CN104988200A - Method for promoting haematococcus pluvialis to produce astaxanthin by use of fulvic acid - Google Patents
Method for promoting haematococcus pluvialis to produce astaxanthin by use of fulvic acid Download PDFInfo
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- CN104988200A CN104988200A CN201510376770.0A CN201510376770A CN104988200A CN 104988200 A CN104988200 A CN 104988200A CN 201510376770 A CN201510376770 A CN 201510376770A CN 104988200 A CN104988200 A CN 104988200A
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- astaxanthin
- haematococcus pluvialis
- algae liquid
- xanthohumic acid
- haematocoocus pluvialls
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- JEBFVOLFMLUKLF-IFPLVEIFSA-N Astaxanthin Natural products CC(=C/C=C/C(=C/C=C/C1=C(C)C(=O)C(O)CC1(C)C)/C)C=CC=C(/C)C=CC=C(/C)C=CC2=C(C)C(=O)C(O)CC2(C)C JEBFVOLFMLUKLF-IFPLVEIFSA-N 0.000 title claims abstract description 67
- 235000013793 astaxanthin Nutrition 0.000 title claims abstract description 67
- MQZIGYBFDRPAKN-ZWAPEEGVSA-N astaxanthin Chemical compound C([C@H](O)C(=O)C=1C)C(C)(C)C=1/C=C/C(/C)=C/C=C/C(/C)=C/C=C/C=C(C)C=CC=C(C)C=CC1=C(C)C(=O)[C@@H](O)CC1(C)C MQZIGYBFDRPAKN-ZWAPEEGVSA-N 0.000 title claims abstract description 67
- 229940022405 astaxanthin Drugs 0.000 title claims abstract description 67
- 239000001168 astaxanthin Substances 0.000 title claims abstract description 67
- 238000000034 method Methods 0.000 title claims abstract description 23
- 241000168517 Haematococcus lacustris Species 0.000 title claims abstract description 21
- 230000001737 promoting effect Effects 0.000 title abstract description 3
- PUKLDDOGISCFCP-JSQCKWNTSA-N 21-Deoxycortisone Chemical compound C1CC2=CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@@](C(=O)C)(O)[C@@]1(C)CC2=O PUKLDDOGISCFCP-JSQCKWNTSA-N 0.000 title abstract 5
- FCYKAQOGGFGCMD-UHFFFAOYSA-N Fulvic acid Natural products O1C2=CC(O)=C(O)C(C(O)=O)=C2C(=O)C2=C1CC(C)(O)OC2 FCYKAQOGGFGCMD-UHFFFAOYSA-N 0.000 title abstract 5
- 229940095100 fulvic acid Drugs 0.000 title abstract 5
- 239000002509 fulvic acid Substances 0.000 title abstract 5
- 230000006698 induction Effects 0.000 claims abstract description 21
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims abstract description 20
- 239000012452 mother liquor Substances 0.000 claims abstract description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 11
- 229910052757 nitrogen Inorganic materials 0.000 claims abstract description 10
- 230000001939 inductive effect Effects 0.000 claims abstract description 5
- 239000002253 acid Substances 0.000 claims description 36
- 241000195493 Cryptophyta Species 0.000 claims description 35
- 239000007788 liquid Substances 0.000 claims description 33
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 18
- 238000009825 accumulation Methods 0.000 claims description 11
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 9
- 238000001556 precipitation Methods 0.000 claims description 9
- 239000006228 supernatant Substances 0.000 claims description 9
- 239000003960 organic solvent Substances 0.000 claims description 8
- 238000000638 solvent extraction Methods 0.000 claims description 8
- 230000001580 bacterial effect Effects 0.000 claims description 7
- 238000005286 illumination Methods 0.000 claims description 7
- 230000000050 nutritive effect Effects 0.000 claims description 6
- 150000003839 salts Chemical class 0.000 claims description 6
- 238000002360 preparation method Methods 0.000 claims description 5
- 241000168525 Haematococcus Species 0.000 claims description 4
- 239000003513 alkali Substances 0.000 claims description 3
- 238000005119 centrifugation Methods 0.000 claims description 3
- 239000011259 mixed solution Substances 0.000 claims description 3
- 238000002525 ultrasonication Methods 0.000 claims description 3
- 238000010790 dilution Methods 0.000 claims description 2
- 239000012895 dilution Substances 0.000 claims description 2
- 238000012258 culturing Methods 0.000 abstract 1
- 238000007865 diluting Methods 0.000 abstract 1
- 239000001963 growth medium Substances 0.000 abstract 1
- 239000008213 purified water Substances 0.000 abstract 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 230000003078 antioxidant effect Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 230000003203 everyday effect Effects 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 230000004151 fermentation Effects 0.000 description 3
- 239000000654 additive Substances 0.000 description 2
- 235000021466 carotenoid Nutrition 0.000 description 2
