CN105925493A - Bamboo parasitic fungus and application thereof in fermentation production of hypocrellin - Google Patents
Bamboo parasitic fungus and application thereof in fermentation production of hypocrellin Download PDFInfo
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- CN105925493A CN105925493A CN201610496366.1A CN201610496366A CN105925493A CN 105925493 A CN105925493 A CN 105925493A CN 201610496366 A CN201610496366 A CN 201610496366A CN 105925493 A CN105925493 A CN 105925493A
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- hypocrellin
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- fermentation
- bamboo parasitic
- parasitic fungus
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- VANSZAOQCMTTPB-SETSBSEESA-N hypocrellin Chemical compound C1[C@@](C)(O)[C@@H](C(C)=O)C2=C(OC)C(O)=C3C(=O)C=C(OC)C4=C3C2=C2C3=C4C(OC)=CC(=O)C3=C(O)C(OC)=C21 VANSZAOQCMTTPB-SETSBSEESA-N 0.000 title claims abstract description 46
- BQJKVFXDDMQLBE-UHFFFAOYSA-N shiraiachrome A Natural products COC1=C2C3=C(OC)C=C(O)C4=C3C3=C5C(CC(C)(O)C(C(C)=O)C3=C(OC)C4=O)=C(OC)C(=O)C(C(O)=C1)=C25 BQJKVFXDDMQLBE-UHFFFAOYSA-N 0.000 title claims abstract description 46
- 238000000855 fermentation Methods 0.000 title claims abstract description 38
- 230000004151 fermentation Effects 0.000 title claims abstract description 38
- KGHNSNSWRMJVND-UHFFFAOYSA-N Hypocrellin Natural products COC1=CC(=O)C2=C3C4C(C(C(=O)C)C(C)(O)Cc5c(OC)c(O)c6C(=O)C=C(OC)C(=C13)c6c45)C(=C2O)OC KGHNSNSWRMJVND-UHFFFAOYSA-N 0.000 title claims abstract description 33
- 235000017166 Bambusa arundinacea Nutrition 0.000 title claims abstract description 30
- 235000017491 Bambusa tulda Nutrition 0.000 title claims abstract description 30
- 241001330002 Bambuseae Species 0.000 title claims abstract description 30
- 235000015334 Phyllostachys viridis Nutrition 0.000 title claims abstract description 30
- 239000011425 bamboo Substances 0.000 title claims abstract description 30
- 241000233866 Fungi Species 0.000 title claims abstract description 28
- 230000003071 parasitic effect Effects 0.000 title claims abstract description 28
- 238000004519 manufacturing process Methods 0.000 title abstract description 7
- 239000001963 growth medium Substances 0.000 claims abstract description 22
- 239000010902 straw Substances 0.000 claims abstract description 21
- 241000277045 Shiraia bambusicola Species 0.000 claims abstract description 17
- 241001465754 Metazoa Species 0.000 claims abstract description 12
- 238000009629 microbiological culture Methods 0.000 claims abstract description 3
- 235000012239 silicon dioxide Nutrition 0.000 claims description 21
- 239000002609 medium Substances 0.000 claims description 20
- 239000007787 solid Substances 0.000 claims description 20
- 238000011218 seed culture Methods 0.000 claims description 19
- 239000007788 liquid Substances 0.000 claims description 17
- 239000000463 material Substances 0.000 claims description 13
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 12
- 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 12
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 12
- 239000008103 glucose Substances 0.000 claims description 12
- 210000000582 semen Anatomy 0.000 claims description 12
- 241000196324 Embryophyta Species 0.000 claims description 9
- 244000046052 Phaseolus vulgaris Species 0.000 claims description 9
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims description 9
- OSGAYBCDTDRGGQ-UHFFFAOYSA-L calcium sulfate Chemical compound [Ca+2].[O-]S([O-])(=O)=O OSGAYBCDTDRGGQ-UHFFFAOYSA-L 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- 239000006071 cream Substances 0.000 claims description 8
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 8
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 8
- 239000000843 powder Substances 0.000 claims description 7
- 239000002994 raw material Substances 0.000 claims description 7
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 6
- 229910052799 carbon Inorganic materials 0.000 claims description 6
- 229910052757 nitrogen Inorganic materials 0.000 claims description 6
- 235000015099 wheat brans Nutrition 0.000 claims description 5
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 claims description 4
- 229930091371 Fructose Natural products 0.000 claims description 4
- 239000005715 Fructose Substances 0.000 claims description 4
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 claims description 4
- 239000007836 KH2PO4 Substances 0.000 claims description 4
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims description 4
- 239000001888 Peptone Substances 0.000 claims description 4
- 108010080698 Peptones Proteins 0.000 claims description 4
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 4
- 229930006000 Sucrose Natural products 0.000 claims description 4
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 4
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 4
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 4
- 229940041514 candida albicans extract Drugs 0.000 claims description 4
- 229910052564 epsomite Inorganic materials 0.000 claims description 4
- 229910017053 inorganic salt Inorganic materials 0.000 claims description 4
- 239000008101 lactose Substances 0.000 claims description 4
- GVALZJMUIHGIMD-UHFFFAOYSA-H magnesium phosphate Chemical compound [Mg+2].[Mg+2].[Mg+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O GVALZJMUIHGIMD-UHFFFAOYSA-H 0.000 claims description 4
- 239000004137 magnesium phosphate Substances 0.000 claims description 4
- 229960002261 magnesium phosphate Drugs 0.000 claims description 4
- 229910000157 magnesium phosphate Inorganic materials 0.000 claims description 4
- 235000010994 magnesium phosphates Nutrition 0.000 claims description 4
