CN102286324B - Method for preparing gingko golden fungus fruit wine - Google Patents
Method for preparing gingko golden fungus fruit wine Download PDFInfo
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Abstract
The invention relates to a method for preparing gingko golden fungus fruit wine, which comprises the following steps that: golden fungus type spore and monoxenous stereum hirsutum parent strains are used for carrying out liquid bottle shaking seed culture of the golden fungus type spores, the seed tank culture of the golden fungus type spores, the bottle shaking seed culture of the autoecious stereum hirsutum, the bottle shaking expansion culture of the autoecious stereum hirsutum and the seed tank culture of the autoecious stereum hirsutum, the seed tank seed of the golden fungus type spores and the seed tank seed of the autoecious stereum hirsutum are subjected to the mixed fermentation culture, ginkgo starch substrates and wine brewing yeasts are added into the mixed fermentation liquid of golden fungus type spores and the autoecious stereum hirsutum for carrying out post fermentation, the gingko golden fungus fruit wine with golden fungus extracellular polysaccharide, stereum hirsutum terpene, ginkgo flavone and bilobalide b can be obtained, the alcohol volume is higher than 3 percent, the ginkgo phenolic acid is smaller than 1ppm, and the hydrocyanic acid is lower than 0.1microgramme per liter. The gingko golden fungus fruit wine obtained by the method has an anti-radiation health care function.
Description
Technical field
The invention belongs to biological technical field and field of food; Specifically the present invention relates to a kind of preparation method of ginkgo gold ear fruit wine; Be through golden ear yeast shape spore and the tough lead fungi co-fermentation of host's pili gingko; Utilize the powerful material decomposition ability of fungi, remove gingko toxic component ginkgolic acid and aldehydes matter.The ginkgo gold ear fruit wine that is obtained by this method has the anti-radiation health care function.
Background technology
The gold ear is called golden auricularia auriculajudae, Tremella mesenterica, Tremella mesenterica Retz ex Fr., golden yellow white fungus, gold and silver ear etc.The Latin formal name used at school
Tremella aurantialba, golden ear medicinal is documented in " Mingyi Bielu " of TAO Hong-Jing the earliest, and Ming Dynasty Compendium of Material Medica dish portion the 28 volume has description to its form, medicinal part and nature,taste and action.Gold remebrance flavor is sweet flat, nourishing Yin and moistening lung, relieving cough and reducing sputum, promotes the production of body fluid.The gold ear colonizes in nerville high mountain oak or light leaf high mountain robur and does, with hair Boreostereum vibrans (
Stereum hirsutum) the association life, wild distribution is rare, is Chinese endemic species.Complete golden ear basidioma brain shape, the section skin is orange-yellow colloid, is the meat district of white in the middle of the sliver, and what have does not enrich, and is the netted of hollow, and the inner white meat is host's hair Boreostereum vibrans silk district.
Gold ear this precious distinct Chinese characteristics medicinal fungi resource forms the Heterogeneous Composite body jointly because the singularity of life has determined it to exist together with host's hair Boreostereum vibrans, and the existence of therefore a small amount of host's hair Boreostereum vibrans is the golden ear basis that normal growth grows of relying.Because the powerful decomposition of hair Boreostereum vibrans and utilize the ability of glucide such as lignocellulose; In the history of growing of golden ear, very easily be in growth vigor; The silk of the hair Boreostereum vibrans often culture that adopts traditional vegetative reproduction means to obtain; But possibly be mistaken as is the tremella silk, adopts this hair Boreostereum vibrans culture to carry out submerged fermentation and is difficult to obtain golden ear product truly.To the golden ear singularity of the life history, Chinese patent CN101225361 B, CN101831369 A, CN101822373 A and CN101182562 A all do not identify the tremella kind of using, and is difficult to guarantee the effect behind the patent working.Chinese patent 89100117.4 has been reported golden fungus substituting stuff cultivation method; This patent is in order to guarantee to obtain effective bacterial classification of golden ear; Adopt and directly the tissue block of golden ear is carried out inoculation culture as seed, but extremely precious difficult acquisition of golden ear resource limited the enforcement of patent.Chinese patent 200410022377.3 has been reported the method for golden ear yeast shape spore fermentative prodn food and foodstuff additive; This patent uses the yeast shape spore of golden ear to make bacterial classification merely; Remove host's hair Boreostereum vibrans thalline, can not represent the complete medicinal kind of golden ear.
Contain in the fungi cultured solution of deep zone of yeast shape spore of gold ear and abundant have the immunizing power of raising, hypoglycemic and delay senility, and the exocellular polysaccharide of anti-inflammatory and multiple pharmacologically active such as antitumor.Contain a series of ethynyl aromatics and tricyclic sesquiterpene alkene classes with the former bacterium of broad-spectrum disease resistance and anti tumor activity in vitro in the orange-yellow bacterium liquid of the tough lead fungi liquid fermentation and culture of gold ear host's pili, these materials have good antitumous effect.
