CN109913444A - It is a kind of using the method for Uv-induced screening selenium enriched Spirulina mutant strain and its bacterial strain and the application of acquisition - Google Patents
It is a kind of using the method for Uv-induced screening selenium enriched Spirulina mutant strain and its bacterial strain and the application of acquisition Download PDFInfo
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Abstract
The present invention relates to a kind of method using Uv-induced screening selenium enriched Spirulina mutant strain and its bacterial strain and the applications of acquisition, are related to breeding and the screening technique field of spirulina.The present invention is that spirulina is broken into unicellular or two cells using ultrasonic wave, then is allowed to mutate under conditions of ultraviolet mutagenesis, obtains the spirulina mutant strain of enduring high-concentration sodium selenite.The present invention calculates lethality using counting method, and compared to traditional solid plate rubbing method, this method is easy to operate, the period is short.Obtained selenium enriched Spirulina mutant strain is strong to the accumulation ability of selenium, industrially has biggish application prospect in selenium-supply.
Description
Technical field
The present invention relates to the breedings of spirulina and screening technique field, and in particular to a kind of preparation side of selenium enriched Spirulina
Method, it is more particularly related to a kind of method and its acquisition using Uv-induced screening selenium enriched Spirulina mutant strain
Bacterial strain and application.
Background technique
Selenium is the essential trace elements of the human body, positioned at the activated centre of a variety of antioxidases, has important physiological function
With extensive pharmacological action.Spirulina Industrialized processing technique is mature, has stronger tolerance to inorganic selenium, is given birth to selenium
The ideal carrier that object organises, thus obtain possess the spirulina of selenium-enriched ability have for the selenium-supply industry in China it is very heavy
The meaning wanted.
Selenium is mainly functioned in the form of selenoprotein in human body, such as maintain body redox equilibrium, improve immunity,
Release heavy metal toxicity etc..Three distinguishing features are presented in China's selenium resource distribution: first is that selenium resources reserve amount is huge, total storage level
Account for 30% or more the whole world；Second is that selenium resource distribution is extremely uneven, about 70% national territorial area belongs to selenium deficiency area；Third is that selenium deficiency people
Mouth substantial amounts, the 72% of Zhan Quanguo population.Selenium is closely related with human health, and selenium deficiency will lead to Keshan disease, Kaschin-Beck disease
Occur.The diseases such as cardiovascular disease, diabetes, cancer also often occur with selenium deficiency together.Therefore rationally using selenium resource to solve
The serious selenium deficiency status in China is one of the significant problem of current national urgent need to resolve.
Selenium in environment is mostly inorganic selenium, and inorganic selenium toxicity is high, human body utilization rate is poor, and in contrast organic selenium is that have safely
The selenium-supply form of effect, organic selenium refer to selenium and protein, carbohydrate and lipid binding at organic compound.Microalgae is tellurian
Carbon dioxide and water can be converted to organic matter by photosynthesis by primary growth person, and spirulina is at present can large-scale farming
One of microalgae, growth cycle is short, the speed of growth is fast, selenoprotein is abundant.
CN1085951 discloses a kind of Tech. for culturing spirulina rich in selenium, and culture, sieve are tamed first in the culture solution of low selenium concentration
The frond for adapting to such culture solution is selected, the screening for carrying out selenium-enriched algae in high selenium concentration culture solution is subsequently placed in, filters out
Algae strain continuously cultivated under identical selenium concentration, or by the algae selected strain be placed in selenium concentration be 700-1000ppm culture solution
In, incubation time 2-24h carries out concentration culture, obtains selenium-rich algal gel and is washed off extra inorganic selenium with clear water, and PH=is arrived in washing
7, it is dried at being 60-80 DEG C in temperature or spray drying becomes selenium-rich algae powder.But the spirulina after concentration is placed in by this method
In superelevation selenium concentration (700-1000ppm) culture solution, its quick selenium-rich in short time (2-24h) is allowed.Firstly, 700-
The selenium concentration working solution of 1000ppm, Se content is very high, is toxic for normal spirulina, secondly, spirulina is by selenium
In absorber and to complete bioconversion be a very long biological process, it is difficult to be completed in 2h, therefore the invention is by spiral
Algae is placed in short time stirring in toxic selenium environment and selenium-enriched algae powder is just made, and selenium-rich effect remains to be confirmed.
