CN106635913B - Method for regulating and controlling nutrient element accumulation of spirulina - Google Patents
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Abstract
The invention provides a method for regulating and controlling the accumulation of nutrient elements in spirulina, which comprises the following steps: 1) inoculating Spirulina into culture medium, and culturing at 20-40 deg.C, 4-12klx, and pH of 8.5-10.5 for 3 days; 2) continuously culturing the spirulina in the step 1) at 15-35 ℃, 2-20klx and pH8.5-11.0, and respectively adding calcium, iron, zinc and selenium elements into the culture medium for 3-7 days to obtain the spirulina with high nutrient elements. The method greatly increases the contents of the four nutrient elements of calcium, iron, zinc and selenium in the spirulina, combines the nutrient elements with organic matters in cells, reduces the irritation to gastrointestinal tracts and improves the absorption and utilization rate of human bodies. The contents of calcium, iron and zinc in the spirulina cells cultured by the method are respectively increased by 97%, 175% and 87% compared with the traditional culture method; the organic degree of calcium, iron, zinc and selenium is 84.17%, 89.98%, 58.28% and 70.54% in sequence.
Description
Technical Field
The invention relates to the technical field of oriented culture and extraction of biological cells, in particular to a method for regulating and controlling the accumulation of nutrient elements in spirulina.
Background
In recent years, the supplement of nutrient elements is receiving more and more attention. According to the third national nutrition survey data, the calcium intake of the nation is only 20% -50%, the zinc deficiency rate of the children is as high as 60%, the average iron deficiency anemia rate in China is about 20%, and the region is about 2/3 is selenium-deficient. In the traditional method for solving the deficiency of the nutrient elements, serious patients are treated by medicaments, light patients are supplemented by diet, the food supplementation usually cannot meet the requirements of human bodies, most of the medicaments are supplemented by inorganic components, such as calcium chloride, zinc sulfate and the like, and the absorption and utilization rate of the human bodies is low. Aiming at the current situation, the spirulina cell nutrient element is taken as a carrier by utilizing the characteristic of absorption and enrichment of the spirulina cell nutrient element, so that the accumulation of the spirulina cell nutrient element is promoted, inorganic ions are organized in the cell, the absorption rate and the utilization rate of a human body are improved, and the aim of supplementing the human body with the nutrient element is fulfilled.
At present, the method for improving the nutrient elements of the algae mainly achieves the aim of element accumulation by increasing the ion concentration in the exogenous culture solution. However, in the process of culturing and developing spirulina by many domestic and foreign enterprises, the influence of the change of the temperature difference between the region, the season, the day and the like on the growth of the spirulina and the accumulation of nutrient substances is very large, but the report of the growth environment on the accumulation of nutrient elements of the spirulina is not seen. The inventor has shown through many years that environmental factors (temperature, environment, pH, etc.) not only affect the growth of spirulina, but also affect its physiological metabolism, substance synthesis, etc., and also have profound effects on its mineral nutrient accumulation. The effective regulation and control of the accumulation of nutrient elements in spirulina cells has great significance for the organization of nutrient elements, the supplement of nutrient elements of human bodies and the widening of the application range of spirulina.
Disclosure of Invention
The invention aims to provide a method for improving the content of nutrient elements (calcium, iron, zinc and selenium) of spirulina.
In order to achieve the purpose of the invention, the method for regulating and controlling the accumulation of nutrient elements in spirulina comprises the following steps:
1) inoculating Spirulina into culture medium, and culturing at 20-40 deg.C under illumination of 4-12klx and pH of 8.5-10.5 for 3 days;
2) continuously culturing the spirulina in the step 1) at the temperature of 15-35 ℃, the illumination of 2-20klx and the pH value of 8.5-11.0, and respectively adding calcium, iron, zinc and selenium elements into the culture medium for 3-7 days to obtain the spirulina with high nutrient elements.
The foregoing method, step 2), is specifically as follows: continuously culturing the spirulina in the step 1), adjusting the temperature to 15-25 deg.C, controlling the illumination at 11-20klx and pH at 8.5-9.0, adding exogenous CaCO into the culture medium3Culturing for 3-7 days to obtain calcium-rich Spirulina product with final concentration of 0.4-0.8 g/L. Preferably, the spirulina of step 1) is continuously cultured, the temperature is adjusted to 20 ℃, the illumination is controlled at 12klx, the pH is controlled at 8.5, and exogenous CaCO is added into the culture medium3Culturing for 7 days to obtain Spirulina rich in calcium to final concentration of 0.6 g/L.
