CN104473145B - A kind of Antrodia camphorata submerged fermentation compound product and preparation method thereof - Google Patents
A kind of Antrodia camphorata submerged fermentation compound product and preparation method thereof Download PDFInfo
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- CN104473145B CN104473145B CN201410791469.1A CN201410791469A CN104473145B CN 104473145 B CN104473145 B CN 104473145B CN 201410791469 A CN201410791469 A CN 201410791469A CN 104473145 B CN104473145 B CN 104473145B
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Abstract
The invention discloses a kind of Antrodia camphorata submerged fermentation compound product and preparation method thereof, with preparation submerged fermentation culture mediums such as Poria cocos powder, yam flour, kudzu-vine root powder, Bee Pollen, soy meal, dusty yeast, glucose, dipotassium hydrogen phosphate, magnesium sulfate, the Antrodia camphorata mycelium fermentation broth of Antrodia camphorata polysaccharide is rich in, with employing supercritical CO through Submerged fermentation
2technology extraction, enzymolysis, separation, filtration, the concentrated obtained extracts of Chinese herbal medicine such as Buddha's hand, sealwort, matrimony vine, fructus amomi, emblic, hoveniae semoveniae semen are formulated.Present invention optimizes the content of Antrodia camphorata polysaccharide isoreactivity composition, have that output is high, the time is short, cost is low, can suitability for industrialized production, be a kind of bioengineering production technology of high-efficiency environment friendly, there is significant development prospect.
Description
Technical field
The invention belongs to Chinese medicine compound prescription goods and biomedicine field, be specifically related to a kind of Antrodia camphorata submerged fermentation compound product and preparation method thereof.
Background technology
Antrodia camphorata is called for short " camphor tree sesame ", and formal name used at school is AntrodiaCamphorata, mushroom, camphor tree mushroom, blood glossy ganoderma etc. in popular name Cinnamomum kanahirai hay mushroom, red camphor tree sesame, camphor tree.Wild Antrodia camphorata is uniquely rely, and the host made a living----Taiwan Cinnamomum kanahirai hay sets (Cinnamomumkanehirai), Cinnamomum kanahirai hay is set as the exclusive primary rare tree in Taiwan, growth is between height above sea level 450 meters to 2000 meters, Antrodia camphorata is grown on the moist surface of the rotten empty internal of aged Cinnamomum kanahirai hay trunk or lodging trunk, growth period is in annual six to October, sporophore shape changeableization, has tabular, bell shape, horse-hof shape or tower-like; Nascent cerise, gradually length becomes white, reddish tan, filbert or khaki.Antrodia camphorata gas fragranced is arduous, flat.Have promoting the circulation of qi of dispeling the wind, change that silt is invigorated blood circulation, the knot that disappears in temperature, removing toxicity for detumescence, sedation-analgesia, antibacterial, antiviral, antitumor, lifting immunity of organisms effect; Unique function is had especially for treatment gastrointestinal distress, diarrhea and vomiting, food poisoning, mushroom poisoning, diabetes, alcoholic liver, fatty liver, cirrhosis, liver cancer etc.Antrodia camphorata has many physiologically active ingredients, as polysaccharide body, triterpene compound, Sudismase, adenosine, protein (containing immune protein), vitamin, trace element, nucleic acid, agglutinin, amino acid, cellulose, blood pressure stabilization material etc.Being the most special with polysaccharide and triterpene compound in the active ingredient of Antrodia camphorata, is that other mushroom bacterium cannot be compared.Camphor tree sesame is at fructification stage polysaccharides content, about 2%, there is the physiologically active such as immune physiologically active, prevention and control tumour, main containing β-D-glucan(callose)) it acts on being through and excites macrophage, T lymphocyte, bone-marrow-derived lymphocyte and natural killer cell etc. to strengthen immunologic function, and then reaches antineoplastic effect.The effects such as cardiac stimulant, hypotensive, hypoglycemic, antithrombotic are had in Antrodia camphorata polysaccharide body.Because Antrodia camphorata to be regarded as the medicinal fungi of uniqueness and preciousness in Taiwan, therefore have high research and commercial value, be also the most expensive wild fungi in current Taiwan, be called as " refreshing sesame " in Hongkong and Macro, Taiwan is among the people to be referred to as " ruby in forest ".
