CN108633717A - A kind of selenium-enriched broccoli soilless culture nutrient fluid formula and its cultural method - Google Patents
A kind of selenium-enriched broccoli soilless culture nutrient fluid formula and its cultural method Download PDFInfo
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Abstract
The invention discloses a kind of selenium-enriched broccoli soilless culture methods, include the following steps：1) yeast selenium is prepared；2) yeast selenium prepared is obtained into hydrolyzate by enzyme hydrolysis broken wall；3) hydrolyzate is centrifuged to obtain supernatant, and nutrient matrix is configured using supernatant；4) nutrient matrix is obtained into selenium-enriched broccoli for broccoli soilless culture.The specific formula of the nutrient solution includes：Potassium nitrate, calcium nitrate, ammonium nitrate, calcium superphosphate, boric acid, EDTA chelated irons, EDTA chelated zincs, yeast selenium broken wall supernatant and water.The present invention by inorganic selenium by saccharomycete culture by way of be converted into Organic Selenium, and cultivate broccoli by the way that yeast selenium is made into nutrient solution, to effectively improve the transformation efficiency of selenium, and selenoprotein is converted by effective selenium ingredient absorbed by the body by plant absorption.
Description
Technical field
The invention belongs to technical fields, and in particular to a kind of to make what matrix and nutrient solution were cultivated using selenium-rich substance
Method.
Background technology
Se-rich farm products, that is, contain the agricultural product of abundant selenium element, and selenium can improve human immunity, promote the increasing of lymphocyte
It grows and the synthesis of antibody and immunoglobulin.Such as selenium-rich rice, selenium-rich agaric, 100 microgram selenium-rich corn powder etc. all belong to
Se-rich farm products.Selenium is a kind of chemical element, and chemical symbol is Se, a kind of nonmetallic.It may be used as light-sensitive material, electrolytic manganese row
The beneficial nutrient etc. of nutrient and plant necessary to industry catalyst, animal body.Selenium is also the essential trace elements of the human body,
Selenium participates in a variety of selenoenzymes and Selenonic protein in synthesized human.Its Glutathione Peroxidase, is catalyzed hydrogen in vivo
Peroxide or lipid peroxide are changed into water or various alcohols, eliminate attack of the free radical to biomembrane, protect biomembrane
From oxidative damage；Selenium participates in constituting the de- iodine enzyme of iodate thyroid gland amino acid.Therefore selenium is referred to as body trace element by scientist
In " king of anti-cancer ".Selenium is divided into as two kinds of vegetable active selenium and inorganic selenium, and inorganic selenium refers generally to sodium selenite and sodium selenate, packet
The remaining yeast selenium of a large amount of inorganic seleniums, malt selenium have been included, has been obtained from the byproduct of metal mineral reserves, inorganic selenium has larger poison
Property, and be not easy to be absorbed, be not suitable for humans and animals and use.Vegetable active selenium is combined by bioconversion and amino acid, and one
As in the form of selenomethionine exist, vegetable active selenium is the selenium source that human and animal allows to use.
And soilless culture, refer to making the matrix of plant root with the media such as water, turf or forest leaf mould, vermiculite to fix and plant
Strain, root system of plant can be in direct contact the cultural method of nutrient solution.Nutrient composition is easily controllable in soilless culture, and can adjust at any time
Section.In illumination, proper temperature without the place of soil, such as desert, seabeach, desert island, as long as having a certain amount of water supply, just
Can carry out.Soilless culture is divided into water planting, mist (gas) training and substrate culture according to the difference of cultivation medium.Water planting refers to root system of plant
It is directly contacted with nutrient solution, does not have to the cultural method of matrix.Earliest water planting is to immerse root system of plant in nutrient solution to grow, this
Kind mode will appear anoxia phenomenon, cause root system dead when serious.Frequently with the water planting mode of nutritional liquid membrane method, though one layer very
Thin nutrition liquid layer, constantly cycle flow through crop root, have not only ensured to be continuing to supply crop water and nutrient, but also be continuing to supply root system
Fresh oxygen.
Present selenium-enriched food is usually using the Organic Selenium that can be absorbed by the body, and directly edible inorganic selenium harm to the human body
It is larger.And existing Organic Selenium usually uses yeast selenium, also there are many products of yeast selenium on the market.But existing technology production
Yeast selenium product because there is yeast selenium thicker cell wall, internal active ingredient to be unable to get preferable utilization, lead
Cause absorption less, effect is smaller.If direct food yeast selenium, a large amount of auxiliary material of addition in need is original to offset yeast selenium
Stench taste, improve mouthfeel.And the selenium content in existing yeast selenium food is less, and it is directly metabolized quilt in human body
The amount that absorption of human body utilizes is less.
