CN108651164A - A kind of salicylic acid culture medium and preparation method thereof suitable for straw mushroom - Google Patents
A kind of salicylic acid culture medium and preparation method thereof suitable for straw mushroom Download PDFInfo
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- CN108651164A CN108651164A CN201811007142.5A CN201811007142A CN108651164A CN 108651164 A CN108651164 A CN 108651164A CN 201811007142 A CN201811007142 A CN 201811007142A CN 108651164 A CN108651164 A CN 108651164A
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- straw mushroom
- salicylic acid
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/20—Culture media, e.g. compost
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Abstract
The salicylic acid culture medium and preparation method thereof that the invention discloses a kind of suitable for straw mushroom, is related to volvaria volvacea cultivation technology field.The salicylic acid culture medium contains salicylic acid.Using the medium culture straw mushroom, the amino acid content of straw mushroom can be improved by salicylic addition.
Description
Technical field
The present invention relates to volvaria volvacea cultivation technology field, in particular to a kind of salicylic acid culture medium suitable for straw mushroom and
Preparation method.
Background technology
Straw mushroom (Volvariella volvacea (Bull.:Fr.) Sing.) also known as Volvariella volvacen, luxuriant foot mushroom originate from Guangdong
In the Nanhua Temple in Shaoguan, 300 China Nian Qian have started artificial cultivation, and countries in the world are passed to by overseas Chinese in the about thirties in 20th century,
It is a kind of important tropical and subtropical zone mushroom class, is that the third-largest culturing edible fungus, China's straw mushroom yield occupy first of the world in the world, it is main
It is distributed in South China.
For straw mushroom because of its unique flavor, nutritional ingredient is high, medicinal effects are good and has very high nutrition and edible value.
Straw mushroom is a kind of good edible mushroom.The height of protein content is usually as evaluation food value height
Important evidence, straw mushroom have the characteristics that high protein and low fat.Protein content accounts for the 2.66%-5.05% of dry weight in fresh grass mushroom,
Compared with the Vegetables of daily consumption, protein content is 2 times of potato and asparagus, 4 times of tomato and carrot, citrus
6 times.In addition, containing 18 kinds of amino acid in straw mushroom fructification, wherein the required ammonia that cannot be synthesized or convert containing 8 kinds of mankind
Base acid, and the content of this 8 kinds of essential amino acids is higher, accounts for the 38.2% of its total amino acid content.
Often edible straw mushroom is with health role to human body.It is well known that the food that edible vitamin content is high, can promote
Into being normally carried out for human metabolism, enhancing body accelerates the healing of wound, and can prevent bad to the resistivity of infectious disease
The generation of blood disease.Containing a kind of substance being known as heterologous protein in straw mushroom fructification, there is antitumaous effect, the nitrogen leaching contained
The growth of cancer cell can be inhibited by going out object and purine base also.In addition, summer, which eats straw mushroom, has the function of sunstroke prevention heat-clearing.
Straw mushroom growth hobby hot and humid environment, is suitble to cultivate in tropical and subtropical region, the unexpected fluctuation pole of temperature is not
Conducive to the growth and development of straw mushroom, the quality and yield of straw mushroom is greatly influenced.In addition, the growth of straw mushroom also need to it is higher wet
Degree, due to China's North of Yangtze River area, most of the time temperature is relatively low, dry, and the time for adapting to straw mushroom plantation is extremely short, no
Conducive to extensive cultivation, this greatlys restrict the cultivation range of straw mushroom, therefore city is also much not achieved in the current yield of straw mushroom
Requirement, especially in straw mushroom the supply of good protein be not met by the growing demand of consumer.But it closes at present
It is seldom in the research for the protein content for improving straw mushroom.The method for improving protein content existing at present has through addition 30
That there are straw mushroom amino acid contents is relatively low for the existing culture volvaria volvacea cultivation technology of alkanol, vitamin B1, potassium dihydrogen phosphate, mycelia is short
It is small.The shortcomings of density is low, the speed of growth is slow.In addition, it is existing detection straw mushroom in amino acid content method accuracy compared with
It is low, it cannot reflect amino acid actual content in straw mushroom.