Poria cocos
Sclerotium accounts for dry weight 93% and triterpene compound Pachymic acid, pachymic acid, 3 beta-hydroxy wool steroid trienic acids containing β-pachyman.Containing much starch, simultaneously containing natural gum, chitin, protein, fat, sterol, lecithin, glucose, adenine, histidine, choline, β-pachyman catabolic enzyme, lipase, protease etc.
Poria cocos medical usage is extremely wide.Poria cocos is flat, slightly sweet flavor, beneficial spleen, calm the nerves, clearing damp and promoting diuresis, the diseases such as splenasthenic diarrhea, palpitation and insomnia, urine are smooth, oedema can be treated.
Chinese yam
Chinese yam is rich in starch, protein, vitamin A
1, B
1, B
2, C, containing the trace element such as calcium, phosphorus, iron, iodine and inorganic salts, and mucilaginous substance, amino acid, cellulose etc.There is effect of strengthening the spleen and stomach, kidney tonifying lung, strengthening by means of tonics.
The root of kudzu vine
Root of kudzu vine isoflavone-containing composition Puerarin, Puerarin xyloside, daizeol, daidzin and cupreol, arachidic acid, again containing volume starch, carbohydrate, vegetable protein, multivitamin and mineral matter, in addition containing Flavonoid substances: Dai, daidzin, also has Dai-4,7-diglucoside, Puerarin, Puerarin-7-xyloside, root of kudzu vine alcohol, root of kudzu vine rattan and daidzin etc.The root of kudzu vine is integration of drinking and medicinal herbs Chinese herbal medicine, cures mainly that typhoid fever is warm, headache Xiang Qiang (neck stiff), dysphoria with smothery sensation are quenched one's thirst, has loose bowels, the card such as dysentery, measles without adequate eruption, hypertension, angina pectoris, deafness.
Bee Pollen
Bee Pollen mainly contains: protein 20%-25%, total amino acid content more than 20%, free amino acid 1%-2%, carbohydrate 40%-50%, fatty 5%-10%; Also has the abundant multiple nutrients material such as natural complex, trace element, enzyme, hormone, bioactivator.Also containing several mineral materials, wherein: potassium 20 ~ 40%, magnesium 1 ~ 20%, calcium 1 ~ 15%, iron 1 ~ 12%, silicon 2 ~ 10%, phosphorus 1 ~ 20%, and a large amount of active protease, nucleic acid, flavone compound and other active material etc.Element in Bee Pollen can prevention and therapy hypertension, diabetes, coronary heart disease and kidney trouble etc.There is anti arteriosclerosis, reduce cholesterol, separate pain, radiation proof, the effect such as antitumor.
Sealwort
Containing Siberian solomonseal rhizome polysaccharide the first and second the third 3 kinds in sealwort, by glucose, mannose, and galacturonic acid composition, its molecular weight is all greater than 200,000; In sealwort, total starches content is 11.74%, containing multiple steroid saponin and lysine, and threonine, isoleucine, serine, leucine, glutamic acid, tyrosine, proline, glycine, alanine, asparatate, homoserine, the necessary element of the several amino acids such as DAB and human body.Pharmacological action has and delays senility, hypotensive, hypoglycemic, reducing blood lipid, atherosclerosis, and delay senility and antibacterially wait effect, Siberian solomonseal rhizome polysaccharide has Immunestimulatory effect.
Buddha's hand
Buddha's hand fresh fruit is about containing protein 1.2%, fat 1.3%, total reducing sugar 8.7%, crude fibre 1.1%, ash content 0.7%, moisture 86.9%, and pericarp inside is containing Coumarins compound, and Limettin, flavone compound also have multivitamin and mineral matter.Wherein Vitamin C content is 0.04%, and mineral element mainly contains calcium, magnesium, iron, zinc, manganese, copper, phosphorus, potassium, sodium, sulphur etc.In carbohydrate, there is the polysaccharide of multiple different molecular weight, there is physiological activity.