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 4
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 4
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 4
- 235000019319 peptone Nutrition 0.000 claims description 4
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 4
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 4
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 claims description 4
- 239000005720 sucrose Substances 0.000 claims description 4
- 239000012138 yeast extract Substances 0.000 claims description 4
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 3
- 229940100486 rice starch Drugs 0.000 claims description 3
- 235000010344 sodium nitrate Nutrition 0.000 claims description 2
- 239000004317 sodium nitrate Substances 0.000 claims description 2
- 210000003608 fece Anatomy 0.000 abstract description 9
- 238000010884 ion-beam technique Methods 0.000 abstract description 8
- 239000000758 substrate Substances 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 3
- 238000004321 preservation Methods 0.000 abstract description 3
- 239000002699 waste material Substances 0.000 abstract description 3
- 238000003912 environmental pollution Methods 0.000 abstract description 2
- 230000002503 metabolic effect Effects 0.000 abstract description 2
- 238000000034 method Methods 0.000 description 16
- YDLBDQPPRTYAIG-UHFFFAOYSA-N hypocrellin A Natural products COC1C2CC(C)(O)C(C(=O)C)C3=C(OC)C(=O)c4c(O)cc(OC)c5c6c(OC)cc(O)c(C1=O)c6c2c3c45 YDLBDQPPRTYAIG-UHFFFAOYSA-N 0.000 description 13
- 239000002361 compost Substances 0.000 description 12
- 230000001580 bacterial effect Effects 0.000 description 11
- 241000894006 Bacteria Species 0.000 description 10
- 240000008042 Zea mays Species 0.000 description 10
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 10
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 8
- 235000005822 corn Nutrition 0.000 description 8
- 239000010871 livestock manure Substances 0.000 description 8
- 239000000243 solution Substances 0.000 description 7
- 230000035772 mutation Effects 0.000 description 6
- 239000000049 pigment Substances 0.000 description 6
- 239000002671 adjuvant Substances 0.000 description 5
- 241000283690 Bos taurus Species 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 241000287828 Gallus gallus Species 0.000 description 4
- 239000012084 conversion product Substances 0.000 description 4
- 238000003113 dilution method Methods 0.000 description 4
- 230000028327 secretion Effects 0.000 description 4
- 235000013339 cereals Nutrition 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- 229960000935 dehydrated alcohol Drugs 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 2
- 240000007594 Oryza sativa Species 0.000 description 2
- 235000007164 Oryza sativa Nutrition 0.000 description 2
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 238000007605 air drying Methods 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000003086 colorant Substances 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 2
- 238000009413 insulation Methods 0.000 description 2
- 235000009973 maize Nutrition 0.000 description 2
- 238000002703 mutagenesis Methods 0.000 description 2
- 231100000350 mutagenesis Toxicity 0.000 description 2
- 230000035479 physiological effects, processes and functions Effects 0.000 description 2
- 238000007747 plating Methods 0.000 description 2
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 230000001373 regressive effect Effects 0.000 description 2
- 235000009566 rice Nutrition 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 230000003248 secreting effect Effects 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 241000143667 Hypocrella Species 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000011953 bioanalysis Methods 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000000576 food coloring agent Substances 0.000 description 1
- 238000000703 high-speed centrifugation Methods 0.000 description 1
- 238000011090 industrial biotechnology method and process Methods 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 239000002932 luster Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 201000003068 rheumatic fever Diseases 0.000 description 1
- 210000003497 sciatic nerve Anatomy 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 230000008736 traumatic injury Effects 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
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- 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
- C12N1/145—Fungal isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/24—Preparation of oxygen-containing organic compounds containing a carbonyl group
- C12P7/26—Ketones
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Abstract
The invention discloses a bamboo parasitic fungus and application thereof in fermentation production of hypocrellin. Three bamboo parasitic fungi are separated from Shiraia bambusicola respectively collected from Huangshan region of Anhui province, Quzhou region of Zhejiang province and Yinbin region of Sichuan province, are cultured by virtue of a basal culture medium and is processed by virtue of ion beams, and therefore, a strain with the highest vigorous activity is obtained, is named as Shiraia bambusicola NJTECH-1 and has been preserved in China General Microbiological Culture Collection Center with a preservation number of CGMCC NO. 11617. The bamboo parasitic fungus has very high metabolic activity and is applied to the fermentation production of the hypocrellin, and the yield of the hypocrellin is high. The field waste straws and animal dung can be taken as fermentation substrates, so that the quantity of incinerated straws can be reduced, and the environmental pollution can be reduced while the cost is saved.
