CN113817649B - Method for producing pseudo-cladophyllin by inducing hair weeds to suspend and culture cells by using solar radiation - Google Patents
Method for producing pseudo-cladophyllin by inducing hair weeds to suspend and culture cells by using solar radiation Download PDFInfo
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- 241000196324 Embryophyta Species 0.000 title claims abstract description 67
- 230000005855 radiation Effects 0.000 title claims abstract description 43
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 18
- 230000001939 inductive effect Effects 0.000 title description 2
- 238000004114 suspension culture Methods 0.000 claims abstract description 27
- 241000195493 Cryptophyta Species 0.000 claims abstract description 25
- 239000001963 growth medium Substances 0.000 claims abstract description 13
- 238000009630 liquid culture Methods 0.000 claims abstract description 11
- 238000000034 method Methods 0.000 claims abstract description 8
- 238000002360 preparation method Methods 0.000 claims abstract description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 34
- 210000004027 cell Anatomy 0.000 claims description 34
- 210000004748 cultured cell Anatomy 0.000 claims description 19
- 238000004108 freeze drying Methods 0.000 claims description 15
- 108010053210 Phycocyanin Proteins 0.000 claims description 8
- 241000308169 Pseudocladosporium Species 0.000 claims description 2
- 238000012258 culturing Methods 0.000 abstract description 12
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 24
- 238000002835 absorbance Methods 0.000 description 13
- 239000002028 Biomass Substances 0.000 description 12
- 229960004799 tryptophan Drugs 0.000 description 12
- 239000000725 suspension Substances 0.000 description 10
- 230000012010 growth Effects 0.000 description 7
- 239000010453 quartz Substances 0.000 description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 230000000977 initiatory effect Effects 0.000 description 5
- 238000005070 sampling Methods 0.000 description 5
- 238000003786 synthesis reaction Methods 0.000 description 5
- 230000000737 periodic effect Effects 0.000 description 4
- 206010019049 Hair texture abnormal Diseases 0.000 description 3
- 238000005265 energy consumption Methods 0.000 description 3
- 230000002411 adverse Effects 0.000 description 2
- 239000002537 cosmetic Substances 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000005286 illumination Methods 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- CGZKSPLDUIRCIO-UHFFFAOYSA-N Scytonemin Natural products OC1=CC=C(C=C1)C=C1C(C(=C2C1=NC=1C=CC=CC2=1)C=1C(C(C2=NC=3C=CC=CC=3C2=1)=CC1=CC=C(C=C1)O)=O)=O CGZKSPLDUIRCIO-UHFFFAOYSA-N 0.000 description 1
- 230000036579 abiotic stress Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000001028 anti-proliverative effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 230000004224 protection Effects 0.000 description 1
- 230000009979 protective mechanism Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- CGZKSPLDUIRCIO-RPCRKUJJSA-N scytonemin Chemical group C1=CC(O)=CC=C1\C=C\1C2=NC3=CC=CC=C3C2=C(C=2C(C(=C/C=3C=CC(O)=CC=3)/C=3C=2C2=CC=CC=C2N=3)=O)C/1=O CGZKSPLDUIRCIO-RPCRKUJJSA-N 0.000 description 1
- 229930000044 secondary metabolite Natural products 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- IFGCUJZIWBUILZ-UHFFFAOYSA-N sodium 2-[[2-[[hydroxy-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyphosphoryl]amino]-4-methylpentanoyl]amino]-3-(1H-indol-3-yl)propanoic acid Chemical compound [Na+].C=1NC2=CC=CC=C2C=1CC(C(O)=O)NC(=O)C(CC(C)C)NP(O)(=O)OC1OC(C)C(O)C(O)C1O IFGCUJZIWBUILZ-UHFFFAOYSA-N 0.000 description 1
- 230000035882 stress Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
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Abstract
The invention relates to a method for producing pseudo-cladophyllin by utilizing sun radiation to induce hair weeds suspension culture cells, which comprises the steps of inoculating the hair weeds suspension culture cells into a liquid culture medium to obtain a culture system A; then the culture system A is radiated for 0.5 to 1 hour under the sunlight to obtain a culture system B; then shake culturing the culture system B at constant temperature by using an LED fluorescent lamp for 1 day, 2 days or 3 days to obtain a culture system C; then treating the culture system C by a mode of treating the culture system A in the second step, and treating the obtained culture system by a mode of treating the culture system B in the third step to obtain a culture system D, so as to form a second cycle treatment on the hair weeds suspension culture cells; the method is used for continuously carrying out circulation treatment on the hair weeds suspension culture cells for 8-14 days, then the algae are collected by the centrifugal culture solution, and the pseudo-branch algae are extracted from the algae, so that the preparation of the pseudo-branch algae is completed.