The fruit of Chinese wolfberry
The fruit of Chinese wolfberry contains betaine, atropine, hyoscyamine, carrotene, thiamine, riboflavin, nicotinic acid, vitamin C.Simultaneously containing amino acid: aspartic acid, proline, alanine, leucine, phenylalanine, serine, glycine, glutamic acid, cysteine, lysine, arginine, isoleucine, threonine, histidine, cruel propylhomoserin, tryptophan, methionine.Again containing taurine, GABA, also containing natural activity elements such as potassium, calcium, sodium, zinc, iron, copper, chromium, strontium, lead, nickel, cadmium, cobalt, magnesium.LBP-X content is 7.09%, has Promote immunity effect.
Emblic
Emblic carbohydrate containing 6.63% ~ 14.10%, fat 0.19% ~ 1.09%, ash content 0.62% ~ 0.70%, protein 0.69% ~ 2.18%, vitamin B1, vitamin B2, the multivitamin such as vitamin C and carrotene, wherein especially high and famous with Vc content, for containing 470 ~ 680mg in 100g pulp, be up to 1814mg/100g, average content is 100 times of oranges and tangerines, 160 times of apple, 3 ~ 5 times of Kiwi berry, containing several amino acids and abundant calcium, iron, potassium, phosphorus, selenium, the macro-and trace-element that the human bodies such as germanium are necessary, wherein iron, zinc is the important element of growth in humans and growth, selenium, magnesium, manganese has anti-oxidant, anti-aging, the function of anti-cancer and anti-cardiovascular disease.Have and promote the production of body fluid to quench thirst, moistening lung for removing phlegm, the stagnant eliminate indigestion that disappears, reduce the effect such as cholesterol.
Trifoliate orange Dulcis
The every hectogram moisture content of trifoliate orange Dulcis 59.1 grams, sugar 28.55 grams, 0.07 gram, fat, 0.03 gram, protein, vitamin C 16.3 milligrams.The sweet acid of trifoliate orange Dulcis nature and flavor is put down, and has heat-clearing, only polydipsia, effect of tonifying middle-Jiao and Qi, moisturizing viscera, relaxing muscles and tendons network.Control drunk, dysphoria with smothery sensation, thirsty, vomiting, difficulty in urination and defecation.Supplement to the Herbal carries, and " relieving restlessness of quenching the thirst, moistens five stolen goods, sharp stool and urine.
According to the feature of above-mentioned raw materials active ingredient, submerged fermentation technology is adopted to have relevant report.But a lot of research just adopts conventional mode to carry out, its goods effective active composition is not high, and extract yield not high, the polysaccharides weight of Antrodia camphorata is low, adds and adopts single raw material, collaborative not wide spectrum of comprehensively tiring, and active ingredient is failed composition and is absorbed by the body.Therefore, adopt the Chinese herbal medicines such as natural plants yam flour, kudzu-vine root powder, Bee Pollen, Poria cocos powder as culture medium, comprehensive nutrition, constantly can supplement nitrogenous source, carbon source, effectively can meet the demand of Antrodia camphorata mycelial growth to nutrition, improve and obtain a large amount of thalline and metabolite, significantly improve the content of the effective active compositions such as polysaccharide body.Because Chinese herbal medicine Buddha's hand, sealwort, matrimony vine, fructus amomi, emblic, hoveniae semoveniae semen and royal jelly have multiple natural active matter, adopt supercritical CO
2extraction, enzymolysis process, the compound product that the extract of extraction and Antrodia camphorata zymotic fluid compound are prepared, there is not yet relevant report.
Summary of the invention
The object of the present invention is to provide a kind of Antrodia camphorata submerged fermentation compound product and preparation method thereof, optimize the content of Antrodia camphorata polysaccharide isoreactivity composition, have that output is high, the time is short, cost is low, can suitability for industrialized production, be a kind of bioengineering production technology of high-efficiency environment friendly, there is significant development prospect.