Description
Technical field
The invention belongs to technical field of biological fermentation, be specifically related to a strain bamboo parasitic fungus and in fermenting and producing hypocrellin
Application.
Background technology
Concretio silicea Bambusae seu schizostachyi is distributed mainly on part province, Southeast China side, it addition, also there is distribution in the Japan in Asia, other countries are then
Have no report.The hypocrellin that bamboo parasitic fungus produces, have good scattered silt to invigorate blood circulation, expelling wind and removing dampness, invigorating the spleen and replenishing QI and promotion new old generation
Thank, the effect such as health invigorating, be used for clinically treating peratodynia, rheumatic arthritis, traumatic injury and sciatic nerve
The diseases such as pain.And this pigment safety non-toxic, color and luster are scarlet, strong coloring force and be rich in health care, as food coloring also
There is wide prospect.
Traditional hypocrellin production method, is generally divided into biological synthesis process and chemical synthesis, and biological synthesis process is main
Being to utilize Semen Maydis powder, wheat bran, the cereal crops such as Semen Tritici aestivi are raw material, utilize bamboo parasitic fungus fermentation to produce hypocrellin, then separate
Purification, although increased relative to yield from the point of view of other modes of production, but the epoch progressively risen at provision price, this side
Method also makes the cost of end product improve many, is unfavorable for the large-scale application of hypocrellin.Another is that chemistry closes
Cheng Fa, utilizes 3, and 5-resorcylic acid is initial reaction thing, through 12 step chemical reactions, has synthesized hypocrellin, but this
Method reactions steps is more and complicated, and side reaction is many, and agents useful for same toxicity is relatively big, and cost is the highest, is not to produce hypocrellin
Good method.
2005, Chinese maize straw reached about 20207 × 104T, and the ratio of straw-returning is the lowest, though
So country prohibites crop straw burning, but still can not effect a radical cure the problem that straw is too much completely.Reasonable utilizes discarded straw
The method of stalk is particularly important.
Summary of the invention
It is an object of the invention to provide a strain bamboo parasitic fungus and the application in fermenting and producing hypocrellin thereof, this bamboo parasitic fungus
Hypocrellin yield is high, and can effectively utilize the fermentation of materials such as waste straw, discarded fowl and animal excrement to produce hypocrellin, reduces raw
Produce cost and safety and environmental protection.
For achieving the above object, the technical solution used in the present invention is as follows:
The present invention will adopt in Mt. Huang in Anhui area respectively, and area, Quzhou, Zhejiang and the Concretio silicea Bambusae seu schizostachyi of Sichuan Yibin Prefecture are isolated to
Three strain bamboo parasitic fungus, carry out base culture base, after ion beam mutagenesis processes, obtain the bamboo parasitic fungus that a strain vigor is the most vigorous,
Its Classification And Nomenclature be Concretio silicea Bambusae seu schizostachyi (Shiraia bambusicola) NJTECH-1, it is preserved in China on November 30th, 2015
General Microbiological Culture preservation administrative center, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Chinese Academy of Sciences's microorganism
Institute, preserving number is CGMCC NO.11617.
Described bacterial strain screens by the following method and obtains: adopt respectively in Mt. Huang in Anhui area, Quzhou, Zhejiang area and
The Concretio silicea Bambusae seu schizostachyi of Sichuan Yibin Prefecture, isolated three strain bamboo parasitic fungus, utilize liquid murphy juice culture medium to carry out basis cultivation, 26 DEG C of trainings
After supporting five days, measuring Biomass and hypocrellin secretory volume, the bacterial strain obtaining a strain vigor the most vigorous is collection simultaneously
From the bacterial strain in Yibin, Sichuan, named Super-3, as dominant strain.