In consideration of it, special propose the present invention.
Invention content
The purpose of the present invention is to provide a kind of salicylic acid culture mediums suitable for straw mushroom, using the salicylic acid medium culture
Straw mushroom can improve the amino acid content of straw mushroom.
The purpose of the present invention is to provide the preparation methods of above-mentioned salicylic acid culture medium.
Another object of the present invention is to provide a kind of method using salicylic acid culture straw mushroom, which can carry
The amino acid content of high straw mushroom.
The invention is realized in this way：
On the one hand, the present invention provides a kind of salicylic acid culture medium suitable for straw mushroom, contain：Salicylic acid.
Further, in some embodiments of the present invention, the salicylic acid in the medium a concentration of
0.00001%-0.01%.
For example, concentration of the salicylic acid in straw mushroom medium can be 0.00001%, 0.0001%, 0.0001%,
In 0.001% and 0.01% any one or both between range.
The present invention's the study found that the salicylic acid of addition suitable concentration can improve straw mushroom in the culture medium of culture straw mushroom
Amino acid content.
Further, in some embodiments of the present invention, the salicylic acid culture medium also contains potato water extract, Portugal
Grape sugar, peptone and dregs of beans water extract.
Further, in some embodiments of the present invention, by weight, institute's salicylic acid culture medium contains：
20-30 parts of 18-22 parts of glucose, 4-8 parts of peptone and dregs of beans water extract, the body of potato water extract and glucose
Product mass ratio is 14-16:1.
Compared to existing ordinary culture medium, salicylic acid culture medium can be with by the way that peptone and dregs of beans water extract is added
Make straw mushroom that good hickory chick growth situation be presented, shows the feature that mycelia is sturdy, density is big and the speed of growth is fast.
Further, in some embodiments of the present invention, the dregs of beans water extract is made as follows：
With the decocting cooked beans dregs of rice, filtering takes filtrate up to the dregs of beans water extract.
Further, in some embodiments of the present invention, when decoction, the mass volume ratio of dregs of beans used and water used
It is 1:22-30.
Further, in some embodiments of the present invention, the dregs of beans water extract is made as follows：25g beans
15-20min is boiled in dregs of rice 500mL boilings, and filtering takes filtrate to get dregs of beans water extract.
Further, in some embodiments of the present invention, the potato water extract is made as follows：By potato
It mixes, boils with water, filter, take filtrate, obtain potato water extract.
Further, in some embodiments of the present invention, the potato water extract is prepared via a method which：It weighs
200g potatoes are cut into the fritter for being about 3cm, and 500ml water is added, boils 15-20min；Then it is filtered using gauze, takes filter
Liquid, as above-mentioned potato water extract.
On the other hand, the present invention provides the preparation methods of above-mentioned salicylic acid culture medium comprising：
Salicylic acid is added into basal medium.
Further, in some embodiments of the present invention, salicylic additive amount is 0.00001%-0.01%.
Further, in some embodiments of the present invention, the basal medium by potato water extract, glucose,
Peptone and dregs of beans water extract mix.
Further, in some embodiments of the present invention, salicylic acid is added to basal medium by the following method
In：Salicylic acid first is dissolved with absolute ethyl alcohol, after then using the dilution of the mixed solution of distilled water and ethyl alcohol, is then added to basic culture
Base.
Further, in some embodiments of the present invention, the dregs of beans water extract is made as follows：
With the decocting cooked beans dregs of rice, filtering takes filtrate up to the dregs of beans water extract.
Further, in some embodiments of the present invention, when decoction, the mass volume ratio of dregs of beans used and water used
(g:Ml it is) 1:22-30.
Further, in some embodiments of the present invention, the dregs of beans water extract is made as follows：25g beans
15-20min is boiled in dregs of rice 500mL boilings, and filtering takes filtrate to get dregs of beans water extract.
Further, in some embodiments of the present invention, the potato water extract is made as follows：By potato
It mixes, boils with water, filter, take filtrate, obtain potato water extract.
Further, in some embodiments of the present invention, the mass volume ratio (g of potato used and water used:ml)
It is 1:2-3.