Ginkgo is commonly called as gingko, is traditional Chinese medicine, also is excellent tonic product, and different name mental eye, Herba Sedi Multicaulis, giant Buddha refer to, Buddhist refers to mandarin orange." Chinese pharmacopoeia (version in 2000) is included medicinal gingko Renhe and is fried gingko benevolence, and the time spent smashs to pieces, and the gas of astringing the lung is coughed surely and breathed heavily, and only is with turbid.The tonifying Qi of " book on Chinese herbal medicine is new again " record gingko is nourished heart, the kidney-nourishing enriching yin, and the cough-relieving relieving restlessness, myogenic is put on flesh, and apocenosis is drawn out pus by applying a plaster to the affected part, the sore that disappears scabies subcutaneous ulcer knurl.Per 100 grams of gingko benevolence contain protein 6.4 grams, fat 2.4 grams, glucide 36 grams, 10 milligrams of calcium, 1 milligram of iron, 218 milligrams in phosphorus, vitamin G 50 micrograms, Serlabo 3.2 micrograms.The ginkgo gold ear health fruit that utilizes the common submerged fermentation gingko of golden ear and hair Boreostereum vibrans to process is not seen same or analogous report in the prior art as yet.
Summary of the invention
The purpose of this invention is to provide a kind of is that matrix is carried out the yeast shape spore of golden ear and the method that host's hair Boreostereum vibrans mixing submerged fermentation prepares ginkgo gold ear fruit wine with the gingko; Prepared ginkgo gold ear fruit wine outward appearance is glittering and translucent; Aroma is elegant, and the wine body is pure, and is mellow and full tasty and refreshing; Pleasant impression is long, has the anti-radiation health care effect.
The preparation method of a kind of ginkgo gold ear fruit wine of the present invention may further comprise the steps:
(1) slant strains with golden ear yeast shape spore inserts in the PDA substratum, and culture temperature 18-28 ℃, incubation time 48-120 hour;
(2) slant strains with the tough lead fungi of host's pili inserts in the PDA substratum, and culture temperature 23-28 ℃, incubation time 120-148 hour;
The slant strains of the golden ear yeast shape spore that (3) step (1) is obtained changes in the triangular flask that the shake-flask seed substratum is housed, cultivates in 23-28 ℃, and rotating speed 50-250 rev/min, incubation time 36-75 hour;
The slant strains of the tough lead fungi of host's pili that (4) step (2) is obtained changes in the triangular flask that the shake-flask seed substratum is housed, cultivates in 23-28 ℃, and rotating speed 50-250 rev/min, incubation time 40-120 hour;
(5) the golden ear yeast shape spore shake-flask seed that step (3) is obtained is transferred by the inoculum size of 5-10% and is carried out seed culture in the seeding tank that the seed tank culture base is housed; Culture temperature 23-28 ℃; Seeding tank pressure 0.05Mpa; Stir speed (S.S.) 50-250 rev/min, ventilation 1: (0.3-1.0) v/v/m, incubation time 36-60 hour;
(6) the tough lead fungi shake-flask seed of host's pili that step (4) is obtained is transferred to be equipped with to shake in bottle triangular flask of enlarged culturing base by the inoculum size of 5-10% and is carried out enlarged culturing, and culture temperature 23-28 ℃, rotating speed 50-250 rev/min, incubation time 60-80 hour;
(7) the tough lead fungi of host's pili that step (6) is obtained shakes bottle and enlarges seed and transfer by the inoculum size of 5-10% and carry out seed culture in the seeding tank that the seed tank culture base is housed; Culture temperature 23-28 ℃; Seeding tank pressure 0.05Mpa, stir speed (S.S.) 50-250 rev/min, inoculum size is per-cent 5-10% by volume; Ventilation 1: (0.3-1.0) v/v/m, incubation time 48-120 hour;
(8) the tough lead fungi seeding tank of the host's pili seed that golden ear yeast shape spore seeding tank seed that step (5) is obtained and step (7) obtain by two kinds of bacterium of inoculum size by volume the inoculum size of each 5-10% of per-cent transfer and carry out mixed culture in the fermentor tank that fermention medium is housed; Culture temperature 23-28 ℃; Fermentor tank pressure 0.05Mpa; Mixing speed 50-250 rev/min, ventilation 1: (0.3-1.0) v/v/m, 72-180 hour;
(9) gingko starch is soaked into mineral water, boiling 25-40 minute, be cooled to 50-65 ℃; The Mi Qu of 0.5%-1.0% adding by mass percentage is incubated saccharification 1-2 hour, and the intensification sterilising and enzyme inactivating is after the saccharification; Be cooled to 15-18 ℃; 2-3 according to saccharification material gross weight extraordinarily goes into golden ear yeast shape spore and the fermented liquid of the tough lead fungi of host's pili after fermention medium mixed fermentation that step (8) obtains, and mixing adds yeast saccharomyces cerevisiae and carries out secondary fermentation; The secondary fermentation temperature is controlled at 15-18 ℃, and the secondary fermentation time is 7-30 days;
(10) mash after the fermentation of step (9) acquisition is carried out squeeze and filter; Carried out germicidal treatment 15 seconds-15 minutes at 65 ℃-80 ℃ then, store, be 20-100 days storage period; The wine of storing after-ripening is colluded accent, carry out sterilization, filtration, packing again and promptly get ginkgo gold ear fruit wine.
