CN1195843C - Solid culture method of antrodia camphorate, its cultured solid substance, and its product and application - Google Patents
Solid culture method of antrodia camphorate, its cultured solid substance, and its product and application Download PDFInfo
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- CN1195843C CN1195843C CNB011236132A CN01123613A CN1195843C CN 1195843 C CN1195843 C CN 1195843C CN B011236132 A CNB011236132 A CN B011236132A CN 01123613 A CN01123613 A CN 01123613A CN 1195843 C CN1195843 C CN 1195843C
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Abstract
The present invention provides a solid culture method of Antrodia camphorate, which comprises the following steps: cultivating mycelia of Antrodia camphorata strains in space bags and cultivating fruiting bodies in the air. The present invention also provides an Antrodia camphorata solid culture substance cultivated by the method, processed products thereof and application thereof.
Description
Technical field
The present invention relates to the cultural method of camphor tree sesame, more particularly, the present invention relates to the solid culture method of camphor tree sesame.In addition, the invention still further relates to camphor tree sesame solid culture and the converted products and the purposes of cultivating gained by described method.
Background technology
Camphor tree sesame (Zang Zhi), its formal name used at school is Antrodia camphorata, have another name called camphor tree mushroom, red camphor tree mushroom, red camphor tree sesame, Cinnamomum kanahirai hay mushroom or Antrodia camphorata, be the peculiar fungi of this province, only be grown in the hollow heartwood inwall of the Cinnamomum kanahirai hay tree (Cinnamonum kanehirai) in Taiwan, sporophore tool intensive camphor tree fragrance, its profile is tabular or mitriform.Popular legend have calm the nerves, timid gas in fashion, stagnation resolvation are invigorated blood circulation, it is long-pending to disappear in the temperature, detoxify and promote the subsdence of swelling, and the effect of sedation-analgesia, the anticorrosion toxinicide for first-class is regarded as the most expensive wild fungus on the market, Taiwan.What is more, improve immunizing power and anticancer indirect action with respect to glossy ganoderma, the camphor tree sesame is real to be the mechanism of action of directly inhibition or kill cancer cell, has cardiac stimulant, immunomodulatory, anti-parasympathetic nerve effect, the active effect of serotonin antagonist simultaneously.Particular words it, the camphor tree sesame can treat that gastrointestinal pain, diarrhea and vomiting, diabetes, gout, ephritis, sacroiliitis, inflammation, allergy, urine protein are too high, uremia, liver cirrhosis, liver cancer and influenza etc.
Existing numerous research units drop into manpower and material resources, be devoted to the research of camphor tree sesame activeconstituents, wherein the monographic study plan of state science commission of Executive Yuan is to utilize chemical process, nuclear magnetic resonance spectroscopy (NMR) reaches and known compound spectrum compares, to set up the structural performance of camphor tree sesame effective constituent compound, the achievement in research of carrying out at camphor tree sesame effective constituent is pointed out again, the water of camphor tree sesame mentions that methanol extract has the supression effect to the growth of streptococcus aureus and sycosis budlet tinea bacterium, and the methanol extract camphor tree mushroom of camphor tree sesame acid A (zhankuic acid) has obvious restraining effect and platelet aggregation effect to the lymph tumor cell of p-388 mouse in addition; And the function of camphor tree mushroom acid faint cholinolytic of B tool and medmain (serotonin).
Height common vetch Master's thesis dawn is pointed out at the triterpenes components research that Ganoderma belongs to novel species camphor tree sesame, the glossy ganoderma novel species that camphor tree sesame (Ganoderma comphoratum Zang et Su) was delivered for nineteen ninety, obtain thick extract with acetone extract camphor tree sesame sporophore, then separate and the means re-crystallization purifying with red, orange, green, blue, yellow (ROGBY) (LC, PLC); Thin layer liquid chromatography scanning (TLCScanning) and high performance liquid chromatography (HPLC) are identified purity.Carry out structure determination via mass spectrum, infrared spectra, UV spectrum, nuclear magnetic resonance spectrometer (H-NMR and C-NMR).Wherein 3,11-dioxy-8, (3,11-dioxo-8 is 23-dien-26-oicacid) for tetracol phenixin (CCl for 23-diene-26-acid
4) bring out that L-glutamic acid-pyruvic acid transaminase (GPT) value has the decline effect in the mice serum of acute hepatitis.
Be distributed in the broad-leaved forest zone, mountain range, platform East Coast, Taiwan because the camphor tree sesame now more, but according to Forest Service's Cinnamomum kanahirai hay child care bulletin, this district has been divided into the wilderness area, seldom and is gathered totally in the camphor tree sesame quantity of child care district outgrowth.Therefore, can't obtain natural wild camphor tree sesame on the market in fact.
Summary of the invention
Main purpose of the present invention provides the solid culture method of a kind of camphor tree sesame, and it can cultivate the camphor tree sesame solids that has with the equal pharmacological effect of natural wild camphor tree sesame in the artificial culture mode.
Another object of the present invention provides the solid culture of a kind of camphor tree sesame, and it is with being deposited at food Industry in Taiwan Institute of Development Studies, and the bacterial classification of numbering CCRC 35398 is the source, and this bacterial classification is commercially available bacterial classification, cultivates the solid culture of gained with solid culture method.
A further object of the present invention provides the converted products of solid culture of the present invention.
Another purpose of the present invention provides the various active purposes of solid culture of the present invention.
Description of drawings
Fig. 1 represents the sporophore profile that cultural method is cultivated the camphor tree sesame according to the present invention.
Fig. 2 represents high-efficient liquid phase chromatogram (HPLC) the analysis comparison of camphor tree sesame composition, comprises that A represents the camphor tree sesame sporophore of the cultural method gained according to the present invention, and B represents to represent wild camphor tree sesame sporophore with other liquid culture mode gained camphor tree sesames with C.
Fig. 3 represents to cause lipid peroxidation composition (TBARS) to increase with the peroxidatic reaction of lipid of ferrous ion stimulation in rats brain homogenizing fluid with the initiation free radical.Cultural method is cultivated three times of sporophore of camphor tree sesame and is improved with its concentration with the octuple concentrated solution according to the present invention, suppresses peroxidation and strengthen.Represent (n=3) to suppress peroxidation per-cent.
On behalf of the camphor tree sesame of different cycles of concentration, Fig. 4 tetracol phenixin is caused liver function biochemical values L-glutamic acid-pyruvic acid transaminase (GPT) enzyme activity change.# represents the difference (P＜0.01) of tool statistical significance between normal group and injury group; * represent feeding group and injury group that the difference (P＜0.01) of statistical significance is arranged.Numerical value is represented with mean+/-standard error.