Description
Technical Field
The invention relates to the technical field of induction synthesis of blue algae secondary metabolites, in particular to a method for producing pseudo-cladophyllin by using sun radiation to induce hair weeds to suspend cultured cells.
Background
The hair weeds are nitrogen-fixing blue algae growing in arid and semiarid regions, can synthesize various bioactive substances, and have high edible and medicinal values. Since long-term exposure to various abiotic stresses, in particular high intensity ultraviolet radiation, hair weeds have formed a variety of protective mechanisms during evolution, with synthesis of pseudocladophyllin being one of the most important pathways for hair weed cells to resist ultraviolet radiation damage.
Pseudo-branch algae element (C) 36 H 20 N 2 O 4 ) The English name is scytonemin, which is a brown yellow fat-soluble pigment small molecule distributed in part of extracellular polysaccharide gum sheaths of blue algae. The pseudo-cladophyllin has strong molecular stability and can resist the influence of various adverse environments such as temperature, oxidative stress and ultraviolet radiation. In addition to its strong ultraviolet absorbing ability, pseudocladophyllin has various biological activities such as antioxidant, anti-inflammatory and antiproliferative activities, and has great application potential in the fields of medicine and cosmetics.
The synthesis of pseudo-cladophyllin is mainly induced by ultraviolet radiation and is also influenced by high light, salt stress and nutritional conditions, but the current method for producing pseudo-cladophyllin has the problems of low yield and high energy consumption.
Disclosure of Invention
Aiming at the problems of high energy consumption and low yield in the production of pseudo-branch phycocyanin in the prior art, the invention provides a method for producing pseudo-branch phycocyanin by utilizing sun radiation to induce the suspension culture cells of the hair weeds.
The invention is realized by the following technical scheme:
a method for producing pseudo-cladophyllin by using sun radiation to induce hair weeds to suspend cultured cells comprises the following steps:
step 1, inoculating a hair weeds suspension culture cell into a liquid culture medium to obtain a culture system A;
step 2, carrying out radiation treatment on the culture system A for 0.5-1 hour in the sun to obtain a culture system B;
step 3, the culture system B is subjected to constant temperature, and the light intensity is 40-45 mu mol of photons m -2 s -1 Carrying out shaking culture for 1 day, 2 days or 3 days on the LED fluorescent lamp to obtain a culture system C;
step 4, treating the culture system C in a mode of treating the culture system A in the step 2, and treating the obtained culture system in a mode of treating the culture system B in the step 3 to obtain a culture system D, so as to form second circulation treatment on the hair weeds suspension culture cells;
and 5, continuously performing circulation treatment on the hair weeds suspension culture cells in the mode of the step 4, wherein the total treatment time is 8-14 days, and then collecting algae by using a centrifugal culture solution, and extracting pseudo-cladophyllin from the algae to finish the preparation of the pseudo-cladophyllin.
Preferably, during the irradiation treatment in step 2, the UV-A, UV-B and visible light intensities are respectively 1.4-23W m -2 、0.1～1.6W m -2 And 76-1298. Mu. Mol of photons m -2 s -1 。
Preferably, the temperature during the radiation treatment in step 2 is 20-32 ℃.
Preferably, step 1 comprises inoculating the hair weeds suspension culture cells in a liquid culture medium, adding L-tryptophan to make the concentration of L-tryptophan in the obtained system be 0.5-5 mmol/L, and then performing the radiation treatment in step 2.
Preferably, step 3 is to perform shaking culture of the culture system B at a constant temperature of 24-26 ℃.
Further, step 1, inoculating the hair weeds suspension culture cells into BG-11 liquid culture medium to obtain a culture system A.
Preferably, the rotation speed is 120-130 rpm when the shake culture is performed in the step 2.
Preferably, step 5 separates the pseudo-cladophyllin in the culture solution as follows:
centrifuging the culture solution to obtain precipitated algae, freeze-drying the algae, extracting with acetone to obtain extractive solution, and freeze-drying the extractive solution to obtain pseudo-branch phycocyanin.