The foregoing method, step 2), is specifically as follows: the spirulina in the step 1) is treatedContinuously culturing, adjusting temperature to 25-35 deg.C, controlling illumination at 2-6klx and pH at 10-11.0, adding exogenous FeSO into culture medium4·7H2Culturing for 3-7 days until the final concentration of O is 30-60mg/L to obtain Spirulina rich in iron. Preferably, the spirulina of step 1) is continuously cultured, the temperature is adjusted to 30 ℃, the illumination is controlled to be 4klx, the pH is controlled to be 10.5, and exogenous iron FeSO4 & 7H is added into the culture medium2And culturing for 3 days to obtain the spirulina rich in iron after the final concentration of O is 40 mg/L.
The foregoing method, step 2), is specifically as follows: continuously culturing the spirulina in the step 1), adjusting the temperature to 25-35 ℃, controlling the illumination to 6-11klx and the pH to 8.5-9.5, and adding exogenous ZnSO into the culture medium4·7H2O to final concentration of 30-70mg/L, and culturing for 3-7 days to obtain zinc-rich Spirulina. Preferably, the spirulina in the step 1) is continuously cultured, the temperature is adjusted to 30 ℃, the illumination is controlled to be 8klx, the pH is controlled to be 8.5, and exogenous zinc ZnSO is added into the culture medium4·7H2And culturing for 3 days to obtain zinc-rich spirulina with final concentration of 40 mg/L.
The foregoing method, step 2), is specifically as follows: continuously culturing the spirulina of step 1), adjusting temperature to 28-35 deg.C, controlling illumination at above 11klx and pH at 9.0-10.0, adding exogenous selenium Na into the culture medium2SeO3Culturing for 3-7 days to obtain selenium-rich Spirulina with final concentration of 80-120 mg/L. Preferably, the spirulina in the step 1) is continuously cultured, the temperature is adjusted to 30 ℃, the illumination is controlled at 12klx, the pH is controlled at 9.5, and exogenous selenium Na is added into the culture medium2SeO3Culturing for 5 days to obtain selenium-rich Spirulina with final concentration of 100 mg/L.
Preferably, step 1) is specifically as follows: spirulina is inoculated into the culture medium, and cultured at 30 deg.C, 12klx, and pH of 9.5 for 3 days.
In one embodiment of the present invention, the method for regulating and controlling the accumulation of nutrient elements in spirulina specifically comprises the following steps:
1) inoculating Spirulina into culture medium, and culturing at 30 deg.C, 12klx and pH of 9.5 for 3 days;
2) continuously culturing the spirulina in the step 1), adjusting the temperature to 20 ℃, controlling the illumination to be 12klx and the pH to be 8.5, and adding exogenous CaCO into the culture medium3Culturing for 7 days to obtain Spirulina rich in calcium with calcium organization degree of 84.17%; alternatively, the first and second electrodes may be,
continuously culturing the spirulina in the step 1), adjusting the temperature to 30 ℃, controlling the illumination to be 4klx and the pH to be 10.5, and adding exogenous iron FeSO4 & 7H into the culture medium2Culturing for 3 days until the final concentration of O is 40mg/L to obtain Spirulina rich in iron, wherein the iron organization degree reaches 89.98%; alternatively, the first and second electrodes may be,
continuously culturing the spirulina in the step 1), adjusting the temperature to 30 ℃, controlling the illumination to be 8klx and the pH to be 8.5, and adding exogenous zinc ZnSO into the culture medium4·7H2Culturing for 3 days until the final concentration of O is 40mg/L to obtain zinc-rich spirulina, wherein the zinc organization degree reaches 58.28%; alternatively, the first and second electrodes may be,
continuously culturing the spirulina in the step 1), adjusting the temperature to 30 ℃, controlling the illumination to be 12klx and the pH to be 9.5, and adding exogenous selenium Na into the culture medium2SeO3Culturing for 5 days to obtain Spirulina rich in selenium with selenium organization degree of 70.54%.
The aforementioned method, steps 1) and 2) the medium is Zarrouk's medium (tables 1 and 2).