Fig. 5 represents that the camphor tree sesame of different cycles of concentration causes liver function biochemical values glutamic-oxaloacetic transaminase (GOT) enzyme activity change to tetracol phenixin.# represents the difference (P＜0.01) of tool statistical significance between normal group and injury group; * represent feeding group and injury group that the difference (P＜0.01) of statistical significance is arranged.Numerical value is represented with mean+/-standard error.
The influence (A represents that normal group, B represent that tetracol phenixin injury group, C represent that three times of feeding camphor tree sesames concentrate group and D represents that feeding camphor tree sesame octuple concentrates group) that on behalf of the camphor tree sesame of different cycles of concentration, Fig. 6 cause the liver organization macroscopic view to change to tetracol phenixin.
The influence that on behalf of the camphor tree sesame of different cycles of concentration, Fig. 7 cause liver organization to change to tetracol phenixin.The camphor tree sesame of the different cycles of concentration of mice group feeding is after five weeks, get its liver, cut into slices and dye, observe liver organization and change (A represents that normal group, B represent that tetracol phenixin injury group, C represent that three times of feeding camphor tree sesames concentrate group and D represents that feeding camphor tree sesame octuple concentrates group).
Fig. 8 represents the influence of camphor tree sesame powder extraction liquid to colon-cancer cell (COLO 320HSR) growth.
Embodiment
The invention provides the solid culture method of a kind of camphor tree sesame, the phase is cultivated composition and the active camphor tree sesame sporophore of the natural wild camphor tree sesame of tool in a large number with manual type.The solid culture method of camphor tree sesame of the present invention comprises and a camphor tree sesame bacterial classification is carried out mycelium with space bag (PP bag) cultivates, and then carries out sporophore and cultivate in air.
Before the mycelium cultivation stage, usually can be earlier through the propagation step, for example cultivate a large amount of bacterial classifications of preparation and cultivate use for the space bag with the following step, can comprise three steps: (1) is preserved the activatory bacterial classification from liquid nitrogen and is got a fritter agar mycelia piece, move in the fresh culture and cultivate down, treat that growth is to vigorous in constant temperature; (2) with this bacterial classification inoculation to the space bag of the fibre object of killing bacterium (for example wheat or wood chip), then cultivate down in constant temperature; When (3) treating that this bacterial classification in the space bag covers with mycelia, remove older mycelia piece topmost, can be seeded in a large number in the space bag then.
Propagation step before the mycelium cultivation stage is fermented with a large amount of cultivation bacterial classifications again or with liquid culture, inoculates to the space bag.The prescription of fermentation culture is 1-3% fructose, 0.01-0.1% sal epsom, 0.1-1% yeast extract, 0.05-0.5% potassium primary phosphate.Preferred group becomes 2% fructose, 0.05% sal epsom, 0.5% yeast extract and 0.1% potassium primary phosphate.
After slant tube is cultivated, the bacterial classification inoculation to 5 that breeding is good is in a large number risen in the fermentation culture, under 24-26 ℃, cultivated about 14 days,, be seeded in 20 liters of liquid fermentation liquids stir culture again 14 days then with about 14 days of 90 rev/mins of round shaking culture with 240 rev/mins of rotational oscillations.
Indication space bag is with dress in the plastics bag by weight percentage herein, the any Mushrooms of 10-70% or the fibre object (being preferably herbal stem, stalk, fruit or fiber wood chip) such as stem, stalk, fruit or wood chip of plant, the starch source (being preferably potato) of 10-30%, and the dairy products of 5-15% (being preferably rice bran), the carbohydrate (being preferably glucose) of 1-10%, 0.5-2% phosphoric acid salt (being preferably potassium primary phosphate), and 0.1-1% vitriol (being preferably sal epsom), described each component concentration sum is 100%.Its humidity is about 60-90%, is preferably 80%, and adjusts its pH value near neutral.
The indication mycelium culture stage is meant that wherein the mycelial growth temperature is preferably 28 ℃ between 5 to 32 ℃ within about 60 days of the bacterial classification implantation space bag herein.The humidity of space bag is preferably between 60-80%, and more preferably 80%.As for its air conditions, then preferably contain the 0.2-1% carbonic acid gas.
After mycelium is cultivated and finished, carry out the sporophore cultivation stage, that is in about 61-90 days, the space bag is removed, make it to be exposed in the air fully.The day and night temperature difference during the sporophore growth must be controlled through strictness, and daytime temperature generally between 20 to 30 ℃, is preferably 25 ℃ ± 1 ℃; Nocturnal temperature is preferably 11 ℃ ± 1 ℃; Day and night the temperature difference is preferably 15 ℃.The atmospheric moisture in sporophore cultivation stage should be controlled between the 90-95%; Sporophore must be cultivated under airiness in addition, and wherein carbon dioxide content can not surpass 1%, the solid culture of generally can gathering in 90-120 days.
According to the present invention, use is deposited at food Industry in Taiwan Institute of Development Studies, the bacterial classification of numbering CCRC35398 is the source, this bacterial classification is commercially available bacterial classification, the solid culture that obtains according to solid culture method as shown in Figure 1, similar taper shape, diameter are about 10-30 centimetre, highly are about 15-35 centimetre and weight is about the 0.2-0.6 kilogram.
The processing treatment mode of utilizing camphor tree sesame solids that above-mentioned cultural method obtains to be familiar with via any known technology, as utilize water to carry or the activeconstituents of this solid culture of alcoholic extract, then, prepare concentrated product as vacuum concentration or freeze concentration with known method of enrichment.
Or giving drying treatment with drying means, for example spraying drying, lyophilize etc. be not wherein to have the spraying drying of heating, the dried powder that can obtain not being influenced by heat fully.Be preferably freeze-drying, between normal temperature, remove the moisture of this solids in the low temperature below 0 ℃, during drying this solid culture also is not subjected to the influence of oxygen and changes its character simultaneously.
Or make the fine particle of spray-dired micropowder particle or other desiccating methods preparation, increase the aggegation and the particle diameter thereof of powder through nodulizing (agglomeration), this agglomerate be configured to porous matter, possess good wetting properties.