Further, in some embodiments of the present invention, the potato water extract is prepared via a method which：It weighs
200g potatoes are cut into the fritter for being about 3cm, and 500ml water is added, boils 15-20min；Then it is filtered using gauze, takes filter
Liquid, as potato water extract.
On the other hand, the present invention provides a kind of methods using salicylic acid culture straw mushroom comprising：Straw mushroom strain is connect
Kind contains to above-mentioned any one cultivates on salicylic salicylic acid culture medium.
Further, in some embodiments of the present invention, the temperature of straw mushroom culture is 36-38 DEG C.
The cultural method, which uses, contains salicylic medium culture straw mushroom, can improve the amino acid content in straw mushroom.
On the other hand, the present invention also provides a kind of straw mushrooms, are obtained by above-mentioned cultural method culture.
Amino acid containing high level in the straw mushroom.
Description of the drawings
In order to illustrate the technical solution of the embodiments of the present invention more clearly, below will be to needed in the embodiment attached
Figure is briefly described, it should be understood that the following drawings illustrates only certain embodiments of the present invention, therefore is not construed as pair
The restriction of range for those of ordinary skill in the art without creative efforts, can also be according to this
A little attached drawings obtain other relevant attached drawings.
Fig. 1 is by the amino in the Mycelia of Straw Mushroom, Volvariel volvacea turned out using the straw mushroom medium of embodiment 2-5 in experimental example 1
Acid content testing result；
In figure：Ordinate indicates the content (mg/kg) of amino acid, abscissa V1-1, V1-2, V1-3, V1-4, V1-5 difference
Indicate that the concentration of calcium sulfate in culture medium is respectively：0%, 0.5%, 1%, 2%, 5%.
Fig. 2 is by the amino in the Mycelia of Straw Mushroom, Volvariel volvacea turned out using the straw mushroom medium of embodiment 6-9 in experimental example 1
Acid content testing result；
In figure：Ordinate indicates the content (mg/kg) of amino acid, abscissa V2-1, V2-2, V2-3, V2-4, V2-5 difference
Indicate that the compound concentration of magnesium sulfate in culture medium is respectively：0%, 1%, 2.5%, 5%, 7.5%.
Fig. 3 is by the ammonia in the Mycelia of Straw Mushroom, Volvariel volvacea turned out using the straw mushroom medium of embodiment 10-13 in experimental example 1
Base acid content testing result；
In figure：Ordinate indicates the content (mg/kg) of amino acid, abscissa V3-1, V3-2, V3-3, V3-4, V3-5 difference
Indicate that salicylic concentration is respectively in culture medium：0%, 0.00001%, 0.0001%, 0.001%, 0.01%.
Fig. 4 is by the ammonia in the Mycelia of Straw Mushroom, Volvariel volvacea turned out using the straw mushroom medium of embodiment 14-16 in experimental example 1
Base acid content testing result；
In figure：Ordinate indicates that the content (mg/kg) of amino acid, abscissa V4-1, V4-2, V4-3, V4-4 indicate respectively
The concentration of methyl jasmonate is respectively in culture medium：0%, 0.000001%, 0.00001%, 0.0001%.
Fig. 5 is to be directed to the amino acid that same sample obtains using different analysis of amino acids methods in experimental example 2
Content detection result.
Fig. 6 is the chromatogram of amino standard acid solution；In figure：A is the standard diagram of various amino acid；B is standard diagram
Relevant parameter.
Fig. 7 is the mycelia life on the culture medium of common PDA culture medium and the embodiment of the present invention 1 during straw mushroom strain is inoculated with respectively
Long figure；In figure：A is common PDA culture medium, and B is the culture medium of embodiment 1.
Specific implementation mode
It in order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below will be in the embodiment of the present invention
Technical solution be clearly and completely described.The person that is not specified actual conditions in embodiment, builds according to normal condition or manufacturer
The condition of view carries out.Reagents or instruments used without specified manufacturer is the conventional production that can be obtained by commercially available purchase
Product.
The feature and performance of the present invention are described in further detail with reference to embodiments.
Embodiment 1
A kind of straw mushroom medium is present embodiments provided, by weight, contains 200 parts of potato, 20 parts of glucose, egg
25 parts of white 5 parts of peptone, 10 parts of agar and dregs of beans water extract.