TABLE 1 Zarrouk culture solution composition
TABLE 2 Zarrouk microelement solution composition
The method comprises the step 1) of inoculating spirulina, and then obtaining the OD of the spirulina liquid560The value was 0.3.
In the method, the pH value is adjusted by using phosphoric acid with the mass concentration of 0.1mol/L, and the spirulina is spirulina platensis.
The invention adds excessive nutrient elements to the spirulina liquid, which aims to improve the organic degree of the nutrient elements and reduce the growth rate without influencing the growth.
The invention also provides the spirulina rich in the nutrient elements of calcium, iron, zinc and selenium prepared by the method.
The invention further provides application of the method in regulating and controlling the accumulation of nutrient elements in spirulina.
According to the invention, the environmental conditions of the absorption and accumulation of the nutrient elements of the spirulina are optimized by researching the relationship between environmental factors (temperature, illumination and pH value) and the absorption and accumulation of the elements of calcium, iron, zinc and selenium and the growth of the spirulina, so that the accumulation of the nutrient elements is maximized on the premise of achieving a certain growth amount of spirulina cells. Namely, the directional accumulation of the nutrient elements of the spirulina cells is realized through the dual-track system of the regulation and control of environmental conditions and the regulation and control of the ion concentration of the external liquid. This is of great value in practical production. The method aims to realize the directional accumulation of nutrient elements by utilizing the change of environmental conditions in different regions and seasons and the photo-thermal combined culture of the spirulina, guides production practice, produces the spirulina powder with high content of different nutrient elements and high organic degree, diversifies spirulina products, greatly improves the economic benefit of enterprises, overcomes the defect of nutrient element supplement at present, and promotes the development of spirulina industry and health industry in China.
The method provided by the invention greatly increases the contents of the four nutrient elements of calcium, iron, zinc and selenium in the spirulina, combines the nutrient elements with organic matters in cells, reduces the irritation to gastrointestinal tracts and improves the absorption and utilization rate of human bodies. The contents of calcium, iron and zinc in the spirulina cells cultured by the method are respectively increased by 97%, 175% and 87% compared with the traditional culture method; the organized degree of the calcium, iron, zinc and selenium is as follows: 84.17%, 89.98%, 58.28% and 70.54%.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are commercially available products.
Example 1
1) Inoculating Spirulina into Z's culture medium, inoculating Spirulina, and collecting the OD of the spirulina liquid560The value was 0.3. The cells were cultured at 30 ℃ under 12klx conditions at pH 9.5 for 3 days.
2) Continuously culturing the spirulina cultured for 3 days in the step 1), adjusting the temperature to 20 ℃, controlling the illumination to be 12klx and the pH to be 8.5, and adding exogenous calcium CaCO into the culture medium3Culturing for 3 days to obtain Spirulina rich in calcium (containing calcium element 2.52 mg/g). The degree of organization of calcium was 53.57%. The growth rate K of spirulina is 0.0465.
Example 2
1) Inoculating Spirulina into Z's culture medium, inoculating Spirulina, and collecting the OD of the spirulina liquid560The value was 0.3. The cells were cultured at 30 ℃ under 12klx conditions at pH 9.5 for 3 days.
2) Continuously culturing the spirulina cultured for 3 days in the step 1), adjusting the temperature to 20 ℃, controlling the illumination to be 12klx and the pH to be 8.5, and adding exogenous calcium CaCO into the culture medium3Culturing for 5 days to obtain Spirulina rich in calcium (the content of calcium element in Spirulina is 5.64mg/g) until the final concentration is 0.6 g/L. The degree of organization of the calcium was 72.14%. The growth rate K of spirulina is 0.0448.
Example 3
1) Inoculating Spirulina into Z's culture medium, inoculating Spirulina, and collecting the OD of the spirulina liquid560The value was 0.3. The cells were cultured at 30 ℃ under 12klx conditions at pH 9.5 for 3 days.
2) Continuously culturing the spirulina cultured for 3 days in the step 1), adjusting the temperature to 20 ℃, controlling the illumination to be 12klx and the pH to be 8.5, and adding exogenous calcium CaCO into the culture medium3Culturing for 7 days to obtain Spirulina rich in calcium (containing calcium element 18.21 mg/g). The degree of organization of the calcium was 84.17%. The growth rate K of spirulina is 0.0411.