For achieving the above object, the present invention adopts following technical scheme:
A kind of Antrodia camphorata submerged fermentation compound product, primarily of Antrodia camphorata mycelium fermentation broth and extracts of Chinese herbal medicine formulated, described Chinese herbal medicine is Buddha's hand, sealwort, Poria cocos, hoveniae semoveniae semen, emblic, matrimony vine and fructus amomi.
The weight ratio of Antrodia camphorata mycelium fermentation broth and extracts of Chinese herbal medicine is the concentration Brix=18.0-20.0% of 1:1, Antrodia camphorata mycelium fermentation broth, the concentration Brix=18.0-20.0% of extracts of Chinese herbal medicine.(Brix is the fluid density that brix hydrometer measures)
Preparation method comprises the following steps:
(1) preparation of Antrodia camphorata mycelium fermentation broth
Culture medium:
Slant medium: potato liquor 1000mL, glucose 20g, agar 20g, peptone 20g, magnesium sulfate 0.2wt.%, pH nature;
Submerged fermentation culture medium: Poria cocos powder 1.5wt.%, yam flour 1.5wt.%, kudzu-vine root powder 1.5wt.%, soy meal 2.0wt.%, Bee Pollen 2.0wt.%, glucose 3.0wt.%, dusty yeast 0.3wt.%, magnesium sulfate 0.2wt.%, dipotassium hydrogen phosphate 0.3wt.%, sodium chloride 0.2wt.%, surplus are water, pH:5-7, powder: 80-100 order;
Preparation technology:
1) 5-7d is cultivated in the inclined-plane of inoculation at 26-28 DEG C;
2) mycelial inclined plane inoculating will be covered with in submerged fermentation culture medium, under shaking table 180r/min rotating speed, 26-28 DEG C of shaken cultivation 2-3d;
3) seeding tank seed culture: by kind of amount 10-15%, a temperature 26-28 DEG C, illumination condition: continuous darkness condition, mixing speed 150-180r/min, filtrated air throughput is 1:0.5v/v per minute, tank pressure 0.04-0.05MPa, cultivates 40-48h, pH and is down to 3.5-4.0;
4) fermented and cultured: inoculum concentration 15-20%, temperature 26-28 DEG C, mixing speed 150-200r/min, filtrated air throughput is 1:0.5v/v per minute, tank pressure 0.04-0.05MPa, and cultivate 40-48h, pH and be down to 3.5-4.0, residual sugar amount is down to 0.3%;
5) enzymolysis: add compound protease 0.5wt.%, cellulase 0.5wt.% and amylase 0.5wt.%, 50-55 DEG C of Enzymolysis 2-3h; The wherein enzyme activity 2 × 10 of protease
5u/kg, diastatic enzyme activity 1.0 × 10
5the enzyme activity 1.5 × 10 of U/kg, cellulase
5u/kg;
6) homogeneous process: the Antrodia camphorata zymotic fluid through enzymolysis processing is first through colloid mill homogeneous, rotating grinding rotating speed 800-1000r/min, then through high pressure homogenizer homogeneous, pressure 40-60MPa, finally crosses 200 order membrane filtrations, obtains Antrodia camphorata mycelium fermentation broth;
(2) preparation of extracts of Chinese herbal medicine
1) pre-treatment: percentage: Buddha's hand 10%, sealwort 10%, Poria cocos 15%, hoveniae semoveniae semen 15%, emblic 20%, matrimony vine 20%, fructus amomi 10%, pulverizes respectively, and fineness is 3-5mm, and mixing is that 1:10 adds water by solid-liquid ratio, soaks 3-5 hour;
2) supercritical CO
2extraction; Extraction conditions is: extraction kettle pressure 24-30MPa, extraction temperature 60-80 DEG C, I grade of separating still pressure 8-10MPa, II grade of separating still pressure 6-8MPa, CO
2flow 120-150L/h;
3) centrifugation obtains extract: rotating speed 2000-3000r/min, time 6-10min;
4) extract remainder enzymolysis and extraction: be that 1:10 adds pure water by solid-liquid ratio by extract remainder, regulates pH=4.5-6.0,60-80 DEG C of digestion 6-12h, is then cooled to 50-55 DEG C, adds cellulase, enzymolysis 1-2h, then add amylase, enzymolysis 1-2h; Be warming up to 100-110 DEG C, go out enzyme 3-5min, filters to obtain enzyme-extracting solution; Enzymolysis process carries out under ultrasonic wave added condition, and ultrasonic frequency is 25-30kHz; The enzyme activity of described cellulase is 2.5 ten thousand u/g, and enzyme dosage is the 1.0-1.2% of extract remainder weight; Described diastatic enzyme activity is 10,000 u/g, and enzyme dosage is the 1.0-1.2% of extract remainder weight;
(3) Antrodia camphorata mycelium fermentation broth, plant liquid extract and Chinese herbal medicine extract remainder enzyme-extracting solution are mixed, through concentrated, add royal jelly, make compound oral liquid goods or compound solid goods.
