CN114836324B - Haematococcus pluvialis high-temperature-resistant mutant strain and application thereof - Google Patents
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Abstract
The invention relates to a Haematococcus pluvialis high-temperature resistant mutant strain named YYAM001, which is preserved in China general microbiological culture collection center with the number of CGMCC No.45099, the preservation date is 2022 years, 4 months and 29 days, and the preservation address is Beijing City Kyowa Yang district Xilu No. 1 Hospital No. 3. The mutant strain is a monoclonal high-temperature resistant alga strain with stable genetic character, which is obtained by using a wild type of Haematococcus Pluvialis 712 as a starting material, mutagenizing Haematococcus Pluvialis by adopting normal-pressure room-temperature plasma, and repeatedly screening at high temperature. The mutant strain has a significantly higher growth rate than the wild type in a short time at a low temperature, is also superior to the wild type in a high-temperature continuous culture, and has a much higher survival rate than the wild type at a higher temperature (34 ℃). Under the conditions of nitrogen deficiency and high salt stress of an m-MCM culture medium, the cells accumulate astaxanthin slightly higher than wild type cells. After the mutant strain is cultured for one year, the high temperature resistance, the growth speed and the pigment accumulation capacity of the mutant strain are stable, which indicates that the genetic characteristic of the mutant strain is stable, and the mutant strain can be used as a high-quality algae strain for producing astaxanthin.
Description
Technical Field
The invention relates to the technical field of microalgae, in particular to a Haematococcus Pluvialis high-temperature-resistant mutant strain of Haematococcus Pluvialis and application thereof.
Background
Astaxanthin is a chain-breaking antioxidant. Has strong oxidation resistance, can eliminate nitrogen dioxide, sulfide, disulfide, etc., can reduce lipid peroxidation, and effectively inhibit lipid peroxidation caused by free radicals. Meanwhile, the traditional Chinese medicine composition has physiological effects in various aspects such as inhibiting tumorigenesis, enhancing immunity, eliminating free radicals in vivo and the like, has a good treatment effect on skin cancer caused by ultraviolet rays, has a prevention and treatment effect on eye diseases caused by diabetes, and has a wide application prospect in the aspects of health care products, medicines, cosmetics, food additives, aquaculture and the like.
Astaxanthin exists in three isomers, namely 3R,3'R and 3R,3' S meso and 3S,3'S, and only astaxanthin with the 3S,3' S configuration has stronger antioxidant activity. However, pure astaxanthin has been expensive so far and is difficult to be reached by more groups. A representative species of microalgae source producing 3S,3' S configuration is Haematococcus pluvialis. Haematococcus pluvialis is a unicellular freshwater green alga which can accumulate a large amount of astaxanthin, is a recognized ideal raw material of high-quality astaxanthin in nature, and is a main industrial source for producing natural astaxanthin at present. Astaxanthin is produced when algae are stressed by nutrient deprivation, increased salinity, or excessive sunlight. Animals fed on algae exhibit a certain orange-red color, i.e., astaxanthin pigmentation, to varying degrees, such as salmon, trout, red sea bream, flamingo, and some crustaceans (shrimp, crab, etc.). Astaxanthin can be used as a dietary supplement for human, animal and aquaculture consumption. Astaxanthin has been approved by the U.S. food and drug administration for specific use as a food pigment (or color additive) for animal and fish food products. Meanwhile, astaxanthin is used as a natural antioxidant, has strong antioxidant activity, can remove free radicals, and improves the anti-aging capability of a human body.
At present, the mainstream haematococcus pluvialis in the market is cultured by an open culture pond mostly, but the open culture pond is easily polluted by diseases and insect pests. In addition, the culture is also carried out by adopting a pipeline type photobioreactor, so that the yield is very easy to be influenced by the external environment, particularly the temperature. In summer, the temperature is hot and the sun is too strong, and meanwhile, bacteria are easy to breed in outdoor large-scale culture, so that the stable and high-quality output of products is difficult to ensure. The stable and high yield of haematococcus pluvialis can be ensured by adopting a pure heterotrophic fermentation technology to culture the haematococcus pluvialis. However, the optimum growth temperature for photoautotrophic haematococcus pluvialis is 25 ℃, and the excessive temperature during the culture process can cause the haematococcus pluvialis to change the form from zoospores to resting spores, namely, the cells begin to stress to generate astaxanthin to resist adverse environments, and the algae hardly proliferate under the conditions. Therefore, a temperature reduction means is required to maintain a proper temperature in the culture process, which greatly increases the culture cost. For example, patents CN105916993B and CN106480155B disclose that more astaxanthin is accumulated in haematococcus pluvialis cells by stressing the algal liquor temperature at 30-40 ℃. However, under the condition of the ultra-high temperature stress, haematococcus pluvialis is immediately converted into resting spores, so that the cell number cannot be multiplied and can be greatly reduced, and although the content of astaxanthin in a single cell is increased, the total astaxanthin yield is not high due to the sudden reduction of the cell number. Therefore, the high-temperature resistant algal strain which still normally grows and propagates at high temperature is obtained, so that the culture cost of haematococcus pluvialis and the production cost of algae-derived astaxanthin can be reduced.