High performance liquid chromatography (HPLC) spectrum data of the camphor tree sesame of camphor tree sesame solid culture of the present invention, known liquid culture method gained and wild camphor tree sesame compares, as Fig. 2.Analyze through HPLC, the sporophore of solid culture method gained of the present invention has and the very similar composition of natural wild camphor tree sesame, should have and identical activity and the purposes of natural wild camphor tree sesame, comprise calm the nerves, timid gas in fashion, stagnation resolvation are invigorated blood circulation, it is long-pending to disappear in the temperature, detoxify and promote the subsdence of swelling, and the effect of sedation-analgesia, and have the ability that suppresses bacterium, virus and tumour cell, reach cardiac stimulant, immunomodulatory, anti-parasympathetic nerve effect, the active effect of serotonin antagonist.Can be too high in order to treatment gastrointestinal pain, diarrhea and vomiting, diabetes, gout, ephritis, sacroiliitis, inflammation, allergy, urine protein, uremia, liver cirrhosis, liver cancer and influenza etc.Even have the ability that skin reproduces, and can be used as the material of human body skin or callus again, can make patch to bedsore, skin injury.
According to the present invention, further concrete real example solid culture of the present invention has Green Tea Extract and anti peroxidation of lipid ability, protects the liver function, and the anticancer effect.Obvious solid culture of the present invention can be used as the usefulness of the heath food of health, or can further be developed as various medicines.
The following example is used the present invention is further illustrated, and is not that modification and application that the content of those skilled in the art's specification sheets according to the present invention is finished all belong to category of the present invention in order to restriction the present invention.
Embodiment 1
To take from Foodstuff Industrial and Development Inst., and deposit the bacterial classification of numbering CCRC 35398, this bacterial classification is commercially available bacterial classification,, inoculates to the space bag with a large amount of cultivation bacterial classifications with the liquid culture fermentation.The prescription of fermentation culture is 2% fructose, 0.05% sal epsom, 0.5% yeast extract and 0.1% potassium primary phosphate.Carrying out mycelium with the space packet mode again cultivates, the space package becomes 65% herbal stem, stalk, fruit and wood fiber thing, 20% potato, 10% rice bran, 3.5% glucose, 1% potassium primary phosphate and 0.5% sal epsom, and adjust its humidity be 80% and the pH value be 7.Its humidity is about 60-90%, is preferably 80%, and adjusts its pH value near neutral.
The indication mycelium culture stage is meant that wherein the mycelial growth temperature is preferably 28 ℃ between 5 to 32 ℃ within about 60 days of the bacterial classification implantation space bag herein.The humidity of space bag is 80%, and air conditions is for containing the 0.2-1% carbonic acid gas.
After mycelium is cultivated and finished, carry out the sporophore cultivation stage, promptly in about 61-90 days the space bag is removed, make it to be exposed to fully in the air, the day and night temperature difference during the sporophore growth must be controlled through strictness, and daytime temperature is generally between 20 to 30 ℃; Nocturnal temperature then is 11 ℃ ± 1 ℃; Day and night the temperature difference maintains 15 ℃.The atmospheric moisture in sporophore cultivation stage is controlled between the 90-95%; Sporophore must be cultivated under airiness in addition, and wherein carbon dioxide content can not surpass 1%.
In 90-120 days, treat that conical sporophore grows up to 15 centimetres of diameters, the height solid culture of can gathering 25 centimetres the time, its weight is about 0.4 kilogram, as Fig. 1.The high performance liquid chromatography of its main active ingredient triterpenes (HPLC) atlas analysis is shown in Fig. 2 A, and its numerical value such as following table 1 are listed.
The high-efficient liquid phase chromatogram spectrum data of the camphor tree sesame sporophore of table 1 cultural method gained of the present invention
Different for camphor tree sesame solids of the present invention relatively and other planting type gained camphor tree sesames, and carry out HPLC, its collection of illustrative plates such as Fig. 2 B and Fig. 2 C, its data such as table 2 and table 3 with the camphor tree sesame and the wild camphor tree sesame of liquid culture method gained.By collection of illustrative plates as seen, the camphor tree sesame sporophore that the present invention's cultivation obtains has the collection of illustrative plates with the identical peak shape of wild camphor tree sesame, also represents that the sporophore that the inventive method is cultivated should have activity and the purposes identical with wild camphor tree sesame.
The high-efficient liquid phase chromatogram spectrum data of table 2 liquid culture method gained camphor tree sesame
Keep the area concentration composition
Numbering area % relative height height % concentration baseline sign indicating number
Time (UV* second) organization
1 1.620 262998 1.2194 38.745 2.2045 0.0000 PV
2 1.857 8294342 38.4242 660.685 37.5912 0.0000 VV
3 2.520 5417476 25.0969 726.398 41.3301 0.0000 VV
4 2.820 1692634 7.8413 99.406 5.6559 0.0000 VV
