CN110257296A - The cultural method and application thereof of chromium-rich spirulina - Google Patents
The cultural method and application thereof of chromium-rich spirulina Download PDFInfo
- Publication number
- CN110257296A CN110257296A CN201910590182.5A CN201910590182A CN110257296A CN 110257296 A CN110257296 A CN 110257296A CN 201910590182 A CN201910590182 A CN 201910590182A CN 110257296 A CN110257296 A CN 110257296A
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- Prior art keywords
- chromium
- spirulina
- rich
- culture medium
- cultural method
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- VYZAMTAEIAYCRO-UHFFFAOYSA-N Chromium Chemical compound [Cr] VYZAMTAEIAYCRO-UHFFFAOYSA-N 0.000 title claims abstract description 130
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Classifications
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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
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/748—Cyanobacteria, i.e. blue-green bacteria or blue-green algae, e.g. spirulina
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- 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
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Abstract
The present invention provides the cultural method and application thereof of chromium-rich spirulina, belongs to microdisk electrode technical field, and logarithmic phase spirulina is inoculated into culture medium and is cultivated, adds chromium chloride in incubation in batches, continues to cultivate to obtain chromium-rich spirulina；Adding in batches is to be inoculated into culture medium to add in batches after culture 4-8d from spirulina, continuously adds chromium chloride 4-6d；Thiomalic acid is added while continuously adding chromium chloride.Cr (III) can be effectively relieved to the coercion of spirulina in cultural method of the present invention, promote the fast-growth of spirulina, chromium is set to obtain trifoliate orange in spirulina cells tired, demand of the spirulina to inorganic matters such as potassium chloride, potassium sulfate, sodium chloride is greatly reduced, the chromium-rich SPIRULINA CULTIVATION object obtained contains the chromium, chlorophyll a and phycobniliprotein of higher amount, and Organic Chromium ratio is high in the chromium of enrichment, can be used for preparing alpha-glucosidase and inhibits, prepares for preventing or treating cardiovascular disease medicine composition and/or health care product.
Description
Technical field
The invention belongs to microdisk electrode technical fields, and in particular to the cultural method and application thereof of chromium-rich spirulina.
Background technique
Spirulina (Spirulina), also referred to as arthrospira are a kind of ancient low water plants, have protokaryon multiple cell subtypes
Filamentous microalgae, belongs to Cyanophyta, Cyanophyceae, Oscillatoriales, Spirullina or Arthrospira, it has now been found that there are about 35 kinds.
Spirulina typically refers to edible spirulina maxim and blunt top spirulina, the two kinds are to be studied most kinds,
It is to be widely used in the kind of industrialization in countries in the world at present.Spirulina because its frond in the shape of a spiral due to gain the name, frond is single-row thin
Born of the same parents arrange, no branch.Most of spirulina is lived in fresh water, and only a small amount of kind life in the seawater, and is used to produce
The spirulina maxim and blunt top spirulina that industry metaplasia produces then mainly are lived in Alkaline waters.Spirulina is rich in a variety of nutriments
And bioactive substance, as protein, amino acid, unsaturated fatty acid, vitamin, mineral element, phycocyanin, polysaccharides and
Beta carotene etc. is currently known nutritional ingredient most one of comprehensive, most balanced wholefood, and medical research shows spiral
Algae has reduction cholesterol, reduces hyperlipidemia, adjust blood glucose, antitumor, anti-cancer tumor suppressor, remove free radical, promote body new
The weight such as old metabolism, activating immune system function, antifatigue, resist oxygen lack and adjustment intestinal microflora, improvement micro-ecological environment
The physiological function wanted.Therefore, it is considered as a kind of ideal nutritional source and potential medicine source, is a kind of novel medicine edible product,
Application prospect is boundless.In addition, spirulina is a kind of very high microalgae of economic value, it has, and photosynthetic efficiency is high, growth is numerous
It grows the features such as fast and strong to environmental suitability, and its Industrialized processing technique is mature, is to carry out biology richness to certain minerals element
The ideal carrier for collecting and organising.Using spirulina as bio-carrier, the raw material containing certain minerals element is added in culture solution
Compound, biological concentration and transformation by spirulina to certain minerals element, obtaining to be rich in has certain minerals element
Functional screw algae.
In recent years, the disease incidence of diabetes constantly rises, it has also become the public health problem of global concern.Study table
Bright, trace element chromium is generated to the insulin of body and energetic supersession all plays particularly important effect, and scarce chromium can cause pancreas
Element effect in island reduces, the utilization of sugar is obstructed, and leads to diabetes；Cause fatty especially cholesterol in blood to increase simultaneously, causes height
Lipoproteinemia, and it is related with incidence of atherosclerosis, and therefore, supplement network appropriate can prevent and treat diabetes, by
In various forms of chromium, combination degree is different in the tissue, causes absorptivity different.Not only toxicity is big for inorganic chromium, and absorbs horizontal
Extremely low is 0.4-3%；And the absorption of Organic Chromium is rapid and safe, absorptivity 10-25%.So how that inorganic chromium is safe
Effectively it is converted into the emphasis that organic network is current research.As the improvement of people's living standards, people increasingly pay attention to food
Green, natural, safety, health care etc., and research and application of the spirulina as the biological concentration absorption carrier of trace element chromium
It is more and more.Prepare Organic Chromium using the bioconcentration of spirulina, be it is a kind of efficiently, at wood richness low, easy to operate
Set method, growth of spirulina platensis is rapid, and biomass is big, the absorptivity and bioavailability to chromium be much higher than inorganic chromium, wherein chromium with
Organic macromolecule form exists, and without any toxicity and side effects, bioavilability is high, good absorption effect, improves biological insulin effect；And
Also contain multiple protein and bioactive substance in spirulina, has good healthcare function to human body.