CN101715986A discloses a kind of preparation method and application of rich selenium haematococcus powder, step: A, screening inorganic
Selenium tolerance is strong, selenoprotein and content astaxanthin are high, fast growing haematococcus pluvialis new lines；B, certain temperature, pH value,
Under illumination, cultivated using haematococcus culture medium；C, according to haematococcus pluvialis kind to the difference of selenium tolerance, using inoculation
Directly add inorganic selenium in culture medium or different amounts of inorganic selenium is added after frustule reaches certain biomass, compounding high concentration contains
Seleno culture medium or low concentration contain seleno culture medium；D, it after frustule is continuously cultivated, is harvested by the method for filtering or being centrifuged, and with clearly
Water rinses fresh algal gel, washes away the inorganic selenium on frustule surface.Drying, spray drying or freeze-drying obtain the raw red ball of selenium-rich rain
Algae powder.For the selenium-rich haematococcus pluvialis algae powder selenium compound content of preparation between 10-2000 μ g/g, organic selenium content reaches 99%
More than.Rich selenium haematococcus product can be applied in preparing food, health food and drug.But the haematococcus pluvialis that the invention uses
It is unicellular eukaryote, growth conditions requirement is harsher, needs aseptically to cultivate, therefore yield is very low, mostly
Number enterprise produces astaxanthin with the algae；And spirulina is the prokaryotes of many cells, the speed of growth is very fast, can be in outdoor big rule
Mould culture.
Based on the above reasons, the application is proposed.
Summary of the invention
The purpose of the present invention is to provide a kind of method using Uv-induced screening selenium enriched Spirulina mutant strain and its obtain
The bacterial strain obtained and application.The present invention screens the spirulina of 1 plant height selenium-rich by the method for domestication and ultraviolet mutagenesis, can be high
Inorganic selenium is converted to organic selenium by effect, this crosses selenium-supply industrial expansion to me and has a very important significance.
To reach said one purpose of the present invention, The technical solution adopted by the invention is as follows:
A method of using Uv-induced screening selenium enriched Spirulina mutant strain, described method includes following steps:
(1) blunt top spirulina is taken, is first cultivated in the culture medium of low concentration sodium selenite, then be successively transferred to concentration successively
In the culture medium of incremental high concentration sodium selenite, until obtaining the blunt top spirulina of tolerance 470mg/L sodium selenite；
(2) by the blunt top spirulina ultrasonication of tolerance 470mg/L sodium selenite obtained by step (1) at unicellular, two thin
Born of the same parents or tricellular algae solution；
(3) precipitating is collected after centrifugation in algae solution obtained by step (2), and is diluted precipitating with culture medium；
(4) algae solution after step (3) dilution is loaded on planar surface, is incident upon lethality > 90% with ultraviolet lighting, then
It is transferred under dark condition, overnight；
(5) collection step (4) overnight after algae solution, centrifugation, by the culture of the supreme concentration of sodium selenite of gained precipitating transfer
It is cultivated in base, the cell survived is selenium enriched Spirulina mutant strain (algae).
Further, above-mentioned technical proposal, the concentration of step (1) the low concentration sodium selenite culture solution is 280~
360mg/L。
Further, above-mentioned technical proposal, step (2) the ultrasonication power are 5~30W, preferably 20W；Ultrasound
The broken time is 20~80s, preferably 20s.
Further, above-mentioned technical proposal, step (4) the ultraviolet light time are 20~60s, preferably 40s.
Further, above-mentioned technical proposal, step (4) described lethality preferably use microscopic counting to count.
Further, above-mentioned technical proposal, the concentration of step (5) the high concentration sodium selenite culture solution is 600~
1500mg/L。
Second object of the present invention is to provide the selenium enriched Spirulina described above filtered out using ultraviolet mutagenesis method
Mutant strain.