- 150000001747 carotenoids Chemical class 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000005648 plant growth regulator Substances 0.000 description 2
- 241000972773 Aulopiformes Species 0.000 description 1
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 1
- 241000180279 Chlorococcum Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- MBLBDJOUHNCFQT-LXGUWJNJSA-N aldehydo-N-acetyl-D-glucosamine Chemical compound CC(=O)N[C@@H](C=O)[C@@H](O)[C@H](O)[C@H](O)CO MBLBDJOUHNCFQT-LXGUWJNJSA-N 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 235000011089 carbon dioxide Nutrition 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 235000003642 hunger Nutrition 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 229910017053 inorganic salt Inorganic materials 0.000 description 1
- 125000000468 ketone group Chemical group 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 239000003375 plant hormone Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 235000019515 salmon Nutrition 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a method for promoting haematococcus pluvialis to produce astaxanthin by use of fulvic acid. The method comprises the following steps: at first, preparing a haematococcus pluvialis solution, wherein when the haematococcus pluvialis is in the later period of the logarithmic phase, the cell concentration reaches 10<6>cells mL<-1>; diluting the haematococcus pluvialis solution by use of a BG-11 culture medium lack of nitrogen till the concentration reaches 2*10<5>cells mL<-1> to obtain the haematococcus pluvialis solution for induction; preparing a 10g L<-1> fulvic acid mother liquor by use of purified water, and adding the fulvic acid mother liquor into the well diluted inducing haematococcus pluvialis solution to enable the concentration of fulvic acid to reach 4.95-5.05 mg L<-1>; culturing the inducing haematococcus pluvialis solution, and when the algal cells become completely red, collecting the algal cells of which the astaxanthin accumulative amount is the highest at the moment. The method is simple and feasible and low in cost, can greatly shorten the time for haematococcus pluvialis to turn red, relatively remarkably increases the quantity of astaxanthin, and accordingly, greatly improves the yield of astaxanthin.
Description
Technical field
The invention belongs to technical field of bioengineering, be specifically related to a kind of method utilizing xanthohumic acid to promote haematococcus pluvialis to produce astaxanthin.
Background technology
Natural astaxanthin (3,3 '-dihydroxyl-4,4 '-diketo-β, β '-carotene) is the strongest antioxidant that current nature finds.Natural astaxanthin is originated in spontaneous husband's yeast, salmon, shrimp shell and Haematocoocus Pluvialls, and containing abundant astaxanthin in Haematocoocus Pluvialls, can reach 4% of its dry weight, be described as the concentrate of natural astaxanthin.It is the highest derivative of carotenoid to the anti-oxidant activity major embodiment of astaxanthin on the one hand; Its conjugated double bond end has undersaturated ketone group and hydroxyl on the other hand, and group forms alpha-alcohol ketone thus, and therefore it is better than the oxidation-resistance of general carotenoid.Natural astaxanthin is without any side effects to human body, and the anti-oxidant activity of astaxanthin makes it have very large application potential at medicine, makeup and healthcare products etc., and astaxanthin also can as the fodder additives of aquatic animal and foodstuff additive in addition.
Haematocoocus Pluvialls belongs to a kind of unicell green alga, and it is only under stress conditions, such as high temperature, intense light irradiation, nitrogen hunger etc., accumulation astaxanthin that can be a large amount of.And mostly concentrate in the physics and chemistry factor about the induction stimulating Haematocoocus Pluvialls to produce astaxanthin, the less research had about plant hormone.Xanthohumic acid, can Promoting plant growth as a plant growth regulators, has very strong degeneration-resistant effect, and Haematocoocus Pluvialls just accumulates astaxanthin down in a large number this coercing just.Foreign aid adds the expression that xanthohumic acid can excite plant defense genes, the chemical defence of inducing plant, the effect similar with insect's food-taking to physical abuse.Therefore xanthohumic acid may induce Haematocoocus Pluvialls to accumulate astaxanthin in a large number.
