Spirulina platensis and its applications

Abstract

The present invention provides Spirulina platensis and its applications, belonging to the field of microbial technology. The Spirulina platensis was deposited at the General Microbiology Center of the China Microbial Culture Collection Center on October 2, 2024. The deposit address is the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The deposit number is CGMCC No. 46152. The present invention isolates, purifies and screens a strain of Spirulina platensis ( Spirulina platensis ) SPPC0024 from the mixed culture solution of Spirulina platensis cultured for a long time. Compared with the existing Spirulina platensis, it has the advantages of rich biomass, easy harvesting, high phycocyanin content and large yield.

Description

Technical Field

[0001] The present invention belongs to the technical field of microbiology, and particularly relates to Spirulina platensis and its applications. Background Art

[0002] C-phycocyanin (C-PC) is a phycobiliprotein with light-harvesting function. Due to its unique fluorescence properties and biological activities such as anti-tumor, antioxidant, and immunomodulatory effects, it has important application values in the fields of food, medicine, cosmetics, and biomedical engineering.

[0003] Spirulina ( Spirulinasp .) is the main commercial source of C-phycocyanin, and its protein content can reach 10% - 20% of the dry cell weight. Among them, Spirulina platensis ( Spirulina platensis ) is a commonly used Spirulina for preparing C-phycocyanin at present. However, in most existing Spirulina platensis, the proportion of C-phycocyanin in the dry weight is generally low, and it is easily affected by fluctuations in culture conditions and has poor adaptability to light intensity changes, resulting in unstable yields. In addition, the existing Spirulina platensis has a low floating rate after cultivation, which is not conducive to subsequent harvesting.

[0004] Therefore, screening out Spirulina platensis with excellent performance is very important for improving the yield and quality of C-phycocyanin. Summary of the Invention

[0005] In view of this, the purpose of the present invention is to provide Spirulina platensis and its applications, aiming to solve at least one technical problem in the background art.

[0006] The present invention is implemented as follows:

[0007] The present invention provides Spirulina platensis ( Spirulina platensis ) SPPC0024, which was deposited on October 2, 2024 at the General Microbiological Center of the China Committee for Culture Collection of Microorganisms, with the deposit address being the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number being CGMCC No. 46152.

[0008] The present invention also provides the application of the above-mentioned Spirulina platensis SPPC0024 in the production of C-phycocyanin.

[0009] The present invention isolated, purified, and screened a strain of Spirulina platensis ( Spirulina platensis ) SPPC0024 from a long-term cultured mixed Spirulina culture solution. Compared with the existing Spirulina platensis, it has the advantages of rich biomass, easy harvesting, high C-phycocyanin content, and large yield.

[0010] The SPPC0024 algal strain of the present invention has a high phycocyanin content under normal culture conditions, and its content can be significantly increased after simple optimized culture, indicating that this algal strain has good industrial application value in phycocyanin production. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 It is an optical microscope observation diagram of Spirulina platensis SPPC0024 in Example 1;

[0012] Figure 2 It is a phylogenetic tree diagram of Spirulina platensis SPPC0024 in Example 1;

[0013] Figure 3 It is a diagram showing the change of biomass of two Spirulina platensis with time under different light intensities in Example 2. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0014] In order to make the objectives, technical solutions and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with embodiments. It should be understood that the specific implementation cases described herein are only used to explain the present invention and are not used to limit the present invention.

[0015] Example 1

[0016] The separation, purification and screening of Spirulina platensis SPPC0024 are carried out as follows:

[0017] Heat the glass capillary with an alcohol lamp and then pull it into an extremely fine capillary tube with tweezers; dilute the long-term cultured Spirulina platensis mixed solution with a basic medium at different gradient concentrations, drop it on a glass slide, and pick the single algal filaments with the same morphology under a stereomicroscope with the capillary tube and place them in an EP tube for purified culture to obtain a Spirulina platensis seed culture solution; transfer the Spirulina platensis seed culture solution to a conical flask for enlarged culture to obtain an algal solution at a transfer ratio of 1:10, set the culture temperature at 30°C ± 0.2°C, and the light intensity at 4000 lux, and select the dominant algal strains with fast growth rate and high phycocyanin content by measuring the biomass and phycocyanin content.

[0018] Among them, the formula of the basal medium is as follows: 16.8 g / L of NaHCO3, 2.5 g / L of NaNO3, 1.0 g / L of NaCl, 0.5 g / L of K2HPO4, 1.0 g / L of K2SO4, 0.2 g / L of MgSO4·7H2O, 0.08 g / L of CaCl2·2H2O, 0.002 g / L of FeSO4·7H2O, 0.016 g / L of Na2EDTA, 2 ml / L of A5 trace element solution and 1 ml / L of B6 trace element solution; among them, the components of the A5 trace element solution are: 2.86 g / L of H3BO3, 1.81 g / L of MnCl2·4H2O, 0.222 g / L of ZnSO4·7H2O, 0.39 g / L of NaMoO4·2H2O, 0.079 g / L of CuSO4·5H2O; the components of the B6 trace element solution are: 22.9 mg / L of NH4VO3, 96.0 mg / L of CrK(SO4)2·12H2O, 47.8 mg / L of NiSO4·6H2O, 17.9 mg / L of NaWO4·2H2O, 44.0 mg / L of Co(NO3)2·6H2O, 40.0 mg / L of Ti(SO4)2. The culture medium and experimental materials are both sterilized.

