CN107739230A - A kind of Hericium erinaceus compost - Google Patents
A kind of Hericium erinaceus compost Download PDFInfo
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- CN107739230A CN107739230A CN201711193950.0A CN201711193950A CN107739230A CN 107739230 A CN107739230 A CN 107739230A CN 201711193950 A CN201711193950 A CN 201711193950A CN 107739230 A CN107739230 A CN 107739230A
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- hericium erinaceus
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- corncob
- calcium carbonate
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05D—INORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C; FERTILISERS PRODUCING CARBON DIOXIDE
- C05D3/00—Calcareous fertilisers
- C05D3/02—Calcareous fertilisers from limestone, calcium carbonate, calcium hydrate, slaked lime, calcium oxide, waste calcium products
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a kind of Hericium erinaceus compost, and it is made up of the component of following weight percent content：The Hericium erinaceus polyoses content that corncob 30 40%, cotton seed hulls 10 35%, wheat bran 5 10%, rice bran 10 30%, soybean skin 1 5%, megasse 1 4%, the compost of the present invention of calcium carbonate 1 2% cultivate to obtain is high, and biological efficiency is high.
Description
Technical field
The present invention relates to field of edible fungus culture, is cultivated more particularly to a kind of Hericium erinaceus of high yield fruitbody polysaccharide
Material.
Background technology
Hericium erinaceus (Hericium erinaceus Pers.) is the famous food medicine dual-purpose bacterium in China, is under the jurisdiction of basidiomycetes
Door, monkey mushroom section, monkey mushroom category, is distributed in many provinces in China.Hericium erinaceus meat tenderness is delicious, nutritious, there is " mountain delicacy from ancient times
The laudatory title of hedgehog hydnum, seafood delights bird's nest ".Hericium erinaceus has multiple efficacies, can treat neurasthenia, gastritis and gastric ulcer etc., has very
High edible and medical value, has been developed that as relevant pharmaceutical formulations.The chemical composition of Hericium erinaceus have polysaccharide, sterols, terpene,
Aliphatic acid, phenols etc., wherein Hericium erinaceus Polysaccharides are most important active components, have improve immunity, it is antitumor, hypoglycemic,
Anti-oxidant, anti-ageing multiple efficacies of waiting for a long time.Hericium erinaceus Polysaccharides are mainly derived from the mycelium of solid culture and the hedgehog hydnum of cultivation at present
Mushroom entity.Because of the degeneration of the unstable and bacterial strain of planting environment, cause its polyoses content not high and unstable, have impact on product
The effect of and quality, therefore the Hericium erinaceus raw material sources of high-quality are particularly important.The Hericium erinaceus of same bacterial strain is in different cultivations
Caused fruitbody polysaccharide has great difference in matrix, and its corresponding bioactivity difference is also very big.Therefore it is more in Hericium erinaceus
During the development and application of sugar, the compost selection of Hericium erinaceus raw material is critically important.
The content of the invention
It is an object of the invention to provide a kind of Hericium erinaceus compost, its by following weight percent content component group
Into：
Corncob 30-40%, cotton seed hulls 10-35%, wheat bran 5-10%, rice bran 10-30%, soybean skin 1-5%, megasse
1-4%, calcium carbonate 1-2%.
A kind of preferable Hericium erinaceus compost of high yield fruitbody polysaccharide is made up of the component of following weight percents content：
Corncob 39%, cotton seed hulls 10%, wheat bran 10%, rice bran 30%, soybean skin 5%, megasse 4%, calcium carbonate
2%.
Another Hericium erinaceus compost is made up of the component of following weight percents content：
Corncob 34%, cotton seed hulls 30%, wheat bran 6%, rice bran 20%, soybean skin 4%, megasse 4%, calcium carbonate 2%；
Another Hericium erinaceus compost is made up of the component of following weight percents content
Corncob 35%, cotton seed hulls 35%, wheat bran 10%, rice bran 10%, soybean skin 5%, megasse 3%, calcium carbonate
2%.
A kind of Hericium erinaceus compost of the present invention, when preparing, each component is mixed, water is added, stirs, contain it
Water is 60~65%, pack (every bag of weight 600g), normal-pressure sterilization, inoculation, fruiting.