Third object of the present invention is to provide the selenium enriched Spirulina described above filtered out using ultraviolet mutagenesis method
The application of mutant strain can be used for preparing selenium enriched Spirulina powder.
A kind of selenium enriched Spirulina powder, is prepared with the following method:
By selenium enriched Spirulina mutant strain 28 DEG C, PH=10, intensity of illumination be 40 μ EM-2·S-1Under the conditions of, it utilizes
Zarrouk culture medium is cultivated；The Na of 1000mg/L is added in Initial stage of culture2SeO3, cultivate 2 weeks, be collected by centrifugation, freezing is true
Sky is dry, and the selenium enriched Spirulina powder is made.
Further, above-mentioned technical proposal, the Zarrouk culture medium main component includes: 13.61g/LNaHCO3,
4.03g/L Na2CO3, 0.5g/LK2HPO4, 1g/LK2SO4, 1g/L NaCl, 0.2g/L MgSO4·7H2O, 2.5g/L NaNO3,
0.04g/L CaCl2·2H2O, 0.01g/L FeSO4·7H2O, 1ml/L microelement A5Solution.
Compared with prior art, a kind of method using Uv-induced screening selenium enriched Spirulina mutant strain of the present invention
And its application has the following beneficial effects:
(1) spirulina is broken into unicellular or two cells using ultrasonic wave by the present invention, then in the condition of ultraviolet mutagenesis
Under be allowed to mutate, obtain the spirulina mutant strain of enduring high-concentration sodium selenite.The present invention calculates cause using counting method
Dead rate, compared to traditional solid plate rubbing method, this method is easy to operate, the period is short.Obtained selenium enriched Spirulina mutant strain
It is strong to the accumulation ability of selenium, industrially there is biggish application prospect in selenium-supply.
(2) principal element that ultraviolet mutagenesis is normally implemented be to ensure that lethality within the scope of suitable, it is lethal
Rate is too low, and mutation effect is not achieved, and lethality is too high, and the cell number that can be survived is very little.Spirulina is the concatenated life of many cells
Object, not on how to calculating the report of spirulina lethality in current document.The present invention is by by the spirulina of many cells
It is broken into unicellular and two cells, is the normal of ultraviolet mutagenesis with the lethality of the method statistic number spirulina of microscopic count
Implement to provide basis.
Detailed description of the invention
Fig. 1 is the microscope photo of the spirulina after 1~16 different condition ultrasonication of the embodiment of the present invention；
Fig. 2 is the OD value comparison diagram of the spirulina after 1~16 different condition ultrasonication of the embodiment of the present invention；
Fig. 3 is that chlorophyll method, OD value method, microscopic counting lethality calculated result is respectively adopted in the embodiment of the present invention 20
Comparison diagram.
Specific embodiment
Below with reference to case study on implementation and attached drawing, invention is further described in detail.The implementation case is with skill of the present invention
Implemented under premised on art, provides detailed embodiment and specific operating process now to illustrate that the present invention has and create
Property, but protection scope of the present invention case study on implementation not limited to the following.
The information for including according to the application, to those skilled in the art can be easily to essence of the invention
Really description carries out various changes, without departing from spirit and scope of the appended claims.It should be understood that the scope of the present invention is not
Process, property defined by being confined to or component, because these embodiments and other descriptions are just for the sake of schematic
Illustrate certain aspects of the present disclosure.In fact, this field or those skilled in the relevant art obviously can be to embodiment party of the present invention
The various changes that formula is made all cover within the scope of the appended claims.
It is not intended to limit the scope of the invention for a better understanding of the present invention, expression dosage used in this application,
All numbers of percentage and other numerical value, are understood to be modified with word " about " in all cases.Therefore,
Unless stated otherwise, otherwise digital parameters listed in specification and appended book are all approximations, may
It can be changed according to the difference for the desirable properties for attempting to obtain.Each digital parameters at least should be considered as according to being reported
Effective digital and obtained by the conventional method of rounding up.