Invention content
In order to solve the problems, such as that Se content of the existing technology is less and absorption of human body is less efficient, the present invention provides one
Kind by by yeast selenium broken wall and as nutrient solution into the method for obtaining selenium-enriched broccoli product by row crop soilless culture.
The technical solution adopted in the present invention is：A kind of selenium-enriched broccoli soilless culture method, includes the following steps：
S1. yeast selenium is prepared；
S2. the yeast selenium prepared is obtained into hydrolyzate by enzyme hydrolysis broken wall；
S3. hydrolyzate is centrifuged to obtain supernatant, and nutrient matrix is configured using supernatant；
S4. nutrient matrix is obtained into selenium-enriched broccoli for broccoli soilless culture.
Wherein, Se-enriched yeast is a kind of organic selenium source that developed using yeast, it is by Se accumulation in growth ferment
The interior production of female cell protein structure, Se-enriched yeast is proved safe and stable more than inorganic selenium, easy and absorbs, is effectively and few dirty
Dye, and there are various healthcare functions, therefore the effect in Animal nutrition was gradually recognized by people in recent years, in herding
The application of industry and fish production is increasingly extensive, the using effect of Se-enriched yeast.That is, by being added with nothing in culture medium
Machine selenium simultaneously allows Yeast Growth breeding absorption selenium and combination to form selenoprotein, then by enzyme hydrolysis broken wall release selenoprotein, and by
Broccoli absorbs obtains selenium-enriched broccoli to be enriched in broccoli.It is absorbable with human body that this, which is selenium in broccoli,
The form of Organic Selenium exists, and relatively stable, it is not easy to aoxidized, compared to the prior art in directly using yeast selenium as having
The product for imitating substance supplement selenium element, has preferable assimilation effect.
Further, the step S1 is as follows：
(1.1) barms high to element sulphur demand are filtered out first；
(1.2) yeast-leavened culture medium is prepared, and selenium-rich culturing base is obtained added with sodium selenite into culture medium；
(1.3) barms screened are accessed into progress fermented and cultured in selenium-rich culturing base and obtains yeast fermentation mixing
Liquid；
(1.4) yeast selenium by the yeast separation in yeast fermentation mixed liquor and is dried to obtain by centrifugal treating.
Wherein, because the property of selenium is similar to the property of sulphur, the higher strain of element sulphur demand is carried out by screening
The absorption culture of selenium has preferable assimilation effect compared to existing yeast selenium training method.
Further, the formula of the culture medium in the step (1.2) is as follows：Including distiller's wort in parts by weight
30-45 parts of waste water, without 8-14 parts of amino yeast nitrogen, 14-28 parts of glucose, 15-26 parts of peptone and sodium selenite 10-25
Part, and agar is added thereto and forms semisolid culturemedium.Wherein, come using distiller's wort waste water as main nutriment
It is cultivated, the dosage therein by adjusting sodium selenite is so as to ensureing the absorption efficiency highest of saccharomycete.Because one
Denier sodium selenite additive amount is excessive, can cause saccharomycete Mortality, and the relatively few selenium for then causing saccharomycete to absorb of additive amount compared with
It is few.
Further, the pH value in culture medium is adjusted into 4.3-4.8 in the step (1.3), and in 25-27
It is cultivated in the environment of DEG C and obtains within 4-6 days yeast fermentation mixed liquor.
Further, the step S2 is as follows：
(2.1) add deionized water into the yeast selenium finished product prepared and be made into bacteria suspension；
(2.2) hydrolyzate of bacteria suspension volume 10-14% is added into bacteria suspension and mixes standing hydrolysis；
(2.3) will stand hydrolysis goes the mixed liquor placement after wall to wait for centrifugal treating.
Further, wherein the hydrolyzate includes 10-16 parts of sodium hydroxide in parts by weight, lauryl sodium sulfate
11-15 parts, 'beta '-mannase 27-35,10-15 parts of buffer solution, 19-24 parts of dextranase, 45-59 parts of sodium chloride and deionization
1500-1800 parts of water.
