CN104839018A - Preparation method for selenium-rich hair weed product - Google Patents
Preparation method for selenium-rich hair weed product Download PDFInfo
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- CN104839018A CN104839018A CN201510206788.6A CN201510206788A CN104839018A CN 104839018 A CN104839018 A CN 104839018A CN 201510206788 A CN201510206788 A CN 201510206788A CN 104839018 A CN104839018 A CN 104839018A
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Abstract
The invention specifically relates to a preparation method for a selenium-rich hair weed product, which belongs to the technical field of food bio-concentration. According to the invention, hair weed cells transforms the inorganic selenium--sodium selenite in a nutrient solution into organic selenium in virtue of the own transformation effect of the cells, so harm to the human body is reduced, and absorption and utilization of selenium by human beings and animals are improved; and the growth rate of the hair weed cells in the process of culture is 30 times of the growth rate of cells of wild hair weed, so rapid growth and reproduction of hair weed are realized. The hair weed cells produced in the invention can be used as a natural biological raw material for selenium supplement and are extensively applied to a variety of health food, beverages and new resource food to meet demands of the human body for selenium. The nutrient solution deprived of the hair weed cells contains abundant hair weed exopolysaccharide which has antiviral activity and can replace hair weed as a raw material for production of functional food. A hair weed selenium enrichment culture method provided by the invention is simple and convenient to operate, poses no pollution to the environment, has low cost and is applicable to industrialized production.
Description
Technical field
The invention belongs to biological food beneficiation technologies field, be specifically related to a kind of rich selenium and deliver vegetables the preparation method of product.
Background technology
Selenium is respectfully called " kindling material of life " by domestic and international the world of medicine and nutrition educational circles, enjoys the good reputation such as " long-lived element ", " anticancer king ", " heart patron saint ", " natural antipoison ".Selenium is 1/10000000th at tissue intensive amount, but it determines the existence of life, is that other materials are irreplaceable to the great function of human health.Scarce selenium can directly cause human immunological competence to decline, and clinical medicine proves, threatens human health and 40 various diseases of life all to lack selenium with human body relevant, as cancer, cardiovascular disease, hepatopathy, cataract, pancreatic disease, diabetes, genital system diseases etc.No matter be zoopery or clinical practice, all show the selenium that constantly should obtain q.s from diet, can not supplement in time, will the ability of getting rid of illness be reduced.
Deliver vegetables, also known as hair-like nostoc, because its look black and elongated, as people hair and gain the name, and deliver vegetables and " making a good deal of money " homophonic, very popular over the past thousands of years.Deliver vegetables rich in nutritive value, containing protein 22.8g in every 100g natural laver, be 1.5 times of egg, fat content is low, is rich in the trace elements such as calcium, iron, iodine, magnesium, zinc, selenium, manganese.In recent years, delivering vegetables excessively gathers causes great harm to ecotope.At present, from frond of delivering vegetables, be separated hair weeds cells carry out liquid suspension culture and be subject to people's attention gradually.Hair weeds cells culture technique is the hair weeds cells that separation and purification goes out to have vigorous splitting ability from frond of delivering vegetables, and makes hair weeds cells amount reproduction by liquid suspension culture, by reprocessing process, realizes hair weeds cells factorial praluction.In incubation, the growth rate of hair weeds cells is 30 times of wild growth rate of delivering vegetables, and achieves the quick growth and breeding of hair-like seaweed resource.