Embodiment 1
A kind of method using Uv-induced screening selenium enriched Spirulina mutant strain of the present embodiment, the method includes as follows
Step:
One, artificial screening
Blunt top spirulina Spirulina platensis (FACHB-882 comes from Chinese Academy of Sciences's fresh water algae library) is taken, is first existed
Cultivated in culture medium containing 360mg/L sodium selenite, then be successively transferred to containing 370,380,390,400,405,410,
415, it 420,425,430,435,440,445,450,455,460,465, in the culture medium of 470mg/L sodium selenite, obtains resistance to
Ultraviolet mutagenesis is carried out by the spirulina of 470mg/L sodium selenite, and as material.
Two, Uv-induced screening
(1) the resulting blunt top spirulina of OD=1.0 artificial screening is taken, with BRANSON ultrasonic cell disruption instrument (model
450D), 1/8 probe, under the conditions of P (ultrasonic power)=40% (20W), ultrasonication 20s.
(2) broken algae solution is centrifuged 5min, revolving speed 1000g.
(3) precipitating is collected, and is diluted to OD=0.3 with culture medium.
(4) take 4ml algae solution loaded on plate, ultraviolet irradiation 40s makes spirulina lethality > 90%, and lethality is using micro-
Counting method calculates.
(5) under dark condition, overnight.
(6) it collects algae solution and is centrifuged 5min, revolving speed 2500g.
(7) gained precipitating is transferred in the culture medium of 1000mg/L concentration of sodium selenite, the cell survived is
Resistance to selenium spirulina algae.
Embodiment 2
The method using Uv-induced screening selenium enriched Spirulina mutant strain of the present embodiment is substantially the same manner as Example 1, area
Be not only that: in Uv-induced screening step (1) under the conditions of ultrasonic power P=40% (20W), the ultrasonication time is 40s.
Embodiment 3
The method using Uv-induced screening selenium enriched Spirulina mutant strain of the present embodiment is substantially the same manner as Example 1, area
Be not only that: in Uv-induced screening step (1) under the conditions of ultrasonic power P=40% (20W), the ultrasonication time is 60s.
Embodiment 4
The method using Uv-induced screening selenium enriched Spirulina mutant strain of the present embodiment is substantially the same manner as Example 1, area
Be not only that: in Uv-induced screening step (1) under the conditions of ultrasonic power P=40% (20W), the ultrasonication time is 80s.
Embodiment 5
The method using Uv-induced screening selenium enriched Spirulina mutant strain of the present embodiment is substantially the same manner as Example 1, area
Be not only that: in Uv-induced screening step (1) under the conditions of ultrasonic power P=30% (15W), the ultrasonication time is 20s.
Embodiment 6
The method using Uv-induced screening selenium enriched Spirulina mutant strain of the present embodiment is substantially the same manner as Example 1, area
Be not only that: in Uv-induced screening step (1) under the conditions of ultrasonic power P=30% (15W), the ultrasonication time is 40s.
Embodiment 7
The method using Uv-induced screening selenium enriched Spirulina mutant strain of the present embodiment is substantially the same manner as Example 1, area
Be not only that: in Uv-induced screening step (1) under the conditions of ultrasonic power P=30% (15W), the ultrasonication time is 60s.
Embodiment 8
The method using Uv-induced screening selenium enriched Spirulina mutant strain of the present embodiment is substantially the same manner as Example 1, area
Be not only that: in Uv-induced screening step (1) under the conditions of ultrasonic power P=30% (15W), the ultrasonication time is 80s.
Embodiment 9
The method using Uv-induced screening selenium enriched Spirulina mutant strain of the present embodiment is substantially the same manner as Example 1, area
Be not only that: in Uv-induced screening step (1) under the conditions of ultrasonic power P=20% (10W), the ultrasonication time is 20s.
Embodiment 10
The method using Uv-induced screening selenium enriched Spirulina mutant strain of the present embodiment is substantially the same manner as Example 1, area
Be not only that: in Uv-induced screening step (1) under the conditions of ultrasonic power P=20% (10W), the ultrasonication time is 40s.