The cell wall outer layer of yeast is the mannosan of phosphorylation, and then internal is mannosan, then in mannosan
Western part is protein layer and glucan layer, and wherein mannosan can carry out itself hydrolysis by the 'beta '-mannase of addition, and
It, being capable of hydroglucan layer when exposing glucan layer after the hydrolysis of mannosan layer by dextranase.And dodecyl sulphate
Sodium is a kind of emulsifier, the substance in hydrolyzate can be made to be distributed more uniform.But reaction is hydrolyzed simply by addition enzyme,
It is incomplete to be easy to cause cell wall hydrolysis, causes internal protein that can not be released effectively, therefore is used for by being added with sodium chloride
Increase the osmotic pressure between cell wall, to enable the glycan not being completely dissolved to rupture so that internal selenoprotein can
It is dropped out from the package of cell wall, to obtain effective selenium ingredient.By centrifugal treating, enable the cell wall residue of peeling
Vessel bottom is sunk to, the supernatant for then drawing upper layer can be obtained the higher effective selenium ingredient of content.
Further, wherein the matrix in the step S3 includes 40-65 parts of organic substrate, inorganic base in parts by weight
8-10 parts of 44-56 parts of matter and broken wall supernatant.
Further, the organic substrate includes stalk clast, sawdust and rice bran.
Further, the inorganic matrix includes the grains of sand, vermiculite, perlite and the peat that diameter is less than 2mm.
A kind of soilless culture nutrient fluid formula of the centrifuged supernatant as adding ingredient in the S3 using step, specifically matches
Side includes calculating in parts by weight：10-15 parts of potassium nitrate, 17-21 parts of calcium nitrate, 11-15 parts of ammonium nitrate, calcium superphosphate 52-58
Part, 1-3 parts of boric acid, 2-6 parts of EDTA chelated irons, 3-5 parts of EDTA chelated zincs, 150-180 parts of yeast selenium broken wall supernatant, water
2000-3000 parts.
Beneficial effects of the present invention are：
(1) present invention can improve the assimilation effect to selenium, phase by filtering out the barms high to element sulphur demand
Compared with existing cultivation method for saccharomycete, while it is a large amount of dead that saccharomycete can be avoided to occur in the environment after sodium selenite is added
The case where dying；
(2) present invention by inorganic selenium by saccharomycete culture by way of be converted into Organic Selenium, and by by yeast selenium
Nutrient solution is made into cultivate broccoli, to effectively improve the transformation efficiency of selenium, and is converted selenoprotein by plant absorption
For effective selenium ingredient absorbed by the body；
(3) present invention is hydrolyzed by using special hydrolyzate, can effectively discharge and be present in inside saccharomycete
Selenoprotein active ingredient, avoid the problem that direct food yeast selenium can not be absorbed by the body and occur.
Specific implementation mode
Embodiment 1：
The present embodiment provides a kind of selenium-enriched broccoli soilless culture methods, mainly convert existing inorganic selenium to
Organic Selenium, and Organic Selenium release culture broccoli can be directly absorbed by the body and to be converted into the stability added
Selenium-rich food.
It is as follows：
1, first, intermediate product yeast selenium is prepared.
Yeast selenium is exactly to be added selenium element during culture yeasts, and selenium has been absorbed and utilized in when yeast growth, make selenium with
Protein and polysaccharide organic in yeast, which combine, is converted into Biological Selenium, to eliminate chemical selenium (such as sodium selenite) to human body
Toxicity and stomach stimulation, enable that selenium is more efficient, be more safely absorbed by the body utilization.Se-enriched yeast is also
Only domestic selenium-replenishing preparation most efficient, most safe, nutrition is most balanced.
Barms, the strain high to sulphur demand obtained here by spontaneons screening are chosen, then all saccharomycete are put
It sets and is cultivated in special culture medium, obtain a large amount of barms.The culture medium containing selenium is prepared again, is added into culture medium
Selenium-rich culturing base is obtained added with sodium selenite；The barms screened are accessed progress fermented and cultured in selenium-rich culturing base to obtain
To yeast fermentation mixed liquor.By the yeast separation in yeast fermentation mixed liquor and it is dried to obtain yeast selenium by centrifugal treating.
Wherein, because the property of selenium is similar to the property of sulphur, the higher strain of element sulphur demand is carried out by screening
The absorption culture of selenium has preferable assimilation effect compared to existing yeast selenium training method.The selenium-rich culturing base
Formula is as follows：Including in parts by weight 37 parts of distiller's wort waste water, without 11 parts of amino yeast nitrogen, 19 parts of glucose, egg
18 parts of 21 parts of white peptone and sodium selenite, and agar is added thereto and forms semisolid culturemedium.Using distiller's wort waste water as
Main nutriment is cultivated, and the dosage therein by adjusting sodium selenite is so as to ensureing the absorption of saccharomycete
Efficiency highest.Because once sodium selenite additive amount is excessive, saccharomycete Mortality can be caused, and additive amount relatively then leads to ferment less
The selenium that female bacterium absorbs is less.PH value in culture medium is adjusted into 4.3-4.8, and is cultivated in the environment of 25-27 DEG C
Obtain within 4-6 days yeast fermentation mixed liquor.