Summary of the invention
It is an object of the present invention to provide a kind of growth logarithmic phases for being able to extend spirulina, and Cr is effectively relieved
(III) to the coercion of spirulina, promote the fast-growth of spirulina, so that Cr (III) is obtained trifoliate orange in spirulina cells and tire out,
The cultural method of the chromium-rich spirulina of demand of the spirulina to inorganic matters such as potassium chloride, potassium sulfate, sodium chloride, the training is greatly reduced
The chromium-rich SPIRULINA CULTIVATION object that the method for supporting obtains contains the chromium, chlorophyll a and phycobniliprotein of higher amount, and organic in the chromium being enriched with
Chromium ratio is high, can be used for preparing alpha-glucosidase inhibit, prepare for prevent or treat cardiovascular disease medicine composition and/
Or health care product.
The technical solution that the present invention is taken to achieve the above object are as follows:
Logarithmic phase spirulina is inoculated into culture medium and cultivates, in incubation in batches by the cultural method of chromium-rich spirulina
Chromium chloride is added, continues to cultivate to obtain chromium-rich spirulina；
Above-mentioned add in batches is to be inoculated into culture medium to add in batches after culture 4-8d from spirulina, continuously adds chromium chloride
3-6d, the spirulina for cultivating 4-8d are in the logarithmic growth phase later period, and cell viability is vigorous, and algae density is big, strong to chromium tolerance,
Metabolic mechanism is active, absorption and enrichment conducive to spirulina frond to chromium.For example, adding chromium chloride in batches after culture 4d, continuously
Add 3d, 4d, 5d or 6d；Or chromium chloride is added in batches after culture 5d, continuously add 3d, 4d, 5d or 6d；Or after culture 6d in batches
Chromium chloride is added, 3d, 4d, 5d or 6d are continuously added；Or culture 7d after add chromium chloride in batches, continuously add 3d, 4d, 5d or
6d；Or chromium chloride is added in batches after culture 8d, continuously add 3d, 4d, 5d or 6d；
Wherein, thiomalic acid is added while continuously adding chromium chloride.
Spirulina has good enrichment to Cr (III), but the growth of spirulina and the addition concentration of Cr (III) have
Certain relationship, the Cr (III) of low concentration can promote the growth of spirulina, and the Cr (III) of high concentration can be to growth of spirulina platensis
It causes certain oxidative stress to act on, destroys the eucaryotic cell structure of spirulina, prevent frustule from completing normal physiological activity, hinder
Hinder the growth and breeding of frustule, or even the death of spirulina cells can be caused, lethal effect is generated to spirulina.So in order to drop
The influence of the low growth to spirulina can be added by gradation and improve spirulina to the tolerance of Cr (III), make spirulina cells
It is adapted under lower Cr (III) concentration, absorb and same by the Cr (III) in culture solution with good growth conditions
Change, so that Cr (III) is obtained trifoliate orange in spirulina cells tired.And other component combines in thiomalic acid and culture medium, it can be significant
SOD and the CAT activity of frustule, oxidative stress caused by resisting because of chromium concn height are reduced, helper cell is resisted chromium toxicity, promoted
It is grown under higher chromium content into spirulina, improves the biomass and chromium enriching quantity of spirulina, and Organic Chromium ratio in the chromium being enriched with
Rate is high；Thiomalic acid can also make the Chlorophyll-a Content in spirulina improve a lot with other component combines in culture medium, mention
The photosynthetic pigments of high Sargassum horneri are horizontal, enhance the optical energy utilization efficiency of frond, and then promote the promotion of its photosynthetic capacity, promote spiral
Algae grows under higher chromium content, improves the biomass and chromium enriching quantity of spirulina；Other groups in thiomalic acid and culture medium
Demand of the spirulina to inorganic matters such as potassium chloride, potassium sulfate, sodium chloride can be also greatly reduced in part combination, improve made chromium-rich spiral
The long-term use safety of algae.
Cultural method of the present invention can promote spirulina to grow under higher chromium content, make Cr (III) in spirulina cells
It is tired inside to obtain trifoliate orange, improves the biomass, chromium enriching quantity and Chlorophyll-a Content of spirulina, spirulina is greatly reduced to potassium chloride, sulphur
The demand of the inorganic matters such as sour potassium, sodium chloride improves the long-term use safety of made chromium-rich spirulina.
In one embodiment, the amount for adding chromium chloride in batches is that daily every liter of culture medium fills into 50-150mg/L, excellent
Select 50-120mg/L, more preferable 50-110mg/L, further preferred 60-100mg/L.