With normal saline, the spore of Super-3 is washed down, be connected in liquid seed culture medium cultivate.By cultured Concretio silicea Bambusae seu schizostachyi
It is 10 that bacterium spore suspension is diluted to cell number by 10 times of dilution methods7/ mL, takes bacterium solution 0.1 mL and is uniformly applied to aseptic empty flat board
In, after air-drying, carry out N+Ion beam mutation, N+Ion beam mutation dosage is (90,135,180,225,270) × 2.6 × 1013N+/cm2, N+Ion beam mutation energy is 15keV.Irradiation terminate after with 1 mL sterilized water washed cell, by 10 times of dilution methods dilutions
After be painted into plating medium, be inverted for 26 DEG C and cultivate 45 d, treat picking list bacterium colony, shaking flask detects, and filters out mycelial growth rate
Fast and that hypocrellin secretion rate is the highest bacterial strain, named Concretio silicea Bambusae seu schizostachyi (Shiraia bambusicola) NJTECH-1。
Described Concretio silicea Bambusae seu schizostachyi (Shiraia bambusicola) condition of culture of NJTECH-1 is:
This strain uses the mode of aerobic cultivation.Can be glucose for cultivating the carbon source of this bacterial strain, sucrose, fructose, lactose,
The materials such as soluble starch, rice, Semen Maydis；Can be Carnis Bovis seu Bubali cream, peptone, yeast extract, nitre for cultivating the nitrogen source of this bacterial strain
The acid material such as sodium, ammonium sulfate.The optimum temperature range of this strain growth is 24-28 DEG C, pH scope 4-9, and optimum PH range is 5-
7.The inorganic salts such as dipotassium hydrogen phosphate, potassium dihydrogen phosphate, magnesium sulfate, magnesium phosphate also can be added during strain culturing.
Described Concretio silicea Bambusae seu schizostachyi (Shiraia bambusicola) physiology and appearance of NJTECH-1 is characterized as:
Form cultivated by flat board: well-grown on PDA plate, cultivation to the 3rd day bacterium colony starts with radial growth；Mycelia is
White fluffy, mid portion relatively both sides mycelia is flourishing；Bacterium colony core culture medium is red, is the pigment of thalline secretion,
Pigment becomes kermesinus in the medium, forms annular concentric circle, and intermediate colors is deep, and ambient color is shallow.Cultivate about five days, cultivate
Base all becomes red, and the back side is substantially it can be seen that a large amount of agglomerating spore of black, and visible tubulose mycelia under microscope, spore is about
About 1.1 μm.
Described Concretio silicea Bambusae seu schizostachyi (Shiraia bambusicola) NJTECH-1 application in fermenting and producing hypocrellin.
Comprise the steps:
1) seed culture: washed down by the spore of bamboo parasitic fungus with normal saline, accesses in liquid seed culture medium, 24 ~ 30 DEG C of cultivations
40~48h；
2) taking in the seed liquor 5mL access solid medium of step 1) and carry out fermentation culture, cultivation temperature is 26 DEG C ~ 30 DEG C, raw
Produce hypocrellin.
Liquid seed culture medium described in step 1) includes the component of following mass percent: carbon source 2% ~ 3%, nitrogen source 0.2%
~ 0.4%, inorganic salt 0.05% ~ 0.1%, pH is natural, and wherein said carbon source is formed sediment selected from glucose, sucrose, fructose, lactose, solubility
At least one in powder, rice, Semen Maydis；Described nitrogen source is selected from Carnis Bovis seu Bubali cream, peptone, yeast extract, sodium nitrate, ammonium sulfate extremely
Few one, at least one in dipotassium hydrogen phosphate, potassium dihydrogen phosphate, magnesium sulfate, magnesium phosphate of described inorganic salt.
