CN110218656A - Ganoderma lucidum-herb residue bidirectional solid fermentation method utilizing air pressure pulsation and application - Google Patents
Ganoderma lucidum-herb residue bidirectional solid fermentation method utilizing air pressure pulsation and application Download PDFInfo
- Publication number
- CN110218656A CN110218656A CN201910470543.2A CN201910470543A CN110218656A CN 110218656 A CN110218656 A CN 110218656A CN 201910470543 A CN201910470543 A CN 201910470543A CN 110218656 A CN110218656 A CN 110218656A
- Authority
- CN
- China
- Prior art keywords
- fermentation
- solid
- ganoderma lucidum
- dregs
- decoction
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000000855 fermentation Methods 0.000 title claims abstract description 90
- 230000004151 fermentation Effects 0.000 title claims abstract description 90
- 239000007787 solid Substances 0.000 title claims abstract description 76
- 241000222336 Ganoderma Species 0.000 title claims abstract description 43
- 238000000034 method Methods 0.000 title claims abstract description 39
- 230000010349 pulsation Effects 0.000 title claims abstract description 35
- 230000002457 bidirectional effect Effects 0.000 title abstract 3
- 238000010563 solid-state fermentation Methods 0.000 claims abstract description 20
- 239000001963 growth medium Substances 0.000 claims description 38
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 19
- 239000007788 liquid Substances 0.000 claims description 18
- 238000002360 preparation method Methods 0.000 claims description 16
- 229920000742 Cotton Polymers 0.000 claims description 15
- 239000000203 mixture Substances 0.000 claims description 14
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 13
- 239000008103 glucose Substances 0.000 claims description 13
- 235000015097 nutrients Nutrition 0.000 claims description 13
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 12
- 238000001035 drying Methods 0.000 claims description 11
- 235000015099 wheat brans Nutrition 0.000 claims description 10
- 230000010355 oscillation Effects 0.000 claims description 8
- 238000011218 seed culture Methods 0.000 claims description 8
- 238000012545 processing Methods 0.000 claims description 7
- 239000001888 Peptone Substances 0.000 claims description 6
- 108010080698 Peptones Proteins 0.000 claims description 6
- 229940041514 candida albicans extract Drugs 0.000 claims description 6
- 239000000110 cooling liquid Substances 0.000 claims description 6
- 239000002054 inoculum Substances 0.000 claims description 6
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 6
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 6
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 6
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 6
- 235000019319 peptone Nutrition 0.000 claims description 6
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 6
- 238000009423 ventilation Methods 0.000 claims description 6
- 239000012138 yeast extract Substances 0.000 claims description 6
- 229940107666 astragalus root Drugs 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 5
- 239000013538 functional additive Substances 0.000 claims description 4
- 239000004615 ingredient Substances 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- 239000000758 substrate Substances 0.000 claims description 4
- 230000001954 sterilising effect Effects 0.000 claims description 2
- 238000004659 sterilization and disinfection Methods 0.000 claims description 2
- 239000006071 cream Substances 0.000 claims 1
- 240000008397 Ganoderma lucidum Species 0.000 abstract description 25
- 235000001637 Ganoderma lucidum Nutrition 0.000 abstract description 25
- 239000013543 active substance Substances 0.000 abstract description 15
- 241000894006 Bacteria Species 0.000 abstract description 11
- 239000003814 drug Substances 0.000 abstract description 11
- 238000005516 engineering process Methods 0.000 abstract description 7
- 230000008901 benefit Effects 0.000 abstract description 4
- 238000009395 breeding Methods 0.000 abstract description 4
- 230000001488 breeding effect Effects 0.000 abstract description 4
- 239000002994 raw material Substances 0.000 abstract description 4
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 abstract description 3
- 238000003912 environmental pollution Methods 0.000 abstract description 3
- 229910052760 oxygen Inorganic materials 0.000 abstract description 3
- 239000001301 oxygen Substances 0.000 abstract description 3
- 230000008859 change Effects 0.000 abstract description 2
- 238000005265 energy consumption Methods 0.000 abstract description 2
- 230000008569 process Effects 0.000 abstract description 2
- 239000002699 waste material Substances 0.000 abstract description 2
- 239000003242 anti bacterial agent Substances 0.000 abstract 1
- 229940088710 antibiotic agent Drugs 0.000 abstract 1
- 238000011109 contamination Methods 0.000 abstract 1
- 238000009776 industrial production Methods 0.000 abstract 1
- 230000003068 static effect Effects 0.000 description 20
- 241000204031 Mycoplasma Species 0.000 description 14
- 229920001282 polysaccharide Polymers 0.000 description 13
- 239000005017 polysaccharide Substances 0.000 description 13
- 239000002028 Biomass Substances 0.000 description 12
- 150000004676 glycans Chemical class 0.000 description 11
- 238000003756 stirring Methods 0.000 description 11
- -1 triterpene compound Chemical class 0.000 description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 10
- 239000000463 material Substances 0.000 description 7
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 6
- 241000238557 Decapoda Species 0.000 description 6
- 235000019441 ethanol Nutrition 0.000 description 6
- 230000004083 survival effect Effects 0.000 description 6
- 241001327110 Macrobrachium rosenbergii Species 0.000 description 5
- 239000000498 cooling water Substances 0.000 description 5
- 238000000605 extraction Methods 0.000 description 5
- 230000004060 metabolic process Effects 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 239000002893 slag Substances 0.000 description 5
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 4
- 102000002322 Egg Proteins Human genes 0.000 description 4
- 108010000912 Egg Proteins Proteins 0.000 description 4
- 241000233866 Fungi Species 0.000 description 4
- 235000003434 Sesamum indicum Nutrition 0.000 description 4
- 241001052560 Thallis Species 0.000 description 4
- 238000005119 centrifugation Methods 0.000 description 4
- 235000014103 egg white Nutrition 0.000 description 4
- 210000000969 egg white Anatomy 0.000 description 4
- 230000036541 health Effects 0.000 description 4
- 238000009630 liquid culture Methods 0.000 description 4
- 230000000638 stimulation Effects 0.000 description 4
- 241001061264 Astragalus Species 0.000 description 3
- 241000371997 Eriocheir sinensis Species 0.000 description 3
- 239000009636 Huang Qi Substances 0.000 description 3
- 241000207961 Sesamum Species 0.000 description 3
- 235000006533 astragalus Nutrition 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000007789 gas Substances 0.000 description 3
- 230000000813 microbial effect Effects 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- AIGAZQPHXLWMOJ-UHFFFAOYSA-N Tanshinone I Chemical compound C1=CC2=C(C)C=CC=C2C(C(=O)C2=O)=C1C1=C2C(C)=CO1 AIGAZQPHXLWMOJ-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 239000011149 active material Substances 0.000 description 2
- 230000003115 biocidal effect Effects 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 230000001151 other effect Effects 0.000 description 2
- 238000003359 percent control normalization Methods 0.000 description 2
- 238000002390 rotary evaporation Methods 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 239000013049 sediment Substances 0.000 description 2