The additive amount of thiomalic acid in cultural method of the invention is 10-20 μm of ol/L, the sulphur in the concentration range
The Chlorophyll-a Content in spirulina made for malic acid is at least equal to 19mg/g.The additive amount of thiomalic acid can be 11 μ
mol/L、12.5μmol/L、13μmol/L、14μmol/L、15.4μmol/L、16μmol/L、17μmol/L、18μmol/L、19μ
Mol/L etc..
In one embodiment, the pH value of culture medium is 8.0-10.0, and temperature is 25-35 DEG C, illuminance 3000-
8000lx, light application time for 24 hours/d.In the growth course of spirulina, pH be influence growth of spirulina platensis metabolism one it is important because
A key factor in son and spirulina enrichment metal process, it influences suction of the cell to nutritious substance and ion
It receives and utilizes.In acid condition, growth of spirulina platensis is very poor and is unfavorable for the enrichment of Cr (III), and excessively high pH value can also press down
The growth rate of spirulina processed.PH value can be 8.5,9.0,9.5 etc..
Contain 2,6- diaminopimelic acid in cultural method of the invention.Cell is in the physiology course for adapting to environment-stress
In, need to adjust the synthesis and degradation of protein, to adapt to new environment, the increase of protein is biology to the one of environment-stress
Kind Physiology and biochemistry stress.2,6- diaminopimelic acid enters after cell the level that can be improved pigment content and enzyme in cell
And promote the growth of cell, so as to accumulate more phycobniliprotein；Meanwhile 2,6- diaminopimelic acid are also used as spirulina
The spare nitrogen source of growth protects the phycobniliprotein of synthesis not to be degraded because of lacking for nitrogen source utilizations, accumulates in promotion cell
More phycobniliprotein.And phycobniliprotein be in spirulina it is a kind of it is important catch photochromic fibroin, can directly capture absorption light
Can, the light energy electronic chain transmitting in chloroplaset is participated in, enhances the adaptive capacity to environment of algae, so that spirulina cells be helped to adapt to
The coercion of Cr (III), promotes the growth of spirulina, improves the biomass of spirulina.
In one embodiment, 10-50 μm of ol/L 2,6- diaminopimelic acid, preferably 15-50 μ are contained in culture medium
Mol/L, more preferable 20-40 μm of ol/L, further preferred 25-35mol/L.The 2,6- diaminopimelic acid of the concentration range can
The growth logarithmic phase for extending spirulina, is effectively relieved Cr (III) to the coercion of spirulina, promotes the fast fast-growing of spirulina
It is long.
Phycobiliprotein Content is at least equal to chromium-rich spirulina dry weight in chromium-rich spirulina obtained by cultural method of the invention
20%.Compared to the range for not applying the 3-12% obtained in the experiment that method of the invention carries out, this is extra high.
Invention additionally discloses the chromium-rich SPIRULINA CULTIVATION object that a kind of above-mentioned cultural method obtains, in chromium-rich SPIRULINA CULTIVATION object
Chromium content >=7.55mg/g, Organic Chromium ratio >=82.60%.The chromium-rich SPIRULINA CULTIVATION object of the present invention contains the chromium of higher amount, leaf
Green element a and phycobniliprotein, and Organic Chromium ratio is high in the chromium being enriched with, and has no toxic side effect, bioavilability is high, good absorption effect,
The protein synthesis of immune function, the biological insulin effect, promotion body that improve body with enhancing body, maintains blood
The balance of middle cholesterol, the hematopoiesis function for improving body, etc. healthcare functions.
The present invention further discloses above-mentioned chromium-rich SPIRULINA CULTIVATION objects to prepare the purposes in alpha-glucosidase inhibition.
Chromium-rich SPIRULINA CULTIVATION object produced by the present invention has excellent inhibiting effect to alpha-glucosidase, can be used for Prevention sugar
Urine disease, will there is vast application prospect in terms of as hypoglycemic medicine and novel healthy food.
The present invention further discloses chromium-rich SPIRULINA CULTIVATION object in preparation for preventing or treating cardiovascular disease medicine
Purposes in composition and/or health care product.Chromium-rich SPIRULINA CULTIVATION object produced by the present invention can stimulate and improve making for body
Blood function, hence it is evident that increase coronary blood flow, while the cholesterol level in blood can be dropped, blood is diluted, thus
Blood is avoided to generate thrombus since cholesterol is excessively high, further protection body internal organs；In addition, can also prevent free radical pair
The infringement of human body, can it is without side-effects to thrombus generate cardiovascular disease treated and prevented and treated, use more secure side
Just.