[0019] The identification of this dominant algal strain specifically includes:

[0020] (1) Take a small amount of diluted algal solution and observe it under a microscope, and collect images. As Figure 1 shown, the results show that this algal strain appears blue-green under an optical microscope, the algal filaments are more than 1300 μm long, the pitch is 110 μm - 130 μm, and the number of spirals is 13 - 16.

[0021] (2) Use PCR amplification to determine the sequence of the nuclear-encoded ribosomal 16s rDNA fragment, and extract the algal genome;

[0022] The PCR reaction system is: 2×PCR Mix, template DNA, forward and reverse primers, ddH2O, with a total volume of 50 μL;

[0023] The reaction program is: denaturation at 95°C for 5 min, and then 34 cycles according to the program of 95°C for 30 s, 55°C for 30 s, 72°C for 1 min, and finally extension at 72°C for 10 min.

[0024] The PCR product is electrophoresed in 1.5% agarose gel; after the PCR product is purified, it is directly sequenced by a sequencing company. Using the Blast function, compare the obtained 16s rDNA fragment sequence of the sample (as shown in SEQ ID NO:1) with the GenBank database, and it is found that the homology of this algal strain with the genus Arthrospira ( Arthrospira ) can reach 100%.

[0025] The phylogenetic tree constructed based on the 16S rDNA sequence of the cell ribosome (as shown in Figure 2 ) shows that the sample is closely related to Spirulina platensis ( Spirulina platensis ) and is on the same evolutionary branch. This algal strain is named Spirulina platensis SPPC0024.

[0026] Spirulina platensis SPPC0024 was deposited on October 2, 2024 at the General Microbiology Center of the China Committee for Culture Collection of Microorganisms. The deposit address is the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The deposit number is CGMCC No. 46152.

[0027] Example 2

[0028] Under different light intensities (4000 lux, 6000 lux, 8000 lux), the algal solution of Spirulina platensis SPPC0024 was inoculated into the basal medium and continuously cultured for 10 days at a temperature of 25°C ± 2°C. The aeration conditions included: a CO2 concentration of 1% and an aeration rate of 15 L / min to 20 L / min.

[0029] At the same time, the existing Spirulina platensis GY-D18 with a high phycocyanin content was used as a comparison object and cultured under the same conditions. The biomass changes of the two algal strains were measured regularly, and the results are as shown in Figure 3 . Figure 3 In Figure 3 , a is the biomass change of Spirulina platensis SPPC0024; Figure 3 In

[0030] , b is the biomass change of the Spirulina platensis GY-D18 algal strain. It can be seen that the Spirulina platensis SPPC0024 of the present invention has better tolerance to high light intensity and a faster biomass accumulation rate.

[0031] Example 3

[0032] Under a temperature of 25°C ± 2°C, a light intensity of 5000 lux ± 1000 lux, and aeration conditions (CO2 concentration of 1%, aeration rate of 15 L / min to 20 L / min), the algal solution of Spirulina platensis SPPC0024 was inoculated into the basal medium and continuously cultured for 10 days.

[0033] At the same time, the existing Spirulina platensis GY-D18 with a high phycocyanin content was used as a comparison object and cultured under the same conditions. At the end of the culture, the biomass, floating rate, protein content, and phycocyanin content of the two algal strains were measured, and the comparison results are shown in Table 1.

[0034] Example 4

[0035] Under the conditions that the temperature is 25°C ± 2°C, the light intensity of the stage light quality strategy is 5000 lux ± 1000 lux, and the aeration conditions (CO2 concentration is 1%, aeration rate is 15 L / min to 20 L / min), the Spirulina platensis SPPC0024 algal solution is inoculated into the basic medium and continuously cultured for 10 days.

[0036] Among them, the stage light quality strategy is: white light in the early stage and blue light in the later stage. In this example, the white light illumination time is 8 days and the blue light illumination time is 2 days.

[0037] At the same time, the existing Spirulina platensis GY-D18 with a high phycocyanin content is used as a comparison object and cultured under the same conditions. At the end of the culture, the biomass, floating rate, protein content and phycocyanin content of the two algal strains are measured, and the comparison results are shown in Table 1.

[0038] Table 1

[0039]

[0040] From the data in Table 1, it can be seen that under normal culture conditions, the phycocyanin content of the SPPC0024 algal strain of the present invention is as high as 29.78%, which is significantly higher than that of the existing Spirulina platensis GY-D18 algal strain with a high phycocyanin content and far exceeds that of general Spirulina platensis. By simply optimizing the culture, the phycocyanin content can be increased by about 3%. In addition, the floating rate of the SPPC0024 algal strain is as high as 90.03%, which is beneficial for subsequent harvesting.

[0041] In summary, the SPPC0024 algal strain of the present invention can be used as a superior algal strain for preparing phycocyanin, and products rich in phycocyanin, such as food, feed, cosmetics, health products or drugs, etc., can be prepared.

[0042] The above embodiments only represent several implementation manners of the present invention. The description is relatively specific and detailed, but it should not be construed as a limitation on the scope of the patent of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present invention, several modifications and improvements can still be made, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention should be subject to the appended claims.

Claims

1. Spirulina platensis ( Spirulina platensis ), SPPC0024, characterized in that The Spirulina platensis SPPC0024 was deposited at the General Microbiological Center of the China Committee for Culture Collection of Microorganisms on October 2, 2024. The deposit address is the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The deposit number is CGMCC No. 46152.

2. Use of the Spirulina platensis SPPC0024 according to claim 1 in the production of phycocyanin.

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