- 210000004233 talus Anatomy 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- XBZYWSMVVKYHQN-MYPRUECHSA-N (4as,6as,6br,8ar,9r,10s,12ar,12br,14bs)-10-hydroxy-2,2,6a,6b,9,12a-hexamethyl-9-[(sulfooxy)methyl]-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carboxylic acid Chemical compound C1C[C@H](O)[C@@](C)(COS(O)(=O)=O)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CCC(C)(C)C[C@H]5C4=CC[C@@H]3[C@]21C XBZYWSMVVKYHQN-MYPRUECHSA-N 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 235000010110 Astragalus glycyphyllos Nutrition 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 241000193388 Bacillus thuringiensis Species 0.000 description 1
- 206010007247 Carbuncle Diseases 0.000 description 1
- 240000008067 Cucumis sativus Species 0.000 description 1
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 description 1
- 241000790917 Dioxys <bee> Species 0.000 description 1
- 206010013786 Dry skin Diseases 0.000 description 1
- 241000371986 Eriocheir Species 0.000 description 1
- 241000207923 Lamiaceae Species 0.000 description 1
- 241000228150 Penicillium chrysogenum Species 0.000 description 1
- 241000207929 Scutellaria Species 0.000 description 1
- 244000000231 Sesamum indicum Species 0.000 description 1
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical class [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 1
- PJANXHGTPQOBST-VAWYXSNFSA-N Stilbene Natural products C=1C=CC=CC=1/C=C/C1=CC=CC=C1 PJANXHGTPQOBST-VAWYXSNFSA-N 0.000 description 1
- 229930183118 Tanshinone Natural products 0.000 description 1
- 241000223259 Trichoderma Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- QMNWISYXSJWHRY-YLNUDOOFSA-N astragaloside IV Chemical compound O1[C@H](C(C)(O)C)CC[C@]1(C)[C@@H]1[C@@]2(C)CC[C@]34C[C@]4(CC[C@H](O[C@H]4[C@@H]([C@@H](O)[C@H](O)CO4)O)C4(C)C)[C@H]4[C@@H](O[C@H]4[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O4)O)C[C@H]3[C@]2(C)C[C@@H]1O QMNWISYXSJWHRY-YLNUDOOFSA-N 0.000 description 1
- QMNWISYXSJWHRY-BCBPIKMJSA-N astragaloside IV Natural products CC(C)(O)[C@@H]1CC[C@@](C)(O1)[C@H]2[C@@H](O)C[C@@]3(C)[C@@H]4C[C@H](O[C@@H]5O[C@H](CO)[C@H](O)[C@@H](O)[C@H]5O)[C@H]6C(C)(C)[C@H](CC[C@@]67C[C@@]47CC[C@]23C)O[C@@H]8OC[C@@H](O)[C@H](O)[C@H]8O QMNWISYXSJWHRY-BCBPIKMJSA-N 0.000 description 1
- 230000002238 attenuated effect Effects 0.000 description 1
- 229940097012 bacillus thuringiensis Drugs 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 235000012343 cottonseed oil Nutrition 0.000 description 1
- PFKIBRPYVNVMRU-UHFFFAOYSA-N cyclosieversioside F Natural products CC(C)(O)C1COC(C)(C1)C2C(O)CC3(C)C4CC(OC5OC(CO)C(O)C(O)C5O)C6C(C)(C)C(CCC67CC47CCC23C)OC8OCC(O)C(O)C8O PFKIBRPYVNVMRU-UHFFFAOYSA-N 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000002481 ethanol extraction Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 235000013402 health food Nutrition 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000000366 juvenile effect Effects 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 230000004089 microcirculation Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000001114 myogenic effect Effects 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000000242 pagocytic effect Effects 0.000 description 1
- 230000000737 periodic effect Effects 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000002787 reinforcement Effects 0.000 description 1
- 239000011435 rock Substances 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- PJANXHGTPQOBST-UHFFFAOYSA-N stilbene Chemical compound C=1C=CC=CC=1C=CC1=CC=CC=C1 PJANXHGTPQOBST-UHFFFAOYSA-N 0.000 description 1
- 235000021286 stilbenes Nutrition 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 1
- 150000003648 triterpenes Chemical class 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 230000003519 ventilatory effect Effects 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
- Y02A40/818—Alternative feeds for fish, e.g. in aquacultures
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Polymers & Plastics (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Animal Husbandry (AREA)
- Biochemistry (AREA)
- Food Science & Technology (AREA)
- Microbiology (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Mycology (AREA)
- Botany (AREA)
- Birds (AREA)
- Marine Sciences & Fisheries (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Insects & Arthropods (AREA)
- Sustainable Development (AREA)
- Molecular Biology (AREA)
- Physiology (AREA)
- Medicines Containing Plant Substances (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to a ganoderma lucidum-decoction dreg bidirectional solid fermentation method utilizing air pressure pulsation and application thereof, and the pressure pulsation solid fermentation is applied to ganoderma lucidum-decoction dreg bidirectional solid fermentation. Compared with the existing solid state fermentation technology of ganoderma lucidum, the invention has the advantages of uniform fermentation environment, sufficient oxygen supply, heat and CO2The discharge is rapid; the ganoderma lucidum grows rapidly and vigorously, the utilization rate of raw materials is high, and the yield of active substances is high; low energy consumption and no contamination to bacteria. The fermentation technology of the invention has simple technological process and condition control, is easy for large-scale industrial production,the method not only can change the waste of the Chinese medicine residues into valuable, reduce the application of antibiotics in the breeding industry, but also can solve the problem of environmental pollution caused by the Chinese medicine residues, and has good application prospect.
Description
Technical field
The two-way solid fermentation method of ganoderma lucidum-dregs of a decoction and the application that the present invention relates to the use of air pressure pulsation, belong to micro- life
Object fermentation technical field.
Background technique
Ganoderma Lucidum is that a kind of nutrition and health care is worth high macro-fungi, have improve immunity of organisms, it is anti-oxidant,
Macrophages phagocytic capacity and the multiple efficacies such as antitumor are improved, is incorporated into Chinese Pharmacopoeia already, is used for health food development.Spirit
Sesame polysaccharide and ganodenic acid are most important active materials in ganoderma lucidum.Radix Astragali is leguminous mongolian scutellaria, Astragalus membranacus or more
The dry root of sequence rock Radix Astragali has QI invigorating qi-restoratives, toxin expelling myogenic and other effects, and market demand is huge, the Huang that the whole nation generates every year
Up to ten thousand tons of 4-6 of the stilbene dregs of a decoction.Many studies have shown that still containing various active substance, such as Astragaloside IV and Huang in astragalus root dregs
Recovery rate of the astragalus polysaccharides in alcohol extracting medicinal material only accounts for the 72.1% and 52.6% of Milkvetch Root.Radix Salviae Miltiorrhizae is Lamiaceae plant Radix Salviae Miltiorrhizae
Dry root and rhizome have activating microcirculation and removing stasis medicinal, cool blood to disappear carbuncle and other effects, and tanshinone is the main active substances in Radix Salviae Miltiorrhizae, at present second
Alcohol extracting efficiency is lower than 10%, therefore, still containing the active constituent of abundant residues in the Radix Salviae Miltiorrhizae dregs of a decoction.Currently, China's Chinese medicine slag is big
It is dropped, this not only causes the wasting of resources, also creates serious environmental pollution more.
Bi-directional fermentation technology be by medicinal fungi (ganoderma lucidum etc.) as fermenting microbe and drug matrices (have certain activity at
The Chinese medicine or the dregs of a decoction divided) fementative composition is constituted, on the one hand change the pharmacological property ingredient of matrix, another party by fungi growth metabolism
Cucumber in the drug matrices of face also can growth to fungi and metabolism have an impact, there is amphicheirality, thus to crude drug/
The dregs of a decoction play synergy, expand the effect used and be attenuated.Bi-directional fermentation is the current most promising approach of dregs of a decoction resource utilization.