Compared with prior art, the invention has the benefit that cultural method of the present invention is able to extend the growth of spirulina
Logarithmic phase is effectively relieved Cr (III) to the coercion of spirulina, promotes the fast-growth of spirulina, makes Cr (III) in spiral
It is tired that trifoliate orange is obtained in frustule, improves the biomass, chromium enriching quantity and Chlorophyll-a Content of spirulina, spirulina is greatly reduced to chlorine
Change the demand of the inorganic matters such as potassium, potassium sulfate, sodium chloride, while improving the long-term use safety of made chromium-rich spirulina.It obtains
Chromium-rich SPIRULINA CULTIVATION object contain the chromium, chlorophyll a and phycobniliprotein of higher amount, and Organic Chromium ratio is high in the chromium being enriched with；
Cultural method of the present invention can promote to accumulate more phycobniliprotein in cell, enhance the adaptive capacity to environment of algae, to help
It helps spirulina cells to adapt to the coercion of Cr (III), promotes the growth of spirulina, improve the biomass of spirulina；The present invention
Chromium-rich SPIRULINA CULTIVATION object contains the chromium, chlorophyll a and phycobniliprotein of higher amount, and Organic Chromium ratio is high in the chromium being enriched with, nothing
Toxic side effect, bioavilability is high, good absorption effect, has the immune function of enhancing body, improves the biological insulin of body
Effect, promote body protein synthesis, maintain Blood Cholesterol balance, improve body hematopoiesis function, etc. health cares function
Energy.
Present invention employs above-mentioned technical proposals to provide the cultural method and application thereof of chromium-rich spirulina, compensates for existing skill
The deficiency of art, reasonable design, easy operation.
Detailed description of the invention
Fig. 1 is absorbance A in test example 1 of the present invention560With the graph of relation of cell product of Spirulina；
Fig. 2 is the growth curve of spirulina in test example 1 of the present invention；
Fig. 3 be in test example 1 of the present invention spirulina to the enrichment total amount and organic degree of chromium；
Fig. 4 is spirulina Determination of Chlorophyll a content in test example 1 of the present invention；
Fig. 5 is the content of phycocyanin and allophycocyanin in spirulina in test example 1 of the present invention.
Specific embodiment
It is next by the following examples that the present invention is furture elucidated.However, it should be understood that the embodiment is merely illustrative
Purpose, be not intended to limit scope and spirit of the present invention.
Embodiment 1:
The cultural method of chromium-rich spirulina, including,
S1: it is 8.0-10.0, containing 25-35 μm of ol/L 2,6- diamino heptan two that logarithmic phase spirulina, which is inoculated into pH value,
Acid Zarrouk culture medium in, temperature be 25-35 DEG C, illuminance 3000-8000lx, light application time for 24 hours/d under conditions of train
Support 4-8d；
S2: adding chromous acid sodium into culture medium in batches, and daily every liter of culture medium fills into 60-100mg/L, continuously adds 4-
6d makes total additional amount 320-460mg/L of chromous acid sodium, by additive amount is 10-20 μm of ol/ while adding chromium chloride the 2nd time
Thiomalic acid is added in L, continues culture after adding and obtains chromium-rich SPIRULINA CULTIVATION object.Above-mentioned cultural method is able to extend
The growth logarithmic phase of spirulina is effectively relieved Cr (III) to the coercion of spirulina, promotes the fast-growth of spirulina, make
Cr (III) obtains trifoliate orange in spirulina cells and tires out, and improves the biomass, chromium enriching quantity and Chlorophyll-a Content of spirulina, substantially drops
Demand of the low spirulina to inorganic matters such as potassium chloride, potassium sulfate, sodium chloride, while improving the long-term use of made chromium-rich spirulina
Safety.Chromium, chlorophyll a and the phycobniliprotein that obtained chromium-rich SPIRULINA CULTIVATION object contains higher amount, and have in the chromium being enriched with
Machine chromium ratio is high, wherein 20% of Phycobiliprotein Content at least equal to chromium-rich spirulina dry weight, Chlorophyll-a Content in spirulina
At least equal to 19mg/g, chromium content >=7.55mg/g, Organic Chromium ratio >=82.60% in chromium-rich SPIRULINA CULTIVATION object；This is chromium-rich
SPIRULINA CULTIVATION object has no toxic side effect, and bioavilability is high, good absorption effect, has the immune function of enhancing body, improves machine
The biological insulin effect of body, the protein synthesis for promoting body maintain the balance of Blood Cholesterol, improve the hematopoiesis of body
Function, etc. healthcare functions, can be used for preparing alpha-glucosidase inhibit, prepare for preventing or treating cardiovascular disease medicine group
Close object and/or health care product.
Phycobiliprotein Content is at least equal to chromium-rich spirulina dry weight in chromium-rich spirulina obtained by cultural method of the invention
20%.Compared to the range for not applying the 3-12% obtained in the experiment that method of the invention carries out, this is extra high.