Wherein, dregs of beans water extract solution is prepared via a method which：
20min is boiled in the 500mL boilings of 25g dregs of beans, and filtering takes filtrate up to dregs of beans water extract solution, about 300mL is spare.
By taking the straw mushroom medium for preparing 1000ml as an example, the preparation method of straw mushroom medium is illustrated, it is as follows：
200g potatoes are weighed, the fritter for being about 3cm is cut into, 500ml water is added, boils 15-20min；Then gauze is used
It is filtered, takes filtrate, as potato water extract solution.Fetch earth beans water extract solution 300ml, then takes 20g glucose, 5g albumen
Peptone, 25g dregs of beans water extract solution and 10g agar are added in filtrate, add water polishing to 1000mL, pH naturally, high pressure sterilization,
121 DEG C, 20min.
Using the straw mushroom medium culture medium inoculated straw mushroom strain of the present embodiment, as a contrast with common PDA culture medium, see
The growth situation for examining straw mushroom, is as a result shown in Fig. 7, it can be seen that relatively common PDA culture medium (Fig. 7 A), using the culture of embodiment 1
Base can make straw mushroom that good hickory chick growth situation be presented, and show the feature that mycelia is sturdy, density is big and the speed of growth is fast
(Fig. 7 B).
Embodiment 2
Straw mushroom medium provided in this embodiment contains 200 parts of potato, 20 parts of glucose, peptone by weight
5 parts, 10 parts of agar, 25 parts of dregs of beans water extract and mass volume ratio (g:Ml) the calcium sulfate for being 0.5%.
Wherein, by taking the straw mushroom medium for preparing 1000ml as an example, the preparation method of straw mushroom medium is illustrated, such as
Under：
Fetch earth beans water extract solution (preparation method is with embodiment 1) 300ml, 20g glucose, 5g peptones, 25g dregs of beans
Water extract, suitable calcium sulphate dihydrate and 10g agar are added in filtrate, add water polishing to 1000mL, and pH is naturally, high pressure is gone out
Bacterium, 121 DEG C, 20min.Wherein, the additive amount of calcium sulphate dihydrate makes it ensure calcium sulfate in the straw mushroom medium of 1000ml
Final concentration of 0.5%.
Embodiment 3
Straw mushroom medium ingredient provided in this embodiment and preparation method are same as Example 2, the difference is that calcium sulfate
A concentration of 1%.
Embodiment 4
Straw mushroom medium ingredient provided in this embodiment and preparation method are same as Example 2, the difference is that calcium sulfate
A concentration of 2%.
Embodiment 5
Straw mushroom medium ingredient provided in this embodiment and preparation method are same as Example 2, the difference is that calcium sulfate
A concentration of 5%.
Embodiment 6
Straw mushroom medium provided in this embodiment contains 200 parts of potato, 20 parts of glucose, peptone by weight
The magnesium sulfate that 5 parts, 10 parts of agar, 25 parts of dregs of beans water extract and mass concentration are 1%.
The preparation method of the straw mushroom medium replaces calcium sulphate dihydrate with embodiment 2, with epsom salt, controls magnesium sulfate
Final concentration of 1% in straw mushroom medium.
Embodiment 7-9
The straw mushroom medium that embodiment 7-9 is provided is substantially the same manner as Example 7, the difference is that the concentration of magnesium sulfate is different,
The magnesium sulfate concentration for the straw mushroom medium that embodiment 7 provides is the magnesium sulfate concentration for the straw mushroom medium that 2.5, embodiment 8 provides
The magnesium sulfate concentration of the straw mushroom medium provided for 5%, embodiment 9 is 7.5%.
Embodiment 10
Straw mushroom medium (i.e. salicylic acid culture medium) provided in this embodiment, by weight, containing 200 parts of potato,
20 parts of glucose, 5 parts of peptone, 10 parts of agar, 25 parts of dregs of beans water extract and final concentration of 0.00001% (mass volume ratio
(g:Ml salicylic acid)).