Further; Among the preparation method of the present invention; Described golden ear yeast shape spore shake-flask seed substratum consists of (unit is a grams per liter): glucose 20-50, wheat bran 50-100, potassium primary phosphate 0.2-2, sal epsom 0.1-2, add water to proper volume, and initial pH is 5.8-7.0; The tough lead fungi shake-flask seed of described host's pili substratum consists of: glucose 20-50, wheat bran 10-50, soya bean 10-80, potassium primary phosphate 0.2-2, sal epsom 0.1-2, add water to proper volume, and initial pH is 5.8-7.0.
Among the preparation method of the present invention; The seed tank culture base of described golden ear yeast shape spore consists of (unit is a grams per liter): glucose 20-50, soya bean 30-60, potassium primary phosphate 1-2, sal epsom 0.1-0.5, soya-bean oil 0.2-0.5; Add water to proper volume, initial pH is 5.8-7.0; The tough lead fungi seed tank culture of described host's pili base consists of: glucose 20-50, soya bean 30-60, Semen Maydis powder 10-50, potassium primary phosphate 1-2, sal epsom 0.1-0.5, soya-bean oil 0.2-0.5, add water to proper volume, and initial pH is 5.8-7.0.
Among the preparation method of the present invention; The tough lead fungi of described host's pili shakes a bottle enlarged culturing base and consists of (unit is a grams per liter): glucose 20-50, soya bean 10-80, Semen Maydis powder 10-50, potassium primary phosphate 1-2, sal epsom 0.1-0.5; Add water to proper volume, initial pH is 5.8-7.0.
Among the preparation method of the present invention, said fermention medium consists of (unit is a grams per liter): gingko 100-200, soya-bean oil 0.2-0.5, add water to proper volume, and initial pH is 5.8-7.0.
Employed golden ear yeast shape spore and hair Boreostereum vibrans were golden ear B101 bacterial strain and the hair Boreostereum vibrans S102 bacterial strains that is obtained by contriver's screening among the following embodiment, and golden ear B101 bacterial strain and hair Boreostereum vibrans S102 submit China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation on April 21st, 2011.The classification called after gold ear of gold ear B101 (
Tremella aurantialba), preserving number is CGMCC No.4785; The classification called after of hair Boreostereum vibrans S102 (
Stereum hirsutum), preserving number is CGMCC No.4786.Depositary institution address: No. 3 Institute of Microorganism, Academia Sinica in Yard 1, BeiChen xi Road, Chaoyang District, Beijing City.
Among the preparation method of the present invention; Described slant medium is a substratum commonly used in the prior art, preferred PDA substratum, the composition of this type of substratum and used referring to China Committee for Culture Collection of Microorganisms agricultural mikrobe center; Chinese agriculture bacterial classification catalogue, 1991 editions: 93 pages.
Among the preparation method of the present invention, described golden ear yeast shape spore and/or the tough lead fungi shake-flask culture of host's pili base can be 3:20 to 5:20 with the ratio of the capacity of triangular flask; The tough lead fungi enlarged culturing of said host's pili base can be 3:20 to 5:20 with the ratio of triangular flask volume; Using than for when the volume of the above-mentioned seeding tank of the present invention during of said golden ear yeast shape spore and/or the tough lead fungi seed culture medium of host's pili as 100L, the amount of the seed culture medium that wherein is equipped with is 50-70L; Using than for when the volume of the above-mentioned fermentor tank of the present invention during as 1000-5000L of said golden ear yeast shape spore and the tough lead fungi mixed fermentive culture medium of host's pili, the amount of the fermention medium that wherein is equipped with should be 500-3500L mutually.
The present invention and existing open source literature relatively have following creativeness and advance:
1) zymotechnique adopts yeast shape spore and two kinds of bacterium co-fermentation of the tough lead fungi of host's pili gingko of golden ear; The meta-bolites advantage that had both kept complete wild golden ear; Overcome technological in the past because a hypha fermentation with lead fungi; And ignored the special life characteristic of golden ear, and can not take into account the defective of golden fungus polysaccharides and hair Boreostereum vibrans ter penoids, brought into play the powerful decomposition and inversion ability of the complementary generation of two bacterium again; Remove the poisonous phenolic acid of gingko, effectively promoted the pharmaceutical use of tunning;
2) the fermentation employing second time in zymotechnique gingko starch is matrix, has both guaranteed the pharmaceutical use of fruit wine, has the difference of essence again with other traditional wine technology of ginkgo, also is creative place of the present invention;
3) in the ginkgo that makes after the fermenting twice gold ear fruit wine enrichment platelet activating factor receptor antagonist Ginkgolide B efficiently; Immunostimulant gold ear glucosides class; And ginkgolic flavone glycoside and hair Boreostereum vibrans terpene with radioprotective, antitumor action, mouthfeel is suitable simultaneously, is easy to absorb; Advanced than other alcohol product technology, market outlook are very wide.