Embodiment 11
The method using Uv-induced screening selenium enriched Spirulina mutant strain of the present embodiment is substantially the same manner as Example 1, area
Be not only that: in Uv-induced screening step (1) under the conditions of ultrasonic power P=20% (10W), the ultrasonication time is 60s.
Embodiment 12
The method using Uv-induced screening selenium enriched Spirulina mutant strain of the present embodiment is substantially the same manner as Example 1, area
Be not only that: in Uv-induced screening step (1) under the conditions of ultrasonic power P=20% (10W), the ultrasonication time is 80s.
Embodiment 13
The method using Uv-induced screening selenium enriched Spirulina mutant strain of the present embodiment is substantially the same manner as Example 1, area
Be not only that: in Uv-induced screening step (1) under the conditions of ultrasonic power P=10% (5W), the ultrasonication time is 20s.
Embodiment 14
The method using Uv-induced screening selenium enriched Spirulina mutant strain of the present embodiment is substantially the same manner as Example 1, area
Be not only that: in Uv-induced screening step (1) under the conditions of ultrasonic power P=10% (5W), the ultrasonication time is 40s.
Embodiment 15
The method using Uv-induced screening selenium enriched Spirulina mutant strain of the present embodiment is substantially the same manner as Example 1, area
Be not only that: in Uv-induced screening step (1) under the conditions of ultrasonic power P=10% (5W), the ultrasonication time is 60s.
Embodiment 16
The method using Uv-induced screening selenium enriched Spirulina mutant strain of the present embodiment is substantially the same manner as Example 1, area
Be not only that: in Uv-induced screening step (1) under the conditions of ultrasonic power P=10% (5W), the ultrasonication time is 80s.
Embodiment 17
The method using Uv-induced screening selenium enriched Spirulina mutant strain of the present embodiment is substantially the same manner as Example 1, area
Be not only that: the concentration of sodium selenite is 600mg/L in Uv-induced screening step (7) culture medium, finally obtains tolerance
The spirulina of 600mg/L sodium selenite.
Embodiment 18
The method using Uv-induced screening selenium enriched Spirulina mutant strain of the present embodiment is substantially the same manner as Example 1, area
Be not only that: the concentration of sodium selenite is 800mg/L in Uv-induced screening step (7) culture medium, finally obtains tolerance
The spirulina of 800mg/L sodium selenite.
Embodiment 19
The method using Uv-induced screening selenium enriched Spirulina mutant strain of the present embodiment is substantially the same manner as Example 1, area
Be not only that: the concentration of sodium selenite is 1500mg/L in Uv-induced screening step (7) culture medium, finally obtains tolerance
The spirulina of 1500mg/L sodium selenite.
Spirulina is multicellular algae, is extracellularly made of inner wall, outer wall and keratinized sheath multilayered structure, has to ultraviolet light
Stronger resistivity.Therefore, it needs for spirulina to be broken into unicellular, two cells or three cells before carrying out ultraviolet mutagenesis.This
Invention using Ultrasonic Cell Disruptor be crushed spirulina, power be 10%, 20%, 30%, 40% under conditions of respectively be crushed 20s,
40s, 60s, 80s observe crushing effect, as shown in Figure 1 under the microscope.
In addition, the present invention evaluates the effect of ultrasonication using two indices: one is that be broken into many cells slender
Born of the same parents, two cells or three cells, this is observed by microscope；The other is keeping broken cell as more as possible, this is logical
It crosses and measures broken OD value to measure.
As seen from Figure 1, in power=30%, 60s, 80s are crushed；Power=40% is crushed 20s, 40s, 60s, 80s
Under conditions of so that spirulina is broken into unicellular, two cells or many cells.Under above-mentioned condition, power=40% is crushed 20s
The OD value highest (Fig. 2) of algae solution is obtained after this condition is broken.Therefore, using power=40%, this condition of 20s is crushed to spiral shell
Rotation algae is crushed.