The preparation of compound oral liquid goods: Antrodia camphorata mycelium fermentation broth, plant liquid extract and Chinese herbal medicine extract remainder enzyme-extracting solution are mixed, under pressure 0.09MPa, being evaporated to soluble solid content is 10-12%, concentration Brix=18.0-20.0%, is cooled to room temperature, add 5wt.% royal jelly, stir, homogeneous process, adds food flavor, allotment pH=5.0-6.0, canned with brown bottle.
The preparation of compound solid goods: Antrodia camphorata mycelium fermentation broth, plant liquid extract and Chinese herbal medicine extract remainder enzyme-extracting solution are mixed, under pressure 0.09MPa, being evaporated to soluble solid content is 12-15%, concentration Brix=20.0-23.0%, freeze drying powder process, add 5wt.% royal jelly dried powder, 10wt.% glucose and 15wt.% maltodextrin, through mixing, granulation, whole grain, make granule, tablet or capsule.
Remarkable advantage of the present invention is: the content that present invention optimizes Antrodia camphorata polysaccharide isoreactivity composition, have that output is high, the time is short, cost is low, can suitability for industrialized production, be a kind of bioengineering production technology of high-efficiency environment friendly, there is significant development prospect.
Detailed description of the invention
A kind of Antrodia camphorata submerged fermentation compound product, primarily of Antrodia camphorata mycelium fermentation broth and extracts of Chinese herbal medicine formulated, described Chinese herbal medicine is Buddha's hand, sealwort, Poria cocos, hoveniae semoveniae semen, emblic, matrimony vine and fructus amomi.
The weight ratio of Antrodia camphorata mycelium fermentation broth and extracts of Chinese herbal medicine is the concentration Brix=18.0-20.0% of 1:1, Antrodia camphorata mycelium fermentation broth, the concentration Brix=18.0-20.0% of extracts of Chinese herbal medicine.
Preparation method comprises the following steps:
(1) preparation of Antrodia camphorata mycelium fermentation broth
Bacterial classification: the Antrodia camphorata bacterial classification provided by Taiwan both sides of the Straits traditional Chinese medicine cooperative development cultural association of China.