Selenium has two kinds of existing forms in hair weeds cells: the ionic state form of first selenite, and it two is the Organic Selenium forms be combined with organic matter.Ionic state selenium can be dissolved in water, is absorbed in cell, and wherein a part and combined with protein change into Organic Selenium further, and this is the prerequisite of hair weeds cells enrichment Organic Selenium.And ionic state selenium soluble in water, especially easily run off in process of production, cause availability not high, economic benefit is not obvious.If ingested for a long time, high-load ionic state selenium, not only causes waste, also there will be selenosis phenomenon.On the contrary, Organic Selenium toxicity is low, and biologically active is high, is easily absorbed by humans and animals, is selenium preparation safely and efficiently.Enrichment Organic Selenium hair weeds cells has that Commercial cultivation rate is high, the feature of edible safety.Organic Selenium hair weeds cells can be used for the various diseases caused because of scarce selenium.Therefore, hair weeds cells is utilized to carry out biological concentration with conversion inorganic selenium for Organic Selenium is the approach at present with development prospect as carrier.
Summary of the invention
The object of this invention is to provide a kind of rich selenium to deliver vegetables the preparation method of product, not only can significantly improve the growth rate of delivering vegetables, can also promote that human body is to the absorption of selenium.
The technical solution used in the present invention is: a kind of rich selenium is delivered vegetables the preparation method of product, comprises the step to separation and purification of delivering vegetables; To the step of the hair weeds cells cultivation that separation and purification obtains; To the step of rich selenium hair weeds cells process; Described to delivering vegetables in the step of separation and purification: delivering vegetables after cleaning and sterilizing is soaked in 8 ~ 11h in minimal medium, then obtain hair weeds cells slurry with glass homogenizer homogenate, hair weeds cells slurry is seeded on minimal medium and carries out the obtained hair weeds cells of purifying cultivation;
In the step that the described hair weeds cells obtained separation and purification is cultivated: aseptically, obtained hair weeds cells is seeded on the selenium-enriched liquid medium in blake bottle, then the blake bottle being connected to hair weeds cells is put into the constant temperature biochemical cultivation case that temperature is 25 DEG C, average intensity of illumination is 2000Lux to cultivate 15 ~ 20 days, in incubation, illumination and dark are every 12h alternately once, filter afterwards and isolate hair weeds cells i.e. obtained rich selenium hair weeds cells;
In the described step to the process of rich selenium hair weeds cells: rich selenium hair weeds cells distilled water is washed, to remove deliver vegetables exocellular polysaccharide and the inorganic selenium of cell surface, richer selenium hair weeds cells to be put into temperature be that the environment freeze drying i.e. obtained rich selenium of-50 ~-55 DEG C is delivered vegetables product;
The preparation method of described selenium-enriched liquid medium is the Na of 1mg/mL by mass concentration
2seO
3join in minimal medium after aqueous solution sterilizing, until Na
2seO
3mass concentration be 2 ~ 40mg/L, adjust ph to 7.5 i.e. obtained selenium-enriched liquid medium.
Described minimal medium is by NaNO
3, MgSO
47H
2o, CaCl
22H
2o, H
3bO
4, MnCl
24H
2o, ZnSO
47H
2o, CuSO
45H
2o, NaMoO
42H
2o, CoCl
2, EDTA-Na
2, FeSO
47H
2o, citric acid, Na
2siO
39H
2o and K
2hPO
4make, and the parts by weight of each component are respectively NaNO
3250 parts, MgSO
47H
2o 75 parts, CaCl
22H
2o 36 parts, H
3bO
42.86 parts, MnCl
24H
2o 1.8 parts, ZnSO
47H
2o 0.2 part, CuSO
45H
2o 0.8 part, NaMoO
42H
2o 0.4 part, CoCl
20.01 part, EDTA-Na
2l part, FeSO
47H
2o 4.8 parts, citric acid 0.6 part, Na
2siO
39H
2o 58 parts, K
2hPO
440 parts.
beneficial effect
1, in the present invention, the inorganic selenium sodium selenite in culture fluid is changed into Organic Selenium by the transformation of self by hair weeds cells, reduces the harm to human body, improves people and animal absorbing selenium.
2, the growth rate of hair weeds cells of the present invention in incubation is 30 times of wild growth rate of delivering vegetables, and achieves the quick growth and breeding of delivering vegetables.
