CN105993584B - A kind of artificial culture method of rich magnesium cicada fungus - Google Patents
A kind of artificial culture method of rich magnesium cicada fungus Download PDFInfo
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- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 title claims abstract description 55
- 239000011777 magnesium Substances 0.000 title claims abstract description 55
- 229910052749 magnesium Inorganic materials 0.000 title claims abstract description 55
- 241000931705 Cicada Species 0.000 title claims abstract description 32
- 241000233866 Fungi Species 0.000 title claims abstract description 32
- 238000012136 culture method Methods 0.000 title claims abstract description 22
- 239000007787 solid Substances 0.000 claims abstract description 47
- 238000000034 method Methods 0.000 claims abstract description 19
- 159000000003 magnesium salts Chemical class 0.000 claims abstract description 9
- 241001625026 Cordyceps cicadae Species 0.000 claims abstract description 8
- 239000007788 liquid Substances 0.000 claims abstract description 5
- 230000003321 amplification Effects 0.000 claims abstract description 3
- 238000003199 nucleic acid amplification method Methods 0.000 claims abstract description 3
- 239000002609 medium Substances 0.000 claims description 57
- 239000001963 growth medium Substances 0.000 claims description 52
- 229940091250 magnesium supplement Drugs 0.000 claims description 51
- 239000004337 magnesium citrate Substances 0.000 claims description 22
- 229960005336 magnesium citrate Drugs 0.000 claims description 22
- 235000002538 magnesium citrate Nutrition 0.000 claims description 22
- PLSARIKBYIPYPF-UHFFFAOYSA-H trimagnesium dicitrate Chemical compound [Mg+2].[Mg+2].[Mg+2].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O PLSARIKBYIPYPF-UHFFFAOYSA-H 0.000 claims description 22
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- 238000003306 harvesting Methods 0.000 claims description 4
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- 238000006243 chemical reaction Methods 0.000 abstract description 2
- 244000005700 microbiome Species 0.000 abstract 1
- 238000009630 liquid culture Methods 0.000 description 15
- 230000008859 change Effects 0.000 description 13
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- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 6
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- JLVVSXFLKOJNIY-UHFFFAOYSA-N Magnesium ion Chemical compound [Mg+2] JLVVSXFLKOJNIY-UHFFFAOYSA-N 0.000 description 3
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- 206010022489 Insulin Resistance Diseases 0.000 description 2
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention relates to a kind of artificial culture methods of rich magnesium cicada fungus, and this method comprises the following steps: step 1, Cordyceps cicadae strain is expanded in liquid medium；Step 2, the strain after step 1 amplification is inoculated into solid medium and carries out solid culture；Step 3, coremium is harvested；It is characterized in that, magnesium salts is added in the solid medium described in step 2, additional amount is 7~15%.Artificial culture method of the present invention in cicada fungus liquid and solid medium using adding quantitative magnesium salts, after microorganism conversion, is made rich magnesium cicada fungus, edible safety, conveniently, can meet requirement of the various people to magnesium elements.
Description
Technical field
The present invention relates to a kind of artificial culture methods of cicada fungus, and in particular to a kind of artificial culture method of rich magnesium cicada fungus,
Belong to biological field.
Background technique
Magnesium is the dominant cation in human body cell, is concentrated in gland plastochondria, is only second to potassium and phosphorus, only secondary in extracellular fluid
Third position is occupied in sodium and calcium.Magnesium is the activator of a variety of enzymes, is played an important role in energy and metabolism.Research in recent years discovery
The generation of magnesium and second messenger cAMP has with the different physiological roles such as hormone and growth factor, cardiac muscle cell and cationic channel
It closes.