Disclosure of Invention
Technical problem to be solved
In view of the above disadvantages and shortcomings of the prior art, the present invention provides a high temperature resistant mutant of Haematococcus pluvialis, which has a survival rate at 34 ℃ much higher than that of wild type Haematococcus pluvialis, has a remarkable high temperature resistant property compared to wild type Haematococcus pluvialis, can be cultured autotrophically or heterotrophically in a high temperature culture environment, eliminates cooling equipment and cost, is beneficial for reducing the culture cost of Haematococcus pluvialis and the production cost of astaxanthin from algae, and can bear a higher stress temperature to promote the accumulation of astaxanthin.
(II) technical scheme
In order to achieve the purpose, the invention adopts the main technical scheme that:
in a first aspect, the invention provides a Haematococcus pluvialis high temperature resistant mutant named YYAM001, which is preserved in China general microbiological culture Collection center with the preservation number of CGMCC No.45099, the preservation date of 2022 years, 4 months and 29 days, and the preservation address is No. 3 of Xilu No. 1 Beijing Kogyo-Naja-Navier.
In a second aspect, the present invention provides a method for preparing astaxanthin derived from algae, the method comprises inoculating and culturing a high temperature-resistant mutant strain YYAM001 of Haematococcus pluvialis, harvesting, and extracting astaxanthin from the harvested Haematococcus pluvialis cells; wherein the culture method is an autotrophic culture, a fermentation culture or a combined culture mode of autotrophic culture and fermentation.
Preferably, the Haematococcus pluvialis high temperature resistant mutant YYAM001 growing to the plateau stage is inoculated in a culture medium with high salt content and is subjected to illumination induced stress culture, so that the accumulation of astaxanthin in cells is promoted.
Preferably, the medium is an m-MCM culture; the light intensity of the induced stress is 6000lux; the culture medium contains NaCl with the mass concentration of 0.8%; the culture time for inducing stress is 5-6 days.
(III) advantageous effects
The mutant strain YYAM001 is obtained by carrying out mutagenesis and screening on wild Haematococcus pluvialis 712 serving as an original strain, has the high temperature resistance, is proved by experiments to be far higher than the wild type in survival rate at 34 ℃, and provides excellent algae seeds for large-scale fermentation culture and astaxanthin production of Haematococcus pluvialis. When the mutant strain YYAM001 is cultured in an m-MCM medium under the nitrogen-deficient high-salt condition, the content of astaxanthin accumulated by cells is slightly higher than that of wild type. The mutant strain is mainly characterized in that: (1) when the biomass is cultured at a lower temperature (18-28 ℃), the biomass growth rate is far higher than that of a wild type; (2) the biomass growth speed of the continuous culture at high temperature (30 ℃) is still obviously higher than that of a wild type; (3) survival at higher temperatures (34 ℃) is much higher than wild type. (4) The time to reach the growth plateau is short, about 6 days. After the YYAM001 mutant strain is cultured for 6 months, the cells can still resist the high temperature of 34 ℃, and the cell growth speed and the astaxanthin accumulation capacity are stable, which shows that the YYAM001 mutant strain has stable genetic characteristics, can be used as a high-quality high-temperature-resistant alga species, and is beneficial to reducing the culture cost of haematococcus pluvialis and the production cost of alga-derived astaxanthin. (5) The strain provided by the invention can bear higher stress temperature to promote the accumulation of astaxanthin in cells because the temperature tolerance of the strain in the logarithmic growth phase is at least 34 ℃ and is at least 9 ℃ higher than that of a wild type.
Drawings
FIG. 1 is a graph comparing the growth curves of mutant YYAM001 and WT algae when cultured for 7 days at 25 ℃ to 18 ℃ in alternating light and dark (14h.
FIG. 2 is a graph showing a comparison of the growth curves of mutant YYAM001 and WT algae when cultured for 7 days at 28 ℃ to 25 ℃ in alternating light and dark (14h.
