CN108587984A - A kind of selenium enriched Spirulina culture, its cultural method and application - Google Patents
A kind of selenium enriched Spirulina culture, its cultural method and application Download PDFInfo
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- CN108587984A CN108587984A CN201810585855.3A CN201810585855A CN108587984A CN 108587984 A CN108587984 A CN 108587984A CN 201810585855 A CN201810585855 A CN 201810585855A CN 108587984 A CN108587984 A CN 108587984A
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- spirulina
- culture
- selenium
- logarithmic phase
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- 239000011669 selenium Substances 0.000 title claims abstract description 75
- 240000002900 Arthrospira platensis Species 0.000 title claims abstract description 60
- 235000016425 Arthrospira platensis Nutrition 0.000 title claims abstract description 60
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 title claims abstract description 59
- 229940082787 spirulina Drugs 0.000 title claims abstract description 59
- 229910052711 selenium Inorganic materials 0.000 title claims abstract description 58
- 238000000034 method Methods 0.000 title claims abstract description 21
- 241000195493 Cryptophyta Species 0.000 claims abstract description 17
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 11
- 239000001963 growth medium Substances 0.000 claims abstract description 5
- 230000001939 inductive effect Effects 0.000 claims abstract description 3
- 229940091258 selenium supplement Drugs 0.000 claims description 53
- AKHNMLFCWUSKQB-UHFFFAOYSA-L sodium thiosulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=S AKHNMLFCWUSKQB-UHFFFAOYSA-L 0.000 claims description 11
- 235000019345 sodium thiosulphate Nutrition 0.000 claims description 11
- BVTBRVFYZUCAKH-UHFFFAOYSA-L disodium selenite Chemical group [Na+].[Na+].[O-][Se]([O-])=O BVTBRVFYZUCAKH-UHFFFAOYSA-L 0.000 claims description 6
- 239000011781 sodium selenite Substances 0.000 claims description 6
- 229960001471 sodium selenite Drugs 0.000 claims description 6
- 235000015921 sodium selenite Nutrition 0.000 claims description 6
- 235000013305 food Nutrition 0.000 claims description 5
- 239000003223 protective agent Substances 0.000 claims description 5
- 230000012010 growth Effects 0.000 abstract description 6
- 239000000654 additive Substances 0.000 description 9
- 230000000996 additive effect Effects 0.000 description 9
- 239000000843 powder Substances 0.000 description 8
- 241000196324 Embryophyta Species 0.000 description 7
- 238000005457 optimization Methods 0.000 description 5
- 238000003306 harvesting Methods 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 3
- 239000000306 component Substances 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000004615 ingredient Substances 0.000 description 3
- 230000001988 toxicity Effects 0.000 description 3
- 231100000419 toxicity Toxicity 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000001704 evaporation Methods 0.000 description 2
- 230000008020 evaporation Effects 0.000 description 2
- 238000005286 illumination Methods 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 239000012533 medium component Substances 0.000 description 2
- 239000003002 pH adjusting agent Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- FDKWRPBBCBCIGA-REOHCLBHSA-N (2r)-2-azaniumyl-3-$l^{1}-selanylpropanoate Chemical compound [Se]C[C@H](N)C(O)=O FDKWRPBBCBCIGA-REOHCLBHSA-N 0.000 description 1
- 235000017060 Arachis glabrata Nutrition 0.000 description 1
- 244000105624 Arachis hypogaea Species 0.000 description 1
- 235000010777 Arachis hypogaea Nutrition 0.000 description 1
- 235000018262 Arachis monticola Nutrition 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 241000195649 Chlorella <Chlorellales> Species 0.000 description 1
- 244000241257 Cucumis melo Species 0.000 description 1
- 235000015510 Cucumis melo subsp melo Nutrition 0.000 description 1
- FDKWRPBBCBCIGA-UWTATZPHSA-N D-Selenocysteine Natural products [Se]C[C@@H](N)C(O)=O FDKWRPBBCBCIGA-UWTATZPHSA-N 0.000 description 1
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 description 1
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- 229910021380 Manganese Chloride Inorganic materials 0.000 description 1