5 3.790 731157 3.3871 23.209 1.3205 0.0000 VB
6 4.123 271974 1.2599 11.713 0.6664 0.0000 BV
7 4.623 154068 0.7137 7.810 0.4444 0.0000 VV
8 5.023 154230 0.7145 4.580 0.2606 0.0000 VV
9 5.897 29822 0.1382 1.652 0.0940 0.0000 VP
10 7.860 150572 0.6975 5.210 0.2964 0.0000 PP
11 9.210 123419 0.5717 3.914 0.2227 0.0000 PP
12 11.930 130807 0.6060 3.563 0.2027 0.0000 PV
13 12.740 37712 0.1747 1.232 0.0701 0.0000 VP
14 15.103 504463 2.3370 11.048 0.6286 0.0000 PP
15 16.890 143477 0.6647 3.512 0.1998 0.0000 PP
16 19.350 56860 0.2634 1.827 0.1039 0.0000 PP
17 23.510 806878 3.7379 11.274 0.6415 0.0000 PP
18 27.167 92042 0.4264 1.224 0.0696 0.0000 PP
19 30.633 144285 0.6684 2.243 0.1276 0.0000 PP
20 33.747 289568 1.3414 3.947 0.2245 0.0000 PP
21 38.727 250488 1.1604 3.231 0.1839 0.0000 PP
22 41.340 61644 0.2856 0.771 0.0438 0.0000 PP
23 43.727 29928 0.1386 0.470 0.0267 0.0000 PP
24 57.047 112074 0.5192 9.096 0.5175 0.0000 PV
25 57.433 175711 0.8140 10.275 0.5846 0.0000 VV
26 57.907 107598 0.4985 4.508 0.2565 0.0000 VV
27 58.137 25810 0.1196 2.279 0.1296 0.0000 VP
28 61.380 23245 0.1077 2.024 0.1152 0.0000 PV
29 62.743 196612 0.9108 21.767 1.2385 0.0000 VV
30 64.223 30411 0.1409 2.094 0.1192 0.0000 PV
31 64.717 441196 2.0439 41.620 2.3681 0.0000 VV
32 64.997 48524 0.2248 4.870 0.2771 0.0000 VV
33 65.420 40354 0.1869 3.529 0.2008 0.0000 VV
34 65.833 67766 0.3139 3.823 0.2175 0.0000 VP
35 66.950 21298 0.0987 1.691 0.0962 0.0000 PP
36 67.773 32193 0.1491 1.708 0.0972 0.0000 PP
37 79.013 29414 0.1363 0.920 0.0523 0.0000 PP
38 97.297 21884 0.1014 0.456 0.0260 0.0000 PP
39 100.053 38474 0.1782 0.738 0.0420 0.0000 PP
40 104.457 126686 0.5869 2.358 0.1342 0.0000 PP
41 107.010 57252 0.2652 2.113 0.1202 0.0000 PP
42 111.397 35854 0.1661 4.236 0.2410 0.0000 VV
43 111.813 28337 0.1313 2.512 0.1429 0.0000 VP
44 113.790 25408 0.1177 1.810 0.1030 0.0000 VP
45 115.883 43858 0.2032 3.808 0.2167 0.0000 PV
46 119.590 25451 0.1179 1.656 0.0942 0.0000 PP
Summation 21,586,254 1757.553 0.0000
The high-efficient liquid phase chromatogram spectrum data of the wild camphor tree sesame of table 3
Keep the area concentration composition
Numbering area % relative height height % concentration baseline sign indicating number
Time (UV* second) organization
1 1.550 1332009 0.5787 162.045 1.1488 0.0000 PV
2 1.913 12708000 5.5210 797.430 5.6535 0.0000 VV
3 2.607 7990504 3.4715 374.149 2.6526 0.0000 VV
4 2.997 3336181 1.4494 151.464 1.0738 0.0000 VV
5 3.480 827969 0.3597 90.536 0.6419 0.0000 VV
6 3.687 2100942 0.9127 89.231 0.6326 0.0000 VV
7 4.110 1665378 0.7235 80.468 0.5705 0.0000 VV
8 4.590 986857 0.4287 61.543 0.4363 0.0000 VV
9 4.867 1300649 0.5651 59.445 0.4214 0.0000 VV
10 5.377 620409 0.2695 37.086 0.2629 0.0000 VV
11 5.623 712514 0.3095 37.666 0.2670 0.0000 VV
12 6.020 617412 0.2582 32.787 0.2324 0.0000 VV
13 6.347 380678 0.1654 24.939 0.1768 0.0000 VV
14 6.660 622333 0.2704 24.053 0.1705 0.0000 VV
15 7.060 557302 0.2421 18.364 0.1302 0.0000 VV
16 7.710 585765 0.2545 27.106 0.1922 0.0000 VV
17 8.353 276221 0.1200 10.957 0.0777 0.0000 VV
18 8.867 379887 0.1650 19.904 0.1411 0.0000 VV
19 9.327 316284 0.1374 21.157 0.1500 0.0000 VV
20 9.623 72843 0.0316 4.914 0.0348 0.0000 VV
21 10.593 120581 0.0524 6.584 0.0467 0.0000 PP
22 11.260 57846 0.0251 2.283 0.0162 0.0000 PV
23 11.747 32476 0.0141 2.199 0.0156 0.0000 VV
24 12.263 209326 0.0909 11.437 0.0811 0.0000 VV
25 12.837 131315 0.0570 6.603 0.0468 0.0000 VV
26 13.733 270732 0.1176 7.438 0.0527 0.0000 VP
27 15.300 156380 0.0679 6.542 0.0464 0.0000 PP
28 17.513 346774 0.1507 10.369 0.0735 0.0000 PV
29 18.270 1011777 0.4596 25.904 0.1837 0.0000 VV
30 19.863 298337 0.1296 7.175 0.0509 0.0000 VP
31 21.567 237547 0.1032 6.715 0.0476 0.0000 VV
32 22.657 281555 0.1136 7.129 0.0505 0.0000 VV
33 23.960 419821 0.1824 8.165 0.0579 0.0000 VP
34 26.780 1014237 0.4406 20.361 0.1444 0.0000 PP
35 23.320 4482109 1.9472 103.960 0.7370 0.0000 PP
36 32.017 6999710 3.0410 156.204 1.1074 0.0000 PV
37 33.343 399151 0.1734 8.733 0.0619 0.0000 VP
38 35.053 97125 0.0422 2.428 0.0172 0.0000 PV
39 37.007 3062612 1.3305 46.150 0.3272 0.0000 VP
40 39.587 832484 0.3617 15.926 0.1129 0.0000 PV
41 40.897 2411303 1.0476 30.354 0.2152 0.0000 VP
42 43.603 94412 0.0410 2.102 0.0149 0.0000 PP
43 46.093 950329 0.4129 12.948 0.0918 0.0000 PP
44 47.820 29686 0.0129 0.697 0.0049 0.0000 PP
45 51.273 22016 0.0096 0.505 0.0036 0.0000 PP
46 53.253 391511 0.1701 6.542 0.0464 0.0000 PP
47 55.177 418679 0.1819 6.082 0.0431 0.0000 PP
48 57.243 3988680 1.7329 240.883 1.7078 0.0000 PV
49 57.500 4155318 1.8053 345.611 2.4503 0.0000 VV
50 57.787 3545656 1.5404 257.829 1.8279 0.0000 VV
51 58.010 1206951 0.5244 175.620 1.2451 0.0000 VV
52 58.257 2986313 1.2974 222.955 1.5807 0.0000 VV
53 58.400 5093407 2.2128 431.601 3.0599 0.0000 VV
54 58.673 2540255 1.1036 286.541 2.0315 0.0000 VV
55 58.943 5661787 2.4597 471.025 3.3394 0.0000 VV
56 59.233 2400643 1.0430 197.347 1.3991 0.0000 VV
57 59.543 3531683 1.5343 200.445 1.4211 0.0000 VV
58 59.850 2228945 0.9684 164.165 1.1639 0.0000 VV
59 60.060 2828869 1.2290 335.781 2.3806 0.0000 VV
60 60.410 17547630 7.6235 994.818 7.0529 0.0000 VV
61 60.743 13833800 6.0101 994.628 7.0516 0.0000 VV
62 60.920 9930270 4.3142 941.167 6.6726 0.0000 VV
63 61.343 3601150 1.5645 265.761 1.8842 0.0000 VV
64 61.723 60485Z3 2.6278 525.238 3.7238 0.0000 VV
65 62.127 2249845 0.9774 160.527 1.1381 0.0000 VV
66 62.357 2849680 1.2380 152.131 1.0786 0.0000 VV
67 63.207 11335680 4.9248 888.220 6.2830 0.0000 VV
68 63.460 3758168 1.6327 214.761 1.5226 0.0000 VV
69 64.087 17403430 7.5609 992.676 7.0377 0.0000 VV