Compared with the prior art, the invention has the following beneficial technical effects:
the invention relates to a method for producing pseudo-branch phycocyanin by utilizing natural sunlight radiation to induce the hair weeds to suspend and culture cells, wherein sunlight is taken as a renewable resource, partial blue algae cells can be induced to synthesize the pseudo-branch phycocyanin, and the sunlight radiation is reasonably utilized to induce the hair weeds to suspend and culture cells to synthesize a large amount of pseudo-branch phycocyanin, so that the problems of high production cost and low yield of the existing method can be solved, and meanwhile, the method also accords with the principles of economy, green and environmental protection. The long-time exposure to sunlight can inhibit the growth and photosynthesis of the hair weeds suspension culture cells, so that the short-term periodic irradiation of sunlight is utilized to irradiate the hair weeds suspension culture cells, the adverse effect of sunlight radiation on the growth of cell biomass can be reduced, and meanwhile, the synthesis of pseudo-cladophyllin can be efficiently induced. The invention saves energy consumption, is simple and easy to operate, is suitable for large-scale production, can obviously improve economic benefit, and can be applied to the fields of medicines and cosmetics.
Furthermore, according to the invention, 0.5-5 mM L-tryptophan is added into the culture medium, and then the process is carried out according to the subsequent process, so that the content of pseudo-cladosporium cucumerinum can be further greatly improved after the periodic irradiation treatment is carried out compared with the culture medium without the L-tryptophan.
Drawings
FIG. 1 is a graph showing the effect of L-tryptophan (Try) on the biomass of hair weeds in example 5 of the invention.
FIG. 2 is a graph showing the effect of L-tryptophan (Try) on the content of arduous sphaericus in example 5 of the present invention.
Detailed Description
The invention will now be described in further detail with reference to specific examples, which are intended to illustrate, but not to limit, the invention.
The invention will be described in detail below with reference to the drawings and the detailed description.
The invention relates to a method for producing pseudo-cladophyllin by using sun radiation to induce hair weeds to suspend cultured cells, which comprises the following steps:
step 1, inoculating a hair weeds suspension culture cell with good growth state into a quartz triangular flask filled with 100mL of BG-11 liquid culture medium, and performing periodic short-term treatment by using natural light to induce efficient synthesis of pseudo-cladophyllin; in addition, L-tryptophan is added into 100mL of BG-11 liquid medium, the concentration is 0.5-5 mmol/L, and the yield of pseudo-cladophyllin can be further improved by the same solar radiation treatment.
Step 2, performing periodic short-term radiation treatment under natural light, wherein each radiation treatment lasts for 0.5-1 hour, and the duration from the beginning of the previous radiation to the beginning of the second radiation in two adjacent radiation lasts for 1 day, 2 days or 3 days; with the addition of L-tryptophan, each irradiation was performed for 0.5 hours, and the time period from the start of the previous irradiation to the start of the second irradiation was 1 day in the two adjacent irradiation.
Step 3, shake culturing for 1 day, 2 days or 3 days at constant temperature under LED fluorescent lamp, short-term radiation and shake culturing according to the above steps for 8-14 days, and light intensity of 40 μmol photons m -2 s -1 The temperature is 25 ℃, and the rotating speed of the shaking table is 120-130 rpm.
And 4, respectively taking 10mL of culture solution during the culture period, centrifuging and collecting materials, freeze-drying, and measuring the cell biomass of the hair weeds and the content of the pseudo-cladophyllin.
Step 5, measuring the content of pseudo-branch phycocyanin, extracting the dried algae body with 100% acetone at 4 ℃ in dark overnight for 12-18 hours to obtain an extracting solution, and measuring the OD in the extracting solution 334 According to molar absorptivity 112.6L g -1 ·cm -1 And calculating the content of pseudo-branch phycin.
And 6, freeze-drying the extracting solution to obtain the pseudo-cladophyllin.
Example 1
The invention relates to a method for producing pseudo-cladophyllin by using sun radiation to induce hair weeds to suspend cultured cells, which comprises the following steps:
step (1), inoculating the hair weeds suspension culture cells with good growth state into a quartz triangular flask filled with 100mL BG-11 liquid culture medium, and initiating OD 730 0.30.