The compost is cultivated for Hericium erinaceus, comprised the following steps that：
The each component of compost is mixed, water is added, stirs, it is 60~65% to make its water content, packs (every bag of weight
600g), normal-pressure sterilization；
The original seed that inoculation is made, cultural hypha (25 DEG C of lucifuge cultures), fructification culture and management (after mycelia purseful,
Cultivating bag is moved into mushroom room, removes the collar and tampon, sack maintains the original state, 15~17 DEG C of indoor temperature of regulation, humidity 85~
95%, below carbon dioxide 800ppm；Daily illumination 5 hours, 50~100lux of intensity；
Later stage fruiting controls 12~14 DEG C of indoor temperature, the 10th day, harvests the first damp mushroom；20th~26 day Repeated Harvesting
Two damp mushrooms.
The Hericium erinaceus polyoses content of harvest is high.
The material of the Hericium erinaceus compost of the present invention is easy to get, cost is low, it is simple to prepare, and can be obtained with its cultivation Hericium erinaceus more
There is more preferable stimulating expression of macrophage to produce NO for the high Hericium erinaceus of sugared content, the Hericium erinaceus polysaccharide for extracting to obtain
Activity, the immunity of body can be strengthened.
Embodiment
The present invention is further illustrated below in conjunction with specific embodiment.
Compost raw material weed tree sawdust used, corncob, cotton seed hulls, corn flour, wheat bran, rice bran, gypsum, calcium carbonate, beet
Slag, soybean skin are ordinary commercial products.
Embodiment 1
A kind of Hericium erinaceus compost, supplementary material used is by following percentage by weight proportioning (%) composition：Corncob 39%, cotton
Seed shell 10%, wheat bran 10%, rice bran 30%, soybean skin 5%, megasse 4%, calcium carbonate 2%.Supplementary material is mixed, adds water
Stirring, its water content is set to be 65%, pH6~7, packed weight in wet base 600g.
Embodiment 2
A kind of Hericium erinaceus compost, supplementary material used is by following percentage by weight proportioning (%) composition：Corncob 34%, cotton
Seed shell 30%, wheat bran 6%, rice bran 20%, soybean skin 4%, megasse 4%, calcium carbonate 2%.Supplementary material is mixed, water is added and stirs
Mix, make its water content be 60%, pH6~7, packed weight in wet base 600g.
Embodiment 3
A kind of Hericium erinaceus compost, supplementary material used is by following percentage by weight proportioning (%) composition：Corncob 35%, cotton
Seed shell 35%, wheat bran 10%, rice bran 10%, soybean skin 5%, megasse 3%, calcium carbonate 2%.Supplementary material is mixed, adds water
Stirring, its water content is set to be 60%, pH6~7, packed weight in wet base 600g.
The culture of the Hericium erinaceus of embodiment 4
Pack：Embodiment 1-3 compost is respectively charged into 15cm*30cm*0.045cm polyethylene plastic bag, every bag
Weight in wet base 600g；
Disinfection inoculation：Autoclaving, 100 DEG C of insulation 60min, 121 DEG C of sterilizing 90min, sterilizes and treats bacterium bag center after terminating
Temperature is cooled to less than 24 DEG C, is inoculated with.Artificial infection (kind monkey mushroom 0605, is edible mushroom research institute of Shanghai City academy of agricultural sciences
Preservation), it is inoculated with using liquid spawn, inoculum concentration 3% (v/v) left and right；
Cultigen culture：24~26 DEG C of bacteria developing period cultivation temperature, lucifuge culture, less than 24 DEG C, faint scattering light easily goes out original
Base.Culture environment condition is：First 3-5 days are setting date, and adjustment culturing room room temperature is 25~26 DEG C, and gas concentration lwevel is less than
4000ppm, relative air humidity is below 65%, dark culturing；Then enter hot stage, culturing room's room temperature is adjusted to 24~25
DEG C, while increase indoor circulation, room gas concentration lwevel is no more than 3000ppm, and humid control is 65% or so, cultivation cycle
13~15d；
Cultivation management：1st day, remove lid, mushroom room temperature is adjusted to 15-18 DEG C, increase interior circulation, dark training
Support；The 2-5 days, initially form small former base.14-15 DEG C of indoor temperature of control, humidity 90-95%, below carbon dioxide 800ppm,
Daily illumination 5 hours, intensity 50-100lux；The 6-8 days, stimulate fruit-body formation.Control 12-14 DEG C of indoor temperature, humidity
85-90%, below carbon dioxide 800ppm, daily illumination 5 hours, intensity 50-100lux；9th day, control indoor temperature 12-
14 DEG C, humidity 80-85%, below carbon dioxide 800ppm；10th day, mushroom body was substantially considerable, solid, and color is white, and bacteria thorn length exists
1.3-1.5 centimetres, spore is not launched, now timely collecting.20th~26 day damp mushroom of Repeated Harvesting second.