Step 2) described in solid medium can be as the culture medium of configuration with grain, it may for example comprise following quality
The component of percentage ratio: Semen Maydis powder 68.9%-75%, wheat bran 5%-10%, Testa oryzae 15%-20%, glucose 1%-3%, KH2PO4
0.1%-0.2% ,MgSO4·7H2O 0.05%-0.07%。
Can also be for weeds, corn straw, fowl and animal excrement (based on chicken manure and cattle manure), Gypsum Fibrosum and a small amount of bean cake being
Fermenting raw materials prepares solid medium, and concrete fermentation process is: first weeds and corn straw are mixed with fowl and animal excrement,
Add the adjuvant such as Gypsum Fibrosum, bean cake after pre-wetted treatment again, fermentation tunnel carries out one time fermentation, treat that temperature rises to 70-80 DEG C
Shi Baowen 4-5 days, then lowers the temperature, and during cooling, turning is several times, obtains the compost that water content is 70%-80%.After carry out two
Secondary fermentation, proceeds to, in ferment in second time tunnel, utilize the fermentable in compost, treat temperature by one time fermentation gained compost
When reaching 55-60 DEG C, insulation 9-11h, then compost is carried out the fermentation stage that becomes thoroughly decomposed of 4-5 days by a definite date, finally give water content
For the culture base-material of 65%-70%, can be as the solid state substrate cultivating bamboo parasitic fungus.
The proportionate relationship of above-mentioned each material can be adjusted according to practical situation.The most first by weeds and corn straw with
Fowl and animal excrement is mixed in about 1:1:0.7 ratio, after pre-wetted treatment again by mixed material and adjuvant with the ratio of about 1:2.5
Add the adjuvant such as Gypsum Fibrosum, bean cake.
Weeds in this culture medium prescription, corn straw, fowl and animal excrement (based on chicken manure and cattle manure), Gypsum Fibrosum and a small amount of bean
The dregs of rice are the raw material being easy to get at a low price, utilize them as the fermentation substrate of bamboo parasitic fungus, are not only substantially reduced the production of hypocrellin
Cost, and straw can be carried out favourable secondary and utilize, protect environment.
The biggest market potential is had for bamboo parasitic fungus solid fermentation to produce hypocrellin after processing with stalk fermentation,
Being mainly manifested in: 2005, Chinese maize straw has reached about 20207*104T, and the ratio of straw-returning is the lowest,
This measure can make full use of waste straw, not only wide material sources, and can also effectively protect environment.Owing to hypocrellin is at medicine
Learning, have the biggest application prospect, but price is the most costly, cause being widely used in biology, this kind of method drops significantly
Low production cost, the use on a large scale for hypocrellin provides possibility.
Beneficial effect: the present invention compared with prior art has the advantage that
1., compared with other strains, the bamboo parasitic fungus of the present invention has the highest metabolic activity, the yield of fermenting and producing hypocrellin
High.
2., compared with traditional bioanalysis, the raw material cultivated is become what field was discarded from cereal crops (Semen Maydis powder etc.)
Straw and fowl and animal excrement, from cost for, obtained the most significantly reducing, and the quantity of crop straw burning can have been reduced,
While cost-effective, reduce environmental pollution.
3., compared with conventional chemical methods, the inventive method is easy, reduces cost further, brings huge economic benefit.
Detailed description of the invention
By following embodiment, the present invention be may be better understood.Then, as it will be easily appreciated by one skilled in the art that reality
Executing the concrete material ratio described by example, process conditions and result thereof are merely to illustrate the present invention, and not should also without limitation on
Claims say described in detail by the present invention.
Embodiment 1 Concretio silicea Bambusae seu schizostachyi (Shiraia bambusicola) mutagenesis screening of NJTECH-1
To adopt in Mt. Huang in Anhui area respectively, area, Quzhou, Zhejiang and the Concretio silicea Bambusae seu schizostachyi of Sichuan Yibin Prefecture, isolated three strain Concretio silicea Bambusae seu schizostachyi
Bacterium, utilizes liquid murphy juice culture medium to carry out basis cultivation, after 26 DEG C are cultivated five days, measures Biomass and hypocrellin simultaneously
Secretory volume, obtains the most vigorous bacterial strain of a strain vigor and is and gathers bacterial strain from Yibin, Sichuan, named Super-3, as
Dominant strain.