- GLFNIEUTAYBVOC-UHFFFAOYSA-L Manganese chloride Chemical compound Cl[Mn]Cl GLFNIEUTAYBVOC-UHFFFAOYSA-L 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- RJFAYQIBOAGBLC-BYPYZUCNSA-N Selenium-L-methionine Chemical compound C[Se]CC[C@H](N)C(O)=O RJFAYQIBOAGBLC-BYPYZUCNSA-N 0.000 description 1
- RJFAYQIBOAGBLC-UHFFFAOYSA-N Selenomethionine Natural products C[Se]CCC(N)C(O)=O RJFAYQIBOAGBLC-UHFFFAOYSA-N 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000005791 algae growth Effects 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 235000021466 carotenoid Nutrition 0.000 description 1
- 150000001747 carotenoids Chemical class 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 229910052927 chalcanthite Inorganic materials 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 229910052564 epsomite Inorganic materials 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 239000004519 grease Substances 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 239000011565 manganese chloride Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 229910052603 melanterite Inorganic materials 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000004792 oxidative damage Effects 0.000 description 1
- 235000020232 peanut Nutrition 0.000 description 1
- 231100000572 poisoning Toxicity 0.000 description 1
- 230000000607 poisoning effect Effects 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- OTYBMLCTZGSZBG-UHFFFAOYSA-L potassium sulfate Chemical compound [K+].[K+].[O-]S([O-])(=O)=O OTYBMLCTZGSZBG-UHFFFAOYSA-L 0.000 description 1
- 229910052939 potassium sulfate Inorganic materials 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- ZKZBPNGNEQAJSX-UHFFFAOYSA-N selenocysteine Natural products [SeH]CC(N)C(O)=O ZKZBPNGNEQAJSX-UHFFFAOYSA-N 0.000 description 1
- 235000016491 selenocysteine Nutrition 0.000 description 1
- 229940055619 selenocysteine Drugs 0.000 description 1
- 229960002718 selenomethionine Drugs 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 1
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 1
- 229910000368 zinc sulfate Inorganic materials 0.000 description 1
- 239000011686 zinc sulphate Substances 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/30—Oligoelements
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/16—Inorganic salts, minerals or trace elements
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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Abstract
The present invention relates to a kind of methods of culture selenium enriched Spirulina, include the following steps:S1:Using full culture medium by SPIRULINA CULTIVATION to logarithmic phase, logarithmic phase SPIRULINA CULTIVATION object;S2:Into the logarithmic phase SPIRULINA CULTIVATION object, addition contains seleno reagent, inducing cycloidic algae selenium-rich, and culture 35 days is to get to selenium enriched Spirulina culture.Method culture spirulina through the invention can make spirulina keep the higher speed of growth, to improve yield, under the premise of ensureing yield, will improve to 800 μ g/g dry weights Se content in spirulina, wherein Organic Selenium accounts for 80% or more.
Description
Technical field
The present invention relates to high added value microdisk electrode and application fields, more specifically it relates to a kind of culture selenium enriched Spirulina
Method, the selenium enriched Spirulina culture that is obtained using this method and its application.
Background technology
Selenium(Se)It is a kind of trace element necessary to animal and people, selenium-supply can prevent a variety of diseases.However, inorganic selenium example
Such as selenate is unfavorable for human body or animal body absorbs.Therefore, it is necessary to Organic Selenium is obtained from food.Some plants can be enriched with
Selenium in environment, synthesizes Organic Selenium in vivo, can be added in food or drug by the way that additive is made in such plant, or
The edible part for directly eating this kind of plant, for supplementing internal selenium.
Has a method that some patents and technical literature disclose culture selenium-enriched plant, such as the tea of selenium-rich, peanut, big
The culture of rice, melon and fruit etc..But the cultivation period of the higher plant of terrestrial is long, breeding condition is not easy to control, and final result is also joined
Difference is uneven, and the Se content of plant is not high.