Magnesium ion participates in all energetic supersessions in vivo as the confactor of enzyme, activates and be catalyzed about more than 350 kinds enzyme systems,
Utilization including glucose, the synthesis of fat, protein and nucleic acid, atriphos metabolism and film ion transport etc., and and line
Plastochondria cell and all membrane structures are related with functional completeness.Magnesium ion keeps intracellular stabilization by the enzyme on activated membrane,
Magnesium ion changes the metabolism for also influencing calcium so magnesium has the special role for maintaining cardiac muscle, neural, muscle normal function.It is existing
Some researches show that magnesium deficiencies can cause for example ventricular premature beat of various arrhythmia cordis, atrial premature beats, tachycardia even room top；Make albumen
Matter synthesis decline, reduces Abwehrkraft des Koepers and immune function, and human tumor incidence increases；Form the high quick synthesis of bronchial asthma
Disease.It is the magnesium due to caused by gastroxia that common enterogastric diseases such as gastric ulcer, duodenal ulcer and gastritis etc. are mostly
Salt has stronger buffer capacity as antacids, can be in Fast Persistence and gastric acid is without causing alkalosis and secondary acid point
The phenomenon that secreting increase；Magnesia mixture can safeguard the stability of liver plasma membrane, promote hepatocyte function to restore, and can promote bile generation
It thanks.Magnesium can promote glycogen and energy storage, improve gastrointestinal function, Early postoperative magnesia mixture restore the function of intestinal canal of patient
Time obviously shifts to an earlier date, and the nutrition condition of patient improves comparatively fast, and post-operative recovery is very fast.Magnesium can treat diarrhea and constipation.
Magnesium is also the main component of group skeletonization, and it is the shape of the cytoplasm of normal bone that magnesium, which promotes bone, tooth and cell to be formed,
At required.Magnesium has crucial adjustment effect in the mineral metabolism of bone, also has relationship with the ca hormone that withers.Magnesium can not stimulate completely
Parathormone (PTH) secretion, promotes bone resorption.Some researches show that it is sclerotin that slight magnesium shortage, which can damage mineral homeostasis,
Loose risk factor.Magnesium lacks and osteoporosis often occurs simultaneously, and treatment osteoporosis not only needs to replenish the calcium and should mend magnesium simultaneously.
Magnesium metabolism influences each other with glycometabolism, and blood sugar concentration is often negatively correlated with blood magnesium levels, diabetes patient's serum magnesium
Lower than normal person, urine magnesium is higher than normal person, and blood magnesium level and insulin sensitivity are negatively correlated, and insulin sensitivity can be improved by mending magnesium
Property and insulin secretion.Diabetes complicated retinopathy and blood magnesium are negatively correlated, and hypomagnesemia is considered as retinopathy
Risk factor.
Summary of the invention
The purpose of the present invention is to provide a kind of cultural methods of rich magnesium cicada fungus.
The purpose of the present invention is what is be achieved by the following scheme:
A kind of artificial culture method of rich magnesium cicada fungus, this method comprises the following steps:
Step 1, Cordyceps cicadae strain is expanded in liquid medium；
Step 2, the strain after step 1 amplification is inoculated into solid medium and carries out solid culture；
Step 3, coremium is harvested；
It is characterized in that, magnesium salts is added in the solid medium described in step 2, additional amount is 7~15%.
Further, the additional amount of magnesium salts is 7~11% in solid medium；It further, is 9~11%.
Further, magnesium salts is added in step 1 fluid nutrient medium, additional amount is 1~3%.
Further, the magnesium salts is magnesium citrate, magnesium acetate, magnesium sulfate, magnesium chloride.Further, the magnesium salts is lemon
Lemon acid magnesium.
Further, step 2 inoculum concentration is 15%~30%；Further, inoculum concentration is 20%~25%；Further it is
25%.
Further, during step 2 solid culture, shading culture is used in the mycelial growth stage；It is originated from coremium
Illumination cultivation is used to harvest stages.
Solid medium of the present invention is this field Conventional solid culture medium, such as grain culture medium, crop stalk training
Support base, crops leather shell culture medium, commodity trees branch culture medium, plant stem culture medium etc.；Wherein preferred grain culture medium；
The cereal can be selected from wheat, corn, rice, millet, soybean powder, buckwheat, barley, oat, brown rice, any one in polished rice
Or it is several.Further, the solid medium is wheat broth or buckwheat culture medium.