By taking the straw mushroom medium for preparing 1000ml as an example, the preparation method of the straw mushroom medium of the present embodiment is said
It is bright, it is as follows：
200g potatoes are weighed, suitable water is added, boils 15-20min；Then it is filtered using gauze, takes filtrate；It takes
20g glucose, 5g peptones, 25g dregs of beans water extract, 0.0001g salicylic acids (salicylic acid needs to be first dissolved in organic solvent herein,
Be then added in culture medium) and 10g agar be added in filtrate, add water polishing to 1000mL, pH is naturally, high pressure sterilization, 121
DEG C, 20min.
Wherein, it is as follows to dissolve salicylic method：
Accurate to weigh 1.0000g salicylic acids, it is 10mL that absolute ethyl alcohol, which is added, and is dissolved to final volume.1mL is drawn with liquid-transfering gun
Above-mentioned lysate 10 is diluted in the distilled water and alcohol mixed solution (V of heating again：V=5:4) in, fully dissolving is (at this point, bigcatkin willow
Acid concentration is 0.01g/mL).According to salicylic additive amount, with 10 times of gradient dilutions in distilled water.Obtain different salicylic acids
The dilution of content.According to salicylic additive amount, the dilution for choosing appropriate dilutions multiple adds it in culture medium.
For example, taking the salicylic acid dilution of a concentration of 0.0001g/10mL of 10mL, being then added to final volume control is
In the culture medium of 1000mL, obtain containing 0.00001% salicylic culture medium.
Embodiment 11
Straw mushroom medium provided in this embodiment contains 200 parts of potato, 20 parts of glucose, peptone by weight
5 parts, 10 parts of agar, 25 parts of dregs of beans water extract and a concentration of 0.0001% salicylic acid.
Embodiment 12
Straw mushroom medium provided in this embodiment contains 200 parts of potato, 20 parts of glucose, peptone by weight
5 parts, 10 parts of agar, 25 parts of dregs of beans water extract and a concentration of 0.001% salicylic acid.
Embodiment 13
Straw mushroom medium provided in this embodiment contains 200 parts of potato, 20 parts of glucose, peptone by weight
5 parts, 10 parts of agar, 25 parts of dregs of beans water extract and a concentration of 0.01% salicylic acid.
Embodiment 14
Straw mushroom medium (i.e. methyl jasmonate culture medium) provided in this embodiment, by weight, contains potato
200 parts, 20 parts of glucose, 5 parts of peptone, 10 parts of agar, 25 parts of dregs of beans water extract and final concentration of 0.000001% (matter
Measure volume ratio (g:Ml methyl jasmonate)).
By taking the straw mushroom medium for preparing 1000ml as an example, the preparation method of the straw mushroom medium of the present embodiment is said
It is bright, it is as follows：
200g potatoes are weighed, suitable water is added, boils 15-20min；Then it is filtered using gauze, takes filtrate；It takes
20g glucose, 5g peptones, 25g dregs of beans water extract, 0.00001g methyl jasmonates (need first to be dissolved in organic solvent, so
After be added in culture medium) and 10g agar be added in filtrate, add water polishing to 1000mL, pH is naturally, high pressure sterilization, 121 DEG C,
20min。
The method for dissolving methyl jasmonate is as follows：
1mL methyl jasmonates are drawn using liquid-transfering gun to be dissolved in 9mL absolute ethyl alcohols, are then diluted to distilled water for 10 times
In, obtain the methyl jasmonate dilution of a concentration of 0.00001g/10mL.Wherein, extension rate is according to being added to culture medium
In jasmonic acid first dosage determine.
For example, the methyl jasmonate dilution of a concentration of 0.00001g/10mL of 10mL is taken all to be added to final volume control
It is made as in the culture medium of 1000mL, both obtains the culture medium containing 0.000001% methyl jasmonate in this way.
Embodiment 15
Straw mushroom medium provided in this embodiment contains 200 parts of potato, 20 parts of glucose, peptone by weight
5 parts, 10 parts of agar, 25 parts of dregs of beans water extract and a concentration of 0.00001% methyl jasmonate.