Solid fermentation is the common method of microbial fermentation, compared with liquid fermentation, has water-saving, energy saving, fermentation process
Advantages, the filamentous fungi such as Ganoderma Lucidums etc. such as rare waste material generates, and environmental pollution is small are particularly suitable for solid fermentation.Ganoderma lucidum at present
The solid fermentation of bacterium uses the methods of tray formula, column type, stirring-type and drum-type substantially.However, static culture there are heat,
Oxygen transmitting is difficult, causes content temperature, humidity and thalli growth seriously uneven, and raw material availability is low, and uses stirring
Mode then causes energy consumption excessive, destroys mycelium, influences thalli growth metabolism.Air pressure oscillation fermenter is with gas
State, solid medium are static state, pass through a kind of technology of the pressure oscillation regulating and controlling microbial solid state fermentation of gas phase, main advantage
Including periodic pressure oscillation can provide sufficient oxygen to the mycelia inside particle and will be present in inside particle
Metabolic heat and the discharge in time such as dioxy salt dissolving, reach the environment uniformity around thallus on small scale, in addition, pressure oscillation can
So that solid state substrate is kept loose, expanded space for mycelial mass propagation, mycelial growth is vigorous inside and outside the layer that ferments, raw material
Utilization rate is high, and is in honeycomb after fermentation substrate drying, easily crushes.So far, pressure fluctuation is sent out as a kind of reinforcement solid-state
Ferment mass transfer, heat transfer effective means, be applied to bacillus thuringiensis, bacillus subtilis, muscardine, P. aeruginosa
The solid state fermentation of the microorganisms such as bacterium, Penicillium notatum, black-koji mould and Trichoderma, however yet there are no report in Ganoderma Lucidum solid state fermentation
Road.
Summary of the invention
The first object of the present invention is in view of the deficiencies of the prior art, to provide a kind of utilization air pressure pulsation solid hair
Ferment produces ganoderma lucidum-dregs of a decoction bi-directional fermentation mycoplasma method, to promote Ganoderma Lucidum to grow, improves main active substances polysaccharide and triterpene
Yield.
To achieve the above object, the invention adopts the following technical scheme:
A method of ganoderma lucidum-dregs of a decoction bi-directional fermentation mycoplasma being produced using air pressure pulsation solid fermentation, including following
Step:
1) preparation of seed liquor: lucid ganoderma fungus slant strains being inoculated in the cooling liquid seed culture medium that sterilized,
In 25~30 DEG C of 4~8d of culture to get lucid ganoderma fungus seed liquor；According to the needs of inoculation, which can be expanded by fermentor
Big culture is secondary seed solution or three-level seed liquor；
2) preparation of solid fermentation culture medium: solid base and nutrient solution are stirred according to the mass ratio of 1:1~1:3
It is even, it puts into pressure fluctuation solid-state fermentation tank, 30~60min of sterilization treatment is under conditions of 121 DEG C~125 DEG C to get solid
Fermentation medium；
3) inoculated and cultured: cooling water and stirring are opened, when fermentation jar temperature is down to 30 DEG C or less, by lucid ganoderma fungus seed
Liquid presses 3%~10% inoculum concentration cut-in pressure pulsation solid-state fermentation tank of solid fermentation culture medium weight, after mixing evenly,
Under the conditions of 25~30 DEG C, 25~30d is to get Zymocyte for air pressure oscillation fermenter culture；
4) processing of Zymocyte: by Zymocyte, drying to constant weight under the conditions of 45~60 DEG C, smashes it through 40 meshes,
Permeate is final products；
Described, the concrete composition of liquid seed culture medium are as follows: glucose 30g/L, peptone 5g/L, potassium dihydrogen phosphate
1.5g/L, magnesium sulfate 1g/L, surplus are water, and initial pH is natural；
In the step 2), solid fermentation culture medium is made of wheat bran, the dregs of a decoction and cotton seed hulls, and the dregs of a decoction are Radix Astragali
The mass ratio of one or two kinds of mixing of the dregs of a decoction and the Radix Salviae Miltiorrhizae dregs of a decoction, the wheat bran and the dregs of a decoction is 5:1~1:1；The cotton
The quality of seed shell is the 1%~5% of wheat bran and dregs of a decoction gross mass；
In the step 2), the composition of nutrient solution are as follows: 10~30g/L of glucose, 1~10g/L of yeast extract, remaining is water；
In the step 3), when air pressure oscillation fermenter culture, ventilatory capacity is 1~5L/min, the air impulsive motion
It is atmospherically limited to 0.15~0.3MPa, the air pressure lower limit of the air pressure pulsation is 0.01~0.05MPa；
In the step 3), when air pressure oscillation fermenter culture 1-14d, the pressure fluctuation period is 8~16h, paddy pressure dimension
Hold 4~8h of time, peak pressure is held time 4~8h, when pressure fluctuation solid fermentation culture 15-25d or 15-30d, pressure fluctuation week
Phase is 12~for 24 hours, paddy pressure is held time 6~12h, and peak pressure is held time 6~12h.
The second object of the present invention is to provide the Zymocyte of above method preparation.The fermentation of method preparation of the invention
The yield of main active substances-polysaccharide and triterpene compound is significantly improved than common static solid fermentation in mycoplasma, wherein more
Candy output can be improved 55~75%, and triterpene compound output increased can 30~50%.
The third object of the present invention is to provide application of the above-mentioned fermentation homogeneous in the functional additive of feed.This hair
The fermentation homogeneous of bright method preparation is used for the functional additive of feed, can promote the growth of shrimp crab, significantly improves the survival of shrimp crab
Rate.
Preferably, additive amount is the 2% of total feed quality when functional additive as feed.
Beneficial effects of the present invention:
1) pressure fluctuation allows the cross-ventilation in base-material gap to spread, and increases the uniformity of humidity and temperature in base-material, promotes
Carbon dioxide and heat discharge generate favorable influence to the growth metabolism of fermenting microbe；
2) during the pressure relief process, because of unexpected Quick air-discharge, solid particle has been loosened, has been expanded for the mass propagation of Ganoderma Lucidum
Space, ferment the bed of material inside and outside thalli growth it is vigorous, improve raw material availability；
3) compared with common static solid fermentation, the addition of cotton seed hulls increases the stream of gas in pressure fluctuation fermentation process
It is dynamic, promote thalli growth and metabolism；
4) Ganoderma Lucidum growth and breeding speed is much higher than common static solid fermentation process in the present invention, and final biomass is about
5-8 times of common static solid state fermentation；
5) yield of main active substances-polysaccharide and triterpene compound is more solid than common static in Zymocyte in the present invention
Body fermentation significantly improves, wherein polysaccharide yield improves 55~75%, triterpene compound output increased 30~50%；
6) zymotechnique of the invention is easy to operate, and condition is easily controllable, is easy to high-volume the factorial production, has good
Application prospect；
7) ganoderma lucidum of the invention-dregs of a decoction mycoplasma can be used as feed addictive and cultivate for crab and Macrobrachium rosenbergii, improve it
Survival rate, but it is not limited to the cultivation of shrimp crab, in the cultivation for applying also for fish and livestock and poultry, extremely agree with big health under the new situation
Industry demand.
Specific embodiment
The present invention and its advantages are described in further detail With reference to embodiment, still, the present invention
Specific embodiment be not limited thereto.
In following embodiments, the measurement of polysaccharide, the extracting method of triterpene compound, ganoderma lucidum bacteria biomass in Zymocyte
Method is specific as follows:
The extraction of ganoderma lucidum-dregs of a decoction Zymocyte polysaccharide: Zymocyte breaks into fritter, and 60 DEG C of drying smash it through 40 meshes,
Obtain Zymocyte sample.Boil water extraction method is taken, Zymocyte sample 5g is accurately weighed, adds water 100mL, boiling waterbath
2h, then supernatant extracting solution is removed in centrifugation, adds 75mL water, and boiling water bath continues to extract 2h, and extracting solution is removed in centrifugation.Merging mentions twice
Liquid is taken, it is 50mL that rotary evaporation, which is concentrated into liquor capacity, and concentrate, which is cooled to room temperature, is slowly added to ethyl alcohol 200mL, and constantly stirs
It mixes, stands overnight, be centrifuged, sediment 10mL distilled water dissolves, and lysate is added in 3500Da bag filter, dialyses in distilled water
Overnight, dialyzate rotary evaporation is concentrated into 10mL, and ethyl alcohol 40mL is added, stands overnight, and is centrifuged, 60 DEG C of dryings of sediment, obtains
Ganoderma lucidum-dregs of a decoction Zymocyte polysaccharide, wherein polyoses content is measured using Phenol sulfuric acid procedure.