The assay of the Hericium erinaceus polysaccharide of embodiment 5
It is prepared by sample prepare liquid：The embodiment 1-3 Hericium erinaceus cultivated is respectively dried, smashes, weighs 0.5g samples
Product, do parallel twice, are accurate to 0.001g, are placed in 50ml tool plug centrifuge tubes.Sample is infiltrated with 5ml pure water, is slowly added to
20ml absolute ethyl alcohols；Shaken up using vortex oscillator vibration, be well mixed sample, be placed in excusing from death extractor and have children outside the state plan extraction
30min；After extraction terminates, 10min, abandoning supernatant are centrifuged in 13000r/min；Precipitation is molten with the ethanol solutions of 10ml 80%
Solution, centrifuges, abandons supernatant again；Precipitation is transferred to centrifuge tube with water, 50ml distilled water is added, is added using constant temperature oscillation instrument
Heat.100 DEG C of design temperature, time setting 2h；Room temperature is cooled to, 10min is centrifuged in 13000r/min, by supernatant plus water constant volume
Into 100ml volumetric flasks, this solution is measure liquid.
Polysaccharide determination：Phend-sulphuric acid determines polyoses content.Measure liquid is diluted 5 times, 1ml is taken in test tube, adds respectively
Enter 0.5ml phenol solutions, mix, add the 2.5ml concentrated sulfuric acids (98%), mix, 100 DEG C of heating water bath 30min.Fully reaction
Absorbance is determined at 490nm afterwards, the polyoses content of sample is calculated using glucan as standard items.
The Hericium erinaceus that embodiment 1-3 compost obtains is carried out to the analysis of polysaccharide as stated above.
The Hericium erinaceus polysaccharide average content of embodiment 1 is 3.28%；The Hericium erinaceus polysaccharide of embodiment 2 is put down
Equal content is 2.21%；The Hericium erinaceus polysaccharide average content of embodiment 3 is 2.07%.
The cultivated fruitbody polysaccharide stimulated in vitro macrophage of embodiment 6 discharges NO determination of activity
1st, preparation of samples：
Weigh the Hericium erinaceus that embodiment 1-3 planting material cultures obtain and dry fructification 0.5g samples, be accurate to 0.001g, put
Have in 50ml in plug centrifuge tube.Sample is infiltrated with 5ml pure water, is slowly added to 20ml absolute ethyl alcohols.Vibrated using vortex oscillator
Shake up, be well mixed sample, be placed in excusing from death extraction 30min in excusing from death extractor.Extraction terminate after, in 11000r/min from
Heart 25min, abandoning supernatant.Precipitation 10ml80% ethanol solutions dissolve, and centrifuge again, abandon supernatant.Precipitation is transferred to water
Centrifuge tube, 50ml distilled water is added, is heated using constant temperature oscillation instrument.100 DEG C of design temperature, time setting 2h.It is cooled to
Room temperature, filtering, supernatant is moved into bag filter, concentrates, is freeze-dried after flowing water dialysis 48h.Obtained sample will be freeze-dried
5mg/ml mother liquor is configured to PBS solution, 12000 × g centrifuges 30min, supernatant is transferred in sterile tube in aseptic operating platform,
It is diluted to 0.5mg/mL, 2mg/mL, 5mg/mL (final concentration of 50, the 200 and 500 μ g/mL of effect) successively according to concentration gradient.This
For sample prepare liquid.