The method that ultraviolet mutagenesis obtains spirulina mutant strain is easy to operate, the period is shorter, but spirulina is multicellular algae,
It is extracellularly made of inner wall, outer wall and keratinized sheath multilayered structure, has stronger resistivity to ultraviolet light, therefore carrying out purple
It needs for spirulina to be broken into unicellular, two cells or three cells before outer mutagenesis.It is to measure ultraviolet lure that whether lethality, which reaches 90%,
The essential condition become takes solid rubbing method to calculate lethality in traditional experiment, but this method period is longer and some
It is broken it is unicellular be not easy to grow on solid plate, calculate lethality using microscopic counting in the present invention, can accurately,
Quickly calculate the lethality of spirulina.
Spirulina just can guarantee the production of more mutant according to needing to count lethality, lethality after ultraviolet when being greater than 90%
It is raw.Therefore the present invention compares painting flat band method, chlorophyll method, OD value method, these four methods of microscopic counting and calculates lethality
Quality.
Embodiment 20
The present embodiment has done parallel laboratory test three times to compare chlorophyll method, OD value method, microscopic counting these three methods, real
Test that steps are as follows:
(1) blunt top spirulina for taking OD=1.0, under 40% power, ultrasonication 20s.
(2) broken algae solution is centrifuged 5min, revolving speed 1000g.
(3) precipitating is collected, and is diluted to OD=0.3 with culture medium.
(4) take 4ml algae solution loaded on plate, respectively ultraviolet irradiation 0s, 5s, 10s, 15s, 20s, 25s, 30s, 35s, 40s,
45s、50s、55s、60s、65s、70s。
(5) dark to place 5d, make dead algae degradation.
(6) it counts under the microscope, and surveys OD value.With these three method statistic lethalities, shown in result figure 3.
The present invention compares painting flat band method, chlorophyll method, OD value method, these four methods of microscopic counting and calculates lethality
Quality.
(1) flat band method is applied:
(2) chlorophyll method:
(3) OD value method:
(4) microscopic counting:
It is to apply flat band method first, broken spirulina is coated on the solid containing high selenium concentration after photograph is ultraviolet
On plate, lethality is calculated after growing up to single colonie.But single celled spirulina grows up to macroscopic bacterium colony on plate to be needed
The time in twenty or thirty day is taken, therefore this method time-consuming is too long, is not suitable for being used to calculate lethality.
As seen from Figure 3, the lethality obtained by chlorophyll method and OD value method is larger as the increase of mutation time has
Fluctuation, analysis the reason is that: both methods is both needed to calculate lethality by measurement absorbance, but algae solution concentration after mutagenesis
Very dilute, the concentration that algae solution is assessed by absorbance can bring biggish error.And OD value method need to measure algae solution under 560nm wavelength
Absorbance, and chlorophyll method needs to measure absorbance of the algae solution under the two wavelength of 665.2nm and 652.0nm, therefore leaf
Green element method bring error is bigger.The lethality obtained by microscopic counting steadily increases with the increase of time of ultraviolet irradiation
Add, illustrates the method that microscopic counting can be used as assessment spirulina ultraviolet mutagenesis lethality, and when time of ultraviolet irradiation is 40s
When spirulina lethality reached 90%.
Application Example 1
Selenium enriched Spirulina mutant strain made from above-described embodiment 1 is used to prepare selenium enriched Spirulina powder, the method is as follows:
(1) by the spirulina screened at 28 DEG C, PH=10, intensity of illumination is 40 μ EM-2·S-1Under the conditions of, it utilizes
Zarrouk culture medium is cultivated.Its culture medium main component includes: 13.61g/LNaHCO3, 4.03g/L Na2CO3, 0.5g/
LK2HPO4, 1g/LK2SO4, 1g/L NaCl, 0.2g/L MgSO4·7H2O, 2.5g/L NaNO3,0.04g/L CaCl2·2H2O,
0.01g/L FeSO4·7H2O, 1ml/L microelement A5Solution.
(2) Na of 1000mg/L is added in Initial stage of culture2SeO3, cultivate 2 weeks, be collected by centrifugation, vacuum freezedrying, be made
Algae powder.