The extraction of triterpene compound in ganoderma lucidum-dregs of a decoction Zymocyte: Zymocyte breaks into fritter, and 60 DEG C of drying crush
40 meshes are crossed afterwards, obtain Zymocyte sample.Ethanol extraction method is taken, Zymocyte sample 10g is accurately weighed, 150mL is added
1.5h is extracted in ethyl alcohol, 75 DEG C of water-baths, and supernatant is removed in centrifugation, adds 150mL ethyl alcohol, and 75 DEG C of water-baths continue to extract 1.5h, and centrifugation is fallen
Remove extracting solution.The dissolution of 100mL chloroform is added in combined extract, rotary evaporated to dryness.60mL saturated sodium bicarbonate solution extraction is added
It takes, takes upper layer of extraction liquid, using salt acid for adjusting pH to 2.5, the extraction of 50mL chloroform is added, takes chloroform layer rotary evaporated to dryness, obtains
Ganoderma lucidum-dregs of a decoction Zymocyte triterpene compound is obtained, wherein triterpenoid content utilizes spectrophotometry.
The measurement of ganoderma lucidum bacteria biomass in ganoderma lucidum-dregs of a decoction Zymocyte: Zymocyte breaks into fritter, and 60 DEG C of drying crush
40 meshes are crossed afterwards, obtain Zymocyte sample.0.25g Zymocyte sample is taken, 5% solution of trichloroacetic acid of 25mL is added, in
25min is extracted in 80 DEG C of water-baths, then ice bath is cooling, is then centrifuged for, supernatant measures OD value at 260nm, with pure thallus bacterium
Amount and nucleic acid ultraviolet absorption curve, using unleavened solid state substrate as blank control, are converted into ganoderma lucidum-dregs of a decoction as reference
The biomass of Ganoderma Lucidum in Zymocyte.
Ganoderma Lucidum used in embodiment is purchased from Chinese agriculture microbial strains preservation administrative center ACCC, deposit number
ACCC50247。
Embodiment 1
Ganoderma lucidum-dregs of a decoction mycoplasma method is produced using air pressure pulsation solid fermentation the present embodiment provides a kind of, including
Following steps:
1) preparation of seed liquor: lucid ganoderma fungus slant strains being inoculated in the cooling liquid seed culture medium that sterilized,
In 28 DEG C of culture 4d to get lucid ganoderma fungus seed liquor；Wherein, the composition of liquid seeds liquid culture medium are as follows: glucose 30g/L, egg
White peptone 5g/L, potassium dihydrogen phosphate 1.5g/L, magnesium sulfate 1g/L, surplus are water, and initial pH is natural；
2) preparation of solid fermentation culture medium: using wheat bran 400g and astragalus root dregs 100g as solid base, cotton is separately added
Seed shell 5g is added 1.5L liquid nutrient solution, stirs evenly to get solid fermentation culture medium, wherein the composition of liquid nutrient solution
Are as follows: glucose 15g/L, yeast extract 1g/L, remaining is water.Solid fermentation culture medium is put into pressure fluctuation solid-state fermentation tank, is led to
Sterilize 40min under conditions of entering 121 DEG C of high-temperature vapor；
3) inoculated and cultured: opening cooling water and stirring, when the tank temperature of pressure fluctuation solid-state fermentation tank is down to 30 DEG C or less,
By Ganoderma Lucidum seed liquor by the 5% inoculum concentration cut-in pressure pulsation solid-state fermentation tank of solid fermentation culture medium weight, in ventilation quantity
Under conditions of 1L/min and tank temperature of fermenting are 28 DEG C, pressure fluctuation stimulation culture 14d, during being somebody's turn to do, air pressure pulsation are carried out
Air pressure upper limit value be 0.25MPa, the air pressure lower limit value of air impulsive motion pressure is 0.01MPa, and the pressure fluctuation period is 8h, wherein
Peak pressure is held time 4h, and paddy pressure is held time 4h.The variable pressure pulsation period is 12h later, and wherein peak pressure is held time 6h, paddy
The 6h that holds time is pressed, continues to cultivate 14d with this condition to get Zymocyte.
4) processing of Zymocyte: by Zymocyte, drying to constant weight under the conditions of 45 DEG C, smashes it through 40 meshes, filtration
Object is final products.
Routinely static fermentation technique using identical solid fermentation culture medium, but does not add cotton seed hulls, carries out Ganoderma Lucidum
Fermented and cultured prepares Ganoderma Lucidum-Chinese medicine slag Zymocyte, compares air pressure pulsation solid fermentation and common static fermentation to spirit
The growth of sesame bacterium and the influence of main active substances are fermented through examining compared to common static, using air pressure pulsation solid hair
Ferment technology, biomass improve 6.4 times, the output increased of main active substances polysaccharide 81.8%, triterpene compound yield
Improve 52.5% (table 1)
Under 1 different fermentations mode of table, the biomass of the two-way solid fermentation of Ganoderma Lucidum-dregs of a decoction and the yield of main active substances
Embodiment 2
Ganoderma lucidum-dregs of a decoction mycoplasma method is produced using air pressure pulsation solid fermentation the present embodiment provides a kind of, including
Following steps:
1) preparation of seed liquor: lucid ganoderma fungus slant strains being inoculated in the cooling liquid seed culture medium that sterilized,
In 28 DEG C of culture 4d to get lucid ganoderma fungus seed liquor；Wherein, the composition of liquid seeds liquid culture medium are as follows: glucose 30g/L, egg
White peptone 5g/L, potassium dihydrogen phosphate 1.5g/L, magnesium sulfate 1g/L, surplus are water, and initial pH is natural；
2) using wheat bran 500g and Radix Salviae Miltiorrhizae dregs of a decoction 100g as solid base, cotton separately the preparation of solid fermentation culture medium: is added
Seed shell 18g is added 600mL liquid nutrient solution, stirs evenly to get solid fermentation culture medium, wherein the composition of liquid nutrient solution
Are as follows: glucose 20g/L, yeast extract 8g/L, remaining is water.Solid fermentation culture medium is put into pressure fluctuation solid-state fermentation tank, is led to
Sterilize 40min under conditions of entering 121 DEG C of high-temperature vapor；
3) inoculated and cultured: opening cooling water and stirring, when the tank temperature of pressure fluctuation solid-state fermentation tank is down to 30 DEG C or less,
By Ganoderma Lucidum seed liquor by the 3% inoculum concentration cut-in pressure pulsation solid-state fermentation tank of solid fermentation culture medium weight, in ventilation quantity
Under conditions of 3L/min and tank temperature of fermenting are 28 DEG C, pressure fluctuation stimulation culture 14d, during being somebody's turn to do, air pressure pulsation are carried out
Air pressure upper limit value be 0.15MPa, the air pressure lower limit value of air impulsive motion pressure is 0.03MPa, and the pressure fluctuation period is 12h,
Middle peak pressure is held time 6h, and paddy pressure is held time 6h.The variable pressure pulsation period is 16h later, and wherein peak pressure is held time 8h,
Paddy pressure is held time 8h, continues to cultivate 16d with this condition to get Zymocyte.
4) processing of Zymocyte: by Zymocyte, drying to constant weight under the conditions of 50 DEG C, smashes it through 40 meshes, filtration
Object is final products.