After the present invention utilizes golden ear and the common submerged fermentation gingko of hair Boreostereum vibrans benevolence, with adding raw materials such as gingko starch and yeast saccharomyces cerevisiae in the beer, the ginkgo gold ear health fruit of processing after the degree of depth fermentation; Be golden yellow low alcohol, removed deleterious composition in the gingko, mouthfeel fragrance is mellow; The wine body is pure, and mellow and full tasty and refreshing, alcoholic strength is greater than 3%; The gingko phenolic acid is less than 1ppm, and prussic acid is rich in golden fungus polysaccharides, sesquiterpene, ginkgolic flavone glycoside, Ginkgolide B etc. less than 0.1 micrograms per litre.The golden ear fruit wine that is obtained by the present invention can be worked in coordination with the double pharmacological action of golden ear of performance and ginkgo, has good antitumor, radioprotective, protection liver, kidney, cns, cardiovascular systems, and the effects such as rejection of anti-organ transplantation.
Embodiment
In order further to set forth the related preparation of technical scheme of the present invention, provided following embodiment.But the scope that these embodiment do not limit the present invention in any way.
Embodiment 1
The preparation method of a kind of ginkgo gold ear fruit wine of the present invention may further comprise the steps:
(1) slant strains with golden ear yeast shape spore inserts in the PDA substratum (China Committee for Culture Collection of Microorganisms agricultural mikrobe center, Chinese agriculture bacterial classification catalogue, 1991 editions: 93 pages) 18 ℃ of culture temperature, incubation time 120 hours;
(2) slant strains with the tough lead fungi of host's pili inserts in the PDA substratum (China Committee for Culture Collection of Microorganisms agricultural mikrobe center, Chinese agriculture bacterial classification catalogue, 1991 editions: 93 pages) 23 ℃ of culture temperature, incubation time 120 hours;
(3) the golden ear yeast shape spore slant strains that step (1) is obtained is transferred and is equipped with in the triangular flask of shake-flask seed substratum, in 23 ℃, and 50 rev/mins of rotating speeds, incubation time 36 hours; Wherein said golden ear yeast shape spore shake-flask seed substratum consists of (unit is a grams per liter): glucose 20, and wheat bran 50 (the boiling water liquor is crossed the leaching clear liquid), potassium primary phosphate 0.2, sal epsom 0.1 adds water to proper volume, and initial pH is 5.8;
(4) the tough lead fungi slant strains of host's pili that step (2) is obtained is transferred and is equipped with in the triangular flask of shake-flask seed substratum, in 23 ℃, and 50 rev/mins of rotating speeds, incubation time 40 hours; The tough lead fungi shake-flask seed of wherein said host's pili substratum consists of (unit is a grams per liter): glucose 20, and wheat bran 10 (the boiling water liquor is crossed the leaching clear liquid), soya bean 10, potassium primary phosphate 1, sal epsom 0.1 adds water to proper volume, and initial pH is 5.8;
The shake-flask seed of the golden ear yeast shape spore that (5) step (3) is obtained is transferred by the inoculum size of 5% (volume percent) and is cultivated in the seeding tank that the seed tank culture base is housed; 23 ℃ of culture temperature; Seeding tank pressure 0.05Mpa; 50 rev/mins of stir speed (S.S.)s, ventilation 1:0.3v/v/m, incubation time 36 hours; The seed culture medium of wherein said golden ear yeast shape spore consists of (unit is a grams per liter): glucose 20, and soya bean 30, potassium primary phosphate 1, sal epsom 0.1, soya-bean oil 0.2 adds water to proper volume, and initial pH is 5.8;
(6) the tough lead fungi of host's pili that step (4) is obtained shakes bottle bacterial classification and transfers to be equipped with to shake in bottle triangular flask of enlarged culturing base by 5% inoculum size and carry out enlarged culturing, 23 ℃ of culture temperature, 50 rev/mins of rotating speeds, incubation time 60 hours; The tough lead fungi of wherein said host's pili shakes a bottle enlarged culturing base and consists of (unit is a grams per liter): glucose 20, and soya bean 10, Semen Maydis powder 10, potassium primary phosphate 1, sal epsom 0.1 adds water to proper volume, and initial pH is 5.8;
The bottle that shakes of the tough lead fungi of host's pili that (7) step (6) is obtained enlarges seed and transfers by the inoculum size of 5% (volume percent) and cultivate in the seeding tank that seed culture medium is housed; 23 ℃ of culture temperature; Seeding tank pressure 0.05Mpa, 50 rev/mins of stir speed (S.S.)s, inoculum size 5% (volume percent); Ventilation 1:0.3v/v/m, incubation time 48 hours; The tough lead fungi seed culture medium of wherein said host's pili consists of (unit is a grams per liter): glucose 20, and soya bean 30, Semen Maydis powder 10, potassium primary phosphate 1, sal epsom 0.1, soya-bean oil 0.2 adds water to proper volume, and initial pH is 5.8;