Example 4
1) Inoculating Spirulina into Z's culture medium, inoculating Spirulina, and collecting the OD of the spirulina liquid560The value was 0.3. The cells were cultured at 30 ℃ under 12klx conditions at pH 9.5 for 3 days.
2) Continuously culturing the spirulina cultured for 3 days in the step 1), adjusting the temperature to 30 ℃, controlling the illumination to be 4klx and the pH to be 10.5, and adding exogenous iron FeSO into the culture medium4·7H2Culturing for 7 days to obtain Spirulina rich in iron (containing 1538.26ppm of iron element in Spirulina) with final concentration of O of 40 mg/L. The degree of organization of the iron was 84.95%. The growth rate K of spirulina is 0.0332.
Example 5
1) Inoculating Spirulina into Z's culture medium, inoculating Spirulina, and collecting the OD of the spirulina liquid560The value was 0.3. The cells were cultured at 30 ℃ under 12klx conditions at pH 9.5 for 3 days.
2) Continuously culturing the spirulina cultured for 3 days in the step 1), adjusting the temperature to 30 ℃, controlling the illumination to be 4klx and the pH to be 10.5, and adding exogenous iron FeSO into the culture medium4·7H2Culturing for 3 days to obtain Spirulina rich in iron (containing 1634.34ppm of iron element in Spirulina) with final concentration of O of 40 mg/L. The degree of organization of the iron was 89.98%. The growth rate K of spirulina is 0.0411.
Example 6
1) Inoculating Spirulina into Z's culture medium, inoculating Spirulina, and collecting the OD of the spirulina liquid560The value was 0.3. The cells were cultured at 30 ℃ under 12klx conditions at pH 9.5 for 3 days.
2) Continuously culturing the spirulina cultured for 3 days in the step 1), adjusting the temperature to 30 ℃, controlling the illumination to be 8klx and the pH to be 8.5, and adding exogenous zinc ZnSO into the culture medium4·7H2Culturing for 7 days to obtain zinc-rich Spirulina (containing zinc element 132.75ppm) with final concentration of O of 40 mg/L. The degree of organization of the zinc was 5.34%. The growth rate K of spirulina is 0.0376.
Example 7
1) Inoculating Spirulina into Z's culture medium, inoculating Spirulina, and collecting the OD of the spirulina liquid560The value was 0.3. The cells were cultured at 30 ℃ under 12klx conditions at pH 9.5 for 3 days.
2) Continuously culturing the spirulina cultured for 3 days in the step 1)Adjusting the temperature to 30 ℃, controlling the illumination to be 8klx and the pH to be 8.5, adding exogenous zinc ZnSO into the culture medium4·7H2Culturing for 3 days to obtain zinc-rich Spirulina (zinc element content of Spirulina is 88.30ppm) with final concentration of O of 40 mg/L. The degree of organization of the zinc was 58.28%. The growth rate K of spirulina is 0.0429.
Example 8
1) Inoculating Spirulina into Z's culture medium, inoculating Spirulina, and collecting the OD of the spirulina liquid560The value was 0.3. The cells were cultured at 30 ℃ under 12klx conditions at pH 9.5 for 3 days to reach the logarithmic growth phase.
2) Continuously culturing the spirulina cultured for 3 days in the step 1), and adding exogenous selenium Na into the culture medium2SeO3Culturing for 7 days to obtain Spirulina rich in selenium (containing selenium element 350.27 ppm). The degree of organisation of selenium was 40.16%. The growth rate K of spirulina is 0.0246.
Example 9
1) Inoculating Spirulina into Z's culture medium, inoculating Spirulina, and collecting the OD of the spirulina liquid560The value was 0.3. The cells were cultured at 30 ℃ under 12klx conditions at pH 9.5 for 3 days to reach the logarithmic growth phase.
2) Continuously culturing the spirulina cultured for 3 days in the step 1), and adding exogenous selenium Na into the culture medium2SeO3Culturing for 3 days to obtain Spirulina rich in selenium (containing selenium element 399.70ppm) with final concentration of 100 mg/L. The degree of organisation of selenium was 58.62%. The growth rate K of spirulina is 0.0277.
Example 10
1) Inoculating Spirulina into Z's culture medium, inoculating Spirulina, and collecting the OD of the spirulina liquid560The value was 0.3. The cells were cultured at 30 ℃ under 12klx conditions at pH 9.5 for 3 days to reach the logarithmic growth phase.