2nd, cell culture：Take the logarithm the RAW264.7 macrophages strain (being purchased from cell institute of the Chinese Academy of Sciences) in growth period, use DMEM
Complete medium (being purchased from Gibco companies) at 37 DEG C, containing 5%CO2 under the conditions of Secondary Culture, with 0.05% pancreatin or 5%EDTA
Solution digestion, suspension 1000rpm/min collect cell after centrifuging 3min, counted standby.
3rd, preparation of reagents and Specification Curve of Increasing
The preparation of Griess reagents：6.25ml H3PO3 are added in beaker, distilled water 250ml is added, is separately added into
2.5g sulfanilamide (4- aminobenzene sulfonamides, Sigma companies) and 0.25g naphthylethylenediamine
Dihydrochloride (hydrochloride naphthodiamide, Sigma companies) magnetic stirring apparatus to whole dissolvings, 4 DEG C of ice of brown reagent bottle
Case preserves.
Specification Curve of Increasing：Be made into the sodium nitrite solution of various concentrations, concentration gradient 0,5,10,15,20,25,30,
35th, 40 μM totally nine；100 μ L are taken to add 50 μ L Griess reagents in 96 orifice bores, determine 543nm light absorption values, standard curve
3 repetitions of each concentration, standard curve is drawn according to light absorption value.
4th, the measure of sample stimulus macrophage release NO amounts：RAW264.7 cells are collected, are trained with colourless RPMI1640
Support base (being purchased from Gibco companies) (antibiotic liquid of 10% hyclone+1%) and cell is diluted to 5 × 105/ mL,
Add 96 orifice plates, per hole add 180 μ L, then add 20 μ L testing samples so that sample final concentration be followed successively by 500 μ g/ml,
200 μ g/ml and 50 μ g/ml, positive control are LPS (final concentration of 1 μ g/mL), and negative control PBS, 37 DEG C are cultivated 48 hours.
100 μ L of supernatant are taken to add 50 μ l Griess reagents, room temperature incubates bath 10 minutes, determines 543nm light absorption values in 96 orifice bores.According to
Standard curve calculates cell NO burst size.
Experimental result：
The polysaccharide of the Hericium erinaceus extraction of acquisition is cultivated by embodiment 1 in 50 μ g/mL of concentration, stimulating expression of macrophage produces NO
Amount (unit：μm oL/5.0 × 105cells) be：19.12；In 200 μ g/mL of concentration, it is 23.14 to produce NO amounts；In concentration
During 500 μ g/mL, it is 25.17 to produce NO amounts；
For the polysaccharide for the Hericium erinaceus extraction that the cultivation of embodiment 2 obtains in 50 μ g/mL of concentration, stimulating expression of macrophage produces NO's
Measure (unit：μm oL/5.0 × 105cells) be：16.15；In 200 μ g/mL of concentration, it is 21.03 to produce NO amounts；In concentration
During 500 μ g/mL, it is 22.19 to produce NO amounts；
For the Hericium erinaceus that the cultivation of embodiment 3 obtains when in 50 μ g/mL of concentration, stimulating expression of macrophage produces NO amount
(unit：μm oL/5.0 × 105cells) be：12.37；In 200 μ g/mL of concentration, it is 14.92 to produce NO amounts；In the μ of concentration 500
During g/mL, it is 20.15 to produce NO amounts；
Negative control PBS 7.42, positive control LPS (the μ g/mL of concentration 1) 24.15.It can be seen that embodiment 1-3 is planted
The Hericium erinaceus polysaccharide obtained is trained, the activity that there is more preferable stimulating expression of macrophage to produce NO, the immune of body can be strengthened
Power.
Claims (2)
1. a kind of Hericium erinaceus compost, it is characterised in that it is made up of the component of following weight percent content：
Corncob 30-40%, cotton seed hulls 10-35%, wheat bran 5-10%, rice bran 10-30%, soybean skin 1-5%, megasse 1-
4%th, calcium carbonate 1-2%.
2. Hericium erinaceus compost according to claim 1, its by following weight percents content groups be grouped into：
Corncob 39%, cotton seed hulls 10%, wheat bran 10%, rice bran 30%, soybean skin 5%, megasse 4%, calcium carbonate 2%.
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| CN201711193950.0A CN107739230B (en) | 2017-11-24 | 2017-11-24 | Method for improving hericium erinaceus polysaccharide yield and enhancing activity of macrophage in producing NO |
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