2, broken wall is hydrolyzed to the yeast selenium prepared
Hydrolyzate include 15 parts of sodium hydroxide in parts by weight, 14 parts of lauryl sodium sulfate, 'beta '-mannase 35,
1700 parts of 11 parts of buffer solution, 20 parts of dextranase, 48 parts of sodium chloride and deionized water.
The cell wall outer layer of yeast is the mannosan of phosphorylation, and then internal is mannosan, then in mannosan
Western part is protein layer and glucan layer, and wherein mannosan can carry out itself hydrolysis by the 'beta '-mannase of addition, and
It, being capable of hydroglucan layer when exposing glucan layer after the hydrolysis of mannosan layer by dextranase.
Lauryl sodium sulfate is a kind of emulsifier, the substance in hydrolyzate can be made to be distributed more uniform.But it is only logical
It crosses addition enzyme and reaction is hydrolyzed, be easy to cause cell wall hydrolysis not exclusively, cause internal protein that can not be released effectively, therefore
By being used for increasing the osmotic pressure between cell wall added with sodium chloride, to enable the glycan not being completely dissolved to rupture,
Internal selenoprotein is dropped out from the package of cell wall, to obtain effective selenium ingredient.By centrifugal treating, make
The cell wall residue of peeling can sink to vessel bottom, and then it is higher effectively to can be obtained content for the supernatant on absorption upper layer
Selenium ingredient.
Add deionized water into the yeast selenium finished product prepared and is made into bacteria suspension；Bacteria suspension volume is added into bacteria suspension
14% hydrolyzate simultaneously mixes standing hydrolysis；Hydrolysis, which will be stood, goes the mixed liquor placement after wall to wait for centrifugal treating, wherein when hydrolyzing
Between be maintained within 8h.
3, using the solution after hydrolysis carry out configuration soilless culture substrate and nutrient solution carry out broccoli plantation, wait for plant at
It can be obtained selenium-rich broccoli product after ripe.
Embodiment 2：
The present embodiment is that restriction is optimized on the basis of above-described embodiment 1, the broccoli soilless culture substrate
Preparation method is as follows：
The clast for obtaining 40-60 mesh that in stalk, sawdust and rice bran addition crusher, will be sieved after broken mixes organic the most
Matrix；The grains of sand, vermiculite, perlite and the peat mixing chosen again, wherein the content of each component is identical or close, and will mix
Material after conjunction, which is put into ball mill, to be ground, and the material after grinding is used as by the particle that 12 mesh screens obtain inorganic
Matrix.By organic substrate and inorganic matrix according to 4:5 ratio is mixed, and the supernatant further taken out is added, the supernatant
Be 1 with the mass ratio of matrix:10.
The broccoli seedling of laboratory culture is transplanted in ready Medium Culture, and according to the soilless culture of normal broccoli
Fertilizer and water management step in method carries out, wherein the nutrient solution prescription is：Including calculate in parts by weight 12 parts of potassium nitrate,
19 parts of calcium nitrate, 12 parts of ammonium nitrate, 57 parts of calcium superphosphate, 2 parts of boric acid, 3 parts of EDTA chelated irons, 4 parts of EDTA chelated zincs, yeast selenium
3000 parts of 150 parts of broken wall supernatant and water.
Trickle irrigation is carried out to the soilless culture broccoli in greenhouse by the way of micro drip-irrigation.
For in above-described embodiment method and formula tested, test experiments group include five groups, respectively to above-mentioned side
Step or formula in method are adjusted, to test the necessity of its step and formula components.
Experimental group 1：
This experimental group provides a kind of selenium-enriched broccoli soilless culture method.
It is as follows：
1, first, intermediate product yeast selenium is prepared.
Barms are chosen, the barms screened, which are accessed progress fermented and cultured in selenium-rich culturing base, obtains yeast
Ferment mixed liquor.By the yeast separation in yeast fermentation mixed liquor and it is dried to obtain yeast selenium by centrifugal treating.
The formula of selenium-rich culturing base is as follows：Including in parts by weight 37 parts of distiller's wort waste water, without amino yeast nitrogen
18 parts of 11 parts of source, 19 parts of glucose, 21 parts of peptone and sodium selenite, and agar is added thereto and forms semisolid culturemedium.