(3) after micro-wave digestion, its Se content is surveyed with inductivity coupled plasma mass spectrometry (ICP-MS), Se content is
3517ppm, 90% or more organic selenium accounting.
Claims (10)
1. a kind of method using Uv-induced screening selenium enriched Spirulina mutant strain, it is characterised in that: the method includes as follows
Step:
(1) it takes blunt top spirulina, is first cultivated in the culture medium of low concentration sodium selenite, then to be successively transferred to concentration incremented by successively
High concentration sodium selenite culture medium in, until obtain tolerance 470mg/L sodium selenite blunt top spirulina；
(2) by the blunt top spirulina ultrasonication of tolerance 470mg/L sodium selenite obtained by step (1) at unicellular, two cells or
Tricellular algae solution；
(3) precipitating is collected after centrifugation in algae solution obtained by step (2), and is diluted precipitating with culture medium；
(4) algae solution after step (3) dilution is loaded on planar surface, is incident upon lethality > 90% with ultraviolet lighting, then shifts
To dark condition, overnight；
(5) collection step (4) overnight after algae solution, centrifugation, will be in the culture medium of the supreme concentration of sodium selenite of gained precipitating transfer
Culture, the cell survived is selenium enriched Spirulina mutant strain.
2. utilizing the method for Uv-induced screening selenium enriched Spirulina mutant strain according to claim 1, it is characterised in that: step
(1) concentration of the low concentration sodium selenite culture solution is 280~360mg/L, step (5) the high concentration sodium selenite culture
The concentration of liquid is 600~1500mg/L.
3. utilizing the method for Uv-induced screening selenium enriched Spirulina mutant strain according to claim 1, it is characterised in that: step
(2) the ultrasonication power is 5~30W；The ultrasonication time is 20~80s.
4. utilizing the method for Uv-induced screening selenium enriched Spirulina mutant strain according to claim 3, it is characterised in that: step
(2) the ultrasonication power is preferably 20W；The ultrasonication time is preferably 20s.
5. utilizing the method for Uv-induced screening selenium enriched Spirulina mutant strain according to claim 1, it is characterised in that: step
(4) the ultraviolet light time is 20~60s.
6. utilizing the method for Uv-induced screening selenium enriched Spirulina mutant strain according to claim 1, it is characterised in that: step
(4) lethality preferably uses microscopic counting to count.
7. any one of claim 1~6 method using Uv-induced screening selenium enriched Spirulina mutant strain is prepared
Selenium enriched Spirulina mutant strain.
8. the selenium enriched Spirulina mutant strain that any one of claim 1~6 is obtained using Uv-induced screening method is in preparation selenium-rich
Application in spirulina powder.
9. a kind of selenium enriched Spirulina powder, it is characterised in that: be prepared with the following method:
By selenium enriched Spirulina mutant strain made from any one of claim 1~6 the method in 28 DEG C, PH=10, intensity of illumination
For 40 μ EM-2·S-1Under the conditions of, it is cultivated using Zarrouk culture medium；It is added 1000mg/L's in Initial stage of culture
Na2SeO3, cultivate 2 weeks, be collected by centrifugation, vacuum freezedrying, the selenium enriched Spirulina powder is made.
10. selenium enriched Spirulina powder according to claim 9, it is characterised in that: the Zarrouk culture medium main component packet
It includes: 13.61g/LNaHCO3, 4.03g/L Na2CO3, 0.5g/LK2HPO4, 1g/LK2SO4, 1g/L NaC l, 0.2g/L
MgSO4·7H2O, 2.5g/L NaNO3, 0.04g/L CaCl2·2H2O, 0.01g/L FeSO4·7H2O, 1ml/L microelement A5
Solution.
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|---|---|---|---|---|
| CN112553191A (en) * | 2020-12-14 | 2021-03-26 | 恩施藻缘硒肽生物科技有限公司 | Method for efficiently extracting selenoprotein and application |
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| CN114073214A (en) * | 2021-11-25 | 2022-02-22 | 云南云硒食用菌开发有限公司 | Cultivation and cultivation method of selenium-rich oyster mushrooms |
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