Microalgae is a kind of unicellular or many cells rudimentary plant.Numerous species are rich in protein, grease, polysaccharide in microalgae
Etc. nutritional ingredients, type also is rich in physiologically active ingredients such as unsaturated fatty acid, carotenoid, vitamins.Spirulina,
The algae such as chlorella dry powder is remarkably improved metabolism and the premunition of human or animal as food or feed addictive.In addition, micro-
The growth cycle of algae is short, and condition of culture is also easy to control, also very convenient in post-production due to being microorganism.However, spiral shell
It is very bad to revolve growth conditions in the solution of high Se content of algae or other microalgaes, the reason is that microalgae is by selenium internalization process
In, the selenomethionine and selenocysteine of synthesis replace methionine and cysteine in peptide chain synthesis, to generate selenium
Toxicity.Therefore, the spirulina and products thereof that there is no high Se content on the market at present needs a kind of new microalgae culture method, comes
Cultivate selenium-rich microalgae.
Invention content
In order to solve the above problem, the present invention provides it is a kind of culture selenium enriched Spirulina method, which is characterized in that including with
Lower step:
S1:Using full culture medium by SPIRULINA CULTIVATION to logarithmic phase, logarithmic phase SPIRULINA CULTIVATION object;
S2:Into the logarithmic phase SPIRULINA CULTIVATION object addition contain seleno reagent, inducing cycloidic algae selenium-rich, culture 3-5 days to get to
Selenium enriched Spirulina culture.
In a specific embodiment, it is sodium selenite solution containing seleno reagent described in S2
In a specific embodiment, which is characterized in that S2 includes the following steps:
S21:Protective agent is added into the logarithmic phase SPIRULINA CULTIVATION object;
S22:It is added to protectant logarithmic phase SPIRULINA CULTIVATION object by described and is placed under bloom high light conditions;
S23:Add in culture to the selenium enriched Spirulina several times daily it is described contain seleno reagent, cultivate 5-8 days to get to
Selenium enriched Spirulina culture.
In a preferred embodiment, in S23, every time addition it is described containing before and after seleno reagent to the logarithmic phase spiral
Algae culture carries out each 20-40 minutes of processing.
In a preferred embodiment, in S23, addition daily 3-5 time is described to contain seleno reagent, the amount added every time for by
The selenium of the logarithmic phase SPIRULINA CULTIVATION volume 100-150 mg/L.
In a preferred embodiment, protective agent described in S21 is sodium thiosulfate, and the amount of addition is by the logarithm
Phase SPIRULINA CULTIVATION volume 500-750 mg/L.
In a preferred embodiment, in S22, the high light-intensity conditions are 350-500 μ Em-2·s-1。
In a specific embodiment, the full culture medium described in S1 is Zarrouk culture mediums.
The present invention also provides a kind of selenium enriched Spirulina cultures, are obtained by above method culture.
The present invention also provides application of the above-mentioned selenium enriched Spirulina culture in preparing food or feed addictive.
Method culture spirulina through the invention can make spirulina keep the higher speed of growth, to improve yield,
Under the premise of ensureing yield, it will improve to 800 μ g/g dry weights Se content in spirulina, wherein Organic Selenium accounts for 80% or more.
Description of the drawings
Fig. 1 is experimental group 1, experimental group 2 and control group during Fiber differentiation spirulina selenium-rich, and the growth of spirulina is bent
Line;
Fig. 2 is the block diagram that algae powder yield and total Se content that spirulina obtains are cultivated under different light intensity;
Fig. 3 is the block diagram that algae powder yield and total Se content that spirulina obtains are cultivated under different sodium selenite additive amounts;
Fig. 4 is the block diagram that algae powder yield and total Se content that spirulina obtains are cultivated under different sodium thiosulfate additive amounts.
Specific implementation mode
The principle and features of the present invention will be described below with reference to the accompanying drawings, and the given examples are served only to explain the present invention, and
It is non-to be used to limit the scope of the present invention.