70 64.807 8379895 2.7717 332.965 2.3606 0.0000 VV
71 65.487 1105522 0.4803 48.756 0.3457 0.0000 VV
72 66.060 1061718 0.4613 41.562 0.2947 0.0000 VV
73 66.490 1903467 0.8270 65.469 0.4642 0.0000 VV
74 67.563 1016818 0.4418 30.915 0.2192 0.0000 VV
75 68.387 1511594 0.6567 37.104 0.2631 0.0000 VV
76 69.390 10743430 4.6675 526.207 3.7306 0.0000 VV
77 70.583 358833 0.1559 13.087 0.0928 0.0000 VV
78 71.257 2732845 1.1873 127.212 0.9019 0.0000 VV
79 72.317 237862 0.1033 7.948 0.0563 0.0000 VV
80 72.730 245290 0.1066 6.784 0.0481 0.0000 VV
81 73.783 87523 0.0380 3.200 0.0227 0.0000 VV
82 74.250 127009 0.0552 4.136 0.0293 0.0000 VV
83 74.890 86115 0.0374 2.980 0.0211 0.0000 VP
84 75.973 28738 0.0125 1.165 0.0083 0.0000 PP
85 77.503 92275 0.0401 1.898 0.0135 0.0000 PP
86 80.060 508744 0.2210 11.408 0.0809 0.0000 PP
87 82.200 28158 0.0122 0.871 0.0062 0.0000 PP
88 84.293 79909 0.0347 1.534 0.0109 0.0000 PP
89 87.500 1092447 0.4746 24.011 0.1702 0.0000 PV
90 87.630 300046 0.1304 24.553 0.1741 0.0000 VV
91 87.893 382295 0.1681 25.603 0.1815 0.0000 VV
92 88.107 224523 0.0975 21.631 0.1534 0.0000 VV
93 88.397 717983 0.3119 37.094 0.2630 0.0000 VV
94 89.010 518852 0.2254 22.242 0.1577 0.0000 VV
95 89.237 307090 0.1334 21.551 0.1528 0.0000 VV
96 89.473 302635 0.1315 19.158 0.1358 0.0000 VV
97 90.007 4050733 1.7598 326.533 2.3150 0.0000 VV
98 91.020 376589 0.1636 16.743 0.1187 0.0000 VV
99 91.427 152969 0.0685 11.423 0.0810 0.0000 VV
100 91.643 205346 0.0892 11.033 0.0782 0.0000 VV
101 92.013 142333 0.0818 11.613 0.0823 0.0000 VV
102 92.157 268901 0.1168 11.479 0.0814 0.0000 VV
103 92.633 124915 0.0543 8.073 0.0572 0.0000 VV
104 93.083 908307 0.3946 88.376 0.6266 0.0000 VV
105 93.690 210904 0.0916 15.423 0.1093 0.0000 VV
106 94.063 323059 0.1404 22.173 0.1572 0.0000 VV
107 94.547 63428 0.0276 3.102 0.0220 0.0000 VV
108 95.123 29222 0.0127 2.059 0.0146 0.0000 VP
109 95.963 45135 0.0196 2.967 0.0210 0.0000 VP
110 98.950 58309 0.0253 3.644 0.0258 0.0000 VP
111 100.273 20525 0.0089 0.713 0.0051 0.0000 PP
112 105.563 38299 0.0166 1.804 0.0128 0.0000 PP
113 112.263 55710 0.0242 1.468 0.0104 0.0000 PP
114 126.417 44605 0.0194 0.881 0.0061 0.0000 PP
Summation 230,177,556 14105.03 0.0000
Following compositions analytical study and pharmacological verification illustrate the composition and the effect of its concentrated product of cultural method gained camphor tree sesame sporophore solids according to the present invention.By analysis, main component comprises triterpene compound, water-soluble polysaccharide and chitin.
Water-soluble polysaccharide
Detect through japanese food analytic centre, find the real sweet wine solids of cultural method gained camphor tree sesame of the present invention tool a large amount water-soluble polysaccharide, per 100 grams contain the water-soluble polysaccharide of 1.2 to 6.5 grams approximately.
Heavy metal content
According to National Tsing Hua University's atomics be the trace analysis laboratory with atomic spectrograph, the camphor tree sesame solids of cultivating according to the inventive method is carried out the heavy metal analysis, the result shows below:
Na：48.7ppm Mn：10.9ppm Cd：ND
Mg：767ppm Fe：100ppm Cr：ND
Al：41.6ppm Zn：16.4ppm
Ca：605ppm As：ND
K：0.560％ Hg：ND
Total antioxidant activity(Total antioxidant activity)
Concentrate powder according to the concentrated powder of camphor tree sesame solids of the present invention and wild camphor tree sesame powder and Ganoderma sporophore and compare, detect the content of every gram powder, it is equivalent to ascorbic milligram number.
The source
The ascorbic milligram of the content of every gram powder/be equivalent to number
Wild camphor tree sesame powder 633 micrograms xitix/gram
Ganoderma sporophore concentrates powder 854 micrograms xitix/gram
The concentrated powder 1180 micrograms xitix/gram of camphor tree sesame solids of the present invention
This result shows that the resistance to oxidation of camphor tree sesame solids of the present invention is far above Ganoderma sporophore and wild camphor tree sesame.
Toxicity test
Use mouse to carry out acute oral toxicity test LD50 according to camphor tree sesame of the present invention, oral one time 2000 mg/kg of experimental mice is according to camphor tree sesame of the present invention, and control group then gives Purified Water and contrasts as solvent.Experimental result shows that unusual or dead example does not appear in experimental mice.Therefore, acute oral toxicity test LD50 value is more than 2000 mg/kg.
Ames test (Ames test)
Camphor tree sesame according to the present invention utilizes dull and stereotyped bulk testing method, carries out Ames test (Ames test).Test strain includes Salmonella typhimurtum TA97, TA98, TA100, TA102 and TA1535 five strains, five concentration analysis, be respectively 312.5,625,1250,2500 and 5000 micrograms/flat board, comprise negative control group and the narrow spectrum positive controls of tool bacterial strain simultaneously, all do three repetitions for every group.Test result shows that the contrary sudden change bacterium colony background that negative control group causes all falls within the acceptable range; And positive controls also causes the increase of contrary sudden change bacterium colony digital display work.No matter whether camphor tree sesame according to the present invention contains under the processing of rat liver enzyme (S9) metabolic system, all test concentrations all significantly do not cause the increase of the contrary sudden change of test strain colony number, and are as shown in table 4.