3, the hair weeds cells that the present invention produces as the natural biological raw material mending selenium, can use, to meet the demand of human body to selenium widely in various health food, beverage and new resource food.
4, containing abundant exocellular polysaccharide of delivering vegetables, there is antiviral activity in the culture fluid after removing hair weeds cells in the present invention, the raw material of delivering vegetables as producing functional food can being substituted.
5, involved in the present invention rich selenium cultural method of delivering vegetables is easy and simple to handle, and do not pollute environment, cost is lower, is applicable to factorial praluction.
Embodiment
Rich selenium is delivered vegetables the preparation method of product, comprises the step to separation and purification of delivering vegetables; To the step of the hair weeds cells cultivation that separation and purification obtains; To the step of rich selenium hair weeds cells process; Described to delivering vegetables in the step of separation and purification: delivering vegetables after cleaning and sterilizing is soaked in 8 ~ 11h in minimal medium, then obtain hair weeds cells slurry with glass homogenizer homogenate, hair weeds cells slurry is seeded on minimal medium and carries out the obtained hair weeds cells of purifying cultivation;
In the step that the described hair weeds cells obtained separation and purification is cultivated: aseptically, obtained hair weeds cells is seeded on the selenium-enriched liquid medium in blake bottle, then the blake bottle being connected to hair weeds cells is put into the constant temperature biochemical cultivation case that temperature is 25 DEG C, average intensity of illumination is 2000Lux to cultivate 15 ~ 20 days, in incubation, illumination and dark are every 12h alternately once, filter afterwards and isolate hair weeds cells i.e. obtained rich selenium hair weeds cells;
In the described step to the process of rich selenium hair weeds cells: rich selenium hair weeds cells distilled water is washed, to remove deliver vegetables exocellular polysaccharide and the inorganic selenium of cell surface, richer selenium hair weeds cells to be put into temperature be that the environment freeze drying i.e. obtained rich selenium of-50 ~-55 DEG C is delivered vegetables product;
The preparation method of described selenium-enriched liquid medium is the Na of 1mg/mL by mass concentration
2seO
3join in minimal medium after aqueous solution sterilizing, until Na
2seO
3mass concentration be 2 ~ 40mg/L, adjust ph to 7.5 i.e. obtained selenium-enriched liquid medium.
Described minimal medium is by NaNO
3, MgSO
47H
2o, CaCl
22H
2o, H
3bO
4, MnCl
24H
2o, ZnSO
47H
2o, CuSO
45H
2o, NaMoO
42H
2o, CoCl
2, EDTA-Na
2, FeSO
47H
2o, citric acid, Na
2siO
39H
2o and K
2hPO
4make, and the parts by weight of each component are respectively NaNO
3250 parts, MgSO
47H
2o 75 parts, CaCl
22H
2o 36 parts, H
3bO
42.86 parts, MnCl
24H
2o 1.8 parts, ZnSO
47H
2o 0.2 part, CuSO
45H
2o 0.8 part, NaMoO
42H
2o 0.4 part, CoCl
20.01 part, EDTA-Na
2l part, FeSO
47H
2o 4.8 parts, citric acid 0.6 part, Na
2siO
39H
2o 58 parts, K
2hPO
440 parts.