1. by SPIRULINA CULTIVATION to logarithmic phase
Using Zarrouk medium culture spirulinas, medium component is as shown in table 1.
The ingredient of 1 Zarrouk culture mediums of table
Component | Concentration(g/L) | Component | Concentration(g/L) |
NaHCO3 | 16.8 | K2HPO4 | 0.50 |
NaNO3 | 2.50 | FeSO4·7H2O | 0.01 |
NaCl | 1.00 | Na2-EDTA | 0.08 |
K2SO4 | 1.00 | CaCl2 | 0.08 |
MgSO4·7H2O | 0.20 | A5 solution | 2 mL/L |
The ingredient of 2 A5 solution of table
Component | Concentration(g/L) |
H3BO3 | 2.86 |
(NH4)6Mo17O24 | 0.02 |
MnCl2·4H2O | 1.80 |
CuSO4·5H2O | 0.08 |
ZnSO4·7H2O | 0.22 |
Spirulina algae is inoculated in the 250 mL triangular flasks containing 150 mL culture mediums, inoculum concentration OD560=0.10.By three
Angle bottle is positioned over constant-temperature table culture at 30-35 DEG C, and intensity of illumination is 25-40 μ Em-2·s-1.It is supplemented and is steamed according to evaporation capacity
Distilled water, and it is properly added pH adjusting agent, so that pH is maintained within the scope of 9.5-10.5.Incubation time is 5-8 days, culture solution
OD560Reach 1-1.5, is in exponential phase.
2. selenium-rich induces
To sodium thiosulfate is added in the spirulina medium of exponential phase, makes its a concentration of 500 mg/L, be positioned over
350 μE·m-2·s-1Light intensity under cultivate, add sodium selenite for 3 times into culture solution per natural gift, each additive amount is 100
Mg/L selenium equivalents.Distilled water is supplemented according to evaporation capacity, and is properly added pH adjusting agent, pH is made to maintain 9.5-10.5 ranges
It is interior.Incubation time is 5 days.As experimental group 1.
With always in intensity of illumination for 25-40 μ Em-2·s-1Under, culture is control group in Zarrouk culture mediums.
After culture 5 days, the spirulina frustule of each group is collected by centrifugation, and pure water repeatedly washs, to remove medium component.Claim
Weight, and detect Se content.The results show that control group obtains the xeraphium of 3.85 g/L, and it is substantially free of selenium, experimental group obtains
The xeraphium of 1.83 ± 0.35 g/L, total Se content are 632 ± 15.82 μ g/g dry weights, and Organic Selenium accounts for 85% or so of total selenium.One
As for, shown in experimental group 1, although high Se culture mediums and bloom according to having certain influence to the growth of spirulina, but still
It more can healthily grow, and there are selenium enriching functions.It can be seen that on the one hand protective agent sodium thiosulfate can help spirulina cells
Selenium toxicity is resisted, the reproducibility of another aspect sodium thiosulfate can resist the oxidative damage caused by bloom is shone.
3. dark processing is added
It on the basis of the condition of culture of experimental group 1, is improved, carries out 20-40 min's before and after adding selenium every time
Dark processing obtains experimental group 2.The results are shown in Figure 1, unexpected, and the micro algae growth state of experimental group 2 is than experimental group 1
Much better, yield of spirulina is significantly increased, and reaches 3.28 ± 0.26 g/L.Total Se content shows that total Se content of experimental group 2 is
852 ± 26.37 μ g/g dry weights, Organic Selenium account for 83% or so of total selenium.It can be seen that dark processing contributes to spirulina cells to resist selenium
Toxicity, and spirulina cells is promoted to absorb selenium.
4. condition optimizing
Based on experimental group 2, successively light intensity, selenium concentration and sodium thiosulfate are optimized.Experiment is with previous step reality every time
The optimal conditions tested carries out next step condition optimizing.