The domestic company having had large-scale production astaxanthin at present; also there has been some relevant technology and patents; such as " the cultivation production method of Haematocoocus Pluvialls " (201210295602) disclose a kind of bioreactor that utilizes and cultivate the method for Haematocoocus Pluvialls, and what comprise different LED light source cultivates the method for green cell with carbonic acid gas adjust ph." a kind of airlift photobioreactor of Haematocoocus Pluvialls high-density culture " (CN200510018203.4) of Hubei Normal University, disclose a kind of airlift photobioreactor of Haematocoocus Pluvialls high-density culture, comprise cultivation tank, water treatment device, means of illumination, gas supply device, realize round-the-clock high-density breeding by regulation and control reaction conditions, and the cultivation of algae and astaxanthin accumulation are combined in same reactor complete." cultivating the method that algae produces astaxanthin with yeast fermentation raffinate " (CN1480524A) of University Of Tianjin, disclose a kind of method utilizing yeast fermentation raffinate to produce astaxanthin, in yeast fermentation raffinate, add inorganic salt be made into substratum, Haematocoocus Pluvialls or Chlorococcum is cultivated, through separation and Extraction astaxanthin in specific container.Although above-mentioned Patents technology is advanced, technique is comparatively complicated, and equipment requirements is high, operates comparatively loaded down with trivial details, this adds increased the difficulty of production cost and technology popularization.
Summary of the invention
The object of this invention is to provide a kind of xanthohumic acid that utilizes and promote the method for haematococcus pluvialis to produce astaxanthin, the method not only simple to operate, cost is low, and the cycle of accumulation astaxanthin can be shortened to a great extent and improve the productive rate of astaxanthin.
The technical solution adopted in the present invention is, a kind of method utilizing xanthohumic acid to promote haematococcus pluvialis to produce astaxanthin, comprises the following steps:
The preparation of step 1, algae liquid: cultivate Haematocoocus Pluvialls to the logarithmic phase later stage, also obtains the algae liquid of induction with the BG-11 substratum dilution algae liquid of nitrogen stress;
Step 2, induction Haematocoocus Pluvialls accumulation astaxanthin: be made into 10g L with pure water
-1xanthohumic acid mother liquor, xanthohumic acid mother liquor is added in the algae liquid of the induction of having diluted and makes xanthohumic acid concentration reach 4.95-5.05mg L
-1; Then above-mentioned algae liquid is cultivated, after frond cell reddens completely, collect frustule;
Step 3, utilize the astaxanthin of organic solvent extraction frustule.
Feature of the present invention is also:
Haematocoocus Pluvialls is Haematocoocus Pluvialls bacterial strain Haematococcus pluvialis LUGU.
In step 1, the preparation of algae liquid is specially: at 24-26 DEG C, light intensity 2900-3000lx, cultivate under continuous light condition, and cultivate Haematocoocus Pluvialls and arrive the logarithmic phase later stage, now cell concn reaches 10
6cells mL
-1, utilize the BG-11 substratum of nitrogen stress to be diluted to 2 × 10
5cells mL
-1as induction algae liquid.
The culture condition of inducing Haematocoocus Pluvialls to accumulate in astaxanthin in step 2 is: 26-28 DEG C, light intensity 9500-10000lx, cold light lamp continuous illumination and hungry nutritive salt Combined Stress.
Utilize the astaxanthin of organic solvent extraction frustule to be specially in step 3: to get the above-mentioned algae liquid of 5ml, the centrifugal 5min of 4000r/min, abandon supernatant and obtain frustule precipitation; Add 2mL30% methyl alcohol and 5%KOH mixed solution, the centrifugal 10min of 60 DEG C of water-bath 15min, 4000r/min respectively in frustule precipitation, except supernatant, precipitation adds pure water 2 times, except alkali lye remains; Then 5mL DMSO is added, 200w ultrasonication 15min, wherein, ultrasonic 1min, 10s interval, in 45 DEG C of water-bath 20min, extracting is turned white to frond, and collected by centrifugation supernatant surveys OD490; And according to formula
calculate the content of astaxanthin, wherein C is content astaxanthin, and A is OD value under 490nm, and Va is DMSO volume, and Vb is that algae liquid amasss.
The invention has the beneficial effects as follows: operation is simple, cost is low in (1) the present invention, starting material are that the Haematocoocus Pluvialls bacterial strain of oneself screening can be cultivated according to a conventional method.