Culture medium:
Slant medium: potato liquor 1000mL, glucose 20g, agar 20g, peptone 20g, magnesium sulfate 0.2wt.%, pH nature;
Submerged fermentation culture medium: Poria cocos powder 1.5wt.%, yam flour 1.5wt.%, kudzu-vine root powder 1.5wt.%, soy meal 2.0wt.%, Bee Pollen 2.0wt.%, glucose 3.0wt.%, dusty yeast 0.3wt.%, magnesium sulfate 0.2wt.%, dipotassium hydrogen phosphate 0.3wt.%, sodium chloride 0.2wt.%, surplus are water, pH:5-7, powder: 80-100 order;
Preparation technology:
1) 5-7d is cultivated in the inclined-plane of inoculation at 26-28 DEG C;
2) mycelial inclined plane inoculating will be covered with in submerged fermentation culture medium, under shaking table 180r/min rotating speed, 26-28 DEG C of shaken cultivation 2-3d;
3) seeding tank seed culture: by kind of amount 10-15%, a temperature 26-28 DEG C, illumination condition: continuous darkness condition, mixing speed 150-180r/min, filtrated air throughput is 1:0.5v/v per minute, tank pressure 0.04-0.05MPa, cultivates 40-48h, pH and is down to 3.5-4.0;
4) fermented and cultured: inoculum concentration 15-20%, temperature 26-28 DEG C, mixing speed 150-200r/min, filtrated air throughput is 1:0.5v/v per minute, tank pressure 0.04-0.05MPa, and cultivate 40-48h, pH and be down to 3.5-4.0, residual sugar amount is down to 0.3%;
5) enzymolysis: add compound protease 0.5wt.%, cellulase 0.5wt.% and amylase 0.5wt.%, 50-55 DEG C of Enzymolysis 2-3h; The wherein enzyme activity 2 × 10 of protease
5u/kg, diastatic enzyme activity 1.0 × 10
5the enzyme activity 1.5 × 10 of U/kg, cellulase
5u/kg;
6) homogeneous process: the Antrodia camphorata zymotic fluid through enzymolysis processing is first through colloid mill homogeneous, rotating grinding rotating speed 800-1000r/min, then through high pressure homogenizer homogeneous, pressure 40-60MPa, finally crosses 200 order membrane filtrations, obtains Antrodia camphorata mycelium fermentation broth;
Natural Antrodia camphorata and Antrodia camphorata zymotic fluid main component detect the table of comparisons
The natural Antrodia camphorata main component of table 1 and content (g/100g dry weight)
Table 2 Antrodia camphorata zymotic fluid main component and content (zymotic fluid concentration Brix=18.0-20.0%, g/100ml)
Table 3 Antrodia camphorata zymotic fluid makes dry powder main component and content (g/100g dry weight)
(2) preparation of extracts of Chinese herbal medicine
1) pre-treatment: percentage: Buddha's hand 10%, sealwort 10%, Poria cocos 15%, hoveniae semoveniae semen 15%, emblic 20%, matrimony vine 20%, fructus amomi 10%, pulverizes respectively, and fineness is 3-5mm, and mixing is that 1:10 adds water by solid-liquid ratio, soaks 3-5 hour;
2) supercritical CO
2extraction; Extraction conditions is: extraction kettle pressure 24-30MPa, extraction temperature 60-80 DEG C, I grade of separating still pressure 8-10MPa, II grade of separating still pressure 6-8MPa, CO
2flow 120-150L/h;
3) centrifugation obtains extract: rotating speed 2000-3000r/min, time 6-10min;
4) extract remainder enzymolysis and extraction: be that 1:10 adds pure water by solid-liquid ratio by extract remainder, regulates pH=4.5-6.0,60-80 DEG C of digestion 6-12h, is then cooled to 50-55 DEG C, adds cellulase, enzymolysis 1-2h, then add amylase, enzymolysis 1-2h; Be warming up to 100-110 DEG C, go out enzyme 3-5min, filters to obtain enzyme-extracting solution; Enzymolysis process carries out under ultrasonic wave added condition, and ultrasonic frequency is 25-30kHz; The enzyme activity of described cellulase is 2.5 ten thousand u/g, and enzyme dosage is the 1.0-1.2% of extract remainder weight; Described diastatic enzyme activity is 10,000 u/g, and enzyme dosage is the 1.0-1.2% of extract remainder weight;
Table 4 extracts of Chinese herbal medicine main component and content (concentration Brix=18.0-20.0%, g/100ml)
(3) Antrodia camphorata mycelium fermentation broth, plant liquid extract and Chinese herbal medicine extract remainder enzyme-extracting solution are mixed, through concentrated, add royal jelly, make compound oral liquid goods or compound solid goods.