Table 4 Ames test
| Experimental subjects | The experimental example number | During the intake | Experimental result |
| Salmonella typhimurtum | Each 5 strain TA97 TA98 TA100 TA102 TA1535 needs the histidine mutant strain | Test concentrations (microgram/flat board) 312.5,625,1250,2500,5000 dull and stereotyped bulk testings heavily cover three 37 ± 1 ℃ of constant temperature culture and calculated the sudden change colony number in 48～72 hours | All test concentrations, under the processing that adds or do not add rat liver enzyme metabolic system (S9), all significantly do not cause the increase of the contrary sudden change of test strain colony number.Test-results is negative |
The test of anti peroxidation of lipid ability
Utilize the peroxidatic reaction of lipid of rat brain homogenizing fluid, to detect Green Tea Extract and resistance of oxidation according to camphor tree sesame solid culture concentrated solution of the present invention.Fig. 3 shows the peroxidatic reaction of lipid that causes free radical with ferrous ion stimulation in rats brain homogenizing fluid, and cause lipid peroxidation composition (TBARS) to increase, and according to three times of camphor tree sesame solid culture of the present invention with the increase of octuple concentrated solution with its concentration, its result shows can significantly suppress peroxidation, and has the Green Tea Extract ability.
Protect the liver functional experiment
Foundation is inquired into the influence of the camphor tree sesame of different cycles of concentration for the mouse chronic hepatic injury with the experimental model of the chronic chemical damage of tetrachloro-methane induction mouse.
24 mouse (ICR strain) of not having a disease are divided into four groups, and are as shown in table 5.
Table 5 camphor tree sesame test dose and animal group distribute
Group tetracol phenixin CCl
4Dosage dosage degree mouse number of elements
A 0 (control group) 0 (control group) 6
(0.3 milliliter/100 gram bodies
Heavy)
The same 50 milligrams of C (three times)/kilogram 6
D the same 50 milligrams (octuple)/kilogram 6
To A and B to D group mouse difference subcutaneous injection sweet oil (0.1 milliliter/10 gram body weight) and 40% tetracol phenixin CCl
4Sweet oil (0.1 milliliter/10 gram body weight), weekly twice, and A and B group mouse irritated with stomach tube give salt solution, organize the camphor tree sesame concentrating sample of cultivating according to the inventive method of then distinguishing two kinds of dosage of oral feeding at C and D, kept for five weeks altogether.All mouse detect liver correlation function biochemical values respectively at the 5th all backs in eye socket blood sampling mode, put to death mouse thereupon, get its liver and fixing maximum lobus dexter hepatic tissue to carry out pathological study, intensity of variations such as for example liver cell is impaired, steatosis, necrosis and fibrosis.Biochemical function is measured and is comprised L-glutamic acid in the blood-pyruvic acid transaminase (GPT) and glutamic-oxaloacetic transaminase (GOT) enzymic activity, and GPT and GOT detection principle are the standard methods according to the federal clinical chemistry in the world.
Experimental result shows that so that GPT and GOT value are in the obviously rising of five week of experiment back in the chronic liver injury of the tetracol phenixin mice plasma down, wherein the GPT value is about 850 units, and this is 20-22 a times of normal value; And the GOT value is about 1700 units, and this is normal value 40-42 times (Fig. 4,5).In addition, three times of feedings and octuple concentrated camphor tree sesame of the present invention obviously suppresses the rising because of GPT that tetracol phenixin causes and GOT numerical value.Especially when GPT and GOT value peak, use three times of concentrated dosage can suppress the rising of about 37%GPT and 65%GOT value.As shown in Figure 6, compared to the liver organization of normal mouse, handle mouse after six weeks with tetracol phenixin, obvious enlargement of its liver organization and surface are greyish white coarse shape; Yet the liver of feeding different concns camphor tree sesame enriched material mouse is not found enlargement or greyish white coarse surface.In addition, the discovery fat particle is obviously piled up under dyeing, near the central vein the obvious enlargement of liver cell, the scavenger cell of iron content metabolite and other inflammatory white cells soak between liver cell, and the visible nodositas of part necrocytosis transforms (nodulartransformation).The liver organization of the mouse of feeding different concns camphor tree sesame does not find that then above-mentioned pathologic changes (see figure 7).In addition, table 6 is illustration mouse liver pathological tissue semi-quantitative analysis (N represents that normal group, D represent that tetracol phenixin injury group, X3 represent that three times of feeding camphor tree sesames concentrate group and X8 represents that feeding camphor tree sesame octuple concentrates group), should confirm that wherein camphor tree sesame enriched material obviously improves pathologic variation.
Table 6 mouse liver pathological tissue semi-quantitative analysis
Liver lipid necrocytosis bile duct increases calcification
The inflammation sex change is given birth to
Local necrosis connects downright bad nodositas and transforms
N 0 0 0-1 0 0 0 0
D 2-3 2-3 2 2 0-1 1 0-1
X3 1 1-2 0-1 1-2 0-1 0-1 0-1
X8 0-1 0-1 0-1 1 0 0-1 0-1
The cancer cells inhibition test
Foodstuff Industrial and Development Inst. accepts commission and carries out camphor tree sesame solid culture concentrated solution of the present invention to cervical cancer (HeLa), cancer of the stomach (AGS), mammary cancer (MCF-7), liver cancer (HepG2) and intestinal cancer cancer cells inhibition tests such as (COLO 320HSR), cell growth inhibiting rate is as shown in table 7, camphor tree sesame solid culture concentrated solution of the present invention is to the potent restraint of tumour cell tool (75%-87%), and wherein the restraint to breast cancer cell (MCF-7) is the strongest.
Table 7 camphor tree sesame solid culture concentrated solution suppresses the test neoplastic cell result
Cell growth inhibiting rate (%)
Cell strain kind mean value (%) ± SD
Hela 83 ± 3.1
AGS 84 ± 2,7
HepG2 75 ± 2.4
MCF-7 87 ± 1.4
*The specimen final concn is 1/10th of a primary sample concentration
In addition, use powdered sample (10 mg/ml) five kinds of (New1, New2, New3, New4 and New5) and liquid sample 11 kinds (20L, New2, New3,520,521,522,523,524,525,526 and 527) and 10 times of dilute samples thereof to cultivate tetrazolium detection (microculture tetrazolium assay) respectively and carry out colon-cancer cell (COLO320HSR) inhibition test with trace.Its result is shown in when final concn is 1/10 stoste and has at least 30% restraint as shown in Figure 8, and the strongest with the restraint of sample 20L, can be up to 90% inhibition colon-cancer cell ability.