2) Continuously culturing the spirulina cultured for 3 days in the step 1), and adding exogenous selenium Na into the culture medium2SeO3Culturing for 5 days to obtain Spirulina rich in selenium (containing selenium element 1011.00ppm) with final concentration of 100 mg/L. The degree of organisation of selenium was 70.54%. Spirulina platensisThe growth rate K is 0.0265.
Comparative example 1
1) Inoculating Spirulina into Z's culture medium, inoculating Spirulina, and collecting the OD of the spirulina liquid560The value was 0.3. The cells were cultured at 30 ℃ under 4klx conditions at pH 9.5 for 3 days.
2) Continuously culturing the spirulina cultured for 3 days in the step 1), adjusting the temperature to 30 ℃, controlling the illumination to be 4klx and the pH to be 9.5, and culturing for 4 days, wherein the accumulation amount of calcium is 244.66 ppm. The growth rate K of spirulina is 0.0852.
Comparative example 2
1) Inoculating Spirulina into Z's culture medium, inoculating Spirulina, and collecting the OD of the spirulina liquid560The value was 0.3. The cells were cultured at 40 ℃ under 8klx conditions at pH8.5 for 3 days.
2) Continuously culturing the spirulina cultured for 3 days in the step 1), adjusting the temperature to 40 ℃, controlling the illumination to be 8klx and the pH to be 8.5, and culturing for 4 days, wherein the accumulation amount of iron is 192.64 ppm. The growth rate K of spirulina is 0.0804.
Comparative example 3
1) Inoculating Spirulina into Z's culture medium, inoculating Spirulina, and collecting the OD of the spirulina liquid560The value was 0.3. The cells were cultured at 40 ℃ under 12klx conditions at pH 9.5 for 3 days.
2) Continuously culturing the spirulina cultured for 3 days in the step 1), adjusting the temperature to 40 ℃, controlling the illumination to be 12klx and the pH to be 9.5, and culturing for 4 days, wherein the accumulation amount of zinc is 30.06 ppm. The growth rate K of spirulina is 0.0944.
The results show that the method provided by the invention can greatly improve the content and the organic degree of several nutrient elements of the spirulina, and provides an effective means for reducing the stimulation to the gastrointestinal tract and improving the absorption and utilization rate of a human body.
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (1)
1. A method for regulating and controlling the accumulation of nutrient elements in spirulina is characterized by comprising the following steps:
1) inoculating Spirulina into Zarrouk culture medium, and inoculating Spirulina to obtain the OD of the algae solution560A value of 0.3; culturing at 30 deg.C, 12klx, pH =9.5 for 3 days to reach logarithmic growth phase;
2) continuously culturing the spirulina in the step 1), adjusting the temperature to 30 ℃, controlling the illumination to be 12klx and the pH to be 9.5, and adding exogenous selenium Na into the Zarrouk's culture medium2SeO3Culturing for 5 days to obtain selenium-rich Spirulina to final concentration of 100 mg/L;
wherein the Spirulina is Spirulina platensis.
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| CN201910261703.2A CN109825461A (en) | 2016-12-31 | 2016-12-31 | A method of regulation nutritious spirulina element zinc element accumulation |
| CN201910261367.1A CN110004103A (en) | 2016-12-31 | 2016-12-31 | A method of regulation nutritious spirulina elemental iron element accumulation |
| CN201910261345.5A CN109837234A (en) | 2016-12-31 | 2016-12-31 | A method of regulation nutritious spirulina element calcium element accumulation |
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| GROWTH AND BIOPIGMENT ACCUMULATION OF CYANOBACTERIUM SPIRULINA PLATENSIS AT DIFFERENT LIGHT INTENSITIES AND TEMPERATURE;Manoj Kumar et al.;《Brazilian Journal of Microbiology》;20111231(第42期);第1128-1135页 * |
| 温度对3种产业化螺旋藻生长及蛋白质含量影响的研究;杨学文 等;《内蒙古农业大学学报》;20060630;第27卷(第2期);第60-64页 * |
Also Published As
| Publication number | Publication date |
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| CN106635913A (en) | 2017-05-10 |
| CN110004103A (en) | 2019-07-12 |
| CN109825461A (en) | 2019-05-31 |
| CN109837234A (en) | 2019-06-04 |
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