Cultivated using distiller's wort waste water as main nutriment, the dosage therein by adjusting sodium selenite to
It can ensure the absorption efficiency highest of saccharomycete.Because once sodium selenite additive amount is excessive, saccharomycete Mortality can be caused,
And additive amount is less compared with few selenium for then causing saccharomycete to absorb.By the control of culture environment temperature between 28-29 DEG C, then cultivate
It is spare that yeast selenium finished product is formed after 5-6 days.
2, yeast selenium is dissolved into aqueous solution and is used for configuring soilless culture substrate and nutrient solution progress broccoli plantation, wait planting
It can be obtained selenium-rich broccoli product after object maturation.
Experimental group 2：
This experimental group provides a kind of selenium-enriched broccoli soilless culture method.
It is as follows：
1, first, intermediate product yeast selenium is prepared.
Barms, the strain high to sulphur demand obtained here by spontaneons screening are chosen, then all saccharomycete are put
It sets and is cultivated in special culture medium, obtain a large amount of barms.The culture medium containing selenium is prepared again, is added into culture medium
Selenium-rich culturing base is obtained added with sodium selenite；The barms screened are accessed progress fermented and cultured in selenium-rich culturing base to obtain
To yeast fermentation mixed liquor.By the yeast separation in yeast fermentation mixed liquor and it is dried to obtain yeast selenium by centrifugal treating.
Wherein, because the property of selenium is similar to the property of sulphur, the higher strain of element sulphur demand is carried out by screening
The absorption culture of selenium has preferable assimilation effect compared to existing yeast selenium training method.The selenium-rich culturing base
Formula is as follows：Including in parts by weight 37 parts of distiller's wort waste water, without 11 parts of amino yeast nitrogen, 19 parts of glucose, egg
18 parts of 21 parts of white peptone and sodium selenite, and agar is added thereto and forms semisolid culturemedium.Using distiller's wort waste water as
Main nutriment is cultivated, and the dosage therein by adjusting sodium selenite is so as to ensureing the absorption of saccharomycete
Efficiency highest.Because once sodium selenite additive amount is excessive, saccharomycete Mortality can be caused, and additive amount relatively then leads to ferment less
The selenium that female bacterium absorbs is less.PH value in culture medium is adjusted into 4.3-4.8, and is cultivated in the environment of 25-27 DEG C
Obtain within 4-6 days yeast fermentation mixed liquor.
2, broken wall is hydrolyzed to the yeast selenium prepared
Hydrolyzate include 15 parts of sodium hydroxide in parts by weight, 14 parts of lauryl sodium sulfate, 'beta '-mannase 35,
1700 parts of 11 parts of buffer solution, 20 parts of dextranase and deionized water.
By centrifugal treating, the cell wall residue of peeling is enable to sink to vessel bottom, then draws the supernatant on upper layer
Liquid can be obtained the higher effective selenium ingredient of content.
Add deionized water into the yeast selenium finished product prepared and is made into bacteria suspension；Bacteria suspension volume is added into bacteria suspension
14% hydrolyzate simultaneously mixes standing hydrolysis；Hydrolysis, which will be stood, goes the mixed liquor placement after wall to wait for centrifugal treating, wherein when hydrolyzing
Between be maintained within 8h.
3, using the solution after hydrolysis carry out configuration soilless culture substrate and nutrient solution carry out broccoli plantation, wait for plant at
It can be obtained selenium-rich broccoli product after ripe.
Experimental group 3：
This experimental group provides a kind of selenium-enriched broccoli soilless culture method.
It is as follows：
1, first, intermediate product yeast selenium is prepared.
Barms, the strain high to sulphur demand obtained here by spontaneons screening are chosen, then all saccharomycete are put
It sets and is cultivated in special culture medium, obtain a large amount of barms.The culture medium containing selenium is prepared again, is added into culture medium
Selenium-rich culturing base is obtained added with sodium selenite；The barms screened are accessed progress fermented and cultured in selenium-rich culturing base to obtain
To yeast fermentation mixed liquor.By the yeast separation in yeast fermentation mixed liquor and it is dried to obtain yeast selenium by centrifugal treating.
Wherein, because the property of selenium is similar to the property of sulphur, the higher strain of element sulphur demand is carried out by screening
The absorption culture of selenium has preferable assimilation effect compared to existing yeast selenium training method.The selenium-rich culturing base
Formula is as follows：Including in parts by weight 37 parts of distiller's wort waste water, without 11 parts of amino yeast nitrogen, 19 parts of glucose, egg
18 parts of 21 parts of white peptone and sodium selenite, and agar is added thereto and forms semisolid culturemedium.Using distiller's wort waste water as
Main nutriment is cultivated, and the dosage therein by adjusting sodium selenite is so as to ensureing the absorption of saccharomycete
Efficiency highest.Because once sodium selenite additive amount is excessive, saccharomycete Mortality can be caused, and additive amount relatively then leads to ferment less
The selenium that female bacterium absorbs is less.PH value in culture medium is adjusted into 4.3-4.8, and is cultivated in the environment of 25-27 DEG C
Obtain within 4-6 days yeast fermentation mixed liquor.