Embodiment 1
Rich selenium is delivered vegetables the preparation method of product, comprises the step to separation and purification of delivering vegetables; To the step of the hair weeds cells cultivation that separation and purification obtains; To the step of rich selenium hair weeds cells process; Described to delivering vegetables in the step of separation and purification: delivering vegetables after cleaning and sterilizing is soaked in 8h in minimal medium, then obtain hair weeds cells slurry with glass homogenizer homogenate, hair weeds cells slurry is seeded on minimal medium and carries out the obtained hair weeds cells of purifying cultivation;
In the step that the described hair weeds cells obtained separation and purification is cultivated: aseptically, obtained hair weeds cells is seeded on the selenium-enriched liquid medium in blake bottle, then the blake bottle being connected to hair weeds cells is put into the constant temperature biochemical cultivation case that temperature is 25 DEG C, average intensity of illumination is 2000Lux to cultivate 15 days, in incubation, illumination and dark are every 12h alternately once, filter afterwards and isolate hair weeds cells i.e. obtained rich selenium hair weeds cells;
In the described step to the process of rich selenium hair weeds cells: rich selenium hair weeds cells distilled water is washed, to remove deliver vegetables exocellular polysaccharide and the inorganic selenium of cell surface, richer selenium hair weeds cells to be put into temperature be that the environment freeze drying i.e. obtained rich selenium of-50 DEG C is delivered vegetables product;
The preparation method of described selenium-enriched liquid medium is the Na of 1mg/mL by mass concentration
2seO
3join in minimal medium after aqueous solution sterilizing, until Na
2seO
3mass concentration be 2mg/L, adjust ph to 7.5 i.e. obtained selenium-enriched liquid medium.
Described minimal medium is by NaNO
3, MgSO
47H
2o, CaCl
22H
2o, H
3bO
4, MnCl
24H
2o, ZnSO
47H
2o, CuSO
45H
2o, NaMoO
42H
2o, CoCl
2, EDTA-Na
2, FeSO
47H
2o, citric acid, Na
2siO
39H
2o and K
2hPO
4make, and the parts by weight of each component are respectively NaNO
3250 parts, MgSO
47H
2o 75 parts, CaCl
22H
2o 36 parts, H
3bO
42.86 parts, MnCl
24H
2o 1.8 parts, ZnSO
47H
2o 0.2 part, CuSO
45H
2o 0.8 part, NaMoO
42H
2o 0.4 part, CoCl
20.01 part, EDTA-Na
2l part, FeSO
47H
2o 4.8 parts, citric acid 0.6 part, Na
2siO
39H
2o 58 parts, K
2hPO
440 parts.
Embodiment 2
Rich selenium is delivered vegetables the preparation method of product, comprises the step to separation and purification of delivering vegetables; To the step of the hair weeds cells cultivation that separation and purification obtains; To the step of rich selenium hair weeds cells process; Described to delivering vegetables in the step of separation and purification: delivering vegetables after cleaning and sterilizing is soaked in 11h in minimal medium, then obtain hair weeds cells slurry with glass homogenizer homogenate, hair weeds cells slurry is seeded on minimal medium and carries out the obtained hair weeds cells of purifying cultivation;
In the step that the described hair weeds cells obtained separation and purification is cultivated: aseptically, obtained hair weeds cells is seeded on the selenium-enriched liquid medium in blake bottle, then the blake bottle being connected to hair weeds cells is put into the constant temperature biochemical cultivation case that temperature is 25 DEG C, average intensity of illumination is 2000Lux to cultivate 20 days, in incubation, illumination and dark are every 12h alternately once, filter afterwards and isolate hair weeds cells i.e. obtained rich selenium hair weeds cells;
In the described step to the process of rich selenium hair weeds cells: rich selenium hair weeds cells distilled water is washed, to remove deliver vegetables exocellular polysaccharide and the inorganic selenium of cell surface, richer selenium hair weeds cells to be put into temperature be that the environment freeze drying i.e. obtained rich selenium of-50 ~-55 DEG C is delivered vegetables product;
The preparation method of described selenium-enriched liquid medium is the Na of 1mg/mL by mass concentration
2seO
3join in minimal medium after aqueous solution sterilizing, until Na
2seO
3mass concentration be 40mg/L, adjust ph to 7.5 i.e. obtained selenium-enriched liquid medium.