(8) with ginkgo nut decapsidate and endothelium, plumule in removing again adds 5 times of water by volume and soaks making beating; The golden ear yeast shape spore that step (5) and step (7) are obtained and the seeding tank seed of the tough lead fungi of host's pili are respectively transferred by the inoculum size of 5% (volume percent) and are carried out mixed culture in the gingko fermention medium; 23 ℃ of culture temperature, fermentor tank pressure 0.05Mpa, 50 rev/mins of mixing speed; Ventilation 1:0.3v/v/m, 72 hours; The substratum of wherein said mixing fermentation culture consists of (unit is a grams per liter): gingko 100, and soya-bean oil 0.2 adds water to proper volume, and initial pH is 5.8;
(9) the gingko starch of the 1kg mineral water with 1L is soaked into, boiling 25 minutes is cooled to 50 ℃; 0.5% adding Mi Qu was incubated saccharification 1 hour by mass percentage, and the intensification sterilising and enzyme inactivating is after the saccharification; Be cooled to 15 ℃, extraordinarily go into golden ear yeast shape spore and the fermented liquid of the tough lead fungi of host's pili after fermention medium mixed fermentation that step (8) obtains, mixing according to 2 of saccharification material gross weight; Add yeast saccharomyces cerevisiae and carry out secondary fermentation, the secondary fermentation temperature is controlled at 15 ℃, and the secondary fermentation time is 7 days;
(10) mash after the fermentation of step (9) acquisition is carried out squeeze and filter, carried out germicidal treatment 15 seconds at 65 ℃ then, store again; Be 20 days storage period; The wine of storing after-ripening is colluded accent, carry out sterilization, filtration, packing again and promptly get ginkgo gold ear fruit wine, alcoholic strength is greater than 3%; The gingko phenolic acid is less than 1ppm, and prussic acid is less than 0.1 micrograms per litre.
Embodiment 2
The preparation method of a kind of ginkgo gold ear fruit wine of the present invention may further comprise the steps:
(1) slant strains with golden ear yeast shape spore inserts in the PDA substratum 28 ℃ of culture temperature, incubation time 48 hours;
(2) slant strains with the tough lead fungi of host's pili inserts in the PDA substratum 28 ℃ of culture temperature, incubation time 148 hours;
(3) the golden ear yeast shape spore slant strains that step (1) is obtained is transferred and is equipped with in the triangular flask of shake-flask seed substratum, in 28 ℃, and 250 rev/mins of rotating speeds, incubation time 75 hours; Wherein said golden ear yeast shape spore shake-flask seed substratum consists of (unit is a grams per liter): glucose 50, and wheat bran 100 (the boiling water liquor is crossed the leaching clear liquid), potassium primary phosphate 2, sal epsom 2 adds water to proper volume, and initial pH is 7.0;
(4) the tough lead fungi slant strains of host's pili that step (2) is obtained is transferred and is equipped with in the triangular flask of shake-flask seed substratum, in 28 ℃, and 250 rev/mins of rotating speeds, incubation time 120 hours; The tough lead fungi shake-flask seed of wherein said host's pili substratum consists of (unit is a grams per liter): glucose 50, and wheat bran 30 (the boiling water liquor is crossed the leaching clear liquid), soya bean 80, potassium primary phosphate 2, sal epsom 0.5 adds water to proper volume, and initial pH is 7.0;
The shake-flask seed of the golden ear yeast shape spore that (5) step (3) is obtained is transferred by the inoculum size of 10% (volume percent) and is cultivated in the seeding tank that seed culture medium is housed; 28 ℃ of culture temperature; Seeding tank pressure 0.05Mpa; 250 rev/mins of stir speed (S.S.)s, ventilation 1:1.0v/v/m, incubation time 60 hours; The seed culture medium of wherein said golden ear yeast shape spore consists of (unit is a grams per liter): glucose 50, and soya bean 60, potassium primary phosphate 2, sal epsom 0.5, soya-bean oil 0.5 adds water to proper volume, and initial pH is 7.0;
(6) the tough lead fungi of host's pili that step (4) is obtained shakes bottle bacterial classification and transfers to be equipped with to shake in bottle triangular flask of enlarged culturing base by 10% inoculum size and carry out enlarged culturing, 28 ℃ of culture temperature, 250 rev/mins of rotating speeds, incubation time 80 hours; The tough lead fungi of wherein said host's pili shakes a bottle enlarged culturing base and consists of (unit is a grams per liter): glucose 50, and soya bean 80, Semen Maydis powder 50, potassium primary phosphate 2, sal epsom 0.5 adds water to proper volume, and initial pH is 7.0;