The ingredient of 1 Zarrouk culture medium of table
| Component | Concentration (g/L) | Component | Concentration (g/L) |
| NaHCO3 | 16.8 | FeSO4·7H2O | 0.01 |
| NaNO3 | 2.50 | Na2EDTA | 0.08 |
| NaNO3 | 1.00 | CaCl2 | 0.08 |
| K2SO4 | 1.00 | A5Solution | 1mL/L |
| MgSO4·7H2O | 0.20 | B5Solution | 1mL/L |
| K2HPO4 | 0.50 |
2 A of table5The ingredient of solution
| Component | H3BO3 | MoO3 | MnCl2·4H2O | ZnSO4·7H2O | CuSO4·5H2O |
| Concentration (g/L) | 2.86 | 0.01 | 1.80 | 0.22 | 0.08 |
3 B of table5The ingredient of solution
| Component | NH4VO | NaWO4 | Ti2(SO4)3 | NiSO3·7H2O | Co(NO3)2·6H2O |
| Concentration (g/L) | 22.90 | 17.90 | 40.00 | 47.80 | 4.40 |
Embodiment 2:
The cultural method of chromium-rich spirulina, including,
S1: it is 8.0, containing 25 μm of ol/L 2,6- diaminopimelic acids that logarithmic phase spirulina, which is inoculated into pH value,
In Zarrouk culture medium, temperature be 25 DEG C, illuminance 3000lx, light application time for 24 hours/d under conditions of cultivate 4d；
S2: adding chromous acid sodium into culture medium in batches, continuously adds 6d, and every liter of culture medium fills into 60mg/L within the 1st day, the
2 days every liter of culture mediums fill into 90mg/L, and the 3rd day every liter of culture medium fills into 90mg/L, and every liter of culture medium fills into 80mg/L within the 4th day,
5th day every liter of culture medium fills into 70mg/L, and the 6th day every liter of culture medium fills into 60mg/L, by adding while adding chromium chloride within the 2nd day
Dosage is that thiomalic acid is added in 10 μm of ol/L, continues culture 2d after adding and obtains chromium-rich SPIRULINA CULTIVATION object.
Embodiment 3:
The cultural method of chromium-rich spirulina, including,
S1: it is 9.5, containing 30 μm of ol/L 2,6- diaminopimelic acids that logarithmic phase spirulina, which is inoculated into pH value,
In Zarrouk culture medium, temperature be 30 DEG C, illuminance 5000lx, light application time for 24 hours/d under conditions of cultivate 5d；
S2: adding chromous acid sodium into culture medium in batches, continuously adds 5d, and every liter of culture medium fills into 60mg/L within the 1st day, the
2 days every liter of culture mediums fill into 80mg/L, and the 3rd day every liter of culture medium fills into 100mg/L, and every liter of culture medium fills into 80mg/L within the 4th day,
5th day every liter of culture medium fills into 80mg/L, adds within the 2nd day thio for 16 μm of ol/L additions by each additive amount while chromium chloride
Malic acid continues 2d culture and obtains chromium-rich SPIRULINA CULTIVATION object after adding.
Embodiment 4:
The cultural method of chromium-rich spirulina, including,
S1: it is 9.5, containing 30 μm of ol/L 2,6- diaminopimelic acids that logarithmic phase spirulina, which is inoculated into pH value,
In Zarrouk culture medium, temperature be 30 DEG C, illuminance 5000lx, light application time for 24 hours/d under conditions of cultivate 6d；
S2: adding chromous acid sodium into culture medium in batches, continuously adds 5d, and every liter of culture medium fills into 60mg/L within the 1st day, the
2 days every liter of culture mediums fill into 80mg/L, and the 3rd day every liter of culture medium fills into 100mg/L, and every liter of culture medium fills into 80mg/L within the 4th day,
4th day every liter of culture medium fills into 60mg/L, adds within the 2nd day thio for 16 μm of ol/L additions by each additive amount while chromium chloride
Malic acid continues 1d culture and obtains chromium-rich SPIRULINA CULTIVATION object after adding.
Embodiment 5:
The cultural method of chromium-rich spirulina, including,
S1: it is 9.5, containing 30 μm of ol/L 2,6- diaminopimelic acids that logarithmic phase spirulina, which is inoculated into pH value,
In Zarrouk culture medium, temperature be 30 DEG C, illuminance 5000lx, light application time for 24 hours/d under conditions of cultivate 7d；
S2: adding chromous acid sodium into culture medium in batches, continuously adds 4d, and every liter of culture medium fills into 60mg/L within the 1st day, the
2 days every liter of culture mediums fill into 80mg/L, and the 3rd day every liter of culture medium fills into 100mg/L, and every liter of culture medium fills into 100mg/ within the 4th day
L is added thiomalic acids by each additive amount while adding within the 2nd day chromium chloride for 16 μm of ol/L, continues 1d after adding
Culture obtains chromium-rich SPIRULINA CULTIVATION object.
Embodiment 6:
The cultural method of chromium-rich spirulina, including,
S1: it is 10.0, containing 35 μm of ol/L 2,6- diaminopimelic acids that logarithmic phase spirulina, which is inoculated into pH value,
In Zarrouk culture medium, temperature be 35 DEG C, illuminance 8000lx, light application time for 24 hours/d under conditions of cultivate 8d；
S2: adding chromous acid sodium into culture medium in batches, and daily every liter of culture medium fills into 80mg/L, continuously adds 4d, and the 1st
It every liter culture medium fills into 80mg/L, and the 2nd day every liter of culture medium fills into 100mg/L, and every liter of culture medium fills into 100mg/L within the 3rd day,
4th day every liter of culture medium fills into 80mg/L, and thio apples are added for 20 μm of ol/L by additive amount while adding within the 2nd day chromium chloride
Acid to get arrive chromium-rich SPIRULINA CULTIVATION object.
Comparative example 1:
With embodiment 3 the difference is that: be added without thiomalic acid while adding chromium chloride within the 2nd day.