For the purpose of Liquid Culture process described in step 1 of the present invention is expanded by strain, conventional strain propagation method can be used,
Combination including any one or a few method in inclined-plane culture, shaking flask culture, seed tank culture, fermentation tank culture；It is used
Culture medium is the fluid nutrient medium of this field routine, such as potato-sucrose-agar medium (PSA), Potato-dextrose-fine jade
Rouge culture medium (PDA), Potato-dextrose-water culture medium (PSB), yeast extract powder-compound amino acid-sucrose culture medium
(SAAY), yeast extract powder-white granulated sugar-soy bean protein hydrolysate culture medium, any one in wheat bran liquor-white sand sugar culture-medium
Or it is several.
Heretofore described cicada fungus (Isaria cicadae Miquel) is dispersed species, is widely distributed in China Qinling Mountains-Huaihe River
18 provinces, city, areas on the south river.Subtropical zone and torrid areas on the south China the Changjiang river, and it is mostly high in low lying areas and Yunnan-Tibet
The river valleys areas such as former Jinsha jiang River, Nujiang, the Lancang River and the Yarlung Zangbo River.As long as, being pressed the annual 6-8 month in its season of growth
It can all be adopted according to the acquisition method of general Chinese medicine, be strain known in the state of the art, and acquisition can be bought from commercial channels.
Cicada fungus bacterial strain used in the present invention discloses that (deposit number is in CN201110178274.6 patent
CGMCC No.3453)。
Artificial culture method of the present invention passes through micro- life using quantitative magnesium salts is added in cicada fungus liquid and solid medium
After object conversion, it is made rich magnesium cicada fungus, edible safety, conveniently can meet requirement of the various people to magnesium elements.
Specific embodiment:
In following embodiment, embodiment 1-9 and comparative examples use deposit number for the Paecilomyces cicadae of CGMCC No.3453
Bacterial strain [Paecilomyces cicadae (Miq.) Samson]；Embodiment 10-13 prepares cicada fungus bacterial strain using embodiment 14.
In following embodiment, magnesium element content measurement is referring to " measurement of GB/T5009.90-200 iron in food, magnesium, manganese "
Method carries out；Contents of Amino Acids is carried out referring to " measurement of amino acid in GB/T 5009.124-2003 food " method；HEA
Assay is referring to " " N in fruiting bodies of cordyceps militaris6(2- ethoxy)-adenosine isolate and purify and antitumor action ", edible mushroom
Journal 2013.20 (1): 62~65 " methods carry out.
Embodiment 1
Inclined-plane culture: Cordyceps cicadae strain is inoculated in slant tube, then the slant tube for being inoculated with strain is put into 22 DEG C of trainings
Case is supported, incubation time is 7 days, covers with test tube to mycelia；
Liquid Culture: 200ml fluid nutrient medium, the high pressure sterilization 30 under 0.11Mpa pressure are packed into 500ml triangular flask
Minute, it is cooled to room temperature, the strain of 1 slant tube is inoculated into 500ml triangular flask culture medium, constant temperature oscillation is placed in
Incubator is cultivated under the conditions of 22 ± 1 DEG C of temperature, 150 revs/min, and incubation time is 3 days；
Solid culture: the strain obtained by 25% inoculum concentration inoculation seed culture, the container after inoculation are put into culturing room
Culture, culture room temperature are 22-24 DEG C, relative air humidity 65%-70%；It first has to culturing room's shading making bacteria-producing room base
This dark.When mycelium covers with charge level (indoor culture 3~5 days), it is transferred to illumination cultivation.It inspects periodically after inoculation, picks in time
Except the culture vessel for growing potential difference, infecting miscellaneous bacteria.It hereafter is induced synthesis coremium period, keeping culture room temperature is 20-22
DEG C, relative air humidity 65%~70%, guarantee has scattering light (100Lx~200Lx) induction coremium to be formed.Culturing room will fit
When ventilation, keep bacterium germination room air it is fresh；
Harvesting and processing: harvesting coremium；
Fluid nutrient medium: SAAY culture medium (yeast extract powder 0.5%, compound amino acid 1%, white granulated sugar 3.5%, water
95%) 3% magnesium citrate, is added；
Solid medium: 7% magnesium citrate is added in wheat broth.
