screening medium.
Step 2: truffle aroma former slant strains is inoculated in seed culture medium, at 20-40 DEG C, shaken cultivation 1-40 hour under rotating speed 10-150rpm condition, obtained truffle aroma former seed liquor; Described seed culture medium is: glucose 1-50g/l, oak leaf juice or Oak Tree endophyte zymotic fluid 1-900ml/l, peptone 1-10g/l, yeast extract 1-10g/l, magnesium sulfate 0.1-5g/l, potassium dihydrogen phosphate 0.1-5g/l, pH4-10.
8. the symbiotic fermentation preparation method of truffle alpha-androstanol microcapsule powder according to claim 6, is characterized in that described Oak Tree endophyte zymotic fluid is prepared by following methods:
Step 1: choosing symbiosis has the skin of the Oak Tree of truffle, leaf, stem, flower, fruit and mycorhiza, first use aseptic water washing surface dirt, then aseptically by the alcohol immersion 30 seconds that volume ratio is 75%, sterilized water washes surperficial ethanol off, 30 seconds are soaked again with the mercuric chloride that mass fraction is 0.1%, aseptic water washing 3-5 time, then be inoculated on PDA culture medium flat plate respectively with the fritter that aseptic cutter is cut into 3 millimeters * 3 millimeters afterwards, 20-50 DEG C of constant temperature culture; Treat obviously to grow bacterium colony around sample, the bacterium colony of picking different shape, forward screening and culturing base inclined-plane to, 20-50 DEG C of purifying cultivates the Oak Tree endophyte that the dominant strain obtained is screening; 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, fresh truffle juice 1-900ml/l, agar 1-30g/l, pH value 4-9;
Step 2: the Oak Tree endophyte of separation is inoculated in activation medium inclined-plane, obtains Oak Tree endophyte slant strains for 1-48 hour in 20-50 DEG C of activation, is inoculated in MS fluid nutrient medium and cultivates 1-50 days, obtain Oak Tree endophyte zymotic fluid; Activation medium is: glucose 20-50g/l, peptone 1-20g/l, magnesium sulfate 1-10g/l, potassium dihydrogen phosphate 1-10g/l, agar 10-30g/l, pH4-8.
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