With normal saline, the spore of Super-3 is washed down, be connected in liquid seed culture medium, cultivate 12 hours for 26 DEG C, will
It is 10 that cultured bamboo parasitic fungus spore suspension is diluted to cell number by 10 times of dilution methods7/ mL, takes bacterium solution 0.1 mL and is uniformly applied to
In aseptic empty flat board, after air-drying, carry out N+Ion beam mutation, N+Ion beam mutation dosage is (90,135,180,225,270)
×2.6×1013N+/cm2, N+Ion beam mutation energy is 15keV.Irradiation terminate after with 1 mL sterilized water washed cell, by 10
Being painted into plating medium after the dilution of times dilution method, be inverted for 26 DEG C and cultivate 4-5 d, treat picking list bacterium colony, shaking flask detects, and filters out
The bacterial strain that mycelial growth rate is fast and hypocrellin secretion rate is the highest, named Concretio silicea Bambusae seu schizostachyi (Shiraia bambusicola)
NJTECH-1。
Embodiment 2 Concretio silicea Bambusae seu schizostachyi (Shiraia bambusicola) cultivation of NJTECH-1 and physiologic character
Described Concretio silicea Bambusae seu schizostachyi (Shiraia bambusicola) condition of culture of NJTECH-1 is:
This strain uses the mode of aerobic cultivation, the carbon source for cultivating this bacterial strain can be glucose, sucrose, fructose, lactose,
The materials such as soluble starch, rice, Semen Maydis；Can be Carnis Bovis seu Bubali cream, peptone, yeast extract, nitre for cultivating the nitrogen source of this bacterial strain
The acid material such as sodium, ammonium sulfate；The optimum temperature range of this strain growth is 24-28 DEG C, pH scope 4-9, and optimum PH range is 5-
7, during strain culturing, also can add the inorganic salts such as dipotassium hydrogen phosphate, potassium dihydrogen phosphate, magnesium sulfate, magnesium phosphate.
Described Concretio silicea Bambusae seu schizostachyi (Shiraia bambusicola) physiology and appearance of NJTECH-1 is characterized as:
Form cultivated by flat board: well-grown on PDA plate, cultivation to the 3rd day bacterium colony starts with radial growth；Mycelia is
White fluffy, mid portion relatively both sides mycelia is flourishing；Bacterium colony core culture medium is red, is the pigment of thalline secretion,
Pigment becomes kermesinus in the medium, forms annular concentric circle, and intermediate colors is deep, and ambient color is shallow.Cultivate about five days, cultivate
Base all becomes red, and the back side is substantially it can be seen that a large amount of agglomerating spore of black, and visible tubulose mycelia under microscope, spore is about
About 1.1 μm.
Embodiment 3 Concretio silicea Bambusae seu schizostachyi (Shiraia bambusicola) (LBR-SB6) fermenting and producing hypocrellin A
With normal saline by bamboo parasitic fungus (LBR-SB6) (by the preservation of key lab of the industrial biotechnology Ministry of Education of Southern Yangtze University)
Spore wash down, access in liquid seed culture medium, 26 DEG C, 120rpm cultivates 40h, takes 5mL seed liquor access solid medium
In, temperature controls 26 DEG C of cultivations.Liquid seed culture medium includes following component: glucose 2g/L, dipotassium hydrogen phosphate 0.1g/L,
Anhydrous magnesium sulfate 0.05g/L, Carnis Bovis seu Bubali cream 0.3g/L, pH are natural.
Seed liquor accesses in solid medium and carries out fermentation culture, and cultivation temperature is 26 DEG C ~ 30 DEG C, produces hypocrellin.
Solid medium includes the composition (%) of following mass ratio: the Semen Maydis powder 68.9 of 60 mesh, wheat bran 9.85, Testa oryzae 20, glucose
1, KH2PO4 0.2 ,MgSO4·7H2O 0.05。
Fermentation results, main conversion product is hypocrellin A, and yield is about 40mg/kg.
Embodiment 4 Concretio silicea Bambusae seu schizostachyi (Shiraia bambusicola) NJTECH-1 fermenting and producing hypocrellin A
Seed culture: washed down by the spore of bamboo parasitic fungus with normal saline, accesses in liquid seed culture medium, cultivates 40h for 26 DEG C；Liquid
Body seed culture medium includes following component: glucose 2g/L, dipotassium hydrogen phosphate 0.1g/L, anhydrous magnesium sulfate 0.05g/L, beef
Cream 0.3g/L, pH are natural.
Seed liquor accesses in solid medium and carries out fermentation culture, and cultivation temperature is 26 DEG C ~ 30 DEG C, produces hypocrellin.
Solid medium includes the composition (%) of following mass ratio: the Semen Maydis powder 68.9 of 60 mesh, wheat bran 9.85, Testa oryzae 20, glucose
1, KH2PO4 0.2 ,MgSO4·7H2O 0.05。
Fermentation results, main conversion product is hypocrellin A, and yield is 78.1mg/kg.