Step (2), the hair weeds suspension cultured cells in the step (1) are cultured in sunlightThe ultraviolet ray UV-A, the ultraviolet ray UV-B, the visible light intensity and the temperature during the irradiation treatment are respectively as follows: 1.4 to 23. 23W m -2 、0.1～1.6W m -2 、76～1298μmol photons m -2 s -1 And 20 to 32 ℃.
And (3) carrying out radiation treatment on the hair weeds suspension culture cells in the step (1) in the sun for 0.5 hour.
Step (4), shake culturing the hair weeds suspension cultured cells after the irradiation in the step (3) under an LED fluorescent lamp and a constant temperature illumination room for 1 day, and then performing short-term irradiation treatment and shake culturing according to the steps, wherein the total treatment time is 12 days, the rotating speed is 130rpm, and the intensity of the LED fluorescent lamp is 40 mu mol photons m -2 s -1 The temperature was 25 ℃.
And (5) sampling the step (4), respectively taking 10mL of culture solution, centrifuging for 20min, collecting samples, freeze-drying, and measuring the cell biomass of the hair weeds and the content of the pseudo-cladophyllin.
Step (6), measuring the content of the pseudo-cladophyllin, extracting the dried algae body with 100% acetone at 4 ℃ in dark overnight, measuring the absorbance of an acetone solution containing the pseudo-cladophyllin at the wavelength of 334nm, and measuring the absorbance according to the molar absorbance coefficient of 112.6L g -1 ·cm -1 Calculating content, and measuring 11.1mg g of pseudo-branch phycin dry hair weeds at 12 days -1 。
And (7) freeze-drying the acetone solution remained in the step (6) to obtain the pseudo-cladophyllin.
Example 2
The invention relates to a method for producing pseudo-cladophyllin by using sun radiation to induce hair weeds to suspend cultured cells, which comprises the following steps:
step (1), inoculating the hair weeds suspension culture cells with good growth state into a quartz triangular flask filled with 100mL BG-11 liquid culture medium, and initiating OD 730 0.30.
Step (2), the hair weeds suspension cultured cells in the step (1) are subjected to radiation treatment in sunlight, and ultraviolet light UV-A, ultraviolet light UV-B, visible light intensity and temperature during the treatment are respectively as follows: 1.4 to 23. 23W m -2 、0.1～1.6W m -2 、76～1298μmol photons m -2 s -1 And 20 to 32 ℃.
And (3) carrying out radiation treatment on the hair weeds suspension culture cells in the step (1) in the sun for 0.5 hour.
Step (4), shake culturing the hair weeds suspension cultured cells after the irradiation in the step (3) for 2 days under the conditions of an LED fluorescent lamp and constant temperature, then performing short-term irradiation treatment and shake culturing according to the steps, wherein the total treatment time is 12 days, the rotating speed is 130rpm, and the intensity of the LED fluorescent lamp is 40 mu mol photons m -2 s -1 The temperature was 25 ℃.
And (5) sampling the step (4), respectively taking 10mL of culture solution, centrifuging for 20min, collecting samples, freeze-drying, and measuring the cell biomass of the hair weeds and the content of the pseudo-cladophyllin.
Step (6), measuring the content of the pseudo-cladophyllin, extracting the dried algae body with 100% acetone at 4 ℃ in dark overnight, measuring the absorbance of an acetone solution containing the pseudo-cladophyllin at the wavelength of 334nm, and measuring the absorbance according to the molar coefficient of 112.6L g -1 cm -1 Calculating content, and measuring 9.05mg g of pseudo-branch phycin dry hair weeds at 12 days -1 。
And (7) freeze-drying the acetone solution remained in the step (6) to obtain the pseudo-cladophyllin.
Example 3
The invention relates to a method for producing pseudo-cladophyllin by using sun radiation to induce hair weeds to suspend cultured cells, which comprises the following steps:
step (1), inoculating the hair weeds suspension culture cells with good growth state into a quartz triangular flask filled with 100mL BG-11 liquid culture medium, and initiating OD 730 0.27.
Step (2), the hair weeds suspension cultured cells in the step (1) are subjected to radiation treatment in sunlight, and ultraviolet light UV-A, ultraviolet light UV-B, visible light intensity and temperature during the treatment are respectively as follows: 1.4 to 23. 23W m -2 、0.1～1.6W m -2 、76～1298μmol photons m -2 s -1 And 20 to 32 ℃.
And (3) carrying out radiation treatment on the hair weeds suspension culture cells in the step (1) in the sun for 0.5 hour.