2, broken wall is hydrolyzed to the yeast selenium prepared
Hydrolyzate include 15 parts of sodium hydroxide in parts by weight, 14 parts of lauryl sodium sulfate, 'beta '-mannase 35,
1700 parts of 11 parts of buffer solution, 48 parts of sodium chloride and deionized water.
By centrifugal treating, the cell wall residue of peeling is enable to sink to vessel bottom, then draws the supernatant on upper layer
Liquid can be obtained the higher effective selenium ingredient of content.
Add deionized water into the yeast selenium finished product prepared and is made into bacteria suspension；Bacteria suspension volume is added into bacteria suspension
14% hydrolyzate simultaneously mixes standing hydrolysis；Hydrolysis, which will be stood, goes the mixed liquor placement after wall to wait for centrifugal treating, wherein when hydrolyzing
Between be maintained within 8h.
3, using the solution after hydrolysis carry out configuration soilless culture substrate and nutrient solution carry out broccoli plantation, wait for plant at
It can be obtained selenium-rich broccoli product after ripe.
Experimental group 4：
This experimental group provides a kind of selenium-enriched broccoli soilless culture method.
It is as follows：
1, first, intermediate product yeast selenium is prepared.
Barms, the strain high to sulphur demand obtained here by spontaneons screening are chosen, then all saccharomycete are put
It sets and is cultivated in special culture medium, obtain a large amount of barms.The culture medium containing selenium is prepared again, is added into culture medium
Selenium-rich culturing base is obtained added with sodium selenite；The barms screened are accessed progress fermented and cultured in selenium-rich culturing base to obtain
To yeast fermentation mixed liquor.By the yeast separation in yeast fermentation mixed liquor and it is dried to obtain yeast selenium by centrifugal treating.
Wherein, because the property of selenium is similar to the property of sulphur, the higher strain of element sulphur demand is carried out by screening
The absorption culture of selenium has preferable assimilation effect compared to existing yeast selenium training method.The selenium-rich culturing base
Formula is as follows：Including in parts by weight 37 parts of distiller's wort waste water, without 11 parts of amino yeast nitrogen, 19 parts of glucose, egg
18 parts of 21 parts of white peptone and sodium selenite, and agar is added thereto and forms semisolid culturemedium.Using distiller's wort waste water as
Main nutriment is cultivated, and the dosage therein by adjusting sodium selenite is so as to ensureing the absorption of saccharomycete
Efficiency highest.Because once sodium selenite additive amount is excessive, saccharomycete Mortality can be caused, and additive amount relatively then leads to ferment less
The selenium that female bacterium absorbs is less.PH value in culture medium is adjusted into 4.3-4.8, and is cultivated in the environment of 25-27 DEG C
Obtain within 4-6 days yeast fermentation mixed liquor.
2, broken wall is hydrolyzed to the yeast selenium prepared
Hydrolyzate includes that 15 parts of sodium hydroxide in parts by weight, 14 parts of lauryl sodium sulfate, 11 parts of buffer solution, Portugal are poly-
1700 parts of 20 parts of carbohydrase, 48 parts of sodium chloride and deionized water.
By centrifugal treating, the cell wall residue of peeling is enable to sink to vessel bottom, then draws the supernatant on upper layer
Liquid can be obtained the higher effective selenium ingredient of content.
Add deionized water into the yeast selenium finished product prepared and is made into bacteria suspension；Bacteria suspension volume is added into bacteria suspension
14% hydrolyzate simultaneously mixes standing hydrolysis；Hydrolysis, which will be stood, goes the mixed liquor placement after wall to wait for centrifugal treating, wherein when hydrolyzing
Between be maintained within 8h.
3, using the solution after hydrolysis carry out configuration soilless culture substrate and nutrient solution carry out broccoli plantation, wait for plant at
It can be obtained selenium-rich broccoli product after ripe.
Experimental group 5：
1, using the yeast selenium product bought in the market
2, broken wall is hydrolyzed to the yeast selenium prepared
Hydrolyzate include 15 parts of sodium hydroxide in parts by weight, 14 parts of lauryl sodium sulfate, 'beta '-mannase 35,
1700 parts of 11 parts of buffer solution, 20 parts of dextranase, 48 parts of sodium chloride and deionized water.