Routinely static fermentation technique using identical solid fermentation culture medium, but does not add cotton seed hulls, carries out Ganoderma Lucidum
Fermented and cultured prepares Ganoderma Lucidum-Chinese medicine slag Zymocyte, compares air pressure pulsation solid fermentation and common static fermentation to spirit
The growth of sesame bacterium and the influence of main active substances are fermented through examining compared to common static, using air pressure pulsation solid hair
Ferment technology, biomass improve 5 times, and the output increased of main active substances polysaccharide 55.4%, triterpene compound yield mentions
High 43% (table 2)
Under 2 different fermentations mode of table, the biomass of the two-way solid fermentation of Ganoderma Lucidum-dregs of a decoction and the yield of main active substances
Embodiment 3
Ganoderma lucidum-dregs of a decoction mycoplasma method is produced using air pressure pulsation solid fermentation the present embodiment provides a kind of, including
Following steps:
1) preparation of seed liquor: lucid ganoderma fungus slant strains being inoculated in the cooling liquid seed culture medium that sterilized,
In 28 DEG C of culture 4d to get lucid ganoderma fungus seed liquor；Wherein, the composition of liquid seeds liquid culture medium are as follows: glucose 10g/L, egg
White peptone 10g/L, potassium dihydrogen phosphate 1.5g/L, magnesium sulfate 1g/L, surplus are water, and initial pH is natural；
2) preparation of solid fermentation culture medium: using wheat bran 400g and astragalus root dregs 200g and Radix Salviae Miltiorrhizae dregs of a decoction 200g as solid
Base-material, it is another that cotton seed hulls 40g is added, 1L liquid nutrient solution is added, stirs evenly to get solid fermentation culture medium, wherein liquid battalion
The composition of nutrient solution are as follows: glucose 15g/L, yeast extract 10g/L, remaining is water.The investment pressure fluctuation of solid fermentation culture medium is consolidated
Body fermentor, sterilize 40min under conditions of being passed through 121 DEG C of high-temperature vapor；
3) inoculated and cultured: opening cooling water and stirring, when the tank temperature of pressure fluctuation solid-state fermentation tank is down to 30 DEG C or less,
By Ganoderma Lucidum seed liquor by the 10% inoculum concentration cut-in pressure pulsation solid-state fermentation tank of solid fermentation culture medium weight, in ventilation quantity
Under conditions of 5L/min and tank temperature of fermenting are 28 DEG C, pressure fluctuation stimulation culture 14d, during being somebody's turn to do, air pressure pulsation are carried out
Air pressure upper limit value be 0.3MPa, the air pressure lower limit value of air impulsive motion pressure is 0.05MPa, and the pressure fluctuation period is 16h, wherein
Peak pressure is held time 8h, and paddy pressure is held time 8h.The variable pressure pulsation period is that for 24 hours, wherein peak pressure is held time 12h later,
Paddy pressure is held time 12h, continues to cultivate 16d with this condition to get Zymocyte.
4) processing of Zymocyte: by Zymocyte, drying to constant weight under the conditions of 60 DEG C, smashes it through 40 meshes, filtration
Object is final products.
Routinely static fermentation technique using identical solid fermentation culture medium, but does not add cotton seed hulls, carries out Ganoderma Lucidum
Fermented and cultured prepares Ganoderma Lucidum-Chinese medicine slag Zymocyte, compares air pressure pulsation solid fermentation and common static fermentation to spirit
The growth of sesame bacterium and the influence of main active substances are fermented through examining compared to common static, using air pressure pulsation solid hair
Ferment technology, biomass improve 5.2 times, the output increased of main active substances polysaccharide 65.4%, triterpene compound yield
Improve 53.9% (table 3)
Under 3 different fermentations mode of table, the biomass of the two-way solid fermentation of Ganoderma Lucidum-dregs of a decoction and the yield of main active substances
Embodiment 4
Ganoderma lucidum-dregs of a decoction mycoplasma method is produced using air pressure pulsation solid fermentation the present embodiment provides a kind of, including
Following steps:
1) preparation of seed liquor: lucid ganoderma fungus slant strains being inoculated in the cooling liquid seed culture medium that sterilized,
In 28 DEG C of culture 4d to get lucid ganoderma fungus seed liquor；Wherein, the composition of liquid seeds liquid culture medium are as follows: glucose 10g/L, egg
White peptone 10g/L, potassium dihydrogen phosphate 1.5g/L, magnesium sulfate 1g/L, surplus are water, and initial pH is natural；
2) preparation of solid fermentation culture medium: using wheat bran 400g and astragalus root dregs 200g and Radix Salviae Miltiorrhizae dregs of a decoction 100g as solid
Base-material, it is another that cotton seed hulls 35g is added, 800mL liquid nutrient solution is added, stirs evenly to get solid fermentation culture medium, wherein liquid
The composition of body nutrient solution are as follows: glucose 15g/L, yeast extract 10g/L, remaining is water.Solid fermentation culture medium is put into pressure arteries and veins
Dynamic solid-state fermentation tank, sterilize 40min under conditions of being passed through 121 DEG C of high-temperature vapor；
3) inoculated and cultured: opening cooling water and stirring, when the tank temperature of pressure fluctuation solid-state fermentation tank is down to 30 DEG C or less,
By Ganoderma Lucidum seed liquor by the 10% inoculum concentration cut-in pressure pulsation solid-state fermentation tank of solid fermentation culture medium weight, in ventilation quantity
Under conditions of 4L/min and tank temperature of fermenting are 28 DEG C, pressure fluctuation stimulation culture 14d, during being somebody's turn to do, air pressure pulsation are carried out
Air pressure upper limit value be 0.3MPa, the air pressure lower limit value of air impulsive motion pressure is 0.03MPa, and the pressure fluctuation period is 12h, wherein
Peak pressure is held time 6h, and paddy pressure is held time 6h.The variable pressure pulsation period is 16h later, and wherein peak pressure is held time 8h, paddy
The 8h that holds time is pressed, continues to cultivate 16d with this condition to get Zymocyte.
4) processing of Zymocyte: by Zymocyte, drying to constant weight under the conditions of 60 DEG C, smashes it through 40 meshes, filtration
Object is final products.
By above-mentioned identical pressure fluctuation fermentation method, using identical solid fermentation culture medium, but cottonseed is not only added
Shell carries out Ganoderma Lucidum fermented and cultured, prepares Ganoderma Lucidum-Chinese medicine slag Zymocyte, compares cotton seed hulls to Ganoderma Lucidum growth and main
The influence of active material, through examining, compared to the fermentation for being not added with cotton seed hulls, after adding cotton seed hulls, biomass improves 1.3
Times, the output increased of main active substances polysaccharide 34.7%, 26.5% (table 4) of triterpene compound output increased
Under 4 different fermentations mode of table, the biomass of the two-way solid fermentation of Ganoderma Lucidum-dregs of a decoction and the yield of main active substances
The raising of 5 Macrobrachium rosenbergii of embodiment
The Macrobrachium rosenbergii for selecting health, specification, weight almost the same, first weight are (0.25 ± 0.03), are tested by feeding
Feed difference is divided into 3 groups, respectively control group, experimental group 1 and experimental group 2.Every group of 3 repetitions, each repetition 30.Control group
Normal shrimp feed is fed, experimental group 1 is Ganoderma Lucidum-dregs of a decoction common static solid fermentation bacterium of the addition 2% in normal shrimp feed
Matter, experimental group 2 are Ganoderma Lucidum-dregs of a decoction air pressure pulsation solid fermentation bacterium that in embodiment 3 2% is added in normal shrimp feed
Matter, incubation do not add any antibiotic, and method for breeding and management routinely carry out, and 35d is fed in experiment.The results show that with
Control group is compared (table 5) with addition common static solid fermentation mycoplasma, and mycoplasma produced by the invention can be shown as feed addictive
It writes and promotes Macrobrachium rosenbergii growth, improve survival rate.
The influence that 5 Zymocyte of table grows Macrobrachium rosenbergii
| Group | Average initial weight/g | Average weight/g eventually | Survival rate/% |
| Control group | 0.27±0.03 | 0.91±0.04 | 61.6 |
| Experimental group 1 | 0.23±0.02 | 1.01±0.04* | 77.9* |
| Experimental group 2 | 0.25±0.02 | 1.11±0.03* | 83.2* |
Note: compared with the control group, " * " indicates p < 0.05, significant difference.