External chromosome structure analysis of variance
Further utilize camphor tree sesame according to the present invention to carry out external chromosome structure analysis of variance again, its method is to handle Chinese hamster ovary cell (Chinesehamster ovary cell) with camphor tree sesame according to the present invention, analyze the cell of metaphase afterwards, observe and calculate the chromosomal frequency of its exception throw kenel.This chromosome structure analysis of variance method can be in order to assess the ability that camphor tree sesame pair cell karyomit(e) according to the present invention damages.This analysis is carried out with following three kinds of processing modes respectively, and first kind is not add under rat liver activating enzymes (S9) system, handles 3 hours with camphor tree sesame according to the present invention; Second kind is to add S9, handles cell 3 hours; The third is not add under the S9, handles cell continuously 20 hours.Its test concentrations and experimental result are then as shown in table 8, camphor tree sesame according to the present invention each test concentrations under three kinds of processing modes, and the frequency that does not all significantly cause chromosomal variation increases.
The external chromosome structure analysis of variance of table 8
| Experimental subjects | The experimental example number | During the intake | Experimental result |
| Chinese hamster ovary cell (ATCC, preserving number CC1-61 CHO-K1) | According to each dull and stereotyped Tissue Culture Dish of implanting 60mm of test concentrations, 2 of every kind of concentration are divided into 3 groups according to the different treatment condition | First and second organizes test concentrations equal 0,312.5,625,1250,2500 and 5000 mcg/ml.The 3rd group of test concentrations is 0,30,100,300,1000 and 3000 mcg/ml are cultivated back selective staining body numbers 18～21 according to indivedual conditions and the cell that evenly scatters is observed | All show three groups of results that process in the examination: camphor tree sesame according to the present invention causes the chromosome abnormality frequency to Chinese hamster ovary cell and there is no remarkable increase before and after the metabolism activation and under the different time processing. So this experimental result is negative. |
Small nut analysis in the animal body
Camphor tree sesame according to the present invention is caused the situation of small nut at the netted red blood corpuscle of mouse, judge by this whether to have in animal body to lure the ability of splitting (clasiogeniciiy), and further provide camphor tree sesame according to the present invention whether the mankind are had genotoxic risk assessment according to camphor tree sesame of the present invention.25 mouse are divided into five groups to be tested, and its test method and experimental result are as shown in table 9.Small nut analysis by mouse shows that camphor tree sesame according to the present invention is to the negative reaction of small nut analytical results of mouse periphery blood.
Small nut analysis in table 9 animal body
| Experimental subjects | The experimental example number | During the intake | Experimental result |
| The ICR mouse | Every group of each 5 male mouse of dosage group 4. low dose group 5. positive controls in the 1. solvent control groups of scoring, 2. high dose group 3. | 1～4 group of each oral throwing and 0, the dosage of 500,1000 and 2000 mg/kg; Positive controls is thrown and 1.0 mg/kg MMC take a blood sample according to condition observes netted red blood corpuscle down in luminescence microscope | 1. body weight and no significant difference 2. netted red blood cell proportion gradings between dosage group and the solvent control group: prove the trend that does not cause mouse bone marrow cells toxicity 3. small nut ratios to there is no dose-response, compare also do not have significantly increase thus camphor tree sesame according to the present invention to the negative reaction of small nut analysis result of mouse periphery blood. |
Should support camphor tree sesame solids according to the present invention not only to have the composition similar based on above-mentioned pharmacological datum to wild shape, as polysaccharide and triterpene compound, and anti peroxidation of lipid ability, Green Tea Extract or the anti-oxidant effect that is equal to, find also in addition that camphor tree sesame solids according to the present invention has to protect the liver and anticancer effect.
Claims (34)
1. method of cultivating solid camphor tree sesame, it comprises that camphor tree sesame bacterial classification is carried out mycelium with the space bag to be cultivated, and removes the space bag, makes it to be exposed to carry out sporophore cultivation for some time to conical sporophore in the air and form; Wherein contain the herbal stem of 10-70%, stalk, fruit or wood chip, 10-30% starch source, 1-10% carbohydrate, 5-15% rice bran, 0.5-2% phosphoric acid salt and 0.1-1% vitriol in this space bag, described each components contents sum is 100%;
Wherein the humidity in space bag cultivation stage is 60-90% again, and adjusts its pH value for neutral.
2. method according to claim 1, wherein this starch source is a potato.
3. method according to claim 1, this space bag is made up of 65% herbal stem, stalk, fruit or wood chip, 20% potato, 10% rice bran, 3.5% glucose, 1% potassium primary phosphate and 0.5% sal epsom.
4. method according to claim 1, the temperature that this mycelium is cultivated is between 5 ℃ to 32 ℃.
5. method according to claim 4, wherein the temperature of mycelium cultivation is 28 ℃.
6. method according to claim 1, wherein space bag humidity is 80%.
7. method according to claim 1, the air conditions when wherein mycelium is cultivated is the carbonic acid gas that contains 0.2-1%.
8. method according to claim 1, its sporophore are cultivated under airiness and are carried out.
9. method according to claim 8, the daytime temperature during its sporophore cultivation is between 20 to 30 ℃, and nocturnal temperature is between 11 ± 1 ℃.
10. method according to claim 9, night its day, the temperature difference was 15 ℃.
11. method according to claim 1, the atmospheric moisture that sporophore is cultivated is between 90-95%.
12. method according to claim 11, wherein Carbon Dioxide in Air content is lower than 1%.
13. method according to claim 1, mycelium are cultivated and were carried out 60 days.
14. method according to claim 1, it comprises a large amount of culturing steps of bacterial classification before the mycelium cultivation stage, comprises (1) and gets a fritter agar mycelia piece from liquid nitrogen preservation activatory bacterial classification, moves in the fresh culture and cultivates down in constant temperature; When (2) treating that growth is extremely vigorous, this bacterial classification inoculation to the space bag of the fibre object of killing bacterium, is then cultivated down in constant temperature; When (3) treating that this bacterial classification in the space bag covers with mycelia, remove older mycelia piece topmost, can be seeded in a large number in the space bag then.