Comparative example 2:
With embodiment 3 the difference is that: in Zarrouk culture medium be free of 2,6- diaminopimelic acid.
Comparative example 3:
With embodiment 3 the difference is that: in Zarrouk culture medium be free of 2,6- diaminopimelic acid；It adds within 2nd day
Thiomalic acid is added without while chromium chloride.
Comparative example 4:
The cultural method of chromium-rich spirulina, including,
Logarithmic phase spirulina is inoculated into the Zarrouk culture medium that pH value is 9.5, is 30 DEG C, illuminance in temperature
5000lx, light application time for 24 hours/d under conditions of culture 12d to get arrive chromium-rich SPIRULINA CULTIVATION object.
Test example 1:
1. test method
The measurement of 1.1 cell product of Spirulina
1.1.1 absorbance A560With the relationship of cell product of Spirulina
3 parts of 1000.00mL algae solutions are taken respectively, with having claimed the filter paper of dry weight to filter, the algal gel filtered with distilled water flushing
For several times, it is put into 105 DEG C of dryings of electric drying oven with forced convection after draining to constant weight, is weighed with electronic analytical balance, 3 parts of test specimens take
The cell product of Spirulina of algae solution is calculated in average value.100.00mL algae solution is taken, is diluted to different times respectively according to doubling dilution
Several serial solutions, using 722S spectrophotometer, is made blank with Zarrouk culture solution, takes the algae of mixing using optical densitometric method
Liquid measures absorbance (A under 560nm wavelength560), with A560For abscissa, cell product of Spirulina W (g/L) is ordinate, is drawn
A560With cell product of Spirulina relational graph (such as Fig. 1), equation of linear regression are as follows: y=-0.02629+0.71068x.
1.2 biomass estimations and growth curve are drawn
Daily timing sampling measures and records A in incubation560Value, is repeated 3 times and is averaged, resulting using 1.1.1
A560With the linear equation of cell product of Spirulina, the algae solution A that will be measured daily560It is converted into biomass W (g/L).With incubation time t
For abscissa, biomass W is ordinate, draws growth of spirulina platensis curve.
Enrichment total amount and organic degree of 1.2 spirulinas to chromium
1.2.1 inorganic chrome determination
Frustule sample is collected by centrifugation, 60 DEG C of oven dryings are for 24 hours.0.002g sample is weighed, 5mL ddH is separately added into2O
In, it is crushed 30 × 5s on Ultrasonic Cell Disruptor and (is intermittently 4s), supernatant is collected by centrifugation, dilute 10 times of upper ICP-OES detections.
1.2.2 total chrome content measures
It weighs 0.004g sample and adds nitration mixture HNO3:HClO4(4:1) 5mL, 180 DEG C of hot digestion 6h on electric hot plate, after cooling
Filtering, with ultrapure water constant volume, upper machine testing after dilution measures total chrome content using ICP-AES.Total chrome subtracts inorganic chromium and measures
To the content of Organic Chromium.
The measurement of 1.3 Chlorophyll-a Contents
It utilizes the content of Extracting method by hot methanol (AzovYetal., 1982) measurement chlorophyll a: pipetting 1ml algae solution and be placed on
In the centrifuge tube of 15ml, 3500rpm is centrifuged 5min, removes supernatant；Then the concussion of 5ml methanol is added to mix, centrifuge tube is placed
The heating extraction 5min in 70 DEG C of water-bath, 3500rpm are centrifuged 5min, and supernatant is taken to measure extinction at 665nm and 750nm
Value.Calculation formula are as follows:
Chl.a (mg/L)=13.9 (OD665-OD750)·U/V；
Wherein, U is methanol volume, and V is algae solution volume.
The measurement of 1.4 Phycobiliprotein Contents
Algae solution to be measured is centrifuged 5min under the conditions of 8000rpm, removes supernatant, twice using aliquots of deionized water cleaning
Afterwards, frond is freeze-dried.Using direct osmometry, 0.06g is taken to do spiral frond, 10mL phosphate buffer is added
(0.15mol/L, pH 7.0) stands 36h at 4 DEG C after mixing, obtains clear indigo plant after being centrifuged 5min under the conditions of 8000rpm
Color supernatant, ultraviolet specrophotometer measure the optical density of liquid at 620nm and 652nm, calculate phycocyanin according to the following formula
Concentration C (g/L):
C=(OD620-0.474×OD652)÷5.34；
Phycocyanin percentage composition Y (%) in frond is calculated according to the following formula:
Y=(C × V ÷ M) × 100%；
Wherein V is phosphate buffer volume (L), and M represents spirulina powder weight (g).