The raising of 6 Eriocheir sinensis of embodiment
The Eriocheir sinensia juvenile crab for selecting health, specification, weight almost the same, first weight are (16.7 ± 0.1), and experiment is by throwing
Feeding material difference is divided into 3 groups, respectively control group, experimental group 1 and experimental group 2.Every group of 3 repetitions, each repetition 20.Control
Group feeds normal crab feed, and experimental group 1 is Ganoderma Lucidum-dregs of a decoction common static solid fermentation of the addition 2% in normal crab feed
Mycoplasma, experimental group 2 are Ganoderma Lucidum-dregs of a decoction air pressure pulsation solid fermentations that in embodiment 3 2% is added in normal crab feed
Mycoplasma, incubation do not add any antibiotic, and method for breeding and management routinely carry out, and 60d is fed in experiment.The results show that
Compared with control group and addition common static solid fermentation mycoplasma (table 6), mycoplasma produced by the invention is as feed addictive energy
The growth of Eriocheir sinensis is remarkably promoted, survival rate is improved.
The influence that 6 Zymocyte of table grows Eriocheir sinensis
| Group | Average initial weight/g | Average weight/g eventually | Survival rate/% |
| Control group | 16.7±0.1 | 42.6±0.04 | 42.1 |
| Experimental group 1 | 16.2±0.1 | 61.7±0.04* | 66.2* |
| Experimental group 2 | 17.5±0.1 | 65.9±0.03* | 70.3* |
Note: compared with the control group, " * " indicates p < 0.05, significant difference.
The foregoing is merely part specific embodiments of the invention, and all those skilled in the art are on basis of the invention
Upper made any conspicuous improvement, replacement or modification all belong to the scope of protection of the present invention.
Claims (10)
1. a kind of two-way solid fermentation method of ganoderma lucidum-dregs of a decoction pulsed using air pressure, which is characterized in that including following step
It is rapid:
1) preparation of seed liquor: lucid ganoderma fungus slant strains being inoculated in the cooling liquid seed culture medium that sterilized, culture
Obtain seed liquor；
2) preparation of solid fermentation culture medium: solid base and nutrient solution are stirred evenly according to the mass volume ratio of 1:1 ~ 1:3,
It puts into pressure fluctuation solid-state fermentation tank, to get solid fermentation culture medium after high-temperature sterilization processing；
The solid base ingredient is wheat bran, the dregs of a decoction and cotton seed hulls；The nutrient solution by glucose, yeast extract and water mix and
At；
3) when fermentation jar temperature is down to 30 DEG C or less, lucid ganoderma fungus seed liquor pressure fluctuation fermented and cultured: is pressed into solid fermentation
The inoculum concentration cut-in pressure pulsation solid-state fermentation tank of the 3% ~ 10% of culture medium weight, after mixing evenly, under the conditions of 25 ~ 30 DEG C,
Air pressure oscillation fermenter 25 ~ 30d of culture is obtained comprising the mixture including mycelia and culture substrate ingredient, i.e. Zymocyte；
4) processing of Zymocyte: by Zymocyte, drying to constant weight under the conditions of 45 ~ 60 DEG C, smashes it through 40 meshes, filtration
Object is final products.
2. the method according to claim 1, wherein the liquid seed culture medium ingredient be glucose 30g/L,
Peptone 5g/L, potassium dihydrogen phosphate 1.5g/L, magnesium sulfate 1g/L, surplus are water, and initial pH is natural；Seed liquor prepares condition of culture
Are as follows: 25 ~ 30 DEG C, cultivate 4-8d.
3. the method according to claim 1, wherein the dregs of a decoction are one of astragalus root dregs and the Radix Salviae Miltiorrhizae dregs of a decoction
Or two kinds of mixing.
4. the method according to claim 1, wherein the mass ratio of the wheat bran and the dregs of a decoction is 5:1 ~ 1:1.
5. the method according to claim 1, wherein the quality of the cotton seed hulls is wheat bran and dregs of a decoction gross mass
1%~5%。
6. the method according to claim 1, wherein the composition of the nutrient solution are as follows: 10 ~ 30g/L of glucose, ferment
Female 1 ~ 10g/L of cream, remaining is water.
7. the method according to claim 1, wherein in the step 3), when pressure fluctuation fermented and cultured, ventilation
Amount is 1 ~ 5L/min, and air impulsive motion is atmospherically limited to 0.15 ~ 0.3MPa, the air pressure lower limit of air pressure pulsation is 0.01 ~
0.05MPa。
8. the method according to claim 1, wherein in the step 3), pressure fluctuation fermented and cultured 1-14d
When, the pressure fluctuation period is 8 ~ 16h, and paddy pressure is held time 4 ~ 8h, and peak pressure is held time 4 ~ 8h, pressure fluctuation solid fermentation culture
When 15-25d or 15-30d, the pressure fluctuation period is 12 ~ for 24 hours, paddy pressure is held time 6 ~ 12h, and peak pressure is held time 6 ~ 12h.
9. the Zymocyte product of any one of claim 1-8 the method preparation.
10. application of the Zymocyte product in the functional additive of feed described in claim 9.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910470543.2A CN110218656B (en) | 2019-05-31 | 2019-05-31 | Ganoderma lucidum-herb residue bidirectional solid fermentation method utilizing air pressure pulsation and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910470543.2A CN110218656B (en) | 2019-05-31 | 2019-05-31 | Ganoderma lucidum-herb residue bidirectional solid fermentation method utilizing air pressure pulsation and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110218656A true CN110218656A (en) | 2019-09-10 |
| CN110218656B CN110218656B (en) | 2022-05-17 |
Family
ID=67819262
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910470543.2A Active CN110218656B (en) | 2019-05-31 | 2019-05-31 | Ganoderma lucidum-herb residue bidirectional solid fermentation method utilizing air pressure pulsation and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110218656B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111034862A (en) * | 2020-01-10 | 2020-04-21 | 曹晓斌 | Production method for producing high-protein feed by deep fermentation of straws |
| CN112586604A (en) * | 2020-08-12 | 2021-04-02 | 淮阴师范学院 | Method and device for preparing feed additive through ganoderma-astragalus bidirectional fermentation based on pressure pulsation technology |
| CN114084967A (en) * | 2021-11-12 | 2022-02-25 | 河南省科学院生物研究所有限责任公司 | Ganoderma lucidum fermented Shuanghuanglian mushroom residue for repairing water environment polluted by polycyclic aromatic hydrocarbon benzo [ a ] pyrene and application thereof |
Citations (32)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030138943A1 (en) * | 2002-01-22 | 2003-07-24 | Hongzhang Chen | Gas dual-dynamic solid state fermentation technique and apparatus |
| CN101376669A (en) * | 2008-10-07 | 2009-03-04 | 南京晓庄学院 | Preparation of 6-O-beta-D- glucosyl-3,6,16,25-tetrahydroxy cycloartane |
| CN101455274A (en) * | 2008-12-29 | 2009-06-17 | 江南大学 | Method for producing feedstuff containing ganoderma lucidum active ingredient and feedstuff using ganoderma lucidum liquid fermentation liquid |