FIG. 3 is a comparison of the growth curves of the mutant YYAM001 and WT algae when cultured at 30 ℃ for 7 days in continuous light.
FIG. 4 is a graph showing a comparison of the bottom-sinking phenomenon of mutant YYAM001 and WT algae cultured in the dark at 30 ℃.
FIG. 5 is a photograph of microscopic algal cells after the mutant strain YYAM001 and wild type WT were placed under m-MCM high salt (0.8% NaCl) 6000lux to induce stress for 5 days.
Detailed Description
For the purpose of better explaining the present invention and to facilitate understanding, the present invention will be described in detail by way of specific embodiments with reference to the accompanying drawings.
The invention provides a Haematococcus pluvialis high-temperature resistant mutant strain named YYAM001, which is preserved in the China general microbiological culture Collection center with the preservation number of CGMCC No.45099, the preservation date of 2022 years, 4 months and 29 days, and the preservation address is No. 3 of Xilu No. 1 Beicheng Yang-oriented district in Beijing. The mutant strain is prepared by using wild type of Haematococcus Pluvialis 712 as material, inducing Haematococcus Pluvialis with Atmospheric pressure Room Temperature Plasma (Atmospheric and Room Temperature Plasma), and repeatedly screening at high Temperature until monoclonal high Temperature resistant algae with stable hereditary character is obtained. The survival rate of the mutant strain at 34 ℃ is far higher than that of the wild type. When the cells are cultured in the m-MCM medium under the nitrogen-deficient high-salt condition, the situation that the cells accumulate astaxanthin is slightly higher than that of the wild type, and the cell survival rate is far higher than that of the wild type. After the Haematococcus pluvialis high-temperature-resistant mutant YYAM001 is cultured for one year, the high-temperature-resistant property, the growth and the pigment content of the Haematococcus pluvialis high-temperature-resistant mutant YYAM001 are stable, so that the Haematococcus pluvialis high-temperature-resistant mutant YYAM has the characteristic of stable inheritance, can be used as a seed to help to ferment Haematococcus pluvialis at low cost and high efficiency, and can also guide the development of a new process for accumulating astaxanthin in Haematococcus pluvialis.
The process of obtaining Haematococcus Pluvialis mutant YYAM001 of the invention is as follows:
1. preparation before mutagenesis
Monoclonal algal colonies of Haematococcus Pluvialis 712 (from freshwater algal species Bank of the Chinese academy of sciences) grown on m-MCM solid medium were picked up in 50mL Erlenmeyer flasks containing 20mL of modified MCM liquid medium at 25 ℃ for 7 days.
The improved MCM liquid culture medium formula (aqueous solution) is as follows: NH (NH) 4 Cl 105.7mg/L、K 2 HPO 4 20mg/L、MgSO 4 ·7H 2 O 48.8mg/L、CaCl 2 40.5mg/L、VB 12 0.004mg/L、EDTA 0.0198mg/L、FeCl 3 ·6H 2 O0.244 mg/L, trace elements 1mL/L, CH 3 COONa 1.2g/L, yeast Extract 2.0g/L, L-Asparagine (L-Asparagine) 0.405g/L.
Preparing a microelement mother solution: 61mg/L of H 3 BO 3 4.86mg/L of ZnSO 4 6mg/L of CuSO 4 ·5H 2 O,4.1mg/L MnCl 2 4H2O,38mg/L (NH) 4 ) 6 Mo 7 O 2 4·4H 2 O。
2. Mutagenesis
Counting algae cells before mutagenesis, centrifuging, concentrating to obtain a product containing 2 × 10 6 The algal solution of each cell was placed in 8 centrifuge tubes, treated with an ARTP mutagenesis system, 0s,10s,20s,30s,40s, 50s,60s,70s, and then transferred to 40mL of m-MCM medium for recovery. And after the algae liquid grows to have an obvious trend, counting cells to calculate the lethality.
Lethality (%) = (number of control cells-number of treated cells)/number of control cells + 100%. The primary screening time is 40-50 s, the experiment is repeated, and finally the lethality of the algae cells under different treatment times is determined (see table 1), and the mutagenesis treatment condition of about 90% lethality is 45s. Mutagenized algae in the centrifuge tubes treated for 45s using the ARTP mutagenesis System were inoculated into 40mL of medium and placed in a 34 ℃ light incubator for further screening. The control group is a group treated for 0s by the ARTP mutagenesis system.