The preparation of compound oral liquid goods: Antrodia camphorata mycelium fermentation broth, plant liquid extract and Chinese herbal medicine extract remainder enzyme-extracting solution are mixed, under pressure 0.09MPa, being evaporated to soluble solid content is 10-12%, concentration Brix=18.0-20.0%, is cooled to room temperature, add 5wt.% royal jelly, stir, homogeneous process, adds food flavor, allotment pH=5.0-6.0, canned with brown bottle.
Table 5 Antrodia camphorata submerged fermentation compound flowing product main component detects (concentration Brix=18.0-20.0%, g/100ml)
The preparation of compound solid goods: Antrodia camphorata mycelium fermentation broth, plant liquid extract and Chinese herbal medicine extract remainder enzyme-extracting solution are mixed, under pressure 0.09MPa, being evaporated to soluble solid content is 12-15%, concentration Brix=20.0-23.0%, freeze drying powder process, add 5wt.% royal jelly dried powder, 10wt.% glucose and 15wt.% maltodextrin, through mixing, granulation, whole grain, make granule or tablet, capsule.Loss on drying≤5.0.
Table 6 Antrodia camphorata submerged fermentation compound solid goods (granule, tablet, capsule) main component detects (g/100g dry weight)
Above-mentioned main component examination criteria and method:
1. polysaccharide body (with glucose meter), phend-sulphuric acid.
2. total triterpene (in ursolic acid), ultraviolet spectrophotometry.
3. protein (in butt, N × 6.25), adopts GB5009-2010 standard detection.
4. vitamin, adopts HPLC high performance liquid chromatography GB5009.82-2003 standard detection.
5. soluble dietary fiber, adopts GB/T5009.88 standard determination method.
The foregoing is only preferred embodiment of the present invention, all equalizations done according to the present patent application the scope of the claims change and modify, and all should belong to covering scope of the present invention.
Claims (3)
1. the preparation method of an Antrodia camphorata submerged fermentation compound product, it is characterized in that: described Antrodia camphorata submerged fermentation compound product primarily of Antrodia camphorata mycelium fermentation broth and extracts of Chinese herbal medicine formulated, described Chinese herbal medicine is Buddha's hand, sealwort, Poria cocos, hoveniae semoveniae semen, emblic, matrimony vine and fructus amomi; The weight ratio of Antrodia camphorata mycelium fermentation broth and extracts of Chinese herbal medicine is the concentration Brix=18.0-20.0% of 1:1, Antrodia camphorata mycelium fermentation broth, the concentration Brix=18.0-20.0% of extracts of Chinese herbal medicine; Its preparation process comprises the following steps:
(1) preparation of Antrodia camphorata mycelium fermentation broth
Culture medium:
Slant medium: potato liquor 1000mL, glucose 20g, agar 20g, peptone 20g, magnesium sulfate 0.2wt.%, pH nature;
Submerged fermentation culture medium: Poria cocos powder 1.5wt.%, yam flour 1.5wt.%, kudzu-vine root powder 1.5wt.%, soy meal 2.0wt.%, Bee Pollen 2.0wt.%, glucose 3.0wt.%, dusty yeast 0.3wt.%, magnesium sulfate 0.2wt.%, dipotassium hydrogen phosphate 0.3wt.%, sodium chloride 0.2wt.%, surplus are water, pH:5-7, powder: 80-100 order;
Preparation technology:
1) 5-7d is cultivated in the inclined-plane of inoculation at 26-28 DEG C;
2) mycelial inclined plane inoculating will be covered with in submerged fermentation culture medium, under shaking table 180r/min rotating speed, 26-28 DEG C of shaken cultivation 2-3d;
3) seeding tank seed culture: by kind of amount 10-15%, a temperature 26-28 DEG C, illumination condition: continuous darkness condition, mixing speed 150-180r/min, filtrated air throughput is 1:0.5v/v per minute, tank pressure 0.04-0.05MPa, cultivates 40-48h, pH and is down to 3.5-4.0;