The bottle that shakes of the tough lead fungi of host's pili that (7) step (6) is obtained enlarges seed and transfers by the inoculum size of 10% (volume percent) and cultivate in the seeding tank that seed culture medium is housed; 28 ℃ of culture temperature; Seeding tank pressure 0.05Mpa, 250 rev/mins of stir speed (S.S.)s, inoculum size 10% (volume percent); Ventilation 1:1.0v/v/m, incubation time 120 hours; Wherein said hair Boreostereum vibrans seed culture medium consists of (unit is a grams per liter): glucose 50, and soya bean 60, Semen Maydis powder 50, potassium primary phosphate 2, sal epsom 0.5, soya-bean oil 0.5 adds water to proper volume, and initial pH is 7.0;
(8) with ginkgo nut decapsidate and endothelium, plumule in removing again adds 10 times of water by volume and soaks making beating; The golden ear yeast shape spore that step (5) and step (7) are obtained and the seeding tank seed of the tough lead fungi of host's pili are respectively transferred by the inoculum size of 10% (volume percent) and are carried out mixed culture in the gingko fermention medium; 28 ℃ of culture temperature, fermentor tank pressure 0.05Mpa, 250 rev/mins of mixing speed; Ventilation 1:1.0v/v/m, 180 hours; The substratum of wherein said mixing fermentation culture consists of (unit is a grams per liter): gingko 200, and soya-bean oil 0.5 adds water to proper volume, and initial pH is 7.0;
(9) the gingko starch of the 1kg mineral water with 1L is soaked into, boiling 40 minutes is cooled to 65 ℃, adds Mi Qu by 1.0% (mass percent) and is incubated saccharification 2 hours, the intensification sterilising and enzyme inactivating.After the saccharification; Be cooled to 18 ℃; Extraordinarily go into golden ear yeast shape spore and the fermented liquid of the tough lead fungi of host's pili after fermention medium mixed fermentation that step (8) obtains according to 3 of saccharification material gross weight, mixing adds yeast saccharomyces cerevisiae and carries out secondary fermentation; The secondary fermentation temperature is controlled at 18 ℃, and the secondary fermentation time is 15 days;
(10) mash after the fermentation of step (9) acquisition is carried out squeeze and filter, carried out germicidal treatment 15 minutes at 80 ℃ then, store again; Be 100 days storage period; The wine of storing after-ripening is colluded accent, carry out sterilization, filtration, packing again and promptly get ginkgo gold ear fruit wine, alcoholic strength is greater than 3%; The gingko phenolic acid is less than 1ppm, and prussic acid is less than 0.1 micrograms per litre.
Embodiment 3
The preparation method of a kind of ginkgo gold ear fruit wine of the present invention may further comprise the steps:
(1) slant strains with golden ear yeast shape spore inserts in the PDA substratum 23 ℃ of culture temperature, incubation time 96 hours;
(2) slant strains with the tough lead fungi of host's pili inserts in the PDA substratum 25 ℃ of culture temperature, incubation time 136 hours;
(3) the golden ear yeast shape spore slant strains that step (1) is obtained is transferred and is equipped with in the triangular flask of shake-flask seed substratum, in 25 ℃, and 150 rev/mins of rotating speeds, incubation time 66 hours; Wherein said golden ear yeast shape spore shake-flask seed substratum consists of (unit is a grams per liter): glucose 40, and wheat bran 75 (the boiling water liquor is crossed the leaching clear liquid), potassium primary phosphate 1, sal epsom 0.5 adds water to proper volume, and initial pH is 6.2;
(4) the tough lead fungi slant strains of host's pili that step (2) is obtained is transferred and is equipped with in the triangular flask of shake-flask seed substratum, in 25 ℃, and 150 rev/mins of rotating speeds, incubation time 76 hours; The tough lead fungi shake-flask seed of wherein said host's pili substratum consists of (unit is a grams per liter): glucose 40, and wheat bran 20 (the boiling water liquor is crossed the leaching clear liquid), soya bean 50, potassium primary phosphate, sal epsom 0.5 adds water to proper volume, and initial pH is 6.2;
The shake-flask seed of the golden ear yeast shape spore that (5) step (3) is obtained is transferred by the inoculum size of 8% (volume percent) and is cultivated in the seeding tank that seed culture medium is housed; 25 ℃ of culture temperature; Seeding tank pressure 0.05Mpa; 150 rev/mins of stir speed (S.S.)s, ventilation 1:0.5v/v/m, incubation time 56 hours; The seed culture medium of wherein said golden ear yeast shape spore consists of (unit is a grams per liter): glucose 40, and soya bean 50, potassium primary phosphate 1, sal epsom 0.5, soya-bean oil 0.3 adds water to proper volume, and initial pH is 6.2;