TABLE 1 lethality of different ARTP treatment times on Haematococcus Pluvialis 712
3. Screening
The mutagenized algae and the control are simultaneously cultured in 34 ℃ environment for 2 weeks (the culture medium is modified MCM liquid culture medium), and then the high-temperature algae liquid is transferred to 25 ℃ environment for recovery. The algae strains in the algae solution capable of being re-green are taken as a screening result, namely a target mutant strain, and are named YYAM001 for verifying the characteristics of the algae strains in the next experiment.
4. Confirmation of growth characteristics of the mutant strains
The obtained mutant strain algae liquid is inoculated into 100mL of m-MCM culture medium for culture, and growth curves under the conditions of 25 ℃,28 ℃,34 ℃ and light and dark are respectively measured. And after the astaxanthin grows to a plateau stage, centrifuging to collect the algae for stress induction of the astaxanthin. The specific process is as follows:
(1) Mutant YYAM001 and WT types were cultured in m-MCM medium at the same time under the conditions of light-dark alternation at 25-18 ℃ (14h, 10h), 28-25 ℃ (14h, 10h) and 30 ℃ continuous illumination, and growth curves were obtained after continuous culture for 7 days.
As shown in FIG. 1, the growth of the wild type and the mutant strain is compared when the wild type and the mutant strain are cultured for 7 days at 25-18 ℃ in light-dark alternation (14h. As shown in FIG. 2, the growth of the wild type and the mutant strain was compared when they were cultured for 7 days at 28 ℃ to 25 ℃ in alternating light and dark (14h. FIG. 3 is a graph showing the comparison between the growth of the wild type and the growth of the mutant strain in continuous light culture at 30 ℃ for 7 days. The experimental results of FIGS. 1-3 show that the mutant YYAM001 has better growth than the wild type under all three conditions. This shows that the mutant YYAM001 provided by the invention has better growth advantages under the conditions of light-dark alternation and certain fluctuation of temperature and under the conditions of continuous high temperature. Especially, the mutant strain YYAM001 has better performance under the high temperature condition, which is shown as follows: referring to fig. 4, the control group (wild type) shows almost all the algae are forced to sink to the bottom (become static spores) under the dark condition at 30 ℃, while the mutant YYAM001 is uniformly distributed in the culture medium, the algae liquid is in a deeper and uniform green color, the phenomenon of sinking is much less than that of the control group (wild type), and the cells can still continue to proliferate.
The above results indicate that when the cultivation time is 5-6 days, the cell proliferation rate of the mutant YYAM001 of the present invention is much higher than that of the wild type at a lower temperature, and the proliferation rate of the mutant is much higher than that of the wild type at a continuous high temperature. The rapid growth at this stage provides a large number of algal cells for later astaxanthin accumulation, which can be transferred into a stress environment to accumulate more astaxanthin.
(2) Stress and astaxanthin content determination:
the mutant strain YYAM001 and wild type WT grown to the plateau stage were centrifuged separately to remove the original m-MCM medium for stress, and the medium was placed in m-MCM high salt (0.8% NaCl) with a light intensity of 6000lux to induce stress, in a system of 50mL stress medium. After 5 days of stress, the algal cells were examined under the microscope as shown in FIG. 5.
As shown in FIG. 5, the bright red areas evident in YYAM001 on the left are mainly produced by the large amount of astaxanthin in the cells, while the WT algal cells on the right are more uniform and darker in color.
After 5 days of stress, microalgae cells were harvested by centrifugation, dried to determine dry weight, and placed in 600 μ L of solvent A (5 g potassium hydroxide dissolved in 30mL methanol and 70mL water). After heating the water bath at 65 ℃ for 15min to destroy chlorophyll, the precipitate was collected by centrifugation at 5000Xg for 5min and washed 3 times with deionized water. Extraction with solvent B (glacial acetic acid HAc and dimethyl sulfoxide DMSO) at 75 deg.C for 5min. The above procedure was repeated until the precipitate was colorless. The astaxanthin content of the extract was measured by spectrophotometry. The absorbance of astaxanthin (A490) was measured spectrophotometrically at 490 nm. The formula is calculated as C (mg/g) =4.5 a490 v/DW. V (mL) represents the volume of solvent B, C is the astaxanthin concentration, and DW is the dry weight of the algal powder.
The results are shown in Table 2, where the mutant group and the wild type group were made 2 replicates, and the astaxanthin content and the mean WT value of YYAM001 were 1.3%. The experiment result shows that under the stress condition, compared with the wild type, the astaxanthin content in the mutant algae cells is slightly higher when the mutant algae cells are induced to turn red to generate astaxanthin. This shows that the mutant YYAM001 of the present invention not only has strong high temperature growth resistance, but also has no reduction in the astaxanthin accumulation capacity of the cells.