Described minimal medium is by NaNO
3, MgSO
47H
2o, CaCl
22H
2o, H
3bO
4, MnCl
24H
2o, ZnSO
47H
2o, CuSO
45H
2o, NaMoO
42H
2o, CoCl
2, EDTA-Na
2, FeSO
47H
2o, citric acid, Na
2siO
39H
2o and K
2hPO
4make, and the parts by weight of each component are respectively NaNO
3250 parts, MgSO
47H
2o 75 parts, CaCl
22H
2o 36 parts, H
3bO
42.86 parts, MnCl
24H
2o 1.8 parts, ZnSO
47H
2o 0.2 part, CuSO
45H
2o 0.8 part, NaMoO
42H
2o 0.4 part, CoCl
20.01 part, EDTA-Na
2l part, FeSO
47H
2o 4.8 parts, citric acid 0.6 part, Na
2siO
39H
2o 58 parts, K
2hPO
440 parts.
Embodiment 3
The deliver vegetables preparation method of product of a kind of rich selenium specifically comprises following step:
Step one: the preparation of minimal medium
1. A liquid: take CaCl respectively
21.36g, MgSO
47H
2o 3.75g, after dissolving with distilled water, be settled to 500mL, normal temperature saves backup;
2. B liquid: take NaMoO respectively
42H
2o 20mg, ZnSO
47H
2o 10mg, H
3bO
4143mg, MnCl
24H
2o 90mg, EDTA-Na
250mg, FeSO
47H
2o 240mg, CuSO
45H
2o 40mg, CoCl
26H
2o 0.5mg, citric acid 30mg, after distilled water dissolves, be settled to 500mL, 4 DEG C of refrigerator cold-storages are for subsequent use;
3. P liquid: take K
2hPO
43H
2o 2g, after distilled water dissolves, be settled to 500mL, normal temperature saves backup;
4. Si liquid: take Na
2siO
39H
2o 2.9g, after distilled water dissolves, be settled to 500mL, normal temperature saves backup;
5. accurately draw A liquid 20mL with pipette, B liquid 2mL mixes, and adding distil water is settled to 1000mL, is dispensed in 10 250mL conical flasks, every bottle of 100mL, wrapping sterilizing.Wet heating sterilizing: temperature 121 DEG C, time 20min.
6. accurately draw P liquid 20mL with pipette, Si liquid 20mL mixes, then adds NaNO3 2.5g, and adding distil water is settled to 1000mL, is dispensed in 10 250mL conical flasks, every bottle of 100mL, wrapping sterilizing.Wet heating sterilizing: temperature 121 DEG C, time 20min.
7. be cooled to room temperature under 5 after sterilizing and 6 being placed on superclean bench aseptic condition, be then mixed in 1 bottle, divide after shaking up again and be filled to two bottles, namely minimal medium prepares.
Step 2: the preparation of rich seleno culture medium
Take Na
2seO
350mg, distilled water is settled to 50mL, is configured to the mother liquor of 1mg/mL after dissolving, and adopts filtration method to carry out degerming to the solution configured.The sodium selenite mother liquor configured is added in minimal medium, makes the concentration of sodium selenite in medium reach 2 ~ 40mg/L, be rich seleno culture medium of delivering vegetables, the pH value to 7.5 of adjustment medium.
Step 3: the separation and purification of hair weeds cells bacterial classification.
Dry delivering vegetables is cleaned 3 ~ 4 times with distilled water before use, dry the alcohol disinfecting 0.5 ~ 1.0min of rear use 75%, distilled water flushing is clean and be soaked in 8 ~ 11h in minimal medium, then uses glass homogenizer homogenate, examines under a microscope and produce a large amount of filament and free single cell.Streak inoculation on minimal medium, is cultivated by repeatedly purifying and obtains hair weeds cells culture.On picking solid culture medium, single algae of purifying falls, and is placed in minimal medium inflation and expands cultivation.The cell that results are cultured to exponential phase is inoculated for this test.