(6) the tough lead fungi of host's pili that step (4) is obtained shakes bottle bacterial classification and transfers to be equipped with to shake in bottle triangular flask of enlarged culturing base by 8% inoculum size and carry out enlarged culturing, 25 ℃ of culture temperature, 150 rev/mins of rotating speeds, incubation time 72 hours; The tough lead fungi of wherein said host's pili shakes a bottle enlarged culturing base and consists of (unit is a grams per liter): glucose 40, and soya bean 50, Semen Maydis powder 30, potassium primary phosphate 1, sal epsom 0.5 adds water to proper volume, and initial pH is 6.2;
The bottle that shakes of the tough lead fungi of host's pili that (7) step (6) is obtained enlarges seed and transfers by the inoculum size of 10% (volume percent) and cultivate in the seeding tank that seed culture medium is housed; 25 ℃ of culture temperature; Seeding tank pressure 0.05Mpa, 150 rev/mins of stir speed (S.S.)s, inoculum size 8% (volume percent); Ventilation 1:0.5v/v/m, incubation time 90 hours; The tough lead fungi seed culture medium of wherein said host's pili consists of (unit is a grams per liter): glucose 40, and soya bean 50, Semen Maydis powder 30, potassium primary phosphate 1, sal epsom 0.5, soya-bean oil 0.3 adds water to proper volume, and initial pH is 6.2;
(8) with ginkgo nut decapsidate and endothelium, plumule in removing again adds 8 times of water by volume and soaks making beating; The golden ear yeast shape spore that step (5) and step (7) are obtained and the seeding tank seed of the tough lead fungi of host's pili are respectively transferred by the inoculum size of 8% (volume percent) and are carried out mixed culture in the gingko fermention medium; 25 ℃ of culture temperature, fermentor tank pressure 0.05Mpa, 150 rev/mins of mixing speed; Ventilation 1:0.5v/v/m, 120 hours; The substratum of wherein said mixing fermentation culture consists of (unit is a grams per liter): gingko 150, and soya-bean oil 0.3 adds water to proper volume, and initial pH is 6.2;
(9) the gingko starch of the 1kg mineral water with 1L is soaked into, boiling 30 minutes is cooled to 55 ℃, adds Mi Qu by 0.8% (mass percent) and is incubated saccharification 1.5 hours, the intensification sterilising and enzyme inactivating.After the saccharification; Be cooled to 16 ℃; Extraordinarily go into golden ear yeast shape spore and the fermented liquid of the tough lead fungi of host's pili after fermention medium mixed fermentation that step (8) obtains according to 2.5 of saccharification material gross weight, mixing adds yeast saccharomyces cerevisiae and carries out secondary fermentation; The secondary fermentation temperature is controlled at 16 ℃, and the secondary fermentation time is 30 days;
(10) mash after the fermentation of step (9) acquisition is carried out squeeze and filter, carried out germicidal treatment 10 minutes at 70 ℃ then, store again; Be 60 days storage period; The wine of storing after-ripening is colluded accent, carry out sterilization, filtration, packing again and promptly get ginkgo gold ear fruit wine, alcoholic strength is greater than 3%; The gingko phenolic acid is less than 1ppm, and prussic acid is less than 0.1 micrograms per litre.
Embodiment 4
The ginkgo gold ear fruit wine (is example with embodiment 1 product) that makes with method of the present invention has radiation-resistant nourishing function, can prove through following function test:
Laboratory animal: Kunming kind inbred mouse, male, body weight 18-22g.
Experimental technique: be divided into normal control group, simple irradiation group, ginkgo gold ear fruit wine low dose group, ginkgo gold ear fruit wine high dose group at random, 10 every group.Ginkgo gold ear fruit wine low dose group and ginkgo gold ear fruit wine high dose group continuous irrigation stomach ginkgo gold ear fruit wine 14 days, every day 1 time, the each 1mL/kg of ginkgo gold ear fruit wine low dose group, the each 5mL/kg of ginkgo gold ear fruit wine high dose group gave mouse to the 14th day
60The total irradiation of Co gamma-rays, irradiation dose are 7Gy, dose rate 0.7Gymin
-1, irradiation time 10min.The normal control group is irritated stomach zero(ppm) water, every day 1 time with simple irradiation group.Observe and respectively to organize mouse 30d survival, statistics 30d survival rate, the average fate of surviving calculate the protection index.The statistical test method adopts the t-check between each group.
The result shows, improves 33% than simple irradiation group for after 30 days the 30d survival rate of ginkgo gold ear fruit wine high dose group mouse, and mean survival time improved 6.9 days, significant difference (
P<0.05), protection index 1.38 (the protection index is effective greater than 1.2).Ginkgo gold ear fruit wine low dose group mean survival time improved 2.6 days, and protection index 1.21 is greater than virtual value.Show that oral ginkgo gold ear fruit wine can alleviate radiation injury, significantly improves the mouse survival rate.