TABLE 2 Haematococcus pluvialis mutant and wild type induced stress astaxanthin production data
(3) Screening mutant single algae colony: cells growing out of the plate under the condition of light at 34 ℃ are picked out for passage, and the monoclonal algae species are activated and the steps are repeated to verify the characteristics.
5. Mutant strain algal species stability verification
After 6 months of culture, the YYAM001 mutant strain was tested to still be able to tolerate a high temperature environment of 34 ℃, and induced stress culture was performed for 5 days at high salt (0.8% nacl) and 6000lux in m-MCM, without a decrease in astaxanthin concentration in the test cells. The result shows that the mutant strain YYAM001 provided by the invention has stable physiological and biochemical characteristics and stable hereditary characteristics, can be used as a high-temperature-resistant haematococcus pluvialis strain, and is used for promoting the high-efficiency large-scale production of haematococcus pluvialis. The mutant strain of the invention belongs to mutation, is different from a transgenic technology, can be used in the domestic food industry without limitation, and has good application prospect in feed, food additive industry and the like in future.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and these modifications or substitutions do not depart from the spirit of the corresponding technical solutions of the embodiments of the present invention.
Claims (4)
1. A high-temperature resistant mutant of Haematococcus pluvialis, named YYAM001, is preserved in China general microbiological culture Collection center with a preservation number of CGMCC No.45099, a preservation day of 2022 years, 4 months and 29 days, and the preservation address is No. 3 of No. 1 Hospital of Xilu Xinyang district of Beijing City.
2. A method for producing astaxanthin derived from algae, comprising inoculating and culturing the Haematococcus pluvialis thermostable mutant YYAM001 of claim 1, harvesting the cultured Haematococcus pluvialis, and extracting astaxanthin from the harvested Haematococcus pluvialis cells; wherein the culture method is an autotrophic culture, a fermentation culture or a combined culture mode of autotrophic culture and fermentation.
3. The method according to claim 2, wherein the Haematococcus pluvialis high temperature resistant mutant YYAM001 growing to the plateau stage is inoculated into a high salt medium and subjected to light induced stress culture to promote the accumulation of astaxanthin in the cells.
4. A method according to claim 3, wherein the medium is an m-MCM culture; the light intensity of the induced stress is 6000lux; the culture medium contains NaCl with the mass concentration of 0.8%; the culture time for inducing stress is 5-6 days.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008141757A1 (en) * | 2007-05-23 | 2008-11-27 | Cognis Ip Management Gmbh | Efficient astaxanthin production strains derived from haematococcus pluvialis |
| CN105916993A (en) * | 2014-04-17 | 2016-08-31 | 高丽大学校产学协力团 | Method for increasing production of astaxanthin in haematococcus pluvialis by mature spore inoculation and iron ion-mediated Harber-Weiss reaction at high temperature |
| CN106480155A (en) * | 2016-12-23 | 2017-03-08 | 山东金晶生物技术有限公司 | A kind of method being suitable for promoting Haematococcus pluvialis production astaxanthin under the high temperature conditions |
| CN106916811A (en) * | 2015-12-24 | 2017-07-04 | 国家开发投资公司 | Rain green blood ball algae mutant strain and its application |
-
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- 2022-05-26 CN CN202210590596.XA patent/CN114836324B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008141757A1 (en) * | 2007-05-23 | 2008-11-27 | Cognis Ip Management Gmbh | Efficient astaxanthin production strains derived from haematococcus pluvialis |
| CN105916993A (en) * | 2014-04-17 | 2016-08-31 | 高丽大学校产学协力团 | Method for increasing production of astaxanthin in haematococcus pluvialis by mature spore inoculation and iron ion-mediated Harber-Weiss reaction at high temperature |
| CN106916811A (en) * | 2015-12-24 | 2017-07-04 | 国家开发投资公司 | Rain green blood ball algae mutant strain and its application |
| CN106480155A (en) * | 2016-12-23 | 2017-03-08 | 山东金晶生物技术有限公司 | A kind of method being suitable for promoting Haematococcus pluvialis production astaxanthin under the high temperature conditions |
Non-Patent Citations (1)
| Title |
|---|
| 温度对雨生红球藻叶绿素荧光特性及虾青素含量的影响;梁英等;《海洋湖沼通报》;20090915(第03期);第114-122页 * |
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