Its cicada fungus coremium magnesium element content cultivated is 300.11mg/100g, has reached Chinese Soclety of Nutrition and has recommended to occupy
The reference value (NRV) of the daily ingestion of magnesium of the people；Compared with Amway calcium and magnesium piece, content of magnesium improves 152%；
In addition, other effective components also significantly improve in the rich magnesium cicada fungus, if total amino acid content is 18.81g/100g, compared with
NRV improves 2.7%；(i.e. N6- (2- ethoxy) adenosine also known as cocoon lavendustin, are the endemic elements of cordyceps sinensis, have been used as cordyceps sinensis HEA
One of quality control index of product) content be 0.078%, compared with NRV improve 56%.
Embodiment 2
Inclined-plane culture, Liquid Culture, solid culture method and minimal medium are with embodiment 1, but culture medium has following change
It is dynamic:
Fluid nutrient medium: 3% magnesium acetate is added in SAAY culture medium；
Solid medium: 7.5% magnesium acetate is added in corn culture medium；
Its cicada fungus coremium magnesium element content cultivated is 157.01mg/100g.
Embodiment 3
Inclined-plane culture, Liquid Culture, solid culture method and minimal medium are with embodiment 1, but culture medium has following change
It is dynamic:
Fluid nutrient medium: 3% magnesium chloride is added in SAAY culture medium；
Solid medium: 8% magnesium chloride is added in barley culture medium；
Its cicada fungus coremium magnesium element content cultivated is 163.21mg/100g.
Embodiment 4
Inclined-plane culture, Liquid Culture, solid culture method and minimal medium are with embodiment 1, but culture medium has following change
It is dynamic:
Fluid nutrient medium: 3% magnesium sulfate is added in SAAY culture medium；
Solid medium: 9% magnesium sulfate is added in soya bean culture medium；
Its cicada fungus coremium magnesium element content cultivated is 125.26mg/100g.
Embodiment 5
Inclined-plane culture, Liquid Culture, solid culture method and minimal medium are with embodiment 1, but culture medium has following change
It is dynamic:
Fluid nutrient medium: 3% magnesium citrate is added in SAAY culture medium；
Solid medium: 10% magnesium citrate is added in buckwheat culture medium；
Its cicada fungus coremium magnesium element content cultivated is 265.66mg/100g.
Embodiment 6
Inclined-plane culture, Liquid Culture, solid culture method and minimal medium are with embodiment 1, but culture medium has following change
It is dynamic:
Fluid nutrient medium: magnesium citrate is not added in SAAY culture medium；
Solid medium: 9% magnesium citrate is added in wheat broth；
Its cicada fungus coremium magnesium element content cultivated is 295.34mg/100g.
Embodiment 7
Inclined-plane culture, Liquid Culture, solid culture method and minimal medium are with embodiment 1, but culture medium has following change
It is dynamic:
Fluid nutrient medium: magnesium citrate is not added in SAAY culture medium；
Solid medium: 9.5% magnesium citrate is added in corn culture medium；
Its cicada fungus coremium magnesium element content cultivated is 275.17mg/100g.
Embodiment 8
Inclined-plane culture, Liquid Culture, solid culture method and minimal medium are with embodiment 1, but culture medium has following change
It is dynamic:
Fluid nutrient medium: 1% magnesium citrate is added in SAAY culture medium；
Solid medium: 10% magnesium citrate is added in barley culture medium；
Its cicada fungus coremium magnesium element content cultivated is 225.62mg/100g.
Embodiment 9
Inclined-plane culture, Liquid Culture, solid culture method and minimal medium are with embodiment 1, but culture medium has following change
It is dynamic:
Fluid nutrient medium: 1% magnesium citrate is added in SAAY culture medium；
Solid medium: 10.5% magnesium citrate is added in soya bean culture medium；
Its cicada fungus coremium magnesium element content cultivated is 225.34mg/100g.
Embodiment 10
Inclined-plane culture, Liquid Culture, solid culture method and minimal medium are with embodiment 1, but culture medium has following change
It is dynamic:
Fluid nutrient medium: 1.5% magnesium citrate is added in SAAY culture medium；
Solid medium: 11% magnesium citrate is added in buckwheat culture medium；
Its cicada fungus coremium magnesium element content cultivated is 295.63mg/100g.