| CN101696430A (en) * | 2009-11-05 | 2010-04-21 | 天津北洋百川生物技术有限公司 | Method for producing beta-polymalic acid by inert support adsorption solid-state fermentation |
| CN101856373A (en) * | 2009-04-10 | 2010-10-13 | 德阳市食用菌专家大院 | Zymocyte mixture containing lucid ganoderma, preparation method and application thereof |
| CN101869303A (en) * | 2009-04-27 | 2010-10-27 | 李全才 | Method for producing biological active calcium |
| CN102187944A (en) * | 2011-06-22 | 2011-09-21 | 湖南农业大学 | Solid fermentation method for Chinese herbal medicament feed additive |
| CN102503647A (en) * | 2011-11-08 | 2012-06-20 | 肖兵南 | Solid fermentation method of lactarius deliciosus mycelium and application of solid fermentation extractive thereof |
| CN103565944A (en) * | 2013-11-20 | 2014-02-12 | 青岛农业大学 | Bi-directional fermentation technology for traditional Chinese medicine |
| CN103908489A (en) * | 2014-03-11 | 2014-07-09 | 中国科学院长春应用化学研究所 | Bidirectional solid fermentation method of ginseng-radix astragali-lucid ganoderma medical mycoplasm |
| CN103948023A (en) * | 2014-04-04 | 2014-07-30 | 浙江省医学科学院 | Health foods for enhancing immunity and improving sleep and two-step fermentation preparation method thereof |
| CN104726467A (en) * | 2015-04-08 | 2015-06-24 | 南京工业大学 | Nucleotide sequence and application thereof in improving acid resistance of strain |
| CN105193905A (en) * | 2014-06-27 | 2015-12-30 | 甘肃农业大学 | Traditional Chinese medicine processing technology of ganoderma-astragalus membranaceus bidirectional fermentation |
| CN105287292A (en) * | 2015-11-18 | 2016-02-03 | 陈洁娣 | Whitening moisture-retention face cream and preparation method thereof |
| CN105602863A (en) * | 2015-12-14 | 2016-05-25 | 南京工业大学 | Bacillus subtilis strain for high yield of lipopeptide antibiotic and poly-gamma-glutamic acid |
| CN106179678A (en) * | 2016-07-27 | 2016-12-07 | 浙江省磐安县外贸药业有限公司 | A kind of drum-type Ganoderma spore powder pulverizer with bactericidal action |
| CN106491663A (en) * | 2016-12-14 | 2017-03-15 | 四川岚晟生物科技有限公司 | A kind of Phellinus igniarius (L. ex Fr.) Quel. LINGZHI JIAONANG and preparation method thereof |
| CN106820134A (en) * | 2017-02-21 | 2017-06-13 | 南京晓庄学院 | A kind of preparation method and purposes of the immunopotentiator based on sesame stilbene mycoplasma |
| CN107022581A (en) * | 2017-05-22 | 2017-08-08 | 东莞理工学院 | A kind of method that utilization air pressure pulsation solid fermentation produces γ polyglutamic acids |
| CN107549449A (en) * | 2017-10-20 | 2018-01-09 | 遵义医学院 | A kind of astragalus root dregs biological feed additive and preparation method and application |
| CN108864319A (en) * | 2018-06-29 | 2018-11-23 | 江苏农林职业技术学院 | The method and matrix used of ganoderma lucidum solid state fermentation production active polysaccharide |
| CN109429900A (en) * | 2018-12-14 | 2019-03-08 | 四川省中医药科学院 | A kind of Wildmimic cultivation method of ganoderma lucidum |
| US10617697B2 (en) * | 2015-01-18 | 2020-04-14 | Gavish-Galilee Bio Applications, Ltd. | Extracts of edible fungi enriched in vitamin D and compostions thereof and their use in the treatment of immune-related disorders |
| US10687482B2 (en) * | 2016-07-14 | 2020-06-23 | Mycoworks, Inc. | Method of producing fungal materials and objects made therefrom |
| US10813960B2 (en) * | 2000-10-04 | 2020-10-27 | Paul Edward Stamets | Integrative fungal solutions for protecting bees and overcoming colony collapse disorder (CCD) |
| CN111991479A (en) * | 2019-05-27 | 2020-11-27 | 东营职业学院 | Bidirectional solid fermentation method of radix isatidis and folium isatidis dregs |
| CN112042469A (en) * | 2020-08-31 | 2020-12-08 | 江苏海昇药业有限公司 | Edible fungus fermentation culture medium containing traditional Chinese medicine residues |
| CN112219638A (en) * | 2020-10-20 | 2021-01-15 | 玉林市益康菌业种植有限公司 | Ganoderma lucidum wild-imitating cultivation method |
| CN112586604A (en) * | 2020-08-12 | 2021-04-02 | 淮阴师范学院 | Method and device for preparing feed additive through ganoderma-astragalus bidirectional fermentation based on pressure pulsation technology |
| CN112662566A (en) * | 2020-12-15 | 2021-04-16 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Ganoderma lucidum spore-less variety with high yield of polysaccharide and artificial cultivation method thereof |
| CN113144023A (en) * | 2021-04-27 | 2021-07-23 | 黄润泽 | Traditional Chinese medicine composition and preparation for improving vascular elasticity and preparation method thereof |
| CN113575754A (en) * | 2021-08-02 | 2021-11-02 | 淮阴师范学院 | Method for preparing functional feed additive by solid state fermentation method |
-
2019
- 2019-05-31 CN CN201910470543.2A patent/CN110218656B/en active Active
Patent Citations (32)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US10813960B2 (en) * | 2000-10-04 | 2020-10-27 | Paul Edward Stamets | Integrative fungal solutions for protecting bees and overcoming colony collapse disorder (CCD) |
| US20030138943A1 (en) * | 2002-01-22 | 2003-07-24 | Hongzhang Chen | Gas dual-dynamic solid state fermentation technique and apparatus |
| CN101376669A (en) * | 2008-10-07 | 2009-03-04 | 南京晓庄学院 | Preparation of 6-O-beta-D- glucosyl-3,6,16,25-tetrahydroxy cycloartane |
| CN101455274A (en) * | 2008-12-29 | 2009-06-17 | 江南大学 | Method for producing feedstuff containing ganoderma lucidum active ingredient and feedstuff using ganoderma lucidum liquid fermentation liquid |
| CN101856373A (en) * | 2009-04-10 | 2010-10-13 | 德阳市食用菌专家大院 | Zymocyte mixture containing lucid ganoderma, preparation method and application thereof |
| CN101869303A (en) * | 2009-04-27 | 2010-10-27 | 李全才 | Method for producing biological active calcium |
| CN101696430A (en) * | 2009-11-05 | 2010-04-21 | 天津北洋百川生物技术有限公司 | Method for producing beta-polymalic acid by inert support adsorption solid-state fermentation |
| CN102187944A (en) * | 2011-06-22 | 2011-09-21 | 湖南农业大学 | Solid fermentation method for Chinese herbal medicament feed additive |
| CN102503647A (en) * | 2011-11-08 | 2012-06-20 | 肖兵南 | Solid fermentation method of lactarius deliciosus mycelium and application of solid fermentation extractive thereof |
| CN103565944A (en) * | 2013-11-20 | 2014-02-12 | 青岛农业大学 | Bi-directional fermentation technology for traditional Chinese medicine |
| CN103908489A (en) * | 2014-03-11 | 2014-07-09 | 中国科学院长春应用化学研究所 | Bidirectional solid fermentation method of ginseng-radix astragali-lucid ganoderma medical mycoplasm |
| CN103948023A (en) * | 2014-04-04 | 2014-07-30 | 浙江省医学科学院 | Health foods for enhancing immunity and improving sleep and two-step fermentation preparation method thereof |
| CN105193905A (en) * | 2014-06-27 | 2015-12-30 | 甘肃农业大学 | Traditional Chinese medicine processing technology of ganoderma-astragalus membranaceus bidirectional fermentation |
| US10617697B2 (en) * | 2015-01-18 | 2020-04-14 | Gavish-Galilee Bio Applications, Ltd. | Extracts of edible fungi enriched in vitamin D and compostions thereof and their use in the treatment of immune-related disorders |
| CN104726467A (en) * | 2015-04-08 | 2015-06-24 | 南京工业大学 | Nucleotide sequence and application thereof in improving acid resistance of strain |