By centrifugal treating, the cell wall residue of peeling is enable to sink to vessel bottom, then draws the supernatant on upper layer
Liquid can be obtained the higher effective selenium ingredient of content.
Add deionized water into the yeast selenium finished product prepared and is made into bacteria suspension；Bacteria suspension volume is added into bacteria suspension
14% hydrolyzate simultaneously mixes standing hydrolysis；Hydrolysis, which will be stood, goes the mixed liquor placement after wall to wait for centrifugal treating, wherein when hydrolyzing
Between be maintained within 8h.
3, using the solution after hydrolysis carry out configuration soilless culture substrate and nutrient solution carry out broccoli plantation, wait for plant at
It can be obtained selenium-rich broccoli product after ripe.
Further include one group of control group and standard group, the standard group is obtained according to the standard method and formula of embodiment 2
Selenium-rich broccoli product, the control group is existing yeast selenium food in the market.Mainly test above-mentioned all experimental groups and
The Se content of control group, including total Se content, organic selenium content and inorganic Se content.Each experimental group and control group respectively take 20 parts
Sample, the broccoli in experimental group samples 6-9g, and grinds squeezing and obtain juice, and juice is collected by filtration, and is washed with distillation
It washs filter residue 2-3 times, the distilled water after washing is added in filtrate, the filtrate of mixing is finally obtained.It is used in combination carbon tetrachloride to carry out more
Secondary extraction, extract liquor is mixed with cleaning solution and is concentrated, and it is 1 that volume ratio, which is added,:The nitration mixture of 3 nitric acid and hydrochloric acid composition
15ml carries out sour digestion and to clarifying, and constant volume is spare to 50ml after cooling.
And the measurement of total selenium is equally to take sample 6-9g, and it is 1 to be directly added into volume ratio:3 nitric acid is formed with hydrochloric acid
Nitration mixture 15ml carry out acid digestion and to clarifying, it is cooling after constant volume it is spare as the digestive juice of total selenium to 50ml.
And above two digestive juice 2ml is taken, the EDTA solution of the 0.1mol/L of 1ml, 10% salt of 1ml are added thereto
Then sour hydroxylamine solution uses concentrated ammonia liquor to adjust pH value, the 0.1%DAN solution of 4ml is added, carbon tetrachloride extraction is used after cooling
The measurement liquid for obtaining inorganic selenium and total selenium is measured, and prepares the standard curve of standard selenium solution, then will be measured liquid and be existed
In the range of 480-600nm to above-mentioned organic phase carry out fluorescence emission spectrum second dervative differential scanning, wavelength width 3nm,
Record spectrogram.Reading the second derivative values d at 516nm and 526nm can be asked by the difference (d516-d526) of two-value from standard curve
Selenium content, the difference of the content of total selenium and inorganic selenium is Organic Selenium.
Testing result is as follows：
| Total selenium (μ g/g) | Inorganic selenium (μ g/g) | Organic Selenium (μ g/g) | |
| Experimental group 1 | 2.18 | 0.81 | 1.37 |
| Experimental group 2 | 3.41 | 0.21 | 3.20 |
| Experimental group 3 | 2.84 | 0.52 | 2.32 |
| Experimental group 4 | 2.45 | 0.41 | 2.04 |
| Experimental group 5 | 5.14 | 1.78 | 3.36 |
| Control group | 1.48 | 0.45 | 1.03 |
| Standard group | 8.47 | 1.48 | 6.99 |
Wherein, it can be obtained by upper table data, standard group is the west orchid planted using the method and formula of embodiment 2
Style sheet, it is higher with higher Se content, including organic selenium content, and there is preferable stability.And experimental group 1 is
Yeast selenium obtained is passed through to hydrolysing step and just directly configures nutrient solution and matrix progress soilless culture, then causes yeast selenium
In effective selenoprotein can not be absorbed by plants so that total Se content of this group of broccoli declines more；And in water in experimental group 2
The sodium chloride in hydrolyzate is removed when solution, no sodium chloride only can not completely carry out broken wall by two kinds of enzyme hydrolysis, also lead
Total Se content is caused to decline more, but the more normal broccoli of content of wherein Organic Selenium still has a distinct increment；And experimental group 3 is
Dextranase is removed, experimental group 4 is to remove 'beta '-mannase, and the cell wall for also resulting in yeast selenium can not complete water
Solution declines so as to cause total Se content；And to be the directly yeast selenium bought on the market be hydrolyzed system to experimental group 5 as raw material
Standby, then can be seen that causes the total selenium of last broccoli also to have decline without targetedly culture yeasts selenium.