Embodiment 16
Straw mushroom medium provided in this embodiment contains 200 parts of potato, 20 parts of glucose, peptone by weight
5 parts, 10 parts of agar, 25 parts of dregs of beans water extract and a concentration of 0.0001% methyl jasmonate.
Embodiment 17
Present embodiments provide a kind of method of culture straw mushroom comprising：
The strain of straw mushroom V23 is seeded in embodiment 2-16 in any one straw mushroom medium, 37 DEG C of trainings are subsequently placed in
It supports, you can obtain Mycelia of Straw Mushroom, Volvariel volvacea.
Opposite ordinary culture medium can make straw mushroom that good hickory chick be presented and grow state using the culture medium of embodiment 2-16
Gesture shows the feature that mycelia is sturdy, density is big and the speed of growth is fast.
Experimental example 1
Detection uses the amino acid content in the Mycelia of Straw Mushroom, Volvariel volvacea that the straw mushroom medium of embodiment 2-16 is cultivated.
1 Mycelia of Straw Mushroom, Volvariel volvacea sample collection, method are as follows：
The straw mushroom medium of the embodiment 2-16 for bacterium of having gone out is poured into sterilized and dry culture dish under gnotobasis, is waited for
It is solidified；
Culture medium after solidification is shown that patch last layer is cut and sterilized glassine paper (pays attention to：Glass paper size with
Being slightly less than culture dish is advisable, excessive or too small all unsuitable)；
It is inoculated with straw mushroom V23, each culture medium connects 5 culture dishes to ensure to be collected into enough mycelia later, in 37 DEG C of trainings
It supports and is cultivated 3 days in case；It is taken out when it covers with culture dish completely, scrapes mycelia, and be collected in 5mL centrifuge tubes, deposit in -70
DEG C freezing with spare.
Analysis of amino acids：
The preparation of 2 reagents
Hydrochloric acid solution (6mol/L):500mL hydrochloric acid is taken to be diluted with water to 1000mL, mixing.
Refrigerant:Commercially available salt is mixed with ice cube by quality 1: 3.
Sodium hydroxide solution (500g/L):50g sodium hydroxides are weighed, are dissolved in 50mL water, it is dilute with water after being cooled to room temperature
It releases to 100mL, mixing.
Sodium citrate buffer [c (Na+)=0.2mol/L]:It weighs 19.6g sodium citrates and 500mL water dissolutions is added,
16.5mL hydrochloric acid is added, is diluted with water to 1000mL, mixing is adjusted with 6mol/L hydrochloric acid solutions or 500g/L sodium hydroxide solutions
PH to 2.2.
The elution buffer solution of different pH and ionic strength:It prepares or buys with reference to instrument specification.
3 standard solution are prepared
1) kilnitamin standard reserving solution (1 μm of ol/mL):Single amino acids standard items are accurately weighed respectively (to be accurate to
0.00001g) in same 50mL beakers, is dissolved, is accurately transferred in 250mL volumetric flasks with 8.3mL 6mol/L hydrochloric acid solutions,
It is diluted with water and is settled to scale, mixing, each amino acid standard weighs quality references value and is shown in Table 1.
The weighing quality references value of amino acid standard when table 1 prepares kilnitamin standard reserving solution
2) kilnitamin standard working solution (100nmol/mL):It is accurate to draw kilnitamin standard reserving solution 1.0mL
In 10mL volumetric flasks, pH2.2 sodium citrate buffers is added to be settled to scale, mixing is machine liquid in standard.
4 analyses
1) sample preparation
The Mycelia of Straw Mushroom, Volvariel volvacea of collection step is crushed using tissue pulverizer or grinder, liquor sample is broken into refiner
Homogenate sealing freezen protective, analysis the used time thawed after use.
2) sample weighing
The good sample of uniformity accurately weighs 1g samples (being accurate to 0.0001g) with balance very much, makes protein in sample
Content is within the scope of 10mg~20mg.It, can protein content in first determination sample for the sample that protein content is unknown.It will claim
Measured sample is placed in hydrolysis pipe.
3) sample hydrolyzes
(a) enter the 8mol/L hydrochloric acid of 1mL in hydrolysis pipe, vortex 3min is added the 7mol/L hydrochloric acid of 1mL volumes, is vortexed
2min is supplemented to about 10mL with 6mol/L hydrochloric acid solutions again, and (sample and the mass volume ratio of final volume are 1:10(g/mL)).