Embodiment 5 Concretio silicea Bambusae seu schizostachyi (Shiraia bambusicola) NJTECH-1 with stalk fermentation produce hypocrellin A
The preparation of solid medium: with weeds, corn straw, fowl and animal excrement (based on chicken manure and cattle manure), Gypsum Fibrosum and a small amount of
Bean cake is raw material, first weeds and corn straw is mixed in about 1:1:0.7 ratio with fowl and animal excrement, presses after pre-wetted treatment again
The ratio of about 1:2.5 adds the adjuvant such as Gypsum Fibrosum, bean cake, carries out one time fermentation, when temperature rises to 70-80 DEG C in fermentation tunnel
Being incubated 4-5 days, then lower the temperature, during cooling, turning is several times, obtains the compost that water content is 70%-80%.After carry out secondary
Fermentation, proceeds to gained compost, in ferment in second time tunnel, utilize the fermentable in compost, treats that temperature reaches 55-60
DEG C time, be incubated 9-11h, then compost carried out the fermentation stage that becomes thoroughly decomposed of 4-5 days by a definite date, finally giving water content is 65%-70%
Culture base-material, as cultivate bamboo parasitic fungus solid state substrate.
Seed culture: washed down by the spore of bamboo parasitic fungus with normal saline, accesses in liquid seed culture medium, 26 DEG C of cultivations
40h；Liquid seed culture medium includes following component: glucose 2g/L, dipotassium hydrogen phosphate 0.1g/L, anhydrous magnesium sulfate 0.05g/
L, Carnis Bovis seu Bubali cream 0.3g/L, pH are natural.
Taking in seed liquor 5mL access solid medium and carry out fermentation culture, cultivation temperature is 26 DEG C ~ 30 DEG C, produces bamboo red
Rhzomorph, measures the content of hypocrellin A, fermentation ends after 168h.Fermentation results, main conversion product is hypocrellin A, and yield is
4mg/kg。
Embodiment 6 Concretio silicea Bambusae seu schizostachyi (Shiraia bambusicola) NJTECH-1 with stalk fermentation produce hypocrellin A
The preparation of solid medium: with weeds, corn straw, fowl and animal excrement (based on chicken manure and cattle manure), Gypsum Fibrosum and a small amount of
Bean cake is raw material, first weeds and corn straw is mixed in about 1:1:1 ratio with fowl and animal excrement, again by 1 after pre-wetted treatment:
The adjuvants such as the ratio of about 3 adds Gypsum Fibrosum, bean cake, carry out one time fermentation in fermentation tunnel, protect when temperature rises to 70-80 DEG C
Temperature 4-5 days, then lowers the temperature, and during cooling, turning is several times, obtains the compost that water content is 70%-80%.After carry out secondary and send out
Ferment, proceeds to gained compost, in ferment in second time tunnel, utilize the fermentable in compost, treats that temperature reaches 55-60 DEG C
Time, insulation 9-11h, then compost is carried out the fermentation stage that becomes thoroughly decomposed of 4-5 days by a definite date, finally giving water content is 65%-70%'s
Culture base-material, as the solid state substrate cultivating bamboo parasitic fungus.
Seed culture: washed down by the spore of bamboo parasitic fungus with normal saline, accesses in liquid seed culture medium, 26 DEG C of cultivations
40h；Liquid seed culture medium includes following component: glucose 2g/L, dipotassium hydrogen phosphate 0.1g/L, anhydrous magnesium sulfate 0.05g/
L, Carnis Bovis seu Bubali cream 0.3g/L, pH are natural.
Seed liquor accesses in solid medium and carries out fermentation culture, and cultivation temperature is 26 DEG C ~ 30 DEG C, produces hypocrellin,
Measuring the content of hypocrellin A after 168h, fermentation ends, in fermentation liquid, main conversion product is hypocrellin A, and yield is 8.5mg/
kg。
In above example, hypocrellin A adopts and measures with the following method:
First determine the standard curve of hypocrellin A: accurately weigh hypocrellin A and mark product 3.6mg, be placed in 10ml volumetric flask,
With anhydrous alcohol solution and be settled to scale, obtaining standard solution concentration is 0.36mg/ml, is diluted to a series of with dehydrated alcohol
Concentration (10,20,30,50,100,120,180,240, unit: μ g/ml), exists the mark product solution of a series of variable concentrations
Surveying absorbance at 464nm, with hypocrellin concentration as abscissa, absorbance is vertical coordinate, carries out regression Calculation, obtains Hypocrella bambusae (Bet Br). Sace
Element standard regressive method is y=0.0403x-0.0283, R=0.9997.