4) fermented and cultured: inoculum concentration 15-20%, temperature 26-28 DEG C, mixing speed 150-200r/min, filtrated air throughput is 1:0.5v/v per minute, tank pressure 0.04-0.05MPa, and cultivate 40-48h, pH and be down to 3.5-4.0, residual sugar amount is down to 0.3%;
5) enzymolysis: add compound protease 0.5wt.%, cellulase 0.5wt.% and amylase 0.5wt.%, 50-55 DEG C of Enzymolysis 2-3h; The wherein enzyme activity 2 × 10 of protease
5u/kg, diastatic enzyme activity 1.0 × 10
5the enzyme activity 1.5 × 10 of U/kg, cellulase
5u/kg;
6) homogeneous process: the Antrodia camphorata zymotic fluid through enzymolysis processing is first through colloid mill homogeneous, rotating grinding rotating speed 800-1000r/min, then through high pressure homogenizer homogeneous, pressure 40-60MPa, finally crosses 200 order membrane filtrations, obtains Antrodia camphorata mycelium fermentation broth;
(2) preparation of extracts of Chinese herbal medicine
1) pre-treatment: percentage: Buddha's hand 10%, sealwort 10%, Poria cocos 15%, hoveniae semoveniae semen 15%, emblic 20%, matrimony vine 20%, fructus amomi 10%, pulverizes respectively, and fineness is 3-5mm, and mixing is that 1:10 adds water by solid-liquid ratio, soaks 3-5 hour;
2) supercritical CO
2extraction; Extraction conditions is: extraction kettle pressure 24-30MPa, extraction temperature 60-80 DEG C, I grade of separating still pressure 8-10MPa, II grade of separating still pressure 6-8MPa, CO
2flow 120-150L/h;
3) centrifugation obtains extract: rotating speed 2000-3000r/min, time 6-10min;
4) extract remainder enzymolysis and extraction: be that 1:10 adds pure water by solid-liquid ratio by extract remainder, regulates pH=4.5-6.0,60-80 DEG C of digestion 6-12h, is then cooled to 50-55 DEG C, adds cellulase, enzymolysis 1-2h, then add amylase, enzymolysis 1-2h; Be warming up to 100-110 DEG C, go out enzyme 3-5min, filters to obtain enzyme-extracting solution; Enzymolysis process carries out under ultrasonic wave added condition, and ultrasonic frequency is 25-30kHz; The enzyme activity of described cellulase is 2.5 ten thousand u/g, and enzyme dosage is the 1.0-1.2% of extract remainder weight; Described diastatic enzyme activity is 10,000 u/g, and enzyme dosage is the 1.0-1.2% of extract remainder weight;
(3) Antrodia camphorata mycelium fermentation broth, plant liquid extract and Chinese herbal medicine extract remainder enzyme-extracting solution are mixed, through concentrated, add royal jelly and make compound oral liquid goods, or add royal jelly dried powder and make compound solid goods.
2. method according to claim 1, it is characterized in that: the preparation of compound oral liquid goods: Antrodia camphorata mycelium fermentation broth, plant liquid extract and Chinese herbal medicine extract remainder enzyme-extracting solution are mixed, under pressure 0.09MPa, being evaporated to soluble solid content is 10-12%, concentration Brix=18.0-20.0%, be cooled to room temperature, add 5wt.% royal jelly, stir, homogeneous process, add food flavor, allotment pH=5.0-6.0, canned with brown bottle.
3. method according to claim 1, it is characterized in that: the preparation of compound solid goods: Antrodia camphorata mycelium fermentation broth, plant liquid extract and Chinese herbal medicine extract remainder enzyme-extracting solution are mixed, under pressure 0.09MPa, being evaporated to soluble solid content is 12-15%, concentration Brix=20.0-23.0%, freeze drying powder process, add 5wt.% royal jelly dried powder, 10wt.% glucose and 15wt.% maltodextrin, through mixing, granulation, whole grain, make granule, tablet or capsule.
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