Finally according to testing result it is found that using the selenium-rich broccoli of the method culture in embodiment 2 with higher completely
Se content, while organic selenium content therein is higher, is convenient for absorption of human body, compared to direct food yeast selenium food, mouth
Sense and absorptivity similarly improve.
Invention is not limited to above-mentioned optional embodiment, anyone can show that other are various under the inspiration of the present invention
The product of form.Above-mentioned specific implementation mode should not be understood the limitation of pairs of protection scope of the present invention, protection model of the invention
It encloses to be subject to and be defined in claims, and specification can be used for interpreting the claims.
Claims (10)
1. a kind of selenium-enriched broccoli soilless culture method, it is characterised in that：Include the following steps：
S1. yeast selenium is prepared；
S2. the yeast selenium prepared is obtained into hydrolyzate by enzyme hydrolysis broken wall；
S3. hydrolyzate is centrifuged to obtain supernatant, and nutrient matrix is configured using supernatant；
S4. nutrient matrix is obtained into selenium-enriched broccoli for broccoli soilless culture.
2. a kind of selenium-enriched broccoli soilless culture method according to claim 1, it is characterised in that：The step S1
It is as follows：
(1.1) barms high to element sulphur demand are filtered out first；
(1.2) yeast-leavened culture medium is prepared, and selenium-rich culturing base is obtained added with sodium selenite into culture medium；
(1.3) barms screened are accessed into progress fermented and cultured in selenium-rich culturing base and obtains yeast fermentation mixed liquor；
(1.4) yeast selenium by the yeast separation in yeast fermentation mixed liquor and is dried to obtain by centrifugal treating.
3. a kind of selenium-enriched broccoli soilless culture method according to claim 2, it is characterised in that：The step
(1.2) formula of the culture medium in is as follows：Including in parts by weight 30-45 parts of distiller's wort waste water, without amino yeast nitrogen
10-25 parts of source 8-14 parts, 14-28 parts of glucose, 15-26 parts of peptone and sodium selenite, and agar is added thereto and forms half
Solid medium.
4. a kind of selenium-enriched broccoli soilless culture method according to claim 2, it is characterised in that：The step
(1.3) pH value in culture medium is adjusted into 4.3-4.8 in, and is cultivated 4-6 days in the environment of 25-27 DEG C and obtains ferment
Mother's fermentation mixed liquor.
5. a kind of selenium-enriched broccoli soilless culture method according to claim 2, it is characterised in that：The step S2
It is as follows：
(2.1) add deionized water into the yeast selenium finished product prepared and be made into bacteria suspension；
(2.2) hydrolyzate of bacteria suspension volume 10-14% is added into bacteria suspension and mixes standing hydrolysis；
(2.3) will stand hydrolysis goes the mixed liquor placement after wall to wait for centrifugal treating.
6. a kind of selenium-enriched broccoli soilless culture method according to claim 5, it is characterised in that：The wherein described water
Solution liquid include 10-16 parts of sodium hydroxide in parts by weight, 11-15 parts of lauryl sodium sulfate, 'beta '-mannase 27-35,
1500-1800 parts of 10-15 parts of buffer solution, 19-24 parts of dextranase, 45-59 parts of sodium chloride and deionized water.
7. a kind of selenium-enriched broccoli soilless culture method according to claim 1, it is characterised in that：The wherein described step
Matrix in rapid S3 includes 44-56 parts of 40-65 parts of organic substrate, inorganic matrix and broken wall supernatant 8-10 in parts by weight
Part.
8. a kind of selenium-enriched broccoli soilless culture method according to claim 7, it is characterised in that：The organic group
Matter includes stalk clast, sawdust and rice bran.
9. a kind of selenium-enriched broccoli soilless culture method according to claim 7, it is characterised in that：The inorganic base
Matter includes the grains of sand, vermiculite, perlite and the peat that diameter is less than 2mm.
10. a kind of selenium-enriched broccoli soilless culture nutrient solution prescription, it is characterised in that：Using step in claim 1
For centrifuged supernatant in S3 as adding ingredient, specific formula includes calculating in parts by weight：10-15 parts of potassium nitrate, nitric acid
17-21 parts of calcium, 11-15 parts of ammonium nitrate, 52-58 parts of calcium superphosphate, 1-3 parts of boric acid, 2-6 parts of EDTA chelated irons, EDTA chelated zincs
3-5 parts, 150-180 parts of yeast selenium broken wall supernatant, 2000-3000 parts of water.
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| CN113773149A (en) * | 2021-10-22 | 2021-12-10 | 武汉富田生物科技有限公司 | Novel fertilizer production process for improving selenium enrichment of crops |
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