2. test result
2.1 Fig. 2 are the growth curve of spirulina, it will thus be seen that in incubation time are 5-7d, the growth of comparative example 3 is better than
Comparative example 4, and be 8-12d in incubation time, the growth of comparative example 3 is worse than comparative example 4；It is to implement in 1-5d in incubation time
The growth trend of 1 spirulina of example 3 and comparative example is better than comparative example 2-4；It is 6-12d, embodiment 3 and comparative example 2 in incubation time
The growth trend of spirulina is better than comparative example 1,3-4；The whole growth curve of 3 spirulina of embodiment is better than comparative example 1-4.This table
Bright chromium there is growth to promote the dual character with toxicity inhibition algae；Other component combines in thiomalic acid and culture medium, can
Oxidative stress caused by resisting because of chromium concn height, helper cell resist chromium toxicity, promote spirulina raw under higher chromium content
It is long；2,6- diaminopimelic acids have facilitation to the growth of spirulina, this is because 2,6- diaminopimelic acids can promote
Into more phycobniliprotein are accumulated in cell, phycobniliprotein can not only enhance the adaptive capacity to environment of algae, and then promote spiral
The growth of algae；And absorption luminous energy can be directly captured, the light energy electronic chain transmitting in chloroplaset is participated in, the environment of algae is enhanced
Adaptability promotes the growth of spirulina so that spirulina cells be helped to adapt to the coercion of Cr (III), improves spirulina
Biomass.
2.2 Fig. 3 are enrichment total amount and organic degree of the spirulina to chromium, it can be seen that rich in embodiment 2-6 spirulina
Collection total amount and organic degree are above comparative example 1-4, and are enriched with total amount and organic degree in 2 spirulina of comparative example and are higher than pair
Ratio 1 and 3.This illustrates that thiomalic acid in culture medium can be improved the chromium enriching quantity of spirulina, and be particularly advantageous for chromium has
Machine chelating improves absorption and enrichment of the spirulina to chromium, obtains high chromium-rich amount spirulina；2,6- diaminopimelic acid is to spirulina
Chromium-rich function influence it is unobvious.
2.3 Fig. 4 are spirulina Determination of Chlorophyll a content, it can be seen that embodiment 2-6 spirulina Determination of Chlorophyll a content is high
In comparative example 1-4, and 2 spirulina Determination of Chlorophyll a content of comparative example is higher than comparative example 1 and 3.This illustrates thio apple in culture medium
Acid can also make the Chlorophyll-a Content in spirulina improve a lot with other component combines in culture medium, improve the photosynthetic of Sargassum horneri
Pigment level, enhances the optical energy utilization efficiency of frond, and then promotes the promotion of its photosynthetic capacity, and spirulina is promoted to contain in higher chromium
The lower growth of amount, improves the biomass and chromium enriching quantity of spirulina.
2.4 Fig. 5 are the content of phycocyanin and allophycocyanin in spirulina, it can be seen that embodiment 2-6 and comparative example
The content of phycobniliprotein is above comparative example 2-4 in 1 spirulina, this explanation 2,6- diaminopimelic acid can mention after entering cell
The level of pigment content and enzyme in high cell and the growth for promoting cell, so as to accumulate more phycobniliprotein；Meanwhile 2,
6- diaminopimelic acid is also used as the spare nitrogen source of growth of spirulina platensis, protects the phycobniliprotein of synthesis will not lacking because of nitrogen source
Be degraded utilization less, promotes to accumulate more phycobniliprotein in cell.
The prior art of routine techniques dawn known to those skilled in the art in above-described embodiment, therefore herein no longer in detail
It repeats.
The above embodiments are only used to illustrate the present invention, and not limitation of the present invention, the ordinary skill people of this field
Member can also make a variety of changes and modification without departing from the spirit and scope of the present invention.Therefore, all equivalent
Technical solution also belong to scope of the invention, scope of patent protection of the invention should be defined by the claims.
Claims (10)
1. the cultural method of chromium-rich spirulina, it is characterised in that: logarithmic phase spirulina is inoculated into culture medium and is cultivated, was cultivated
Chromium chloride is added in journey in batches, continues to cultivate to obtain chromium-rich spirulina；
Described adding in batches is to be inoculated into culture medium to add in batches after culture 4-8d from spirulina, continuously adds chromium chloride 4-
6d；
Described is added thiomalic acid while continuously adding chromium chloride.
2. the cultural method and application thereof of chromium-rich spirulina according to claim 1, it is characterised in that: the benefit in batches
The amount for adding chromium chloride is that daily every liter of culture medium fills into 50-150mg/L.
3. the cultural method and application thereof of chromium-rich spirulina according to claim 1, it is characterised in that: the thio apple
The additive amount of tartaric acid is 10-20 μm of ol/L.
4. the cultural method and application thereof of chromium-rich spirulina according to claim 1, it is characterised in that: the culture medium
PH value be 8.0-10.0, temperature be 25-35 DEG C, illuminance 3000-8000lx, light application time for 24 hours/d.
5. the cultural method of chromium-rich spirulina according to claim 1-4, it is characterised in that: the culture medium
In contain 2,6- diaminopimelic acid.
6. the cultural method of chromium-rich spirulina according to claim 5, it is characterised in that: contain in the culture medium
10-50 μm of ol/L2,6- diaminopimelic acid.
7. the cultural method of chromium-rich spirulina according to claim 1, it is characterised in that: algae in the chromium-rich spirulina
20% of biliprotein content at least equal to the chromium-rich spirulina dry weight.