Step 4: the cultivation of rich selenium hair weeds cells
Aseptically, hair weeds cells through separation and purification is seeded in selenium-enriched liquid medium, then the blake bottle being connected to hair weeds cells is put into the constant temperature biochemical cultivation case of 25 DEG C, condition of culture is that periodicity of illumination 12h illumination 12h is dark, intensity of illumination is 2000Lux, cultivate 15 ~ 20 days, quiescent culture liquid, filters to isolate hair weeds cells for subsequent use;
Step 5: the preparation of rich selenium hair weeds cells
By isolated hair weeds cells distilled water washing two to three times, to remove the inorganic selenium of delivering vegetables in exocellular polysaccharide and culture fluid of cell surface.Hair weeds cells after washing is poured in flat board, through freeze drying, namely obtains the hair weeds cells of rich selenium.
Result shows: the growth rate of hair weeds cells of the present invention in incubation is 30 times of wild growth rate of delivering vegetables, and achieves the quick growth and breeding of delivering vegetables.
Claims (2)
1. rich selenium is delivered vegetables a preparation method for product, comprises the step to separation and purification of delivering vegetables; To the step of the hair weeds cells cultivation that separation and purification obtains; To the step of rich selenium hair weeds cells process; It is characterized in that:
Described to delivering vegetables in the step of separation and purification: delivering vegetables after cleaning and sterilizing is soaked in 8 ~ 11h in minimal medium, then obtain hair weeds cells slurry with glass homogenizer homogenate, hair weeds cells slurry is seeded on minimal medium and carries out the obtained hair weeds cells of purifying cultivation;
In the step that the described hair weeds cells obtained separation and purification is cultivated: aseptically, obtained hair weeds cells is seeded on the selenium-enriched liquid medium in blake bottle, then the blake bottle being connected to hair weeds cells is put into the constant temperature biochemical cultivation case that temperature is 25 DEG C, average intensity of illumination is 2000Lux to cultivate 15 ~ 20 days, in incubation, illumination and dark are every 12h alternately once, filter afterwards and isolate hair weeds cells i.e. obtained rich selenium hair weeds cells;
In the described step to the process of rich selenium hair weeds cells: rich selenium hair weeds cells distilled water is washed, to remove deliver vegetables exocellular polysaccharide and the inorganic selenium of cell surface, richer selenium hair weeds cells to be put into temperature be that the environment freeze drying i.e. obtained rich selenium of-50 ~-55 DEG C is delivered vegetables product;
The preparation method of described selenium-enriched liquid medium is the Na of 1mg/mL by mass concentration
2seO
3join in minimal medium after aqueous solution sterilizing, until Na
2seO
3mass concentration be 2 ~ 40mg/L, adjust ph to 7.5 i.e. obtained selenium-enriched liquid medium.
2. a kind of rich selenium according to claim 1 is delivered vegetables the preparation method of product, it is characterized in that: described minimal medium is by NaNO
3, MgSO
47H
2o, CaCl
22H
2o, H
3bO
4, MnCl
24H
2o, ZnSO
47H
2o, CuSO
45H
2o, NaMoO
42H
2o, CoCl
2, EDTA-Na
2, FeSO
47H
2o, citric acid, Na
2siO
39H
2o and K
2hPO
4make, and the parts by weight of each component are respectively NaNO
3250 parts, MgSO
47H
2o 75 parts, CaCl
22H
2o 36 parts, H
3bO
42.86 parts, MnCl
24H
2o 1.8 parts, ZnSO
47H
2o 0.2 part, CuSO
45H
2o 0.8 part, NaMoO
42H
2o 0.4 part, CoCl
20.01 part, EDTA-Na
2l part, FeSO
47H
2o 4.8 parts, citric acid 0.6 part, Na
2siO
39H
2o 58 parts, K
2hPO
440 parts.
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Cited By (1)
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CN112624807A (en) * | 2021-01-14 | 2021-04-09 | 江西省农业科学院土壤肥料与资源环境研究所 | Cadmium-reducing agent for rice, seed soaking liquid, preparation method and seed soaking method |
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