Step (4), shake culturing the hair weeds suspension cultured cells after the irradiation in the step (3) for 3 days under the conditions of an LED fluorescent lamp and constant temperature, then performing short-term irradiation treatment and shake culturing according to the steps, wherein the total treatment time is 12 days, the rotating speed is 130rpm, and the intensity of the LED fluorescent lamp is 40 mu mol photons m -2 s -1 The temperature was 25 ℃.
And (5) sampling the step (4), respectively taking 10mL of culture solution, centrifuging for 20min, collecting samples, freeze-drying, and measuring the cell biomass of the hair weeds and the content of the pseudo-cladophyllin.
Step (6), measuring the content of the pseudo-cladophyllin, extracting the dried algae body with 100% acetone at 4 ℃ in dark overnight, measuring the absorbance of an acetone solution containing the pseudo-cladophyllin at the wavelength of 334nm, and measuring the absorbance according to the molar absorbance coefficient of 112.6L g -1 ·cm -1 Calculating content, and measuring pseudo-branch algae content in dried hair weeds at 12 days to obtain pseudo-branch algae content of 7.61mg g -1 。
And (7) freeze-drying the acetone solution remained in the step (6) to obtain the pseudo-cladophyllin.
Example 4
The invention relates to a method for producing pseudo-cladophyllin by using sun radiation to induce hair weeds to suspend cultured cells, which comprises the following steps:
step (1), inoculating the hair weeds suspension culture cells with good growth state into a quartz triangular flask filled with 100mL BG-11 liquid culture medium, and initiating OD 730 0.28.
Step (2), the hair weeds suspension cultured cells in the step (1) are subjected to radiation treatment in sunlight, and ultraviolet light UV-A, ultraviolet light UV-B, visible light intensity and temperature during the treatment are respectively as follows: 1.4 to 23. 23W m -2 、0.1～1.6W m -2 、76～1298μmol photons m -2 s -1 And 20 to 32 ℃.
And (3) carrying out radiation treatment on the hair weeds suspension culture cells in the step (1) in the sun for 1 hour.
Step (4), shake culturing the hair weeds suspension cultured cells after the irradiation of the step (3) for 1 day, and then performing short-term culture according to the stepsRadiation treatment and shaking culture, total treatment time of 12 days, rotation speed of 130rpm, and LED fluorescent lamp intensity of 40 μmol photons m -2 s -1 The temperature was 25 ℃.
And (5) sampling the step (4), respectively taking 10mL of culture solution, centrifuging for 20min, collecting samples, freeze-drying, and measuring the cell biomass of the hair weeds and the content of the pseudo-cladophyllin.
Step (6), measuring the content of the pseudo-cladophyllin, extracting the dried algae body with 100% acetone at 4 ℃ overnight in a dark place, measuring the absorbance of an acetone solution containing the pseudo-cladophyllin at the wavelength of 334nm, and measuring the absorbance according to the molar coefficient of 112.6L g -1 ·cm -1 Calculating content, and measuring 3.58mg g of pseudo-branch phycin dry hair weeds at 12 days -1 。
And (7) freeze-drying the acetone solution remained in the step (6) to obtain the pseudo-cladophyllin.
Example 5
The invention relates to a method for producing pseudo-cladophyllin by using sun radiation to induce hair weeds to suspend cultured cells, which comprises the following steps:
step (1), inoculating the hair weeds suspension culture cells with good growth state into a quartz triangular flask filled with 100mL BG-11 liquid culture medium, and initiating OD 730 0.28, with the addition of 5mM L-tryptophan.
Step (2), the hair weeds suspension cultured cells in the step (1) are subjected to radiation treatment in sunlight, and ultraviolet light UV-A, ultraviolet light UV-B, visible light intensity and temperature are respectively as follows: 1.2 to 35. 35W m -2 、0.1～2.6W m -2 、51～1720μmol photons m -2 s -1 And 27 to 39 ℃.
And (3) carrying out radiation treatment on the hair weeds suspension culture cells in the step (1) in the sun for 0.5 hour.
Step (4), shake culturing the hair weeds suspension cultured cells after the irradiation of the step (3) for 1 day, then performing short-term irradiation treatment and shake culturing according to the steps, wherein the total treatment time is 12 days, the rotating speed is 130rpm, and the illumination intensity is 40 mu mol photons m -2 s -1 The temperature was 25 ℃.