15. method according to claim 1, it comprises a large amount of culturing steps of bacterial classification before the mycelium cultivation stage, comprise with the liquid culture fermentation with a large amount of cultivation bacterial classifications, after slant tube is cultivated, the bacterial classification inoculation to 5 that breeding is good is in a large number risen in the fermentation culture, under 24-26 ℃, cultivated about 14 days,, be seeded in 20 liters of liquid fermentation liquids stir culture again 14 days then with about 14 days of 90 rev/mins of round shaking culture with 240 rev/mins of rotational oscillations; Wherein the prescription of fermentation culture is 1-3% fructose, 0.01-0.1% sal epsom, 0.1-1% yeast extract, 0.05-0.5% potassium primary phosphate.
16. method according to claim 15, wherein the prescription of fermentation culture is 2% fructose, 0.05% sal epsom, 0.5% yeast extract and 0.1% potassium primary phosphate.
17. the camphor tree sesame sporophore solids that the method for claim 1 is cultivated is characterized in that having the physiologically active of the triterpenes components consistent with wild shape camphor tree sesame.
18. camphor tree sesame sporophore solids, it is that to deposit numbering CCRC 35398 with Foodstuff Industrial and Development Inst. be that bacterial classification is originated, this bacterial classification is commercially available bacterial classification, and the method for claim 1 cultivation and the camphor tree sesame sporophore solids that obtains, it is characterized in that it has the physiologically active of the triterpenes components consistent with wild shape camphor tree sesame, and its profile is that coniform, diameter is 10-30 centimetre, highly is the 0.2-0.6 kilogram for 15-35 centimetre and weight.
19. the product of a camphor tree sesame sporophore solids is characterized in that the described camphor tree sesame of claim 18 sporophore solids through pulverizing, and be concentrated into alcohol or water extraction thick, with concentrated solution.
20. product according to claim 19, it can further make dry powdered with drying treatment.
21. product according to claim 20, its dried powder can row be dry again through the water damping, to get agglomerate.
22. the purposes of the described product of claim 19 in producing a kind of food compositions.
23. the described camphor tree sesame of claim 18 sporophore solids or its converted products have purposes in the food compositions of the function of protecting the liver in production.
24. the described camphor tree sesame of claim 18 sporophore solids or its converted products purposes in the food compositions that production is used for the treatment of or anticancer is grown.
25. the described camphor tree sesame of claim 18 sporophore solids or its converted products are used for suppressing the purposes of the food compositions of peroxidation in production.
26. the described camphor tree sesame of claim 18 sporophore solids or its converted products are used for the purposes of the food compositions of Green Tea Extract or anti peroxidation of lipid reaction in production.
27. the described camphor tree sesame of claim 18 sporophore solids or its converted products are used for protecting the liver the purposes of the medical composition of function in production.
28. the described camphor tree sesame of claim 18 sporophore solids or its converted products purposes in the medical composition that production is used for the treatment of or anticancer is grown.
29. the described purposes of claim 28, wherein cancer cells is meant cervical cancer cell (HeLa), stomach cancer cell (AGS), breast cancer cell (MCF-7), liver cancer cell (HepG2) and colon-cancer cell (COLO 320HSR).
30. the described camphor tree sesame of claim 18 sporophore solids or its converted products are used for suppressing the purposes of the medical composition of peroxidation in production.
31. the described camphor tree sesame of claim 18 sporophore solids or its converted products are used for the purposes of the medical composition of Green Tea Extract or anti peroxidation of lipid reaction in production.
32. the described camphor tree sesame of claim 18 sporophore solids is used to calm the nerves in production, timid gas in fashion, stagnation resolvation is invigorated blood circulation, it is long-pending to disappear in the temperature, detoxify and promote the subsdence of swelling, and the medicine of sedation-analgesia, or produce a kind of anticorrosion toxinicide, or production is used to suppress bacterium, the medicine of virus and tumour cell, or production is used for cardiac stimulant, immunomodulatory, anti-parasympathetic nerve effect, the active medicine of serotonin antagonist, or production is used for the treatment of gastrointestinal pain, diarrhea and vomiting, diabetes, gout, ephritis, sacroiliitis, inflammation, irritated, urine protein is too high, uremia, liver cirrhosis, purposes in liver cancer and the grippal medicine.
33. the described camphor tree sesame of claim 18 sporophore solids is being produced in order to as the purposes in the material of human body skin or callus.
34. the described purposes of claim 32, medicine wherein are the patches of treatment bedsore, skin injury.
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| CNB011236132A CN1195843C (en) | 2001-08-10 | 2001-08-10 | Solid culture method of antrodia camphorate, its cultured solid substance, and its product and application |
| HK03103307A HK1051217A1 (en) | 2001-08-10 | 2003-05-13 | Incubation method for obtaining solid culture of zhang zhi, solid culture obtained therefrom, processed products and use thereof. |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101803534A (en) * | 2010-05-05 | 2010-08-18 | 深圳市中科海外科技有限公司 | Large-scale solid cultural method of antrodia camphorata |
| CN101785402B (en) * | 2009-01-23 | 2012-05-30 | 伟诚生物科技有限公司 | Method for preparing antradiacomphora extract with high contents of polysaccharides and triterpenoids |
| CN103828598A (en) * | 2013-11-18 | 2014-06-04 | 叶宗铭 | Solid state fermentation fruiting body cultivation transfer fusion method and device |
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| CN102523910B (en) * | 2010-12-28 | 2014-01-08 | 光晟生物科技股份有限公司 | Novel antrodia cinnamomea bacteria for generating high-content triterpenoid |
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| CN106852496A (en) * | 2015-12-09 | 2017-06-16 | 黄建钧 | The method of the triterpenes synthesis contained by induction mushroom gill fungus |
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-
2001
- 2001-08-10 CN CNB011236132A patent/CN1195843C/en not_active Expired - Lifetime
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101785402B (en) * | 2009-01-23 | 2012-05-30 | 伟诚生物科技有限公司 | Method for preparing antradiacomphora extract with high contents of polysaccharides and triterpenoids |
| CN101803534A (en) * | 2010-05-05 | 2010-08-18 | 深圳市中科海外科技有限公司 | Large-scale solid cultural method of antrodia camphorata |
| CN101803534B (en) * | 2010-05-05 | 2011-06-29 | 深圳市中科海外科技有限公司 | Large-scale solid cultural method of antrodia camphorata |
| CN103828598A (en) * | 2013-11-18 | 2014-06-04 | 叶宗铭 | Solid state fermentation fruiting body cultivation transfer fusion method and device |
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| CN1379082A (en) | 2002-11-13 |
| HK1051217A1 (en) | 2003-07-25 |
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