It is acid protein hydrolysats matter that the effect of hydrochloric acid is added during analysis of amino acids, passes through the action breaks down albumen of acid
Peptide bond among matter, due to the mycelium that this sample is straw mushroom, which has moisture compared with other dry products or food
Content is higher, the relatively low feature of protein content, and in order to increase the extraction efficiency of amino acid, the present invention is special to select ladder
Spend acid hydrolysis method, the advantages of this method be for straw mushroom mycelia cell wall in the component contents such as chitin and cellulose compared with
Moisture is high in high and sample, therefore first with high concentrated acid cell wall hydrolysis ingredient, protein is discharged, while keeping effective water
The acid concentration for solving peptide bond, improves the extraction efficiency of amino acid.
(b)~4 drop of the drop of phenol 3 is added into hydrolysis pipe.Hydrolysis pipe is put into refrigerant, 3min~5min is freezed, connects
It onto the exhaust tube of vacuum pump, vacuumizes (close to 0Pa), is then charged with nitrogen, after repetition vacuumizes-be filled with nitrogen 3 times, filling
Nitrogen state lower sealing tightens screw cap.
(c) the hydrolysis pipe sealed is placed in 110 DEG C ± 1 DEG C of electric heating air blast insulating box or hydrolysis stove, hydrolyzes 22h
Afterwards, it takes out, is cooled to room temperature.
(d) hydrolysis pipe is opened, hydrolyzate is filtered to 50mL volumetric flasks, hydrolysis pipe, washing are repeatedly rinsed with a small amount of water
Liquid moves into same 50mL volumetric flasks, is finally settled to scale with water, vibrates mixing.
Accurate absorption 1.0mL filtrates are moved into 15mL or 25mL test tubes, with test tube concentrating instrument or Parallel evaporator 45
Be dried under reduced pressure under DEG C heating environment, it is dry after residue 1mL~2mL water dissolutions, then be dried under reduced pressure, be finally evaporated.
(e) it dissolves in test tube after being added to drying with 1.5mL pH2.2 sodium citrate buffers, after vibrating mixing, inhales
Solution is taken, after 0.22 μm of filter membrane, is transferred to instrument sample injection bottle, liquid is measured for sample, for Instrument measuring.
4) it detects
Kilnitamin standard working solution and sample measure liquid and inject amino-acid analyzer respectively with same volume,
(Hitachi L-8900Amino Acid Analyzer (Tokyo, Japan)) by external standard method by peak area in terms of
Calculate the concentration that sample measures amino acid in liquid.
Chromatography reference conditions：
Chromatographic column：Sulfonic acid cation resin；
Detection wavelength：570nm and 440nm；
Standard solution：
Amino Acids Mixture Standard Solution,Type H(lot TWK6209Wako Pure
Chemical Industries,Ltd.)；
Standard solution is prepared：0.400mL amino acid mixed standard solutions are taken to be settled to 10.00mL with water.
Amino-acid analyzer splitter：
4.6mmI.D.×60mm L packed with Hitachi custom ion exchange resin
(Partical size:3 μm), sodium form (product type #2622)；
Deamination column：4.6mm I.D. × 40mm L (product type #2650L)；
Reaction column：4.6mm I.D.x40mm L, built-in inert silicon sand little particle；
Sample size：20μL；
Flow velocity：0.4ml/min；Column temperature：57℃；React flow velocity：0.35ml/min；Reaction temperature：135℃.5) result
It the results are shown in Table 2- tables 5 and Fig. 1-Fig. 4, Fig. 6.