8. a kind of chromium-rich SPIRULINA CULTIVATION object that the described in any item cultural methods of claim 1-7 obtain, it is characterised in that: institute
Chromium content >=7.55mg/g, Organic Chromium ratio >=82.60% in the chromium-rich SPIRULINA CULTIVATION object stated.
9. chromium-rich SPIRULINA CULTIVATION object according to any one of claims 8 is preparing the purposes in alpha-glucosidase inhibition.
10. chromium-rich SPIRULINA CULTIVATION object according to any one of claims 8 is in preparation for preventing or treating cardiovascular disease medicine combination
Purposes in object and/or health care product.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| IT202000002206A1 (en) * | 2020-02-05 | 2021-08-05 | Medical Network Liaison Srl | PROCEDURE FOR OBTAINING ENRICHED ALGAE |
| CN115521890A (en) * | 2022-11-11 | 2022-12-27 | 江苏省沿海开发投资有限公司 | Method for promoting spirulina to produce phycocyanin by adding exogenous substances |
| CN115666610A (en) * | 2020-05-19 | 2023-01-31 | 瓦克萨科技有限公司 | Photosynthetic controlled spirulina extracts for the treatment of cytokine storm syndrome |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2144078C1 (en) * | 1998-02-17 | 2000-01-10 | Акционерное общество открытого типа "Биохиммаш" | Method of preparing blue-green alga spirulina enriched with trace elements |
| RU2002115679A (en) * | 2002-06-11 | 2004-01-27 | Объединенный институт дерных исследований (RU) | A method of obtaining a biomass of spirulina |
| US20080260692A1 (en) * | 2005-03-16 | 2008-10-23 | Jean-Louis Vidalo | Spirulin Composition Rich in Active Principles, Method for Obtaining Same and Use Thereof |
| CN102845780A (en) * | 2012-09-01 | 2013-01-02 | 山东博然螺旋藻生物股份有限公司 | Preparation method of chrome-rich quick-frozen spiral algae |
| WO2013058432A1 (en) * | 2011-10-18 | 2013-04-25 | 한국해양연구원 | Method for manufacturing a culture medium of algae using natural seawater |
| CN104450581A (en) * | 2014-12-10 | 2015-03-25 | 安徽大学 | Cultivation method of hpyerglycemic spirulina |
| US20150353982A1 (en) * | 2012-12-28 | 2015-12-10 | University Of Newcastle Upon Tyne | Micro-evolution of microbes |
| CN105483062A (en) * | 2016-02-02 | 2016-04-13 | 杭州优普西生物科技有限公司 | Microelement-rich spirulina culture medium |
| CN106085943A (en) * | 2016-08-04 | 2016-11-09 | 湖南新生活螺旋藻生物科技有限公司 | A kind of Spirulina powder being enriched with rare element and its preparation method and application |
-
2019
- 2019-07-02 CN CN201910590182.5A patent/CN110257296B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2144078C1 (en) * | 1998-02-17 | 2000-01-10 | Акционерное общество открытого типа "Биохиммаш" | Method of preparing blue-green alga spirulina enriched with trace elements |
| RU2002115679A (en) * | 2002-06-11 | 2004-01-27 | Объединенный институт дерных исследований (RU) | A method of obtaining a biomass of spirulina |
| US20080260692A1 (en) * | 2005-03-16 | 2008-10-23 | Jean-Louis Vidalo | Spirulin Composition Rich in Active Principles, Method for Obtaining Same and Use Thereof |
| WO2013058432A1 (en) * | 2011-10-18 | 2013-04-25 | 한국해양연구원 | Method for manufacturing a culture medium of algae using natural seawater |
| CN102845780A (en) * | 2012-09-01 | 2013-01-02 | 山东博然螺旋藻生物股份有限公司 | Preparation method of chrome-rich quick-frozen spiral algae |
| US20150353982A1 (en) * | 2012-12-28 | 2015-12-10 | University Of Newcastle Upon Tyne | Micro-evolution of microbes |
| CN104450581A (en) * | 2014-12-10 | 2015-03-25 | 安徽大学 | Cultivation method of hpyerglycemic spirulina |
| CN105483062A (en) * | 2016-02-02 | 2016-04-13 | 杭州优普西生物科技有限公司 | Microelement-rich spirulina culture medium |
| CN106085943A (en) * | 2016-08-04 | 2016-11-09 | 湖南新生活螺旋藻生物科技有限公司 | A kind of Spirulina powder being enriched with rare element and its preparation method and application |
Non-Patent Citations (1)
| Title |
|---|
| 何颖敏等: "螺旋藻富铬及其含铬蛋白组份对 α-葡萄糖苷酶抑制的研究", 《安徽农业大学学报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| IT202000002206A1 (en) * | 2020-02-05 | 2021-08-05 | Medical Network Liaison Srl | PROCEDURE FOR OBTAINING ENRICHED ALGAE |
| CN115666610A (en) * | 2020-05-19 | 2023-01-31 | 瓦克萨科技有限公司 | Photosynthetic controlled spirulina extracts for the treatment of cytokine storm syndrome |
| CN115521890A (en) * | 2022-11-11 | 2022-12-27 | 江苏省沿海开发投资有限公司 | Method for promoting spirulina to produce phycocyanin by adding exogenous substances |
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