And (5) sampling the step (4), respectively taking 10mL of culture solution, centrifuging for 20min, collecting samples, freeze-drying, and measuring the cell biomass of the hair weeds and the content of the pseudo-cladophyllin.
Step (6), measuring the content of the pseudo-cladophyllin, extracting the dried algae body with 100% acetone at 4 ℃ in dark overnight, measuring the absorbance of an acetone solution containing the pseudo-cladophyllin at the wavelength of 334nm, and measuring the absorbance according to the molar absorbance coefficient of 112.6L g -1 ·cm -1 And calculating the content.
As shown in FIG. 1, the biomass after the addition of 5mM L-tryptophan was 1.23. 1.23g L -1 Slightly lower than the untreated group (1.43 g L) -1 ) The biomass impact is less. As shown in FIG. 2, the content of pseudo-cladophyllin in dried hair weeds was 8.78mg g as measured by adding 5mM L-tryptophan -1 Is significantly higher than the treatment group without L-tryptophan (3.44 mg g -1 )。
As can be seen from Table 1, the different treatment frequencies were used, the different levels of hair weeds biomass and pseudo-cladophylline. The treatment frequency is 1 time per day when the irradiation time is 0.5 hr, and the content of pseudo-branch phycin in dried hair weeds can reach 11.11mg g -1 The effect is best. If L-tryptophan substrate is added to the medium, the production of pseudo-cladophyllin is further increased.
TABLE 1 Hair weed suspension culture cell biomass and pseudo-cladophyllin content at different treatment frequencies
Claims (3)
1. A method for producing pseudo-cladophyllin by using sun radiation to induce hair weeds to suspend cultured cells, which is characterized by comprising the following steps:
step 1, inoculating a hair weeds suspension culture cell into a BG-11 liquid culture medium to obtain a culture system A;
step 2, the culture system A is subjected to radiation treatment for 0.5 hour in the sun, the temperature is 20-32 ℃ during the radiation treatment, and the intensity of UV-A, UV-B and visible light is 1.4-23W m respectively -2 、0.1~1.6 W m -2 And 76-1298 mu mol of photons m -2 s -1 Obtaining a culture system B;
step 3, the culture system B is subjected to light intensity of 40-45 mu mol of photons m at the constant temperature of 24-26 DEG C -2 s -1 Carrying out shaking culture for 1 day or 2 days on the LED fluorescent lamp to obtain a culture system C;
step 4, treating the culture system C in a mode of treating the culture system A in the step 2, and treating the obtained culture system in a mode of treating the culture system B in the step 3 to obtain a culture system D, so as to form second circulation treatment on the hair weeds suspension culture cells;
and 5, continuously performing circulation treatment on the hair weeds suspension culture cells in the mode of the step 4, wherein the total treatment time is 12 days, and then, collecting algae by using a centrifugal culture solution, and extracting pseudo-cladophyllin from the algae to finish the preparation of the pseudo-cladophyllin.
2. The method for preparing pseudo-cladosporium cucumerinum by using solar radiation according to claim 1, wherein the rotation speed is 120-130 rpm when the shaking culture is performed in the step 3.
3. The method for preparing pseudo-branch phycin using solar radiation according to claim 1, wherein step 5 separates pseudo-branch phycin in the culture solution according to the following procedure:
centrifuging the culture solution to obtain precipitated algae, freeze-drying the algae, extracting with acetone to obtain extractive solution, and freeze-drying the extractive solution to obtain pseudo-branch phycocyanin.
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Non-Patent Citations (5)
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| Biotechnological Production of the Sunscreen Pigment Scytonemin in Cyanobacteria: Progress and Strategy;Xiang Gao;Mar Drugs;第19卷(第3期);摘要 * |
| 地木耳人工培养最适光温条件及功能性成分Scytonemin的变化;赵丽娟;上官旖旎;陈娟娟;严小军;;食品科学(第09期);第208-212页 * |
| 赖永忠.阳光辐射和干出对水培发状念珠藻生理特性的影响.中国优秀硕士学位论文全文数据库 基础科学辑.2008,第2.2、2.3、3.2.1节. * |
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| 阳光辐射和干出对水培发状念珠藻生理特性的影响;赖永忠;中国优秀硕士学位论文全文数据库 基础科学辑;第2.2、2.3、3.2.1节 * |
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