| CN105287292A (en) * | 2015-11-18 | 2016-02-03 | 陈洁娣 | Whitening moisture-retention face cream and preparation method thereof |
| CN105602863A (en) * | 2015-12-14 | 2016-05-25 | 南京工业大学 | Bacillus subtilis strain for high yield of lipopeptide antibiotic and poly-gamma-glutamic acid |
| US10687482B2 (en) * | 2016-07-14 | 2020-06-23 | Mycoworks, Inc. | Method of producing fungal materials and objects made therefrom |
| CN106179678A (en) * | 2016-07-27 | 2016-12-07 | 浙江省磐安县外贸药业有限公司 | A kind of drum-type Ganoderma spore powder pulverizer with bactericidal action |
| CN106491663A (en) * | 2016-12-14 | 2017-03-15 | 四川岚晟生物科技有限公司 | A kind of Phellinus igniarius (L. ex Fr.) Quel. LINGZHI JIAONANG and preparation method thereof |
| CN106820134A (en) * | 2017-02-21 | 2017-06-13 | 南京晓庄学院 | A kind of preparation method and purposes of the immunopotentiator based on sesame stilbene mycoplasma |
| CN107022581A (en) * | 2017-05-22 | 2017-08-08 | 东莞理工学院 | A kind of method that utilization air pressure pulsation solid fermentation produces γ polyglutamic acids |
| CN107549449A (en) * | 2017-10-20 | 2018-01-09 | 遵义医学院 | A kind of astragalus root dregs biological feed additive and preparation method and application |
| CN108864319A (en) * | 2018-06-29 | 2018-11-23 | 江苏农林职业技术学院 | The method and matrix used of ganoderma lucidum solid state fermentation production active polysaccharide |
| CN109429900A (en) * | 2018-12-14 | 2019-03-08 | 四川省中医药科学院 | A kind of Wildmimic cultivation method of ganoderma lucidum |
| CN111991479A (en) * | 2019-05-27 | 2020-11-27 | 东营职业学院 | Bidirectional solid fermentation method of radix isatidis and folium isatidis dregs |
| CN112586604A (en) * | 2020-08-12 | 2021-04-02 | 淮阴师范学院 | Method and device for preparing feed additive through ganoderma-astragalus bidirectional fermentation based on pressure pulsation technology |
| CN112042469A (en) * | 2020-08-31 | 2020-12-08 | 江苏海昇药业有限公司 | Edible fungus fermentation culture medium containing traditional Chinese medicine residues |
| CN112219638A (en) * | 2020-10-20 | 2021-01-15 | 玉林市益康菌业种植有限公司 | Ganoderma lucidum wild-imitating cultivation method |
| CN112662566A (en) * | 2020-12-15 | 2021-04-16 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Ganoderma lucidum spore-less variety with high yield of polysaccharide and artificial cultivation method thereof |
| CN113144023A (en) * | 2021-04-27 | 2021-07-23 | 黄润泽 | Traditional Chinese medicine composition and preparation for improving vascular elasticity and preparation method thereof |
| CN113575754A (en) * | 2021-08-02 | 2021-11-02 | 淮阴师范学院 | Method for preparing functional feed additive by solid state fermentation method |
Non-Patent Citations (11)
| Title |
|---|
| I-CHUAN SHEIH等: ""Purification and Properties of a Novel Phenolic Antioxidant from Radix astragali Fermented by Aspergillus oryzae M29"", 《JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY》 * |
| WEI WEI等: ""Anti-fatigue activity of extract form the submerged fermentation of Ganoderma Lucidum using Radix astragali as substrate"", 《JOURNAL OF ANIMAL & PLANT SCIENCES》 * |
| 何斌: ""灵芝-丹参双向固体发酵工艺的研究"", 《中国优秀博硕士学位论文全文数据库(硕士) 医药卫生科技辑》 * |
| 徐福建等: "纤维素酶气相双动态固态发酵", 《环境科学》 * |
| 李宏强等: "固态发酵的参数周期变化及对微生物发酵的影响", 《生物工程学报》 * |
| 杨宗渠等: "灵芝固态发酵研究进展", 《北方园艺》 * |
| 裴智鹏等: "灵芝-黄芪双向发酵菌质多糖的分离纯化及生物活性研究", 《生物加工过程》 * |
| 裴海生等: "木质素对灵芝菌丝体生长的影响", 《农业工程学报》 * |
| 许晓燕等: "灵丹菌质双向发酵工艺及药效品质研究", 《中国食用菌》 * |
| 谷睿等: "14个灵芝菌株的新型(双向型)固体发酵及菌质多糖含量测定", 《时珍国医国药》 * |
| 陈合等: "中药渣用于培养灵芝菌及功能成分分析", 《农业工程学报》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111034862A (en) * | 2020-01-10 | 2020-04-21 | 曹晓斌 | Production method for producing high-protein feed by deep fermentation of straws |
| CN112586604A (en) * | 2020-08-12 | 2021-04-02 | 淮阴师范学院 | Method and device for preparing feed additive through ganoderma-astragalus bidirectional fermentation based on pressure pulsation technology |
| CN114084967A (en) * | 2021-11-12 | 2022-02-25 | 河南省科学院生物研究所有限责任公司 | Ganoderma lucidum fermented Shuanghuanglian mushroom residue for repairing water environment polluted by polycyclic aromatic hydrocarbon benzo [ a ] pyrene and application thereof |
| CN114084967B (en) * | 2021-11-12 | 2024-01-23 | 河南省科学院生物研究所有限责任公司 | Lucid ganoderma fermented Shuanghuanglian mushroom dreg for repairing polycyclic aromatic hydrocarbon benzo [ a ] pyrene polluted water environment and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110218656B (en) | 2022-05-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101695257B (en) | Solid fermentation method for cordyceps sinensis | |
| CN103333022B (en) | Cultivation material compatibility of agrocybe cylindracea and manufacture method of cultivation material | |
| CN103553834B (en) | Method for preparing tremella cultivation material from tea cattail and tea seed shells | |
| CN110218656A (en) | Ganoderma lucidum-herb residue bidirectional solid fermentation method utilizing air pressure pulsation and application | |
| CN102976801A (en) | Method for producing functional microorganism organic fertilizer by using food residue | |
| CN102154407B (en) | Corayceps militaris polysaccharide two-stage fermentation synthesis process | |
| CN101139562A (en) | Process for preparing rare ginsenoside Compound K by fermenting panax notoginseng saponins with streptomycete | |
| CN108823261A (en) | A kind of Ultra-low molecular weight Dendrobium officinale polysaccharide and its preparation and application | |
| CN1723758A (en) | Cordyceps sinensis mycelium and its prepn. method | |
| CN1994116A (en) | Pure breed fermentation process for producing tasteless preserved soybean | |
| CN102408991B (en) | Bioreactor system for large scale culture of medicinal plants | |
| CN105533707A (en) | Ginseng small molecule nutritional ingredient preparation and preparation method thereof | |
| CN101397542A (en) | Artificial culture method of zinc-enriched Chinese caterpillar fungus mycelia and culture medium thereof | |
| CN103484421A (en) | Pilot scale production method for gliocladium roseum chlamydospore by liquid fermentation | |
| CN101113409A (en) | Method for cultivating antler mythic fungus by using bacterium glass | |
| CN105861327A (en) | Armillaria-mellea repeated segmentation feeding fermentation technology | |
| CN110684672B (en) | Fermentation method of antioxidant cordyceps sobolifera mycelium | |
| CN103289937A (en) | Method for producing bacillus laterosporus live bacteria by high density solid fermentation | |
| CN109429911A (en) | A kind of fermentation cordyceps sinensis culture solution and preparation method thereof prepared with jerusalem artichoke | |
| CN106755179A (en) | A kind of culture medium for being suitable to bacteria cellulose fermentation | |
| CN106434368A (en) | Culture method of ganoderma leucocontextum liquid strain | |
| CN104756767A (en) | High-density pleurotus eryngii liquid strain and fermentation method thereof | |
| CN102219567A (en) | Method for producing biological organic fertilizer by using methane liquid as basic culture medium through fermentation | |
| CN106883010A (en) | Antrodia growth promoter and the method with its quick fermentation production antrodia | |
| CN108796027A (en) | A method of producing carotenoid |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |