CN105907644A - Solid-state culture medium for benefitting long-term storage of haematococcus pluvialis algae and preparation method thereof - Google Patents
Solid-state culture medium for benefitting long-term storage of haematococcus pluvialis algae and preparation method thereof Download PDFInfo
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- CN105907644A CN105907644A CN201610369738.4A CN201610369738A CN105907644A CN 105907644 A CN105907644 A CN 105907644A CN 201610369738 A CN201610369738 A CN 201610369738A CN 105907644 A CN105907644 A CN 105907644A
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Abstract
The invention discloses a solid-state culture medium for benefiting long-term storage of haematococcus pluvialis algae. The solid-state culture medium is characterized by being prepared from the following components in every liter of the culture medium: 1.3-1.4g of sodium nitrate, 0.13-0.15g of sodium carbonate, 0.15-0.25g of magnesium sulfate, 0.05-0.10g of dipotassium phosphate, 0.009-0.019g of ammonium ferric citrate, 0.010-0.050g of citric acid, 0.025-0.035g of potassium sorbate, 0.001-0.010g of ethylene diamine tetraacetic acid, 0.000222-0.001000g of zinc sulfate, 0.00286-0.00586g of 0.00286-0.00586g of boric acid, 0.0018-0.0036g of manganese chloride, 0.000494-0.000894g of cobalt nitrate, 0.00039-0.00089g of sodium molybdate, 0.000079-0.000150g of copper sulfate, 0.1-1.0g of sodium acetate, 15-17g of agar powder, 0.025-0.050g of biotin, 2-5ml of vitamin B1 and 5-10ml of vitamin B12. The invention aims to provide a novel formula and the preparation method of the haematococcus pluvialis solid-state seed-conservation culture medium to solve the problems that reproduction of haematococcus pluvialis is inhibited by the existing formula, growth factors are not sufficient or excessive and are easily polluted by bacteria, and the like, so that the effect of prolonging the preservation period of haematococcus pluvialis algae can be achieved.
Description
Technical field
The present invention relates to culture medium prescription technical field, particularly relate to a kind of solid medium being conducive to long term storage Haematocoocus Pluvialls algae kind, and the preparation method of this culture medium.
Background technology
Haematocoocus Pluvialls is one way of life Shan Hong born of the same parents chlorella in fresh water, also known as the raw Haematococcus Pluvialis in lake or lake hemopoietic ball algae, is a kind of chlorella generally existed, this algae can in a large number accumulation astaxanthin and present redness, therefore named Haematococcus Pluvialis, because being the rain non-hibernating eggs in chlorella, also known as Haematocoocus Pluvialls.Astaxanthin in Haematocoocus Pluvialls is the one of carotenoid, is also the highest level product of the carotenogenesis found so far, has the strongest non-oxidizability at nature, is therefore described as 21 century antioxidative revolutionary character product.The aging of human body, the oxidation caused mainly due to free radical institute is extremely.And the molecular configuration that Haematococcus Pluvialis extract is special, human body cell outer wall, the oxygen-derived free radicals in direct scavenger cell can be passed through, strengthen cell regeneration ability, maintain function of human body balance and reduce the accumulation of senile cell, protection cell and NDA are healthy from inside to outside.Antioxidative composition is the most all improving: first generation antioxidative composition is vitamins, second filial generation antioxidation is B-carotene, coenzyme Q10, SOD etc, third generation antioxidation is anthocyanidin, Semen Vitis viniferae, blueberry extract, green tea element, thioctic acid, lycopene etc, and forth generation antioxidant content is exactly natural astaxanthin.Research shows, the antioxidant activity of Determination of Astaxanthin in Haematococcus Pluvialis is 10 times of B-carotene, 17 times of Semen Vitis viniferae, 75 times of thioctic acid, 200 times of phylloxanthin, 1000 times of vitamin E.Equally, Haematocoocus Pluvialls is such as other algae, rich in several amino acids, unsaturated fatty acid, vitamin, mineral etc..But, the orange system pigment such as its distinctive astaxanthin, is not available for other microalgae, and this is the most real healthy is worth place！Along with under modern society, the mankind's high pursuit to healthy living, the product of the health care theories such as opposing is old and feeble, remain youthful forever increasingly receives the favor of consumers in general, and as the strongest biological metabolic product of oxidation resistance in nature, the economic worth of the natural astaxanthin that Haematocoocus Pluvialls content is high will be following immeasurable great wealth！Increase along with Haematocoocus Pluvialls breeding enterprise quantity, a outstanding algae kind will produce tremendous influence to the yield of an enterprise, so screening and preservation to Haematocoocus Pluvialls algae kind increasingly come into one's own, at present existing enterprise to the preservation of algae kind mainly by solid medium conservation mode, this mode conservation is rapid, recover expanding species convenient, but under technology, the design of its culture medium prescription is not reasonable at present, it is commonly present suppression Haematocoocus Pluvialls breeding, the drawbacks such as somatomedin is inadequate or excessive, easy polluted bacteria.
Summary of the invention
It is an object of the invention to provide the new formula of a kind of Haematocoocus Pluvialls solid-state culture medium for preserving, solve the suppression Haematocoocus Pluvialls breeding of existing formula, the drawbacks such as somatomedin is inadequate or excessive, easy polluted bacteria, reach to extend the effect of Haematocoocus Pluvialls algae kind holding time.
The technical scheme is that a kind of solid medium being conducive to long term storage Haematocoocus Pluvialls algae kind, its each component ratio is as follows by content in every 1 liter of culture medium: sodium nitrate 1.3g-1.4g；Sodium carbonate 0.13g-0.15g；Magnesium sulfate 0.15g-0.25g；Dipotassium hydrogen phosphate 0.05g-0.10g；Ferric ammonium citrate 0.009g-0.019g；Citric acid 0.010g-0.050g；Potassium sorbate 0.025g-0.035g；Disodiumedetate 0.001g-0.010g；Zinc sulfate 0.000222g-0.001000g；Boric acid 0.00286g-0.00586g manganese chloride 0.0018g-0.0036g；Cobalt nitrate 0.000494g-0.000894g；Sodium molybdate 0.00039g-0.00089g；Copper sulfate 0.000079g-0.000150g；Sodium acetate 0.1g-1.0g；Agar powder 15g-17g；Biotin 0.025g-0.050g；Vitamin B1 2-5ml；Vitamin B12 5-10ml.
In the above-mentioned solid medium being conducive to long term storage Haematocoocus Pluvialls algae kind, also include the ultra-pure water of surplus.
Meanwhile, the invention also discloses the preparation method of the above-mentioned solid medium being conducive to long term storage Haematocoocus Pluvialls algae kind, comprise the following steps:
Step 1: measure 1 liter of ultra-pure water；
Step 2: above-mentioned component is joined in ultra-pure water, and make each component dissolve；
Step 3: solution step 2 obtained joins in high-pressure sterilizing pot sterilizing 18-22min under conditions of 115-125 DEG C, is then saved in gnotobasis, and storage temperature is 38-42 DEG C.
Beneficial effects of the present invention is as follows:
The long term growth breeding that the new formula of a kind of Haematocoocus Pluvialls solid-state culture medium for preserving disclosed by the invention provides enough nitrogen source, phosphorus source, carbon source are frustule provides enough growth conditionss, its pH value can long term maintenance between 6.5-8.0, it is suitable for the merisis of microalgae, the citric acid, the potassium sorbate that add can be in the case of not affecting micro algae growth, the effectively growth of suppression antibacterial.The solid-state culture medium for preserving made of this formula can improve about preservation Haematocoocus Pluvialls algae kind 6-10 month, because the convenience of solid conservation, convenient and swift can carry out microalgae aquaculture when using algae kind, and put into production line rapidly.
Detailed description of the invention
Below in conjunction with detailed description of the invention, technical scheme is described in further detail, but does not constitute any limitation of the invention.
Embodiment
1
The preparation method of a kind of solid medium being conducive to long term storage Haematocoocus Pluvialls algae kind, comprises the following steps:
Step 1: measure 1 liter of ultra-pure water；
Step 2: by sodium nitrate 1.4g；Sodium carbonate 0.13g；Magnesium sulfate 0.25g；Dipotassium hydrogen phosphate 0.05g；Ferric ammonium citrate 0.019g；Citric acid 0.010gg；Potassium sorbate 0.035g；Disodiumedetate 0.010g；Zinc sulfate 0.000222g；Boric acid 0.00586g manganese chloride 0.0018g；Cobalt nitrate 0.000894g；Sodium molybdate 0.00039g；Copper sulfate 0.000150g；Sodium acetate 0.1g；Agar powder 17g；Biotin 0.025g；Vitamin B1 5ml；Vitamin B12 5 ml joins in ultra-pure water, and makes each component dissolve；
Step 3: solution step 2 obtained joins in high-pressure sterilizing pot sterilizing 20min under conditions of 120 DEG C, is then saved in gnotobasis, and storage temperature is 40 DEG C.
Embodiment
2
The preparation method of a kind of solid medium being conducive to long term storage Haematocoocus Pluvialls algae kind, comprises the following steps:
Step 1: measure 1 liter of ultra-pure water；
Step 2: by sodium nitrate 1.35g；Sodium carbonate 0.14g；Magnesium sulfate 0.2g；Dipotassium hydrogen phosphate 0.08g；Ferric ammonium citrate 0.014g；Citric acid 0.03g；Potassium sorbate 0.03g；Disodiumedetate 0.005g；Zinc sulfate 0.0007g；Boric acid 0.0045g manganese chloride 0.0028g；Cobalt nitrate 0.0006g；Sodium molybdate 0.0006g；Copper sulfate 0.00012g；Sodium acetate 0.8g；Agar powder 16g；Biotin 0.035g；Vitamin B1 4ml；Vitamin B12 6ml joins in ultra-pure water, and makes each component dissolve；
Step 3: solution step 2 obtained joins in high-pressure sterilizing pot sterilizing 20min under conditions of 120 DEG C, is then saved in gnotobasis, and storage temperature is 40 DEG C.
Embodiment
3
The preparation method of a kind of solid medium being conducive to long term storage Haematocoocus Pluvialls algae kind, comprises the following steps:
Step 1: measure 1 liter of ultra-pure water；
Step 2: by sodium nitrate 1.3g；Sodium carbonate 0.15g；Magnesium sulfate 0.15g；Dipotassium hydrogen phosphate 0.10g；Ferric ammonium citrate 0.009g；Citric acid 0.050g；Potassium sorbate 0.025g；Disodiumedetate 0.001g；Zinc sulfate 0.001000g；Boric acid 0.00286g manganese chloride 0.0036g；Cobalt nitrate 0.000494g；Sodium molybdate 0.00089g；Copper sulfate 0.000079g；Sodium acetate 1.0g；Agar powder 15g；Biotin 0.050g；Vitamin B1 2ml；Vitamin B12 10ml joins in ultra-pure water, and makes each component dissolve；
Step 3: solution step 2 obtained joins in high-pressure sterilizing pot sterilizing 20min under conditions of 120 DEG C, is then saved in gnotobasis, and storage temperature is 40 DEG C.
The long term growth breeding that embodiment 1-3 provides enough nitrogen source, phosphorus source, carbon source are frustule provides enough growth conditionss, its pH value can long term maintenance between 6.5-8.0, it is suitable for the merisis of microalgae, the citric acid, the potassium sorbate that add can be in the case of not affecting micro algae growth, the effectively growth of suppression antibacterial.The solid-state culture medium for preserving made of this formula can improve about preservation Haematocoocus Pluvialls algae kind 6-10 month, because the convenience of solid conservation, convenient and swift can carry out microalgae aquaculture when using algae kind, and put into production line rapidly.
Above-described only presently preferred embodiments of the present invention, all any amendment, equivalent and improvement etc. made in the range of the spirit and principles in the present invention, should be included within the scope of the present invention.
Claims (3)
1. the solid medium being conducive to long term storage Haematocoocus Pluvialls algae kind, it is characterised in that: its each component ratio is as follows by content in every 1 liter of culture medium: sodium nitrate
1.3g-1.4g；Sodium carbonate 0.13g-0.15g；Magnesium sulfate 0.15g-0.25g；Dipotassium hydrogen phosphate 0.05g-0.10g；Ferric ammonium citrate 0.009g-0.019g；Citric acid 0.010g-0.050g；Potassium sorbate 0.025g-0.035g；Disodiumedetate 0.001g-0.010g；Zinc sulfate 0.000222g-0.001000g；Boric acid 0.00286g-0.00586g manganese chloride 0.0018g-0.0036g；Cobalt nitrate 0.000494g-0.000894g；Sodium molybdate 0.00039g-0.00089g；Copper sulfate 0.000079g-0.000150g；Sodium acetate 0.1g-1.0g；Agar powder 15g-17g；Biotin 0.025g-0.050g；Vitamin B1 2-5ml；Vitamin B12 5-10ml.
The solid medium being conducive to long term storage Haematocoocus Pluvialls algae kind the most according to claim 1, it is characterised in that also include the ultra-pure water of surplus.
3. the preparation method of the solid medium being conducive to long term storage Haematocoocus Pluvialls algae kind as claimed in claim 1, it is characterised in that: comprise the following steps:
Step 1: measure 1 liter of ultra-pure water；
Step 2: the component described in claim 1 is joined in ultra-pure water, and make each component dissolve；
Step 3: solution step 2 obtained joins in high-pressure sterilizing pot sterilizing 18-22min under conditions of 115-125 DEG C, is then saved in gnotobasis, and storage temperature is 38-42 DEG C.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106479898A (en) * | 2016-12-29 | 2017-03-08 | 青岛藻蓝生物有限公司 | A kind of culture medium of Haematocoocus Pluvialls and its application |
| CN106701582A (en) * | 2016-12-29 | 2017-05-24 | 中国科学院广州能源研究所 | Long-term microalgae species preserving method |
| CN109182132A (en) * | 2018-08-16 | 2019-01-11 | 浙江海洋大学 | Produce the cultural method of the Alexandrium mimutum Halim of gonyatoxin |
| CN109294919A (en) * | 2018-10-17 | 2019-02-01 | 云南博欣生物科技股份有限公司 | A kind of algae update purification process of haematococcus pluvialis |
| CN109971649A (en) * | 2019-04-28 | 2019-07-05 | 哈尔滨师范大学 | It is a kind of for cultivating the culture medium and intelligent checking system of haematococcus pluvialis |
| CN110577898A (en) * | 2019-10-25 | 2019-12-17 | 四川轻化工大学 | Method for producing astaxanthin by culturing haematococcus pluvialis through discontinuous two-step method |
| CN114621874A (en) * | 2021-12-28 | 2022-06-14 | 宁波浮田生物技术有限公司 | Microalgae culture medium and application |
-
2016
- 2016-05-30 CN CN201610369738.4A patent/CN105907644A/en active Pending
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106479898A (en) * | 2016-12-29 | 2017-03-08 | 青岛藻蓝生物有限公司 | A kind of culture medium of Haematocoocus Pluvialls and its application |
| CN106701582A (en) * | 2016-12-29 | 2017-05-24 | 中国科学院广州能源研究所 | Long-term microalgae species preserving method |
| CN109182132A (en) * | 2018-08-16 | 2019-01-11 | 浙江海洋大学 | Produce the cultural method of the Alexandrium mimutum Halim of gonyatoxin |
| CN109182132B (en) * | 2018-08-16 | 2021-09-14 | 浙江海洋大学 | Method for culturing Alexandrium micranthum producing gonyautoxin |
| CN109294919A (en) * | 2018-10-17 | 2019-02-01 | 云南博欣生物科技股份有限公司 | A kind of algae update purification process of haematococcus pluvialis |
| CN109971649A (en) * | 2019-04-28 | 2019-07-05 | 哈尔滨师范大学 | It is a kind of for cultivating the culture medium and intelligent checking system of haematococcus pluvialis |
| CN110577898A (en) * | 2019-10-25 | 2019-12-17 | 四川轻化工大学 | Method for producing astaxanthin by culturing haematococcus pluvialis through discontinuous two-step method |
| CN110577898B (en) * | 2019-10-25 | 2023-01-31 | 四川轻化工大学 | Method for producing astaxanthin by culturing haematococcus pluvialis through discontinuous two-step method |
| CN114621874A (en) * | 2021-12-28 | 2022-06-14 | 宁波浮田生物技术有限公司 | Microalgae culture medium and application |
| CN114621874B (en) * | 2021-12-28 | 2023-09-29 | 宁波浮田生物技术有限公司 | Microalgae culture medium and application thereof |
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