Embodiment 11
Inclined-plane culture, Liquid Culture, solid culture method and minimal medium are with embodiment 1, but culture medium has following change
It is dynamic:
Fluid nutrient medium: 1.5% lemon is added in yeast extract powder 3%, white granulated sugar 3%, soy bean protein hydrolysate 0.5%
Sour magnesium；
Solid medium: 11% magnesium citrate is added in buckwheat culture medium；
Its cicada fungus coremium magnesium element content cultivated is 320.53mg/100g.
Embodiment 12
Inclined-plane culture, Liquid Culture, solid culture method and minimal medium are with embodiment 1, but culture medium has following change
It is dynamic:
Fluid nutrient medium: 3% magnesium sulfate is added in wheat bran liquor 4%, white granulated sugar 3%；
Solid medium: 9% magnesium sulfate is added in wheat broth；
Its cicada fungus coremium magnesium element content cultivated is 315.63mg/100g.
Embodiment 13
Inclined-plane culture, Liquid Culture, solid culture method and minimal medium are with embodiment 1, but culture medium has following change
It is dynamic:
Fluid nutrient medium: 1% magnesium citrate is added in PSB culture medium (potato 20%, glucose 2%)；
Solid medium: 10% magnesium citrate is added in wheat broth；
Its cicada fungus coremium magnesium element content cultivated is 300.16mg/100g.
Embodiment 14
Subtropical zone and torrid areas low lying areas on the south China the Changjiang river, are adopted July according to the method for general Chinese medicine
Collection.It is aseptically, the fresh cordyceps sinensis for adopting next is clean with aseptic water washing on super mirror workbench, it is placed in sterilized
Diameter is interior sclerotium part (polypide) the absorbent cotton package drenched of cordyceps sinensis, with moisturizing in the culture dish of 15cm.The son of cordyceps sinensis
The glass slide of a sterilizing is placed below entity, aweto sclerotium part is paved with small slide so that can pregnant part not contact directly
Glass slide, distance about 0.5cm or so.Then culture dish is placed in 20 DEG C ± 0.5 DEG C of illumination box culture, to cicada fungus
Spore launches when on glass slide, and it is a little to be added dropwise the PDA culture medium just dissolved around spore, removes slide, is placed in another go out
Moisturizing culture in the culture dish of bacterium turns another plate (SDAY culture medium) training with a small amount of mycelia of transfer needle picking after growing mycelia
It supports (ibid), until the bacterium colony for growing single purifying.And verified through morphology or molecular biology method, which is
Periostracum cicadae (Isaria cicadae Miquel) phorozoon.
Comparative examples
Inclined-plane culture, Liquid Culture, solid culture method and minimal medium are with embodiment 1, but culture medium has following change
It is dynamic:
Fluid nutrient medium: SAAY culture medium；It is added without magnesium citrate；
Solid medium: wheat broth is added without magnesium citrate.
Its cicada fungus coremium magnesium element content cultivated is 59.2mg/100g.
Claims (6)
1. a kind of artificial culture method of rich magnesium cicada fungus, this method comprises the following steps:
Step 1, Cordyceps cicadae strain is expanded in liquid medium；
Step 2, the strain after step 1 amplification is inoculated into solid medium and carries out solid culture；
Step 3, coremium is harvested；
It is characterized in that, magnesium citrate is added in the solid medium described in step 2, additional amount is 7~15%；The solid
Culture medium is wheat broth or buckwheat culture medium.
2. the method as described in claim 1, which is characterized in that the additional amount of magnesium salts is 7~11% in solid medium.
3. the method as described in claim 1, which is characterized in that magnesium citrate is added in the fluid nutrient medium described in step 1, adds
Entering amount is 1~3%.
4. the method as described in claim 1, which is characterized in that step 2 inoculum concentration is 15%~30%.
5. the method as described in claim 1, which is characterized in that during step 2 solid culture, adopted in the mycelial growth stage
With shading culture；It originates to harvest stages from coremium using illumination cultivation.
6. such as the rich magnesium cicada fungus of any the method culture of Claims 1 to 5.
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