Amino acid content detection in the Mycelia of Straw Mushroom, Volvariel volvacea that table 2 is turned out using the straw mushroom medium of embodiment 2-5 is tied
Fruit (unit：mg/kg)
Amino acid content detection in the Mycelia of Straw Mushroom, Volvariel volvacea that table 3 is turned out using the straw mushroom medium of embodiment 6-9 is tied
Fruit (unit：mg/kg)
Amino acid content in the Mycelia of Straw Mushroom, Volvariel volvacea that table 4 is turned out using the straw mushroom medium of embodiment 10-13 detects
As a result (unit：mg/kg)
Amino acid content in the Mycelia of Straw Mushroom, Volvariel volvacea that table 5 is turned out using the straw mushroom medium of embodiment 14-16 detects
As a result (unit：mg/kg)
Addition calcium sulfate, magnesium sulfate, salicylic acid or methyl jasmonate are can be seen that from table 2- tables 5 and Fig. 1-Fig. 4 results
The amino acid content in Mycelia of Straw Mushroom, Volvariel volvacea can be improved, the methyl jasmonate of 0.0001% concentration, amino acid are especially added
Total amount reaches 33801mg/kg, i.e., contains 33801mg total amino acids per kg Mycelia of Straw Mushroom, Volvariel volvacea, this knot all high compared with other embodiment
Fruit is that the present inventor is unanticipated.
Experimental example 2
Experimental group：The Mycelia of Straw Mushroom, Volvariel volvacea obtained using embodiment 1 analyzes its amino acid content, the analysis method of experimental group
It is essentially identical with the step 2- steps 4 in experimental example 1；
Control group：It is identical as experimental group, the difference is that be handled by following in (a) step in sample hydrolysing step：
Disposably 10mL 6mol/L hydrochloric acid solutions is added to be hydrolyzed in hydrolysis pipe.
As a result see Fig. 5, it can be seen that the amino acid content of experimental group is higher than control group, the ammonia being indicated above in experimental example 1
Base acid content analysis method can be such that the protein in sample more fully dissolves out in sample measurement solution, improve testing result
Accuracy.
The foregoing is only a preferred embodiment of the present invention, is not intended to restrict the invention, for the skill of this field
For art personnel, the invention may be variously modified and varied.All within the spirits and principles of the present invention, any made by repair
Change, equivalent replacement, improvement etc., should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of salicylic acid culture medium suitable for straw mushroom, which is characterized in that it contains：Salicylic acid.
2. salicylic acid culture medium according to claim 1, which is characterized in that the salicylic acid in the medium a concentration of
0.00001%-0.01%.
3. salicylic acid culture medium according to claim 2, which is characterized in that the salicylic acid culture medium also contains potato water
Extract, glucose, peptone and dregs of beans water extract.
4. salicylic acid culture medium according to claim 3, which is characterized in that by weight, the salicylic acid culture medium
Contain：
20-30 parts of 18-22 parts of glucose, 4-8 parts of peptone and dregs of beans water extract, the volume matter of potato water extract and glucose
Amount is than being 14-16:1.
5. salicylic acid culture medium according to claim 4, which is characterized in that the dregs of beans water extract is made as follows
：
With the decocting cooked beans dregs of rice, filtering takes filtrate up to the dregs of beans water extract.
6. salicylic acid culture medium according to claim 5, which is characterized in that when decoction, the matter of dregs of beans used and water used
It is 1 to measure volume ratio:22-30.
7. salicylic acid culture medium according to claim 4, which is characterized in that the potato water extract is made as follows
：Potato is mixed with water, is boiled, is filtered, is taken filtrate, obtain potato water extract.
8. the preparation method of salicylic acid culture medium as described in claim 1, which is characterized in that it includes：
Salicylic acid is added into basal medium.
9. preparation method according to claim 8, which is characterized in that salicylic additive amount is 0.00001%-
0.01%.
10. preparation method according to claim 8 or claim 9, which is characterized in that the basal medium by potato water extract,
Glucose, peptone and dregs of beans water extract mix.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111418443A (en) * | 2020-05-08 | 2020-07-17 | 青海大学 | Application of salicylic acid or proline in high-temperature cultivation of lentinus edodes |
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2018
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111418443A (en) * | 2020-05-08 | 2020-07-17 | 青海大学 | Application of salicylic acid or proline in high-temperature cultivation of lentinus edodes |
| CN111418443B (en) * | 2020-05-08 | 2021-11-23 | 青海大学 | Application of proline in high-temperature cultivation of lentinus edodes |
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Application publication date: 20181016 |