Claims (1)
1. the preparation method of ginkgo gold ear fruit wine may further comprise the steps:
(1) slant strains with golden ear yeast shape spore inserts in the PDA substratum, and culture temperature 18-28 ℃, incubation time 48-120 hour;
(2) slant strains with the tough lead fungi of host's pili inserts in the PDA substratum, and culture temperature 23-28 ℃, incubation time 120-148 hour;
The slant strains of the golden ear yeast shape spore that (3) step (1) is obtained changes in the triangular flask that the shake-flask seed substratum is housed; Cultivate in 23-28 ℃, rotating speed 50-250 rev/min, incubation time 36-75 hour; With the grams per liter is unit; Described golden ear yeast shape spore shake-flask seed substratum consists of: glucose 20-50, wheat bran 50-100, potassium primary phosphate 0.2-2, sal epsom 0.1-2, add water to proper volume, and initial pH is 5.8-7.0;
The slant strains of the tough lead fungi of host's pili that (4) step (2) is obtained changes in the triangular flask that the shake-flask seed substratum is housed, cultivates in 23-28 ℃, and rotating speed 50-250 rev/min, incubation time 40-120 hour; With the grams per liter is unit, and the tough lead fungi shake-flask seed of described host's pili substratum consists of: glucose 20-50, wheat bran 10-50, soya bean 10-80, potassium primary phosphate 0.2-2, sal epsom 0.1-2, add water to proper volume, and initial pH is 5.8-7.0;
(5) the golden ear yeast shape spore shake-flask seed that step (3) is obtained is transferred by the inoculum size of 5-10% and is carried out seed culture in the seeding tank that the seed tank culture base is housed; Culture temperature 23-28 ℃; Seeding tank pressure 0.05Mpa; Stir speed (S.S.) 50-250 rev/min, ventilation 1: (0.3-1.0) v/v/m, incubation time 36-60 hour; With the grams per liter is unit, and the seed tank culture base of described golden ear yeast shape spore consists of: glucose 20-50, soya bean 30-60, potassium primary phosphate 1-2, sal epsom 0.1-0.5, soya-bean oil 0.2-0.5, add water to proper volume, and initial pH is 5.8-7.0;
(6) the tough lead fungi shake-flask seed of host's pili that step (4) is obtained is transferred to be equipped with to shake in bottle triangular flask of enlarged culturing base by the inoculum size of 5-10% and is carried out enlarged culturing, and culture temperature 23-28 ℃, rotating speed 50-250 rev/min, incubation time 60-80 hour; With the grams per liter is unit, and the tough lead fungi of described host's pili shakes a bottle enlarged culturing base and consists of: glucose 20-50, soya bean 10-80, Semen Maydis powder 10-50, potassium primary phosphate 1-2, sal epsom 0.1-0.5, add water to proper volume, and initial pH is 5.8-7.0;
(7) the tough lead fungi of host's pili that step (6) is obtained shakes bottle and enlarges seed and transfer by the inoculum size of 5-10% and carry out seed culture in the seeding tank that the seed tank culture base is housed; Culture temperature 23-28 ℃; Seeding tank pressure 0.05Mpa, stir speed (S.S.) 50-250 rev/min, inoculum size is per-cent 5-10% by volume; Ventilation 1: (0.3-1.0) v/v/m, incubation time 48-120 hour; With the grams per liter is unit; The tough lead fungi seed tank culture of described host's pili base consists of: glucose 20-50, soya bean 30-60, Semen Maydis powder 10-50, potassium primary phosphate 1-2, sal epsom 0.1-0.5, soya-bean oil 0.2-0.5; Add water to proper volume, initial pH is 5.8-7.0;
(8) the tough lead fungi seeding tank of the host's pili seed that golden ear yeast shape spore seeding tank seed that step (5) is obtained and step (7) obtain by two kinds of bacterium of inoculum size by volume the inoculum size of each 5-10% of per-cent transfer and carry out mixed culture in the fermentor tank that fermention medium is housed; Culture temperature 23-28 ℃; Fermentor tank pressure 0.05Mpa; Mixing speed 50-250 rev/min, ventilation 1: (0.3-1.0) v/v/m, 72-180 hour; With the grams per liter is unit, and said fermention medium consists of: gingko 100-200, soya-bean oil 0.2-0.5, add water to proper volume, and initial pH is 5.8-7.0;
(9) gingko starch is soaked into mineral water, boiling 25-40 minute, be cooled to 50-65 ℃; The Mi Qu of 0.5%-1.0% adding by mass percentage is incubated saccharification 1-2 hour, and the intensification sterilising and enzyme inactivating is after the saccharification; Be cooled to 15-18 ℃; 2-3 according to saccharification material gross weight extraordinarily goes into golden ear yeast shape spore and the fermented liquid of the tough lead fungi of host's pili after fermention medium mixed fermentation that step (8) obtains, and mixing adds yeast saccharomyces cerevisiae and carries out secondary fermentation; The secondary fermentation temperature is controlled at 15-18 ℃, and the secondary fermentation time is 7-30 days;
(10) mash after the fermentation of step (9) acquisition is carried out squeeze and filter; Carried out germicidal treatment 15 seconds-15 minutes at 65 ℃-80 ℃ then, store, be 20-100 days storage period; The wine of storing after-ripening is colluded accent, carry out sterilization, filtration, packing again and promptly get ginkgo gold ear fruit wine.
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| CN103451250B (en) * | 2013-09-02 | 2015-08-26 | 张志才 | White fungus thalline is preparing the application in Ginkgolide B |
| CN103450217B (en) * | 2013-09-02 | 2015-12-02 | 北京绿科天成生物科技有限公司 | Thick hair Boreostereum vibrans thalline is preparing the application in Ginkgolide B |
| CN103468760B (en) * | 2013-09-02 | 2016-07-06 | 北京绿科天成生物科技有限公司 | Utilize Tremella thalline as the method for catalyst preparing ginkalide B |
| CN103451251B (en) * | 2013-09-02 | 2016-06-01 | 北京绿科天成生物科技有限公司 | Utilize thick hair Boreostereum vibrans thalline as the method for catalyst preparing Ginkgolide B |
| CN103882075A (en) * | 2014-01-28 | 2014-06-25 | 江苏河海科技工程集团有限公司 | Process for obtaining ginkgolide B by utilizing tremella aurantialba strains to transform ginkgo biloba extracts |
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