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 PDF

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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
selenium enriched
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CN108587984B (en
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王强
陈为先
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Wuhan Algae Excellence Bio Technology Co Ltd
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    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K10/00Animal feeding-stuffs
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    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, 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
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135Bacteria or derivatives thereof, e.g. probiotics
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    • A23LFOODS, 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
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/16Inorganic salts, minerals or trace elements
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00Food 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

A kind of selenium enriched Spirulina culture, its cultural method and application
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.
CN201810585855.3A 2018-06-08 2018-06-08 Selenium-rich spirulina culture, and culture method and application thereof Expired - Fee Related CN108587984B (en)

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CN110218686A (en) * 2019-07-02 2019-09-10 浙江海洋大学 The cultural method of selenium enriched Spirulina

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110218686A (en) * 2019-07-02 2019-09-10 浙江海洋大学 The cultural method of selenium enriched Spirulina
CN110218686B (en) * 2019-07-02 2023-06-06 浙江海洋大学 Method for culturing selenium-enriched spirulina

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