The optimization of 4.1 light intensity
Based on experimental group 2, only spirulina is cultivated respectively in 50,100,200,300,350,400,450,500,550 μ
E·m-2·s-1Light intensity under cultivated, explore optimal light intensity.The results are shown in Figure 2, when light intensity is in 350 μ Em-2·
s-1When above, total Se content is higher, reaches 800-1500 μ g/g dry weights.But with the raising of light intensity, the algae dry weight of harvest
It is gradually lowered, when light intensity reaches 550 μ Em-2·s-1When, the algae powder of harvest is only 1.22 ± 0.37 g/L.Therefore, suitable
Light intensity be 350-500 μ Em-2·s-1.In next experiment, 450 μ Em are chosen-2·s-1As culture light intensity into
The further optimization of row.
The optimization of 4.2 selenium additive amounts
Under conditions of identified above, the selenium additive amount of addition is optimized, adds 50,75,100,125,150 respectively every time
The sodium selenite solution of mg/L selenium equivalents.The results are shown in Figure 3, and when selenium concentration is in 100 mg/L or more, total selenium of algae powder contains
Amount is higher, but at the same time, the content of the algae powder of harvest also reduces, it is seen that the raising of selenium concentration has certain murder by poisoning to algae.
In the experiment of next step, the sodium selenite of 100 mg/L selenium equivalents of addition is chosen to optimize the additive amount of sodium thiosulfate.
The optimization of 4.3 sodium thiosulfate additive amounts
Under conditions of identified above, the sodium thiosulfate additive amount of addition is optimized, respectively add 500,600,700,
The hypo solution of 750 mg/L.The results are shown in Figure 4, with the raising of concentration of sodium thiosulfate, the dry algae powder of harvest
Amount be gradually increased, and total Se content is unaffected.
In the above optimization process, the content of Organic Selenium maintains 80% or more of total Se content always.
The foregoing is merely presently preferred embodiments of the present invention, is not intended to limit the invention, it is all the present invention spirit and
Within principle, any modification, equivalent replacement, improvement and so on should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of method of culture selenium enriched Spirulina, which is characterized in that include the following steps:
S1:Using full culture medium by SPIRULINA CULTIVATION to logarithmic phase, logarithmic phase SPIRULINA CULTIVATION object;
S2:Into the logarithmic phase SPIRULINA CULTIVATION object addition contain seleno reagent, inducing cycloidic algae selenium-rich, culture 5-8 days to get to
Selenium enriched Spirulina culture.
2. according to the method described in claim 1, it is characterized in that, it is sodium selenite solution to contain seleno reagent described in S2.
3. according to the method described in claim 1, it is characterized in that, S2 includes the following steps:
S21:Protective agent is added into the logarithmic phase SPIRULINA CULTIVATION object;
S22:It is added to protectant logarithmic phase SPIRULINA CULTIVATION object by described and is placed under bloom high light conditions;
S23:Add in culture to the selenium enriched Spirulina several times daily it is described contain seleno reagent, cultivate 5-8 days to get to
Selenium enriched Spirulina culture.
4. according to the method described in claim 3, it is characterized in that, in S23, every time addition it is described containing before and after seleno reagent to institute
It states logarithmic phase SPIRULINA CULTIVATION object and carries out each 20-40 minutes of processing.
5. according to the method described in claim 3, it is characterized in that, in S23, addition daily 3-5 time is described containing seleno reagent, often
The amount of secondary addition is the selenium by the logarithmic phase SPIRULINA CULTIVATION volume 100-150 mg/L.
6. according to the method described in claim 3, it is characterized in that, protective agent described in S21 be sodium thiosulfate, addition
Amount is by the logarithmic phase SPIRULINA CULTIVATION volume 500-750 mg/L.
7. according to the method described in claim 3, it is characterized in that, in S22, the high light-intensity conditions are 250-450 μ Em-2·s-1。
8. according to the described method of any one of claim 1-7, which is characterized in that the full culture medium described in S1 is
Zarrouk culture mediums.
9. a kind of selenium enriched Spirulina culture, which is characterized in that obtained by the method culture described in any one of claim 1-8
It arrives.
10. application of the selenium enriched Spirulina culture in preparing food or feed addictive described in claim 9.
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