The mensuration of hypocrellin content: solid medium is put into drying baker and is dried, pulverizes by food processor, takes 0.1g
Ground product mix with 50ml dehydrated alcohol, stirring and leaching 2h, take lixiviating solution high speed centrifugation after extraction, take supernatant, obtain clear
Clear pigment solution, after dehydrated alcohol dilution several times, uses spectrophotometer method to measure trap at 464nm wavelength,
The content of hypocrellin is calculated according to standard regressive method.
Claims (6)
1. a strain bamboo parasitic fungus, its Classification And Nomenclature be Concretio silicea Bambusae seu schizostachyi (Shiraia bambusicola) NJTECH-1, it is preserved in China
Microbiological Culture Collection administration committee common micro-organisms center, preserving number is CGMCC NO.11617.
2. the application in fermenting and producing hypocrellin of the bamboo parasitic fungus described in claim 1.
Application the most according to claim 2, it is characterised in that comprise the steps:
1) seed culture: washed down by the spore of bamboo parasitic fungus with normal saline, accesses in liquid seed culture medium, 24 ~ 30 DEG C of cultivations
40~48h；
2) taking in the seed liquor access solid medium of step 1) and carry out fermentation culture, cultivation temperature is 26 DEG C ~ 30 DEG C, produces bamboo
Red rhzomorph.
Application the most according to claim 3, it is characterised in that liquid seed culture medium described in step 1) includes following matter
The component of amount percentage ratio: carbon source 2% ~ 3%, nitrogen source 0.2% ~ 0.4%, inorganic salt 0.05% ~ 0.1%, pH is natural, and wherein said carbon source is selected
At least one in glucose, sucrose, fructose, lactose, soluble starch, rice, Semen Maydis；Described nitrogen source selected from Carnis Bovis seu Bubali cream,
At least one in peptone, yeast extract, sodium nitrate, ammonium sulfate, described inorganic salt selected from dipotassium hydrogen phosphate, potassium dihydrogen phosphate,
At least one in magnesium sulfate, magnesium phosphate.
Application the most according to claim 3, it is characterised in that step 2) described in solid medium for weeds, Semen Maydis
At least one in straw, fowl and animal excrement, Gypsum Fibrosum or bean cake is raw material, and fermentation obtains the culture medium that water content is 65%-70%
Material.
Application the most according to claim 3, it is characterised in that step 2) described in solid medium include following quality hundred
The component of proportion by subtraction: Semen Maydis powder 68.9% ~ 75%, wheat bran 5% ~ 10%, Testa oryzae 15% ~ 20%, glucose 1% ~ 3%, KH2PO4 0.1%~
0.2% ,MgSO4·7H2O 0.05%~0.07%。
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| CN107668374A (en) * | 2017-10-03 | 2018-02-09 | 长沙仲善新能源科技有限公司 | A kind of single cell protein biological feedstuff and preparation method thereof |
| CN110172409A (en) * | 2019-04-26 | 2019-08-27 | 华南理工大学 | One plant height produces tabasheer bacterial strain and its application of hypocrellin A |
| CN111218407A (en) * | 2020-01-15 | 2020-06-02 | 山东国力生物科技有限公司 | Shiraia bambusicola with high yield of hypocrellin and culture method and application thereof |
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| CN102703328A (en) * | 2012-04-27 | 2012-10-03 | 苏州维因生物科技有限公司 | Hypocrellin high-yielding Shiraia bambusicola strain |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107099489A (en) * | 2017-06-30 | 2017-08-29 | 苏州大学 | One plant raising hypocrellin fermentation production rate associated bacteria bacterial strain and its application |
| CN107099489B (en) * | 2017-06-30 | 2020-10-09 | 苏州大学 | Associated bacterial strain for improving hypocrellin fermentation yield and application thereof |
| CN107668374A (en) * | 2017-10-03 | 2018-02-09 | 长沙仲善新能源科技有限公司 | A kind of single cell protein biological feedstuff and preparation method thereof |
| CN110172409A (en) * | 2019-04-26 | 2019-08-27 | 华南理工大学 | One plant height produces tabasheer bacterial strain and its application of hypocrellin A |
| CN110172409B (en) * | 2019-04-26 | 2020-04-28 | 华南理工大学 | Shiraia bambusicola strain capable of highly producing hypocrellin A and application thereof |
| CN111218407A (en) * | 2020-01-15 | 2020-06-02 | 山东国力生物科技有限公司 | Shiraia bambusicola with high yield of hypocrellin and culture method and application thereof |
| CN111218407B (en) * | 2020-01-15 | 2020-12-08 | 山东国力生物科技有限公司 | Shiraia bambusicola with high yield of hypocrellin and culture method and application thereof |
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