(2) the present invention significantly improves the productive rate of astaxanthin, and experiment proves, adds 4.95-5.05mg L
-1the induction group of xanthohumic acid than the cycle time 20-30% of control group accumulation astaxanthin, the yield increased group of astaxanthin improves 82.3-86.9%, reach 20.3-20.817mg L
-1.
Embodiment
Below in conjunction with embodiment, the present invention is described in detail.
The invention provides a kind of method utilizing xanthohumic acid to promote haematococcus pluvialis to produce astaxanthin, comprise the following steps:
The preparation of step 1, algae liquid: Haematocoocus Pluvialls adopts Haematocoocus Pluvialls bacterial strain Haematococcuspluvialis LUGU (NCBI:KM115647), utilize BG-11 substratum, at 24-26 DEG C, light intensity 2900-3000lx, cultivate under continuous light condition, cultivate Haematocoocus Pluvialls and arrive the logarithmic phase later stage, now frustule concentration reaches 10
6cells mL
-1, utilize the BG-11 substratum of nitrogen stress to be diluted to 2 × 10
5cells mL
-1as induction algae liquid;
Step 2, induction Haematocoocus Pluvialls accumulation astaxanthin: be made into 10g L with pure water
-1xanthohumic acid mother liquor, xanthohumic acid mother liquor is added in the induction algae liquid diluted and makes xanthohumic acid concentration reach 4.95-5.05mg L
-1; Then by above-mentioned algae liquid 26-28 DEG C, light intensity 9500-10000lx, cold light lamp continuous illumination and hungry nutritive salt Combined Stress under cultivate, after frond cell reddens completely, collect frustule;
Step 3, utilize the astaxanthin of organic solvent extraction frustule, the extracting method of astaxanthin:
Within every two days, get the above-mentioned algae liquid of 5ml, the centrifugal 5min of 4000r/min, abandon supernatant and obtain frustule precipitation; Add 2mL 30% methyl alcohol and 5%KOH mixed solution, the centrifugal 10min of 60 DEG C of water-bath 15min, 4000r/min respectively in frustule precipitation, except supernatant, precipitation adds pure water 2 times, except alkali lye remains; Then add 5mL DMSO, 200w ultrasonication 15min (ultrasonic 1min, 10s interval), in 45 DEG C of water-bath 20min, extracting is turned white to frond, and collected by centrifugation supernatant surveys OD490; And according to formula
calculate the content of astaxanthin, wherein C is content astaxanthin, and A is OD value under 490nm, and Va is DMSO volume, and Vb is that algae liquid amasss.
Embodiment 1
(1) Haematocoocus Pluvialls adopts Haematocoocus Pluvialls bacterial strain Haematococcus pluvialisLUGU, utilize BG-11 substratum, at 24 DEG C, light intensity 2900lx, cultivate under continuous light condition, cultivate Haematocoocus Pluvialls and arrive logarithmic phase, now frustule concentration reaches 10
6cells mL
-1, utilize the BG-11 substratum of nitrogen stress to be diluted to 2 × 10
5cells mL
-1as induction algae liquid.
(2) 10gL is made into pure water
-1xanthohumic acid mother liquor, xanthohumic acid mother liquor is added in the induction algae liquid diluted and makes xanthohumic acid concentration reach 4.95mg L
-1; Then by above-mentioned algae liquid 26 DEG C, light intensity 9500lx, cold light lamp continuous illumination and hungry nutritive salt Combined Stress under cultivate, regularly sample every day and observe frustule colour-change, after finding that after 7 days, frond cell reddens completely, collect the astaxanthin that frustule utilizes organic solvent extraction frustule, now the accumulation volume of its astaxanthin has reached the highest.
Embodiment 2
(1) Haematocoocus Pluvialls adopts Haematocoocus Pluvialls bacterial strain Haematococcus pluvialisLUGU, utilize BG-11 substratum, at 25 DEG C, light intensity 3000lx, cultivate under continuous light condition, be cultured to Haematocoocus Pluvialls and arrive logarithmic phase, now frustule concentration reaches 10
6cells mL
-1, utilize the BG-11 substratum of nitrogen stress to be diluted to 2 × 10
5cells mL
-1as induction algae liquid.
(2) 10g L is made into pure water
-1xanthohumic acid mother liquor, xanthohumic acid mother liquor is added in the induction algae liquid diluted and makes xanthohumic acid concentration reach 5mg L
-1; Then by above-mentioned algae liquid 27 DEG C, light intensity 9800lx, cold light lamp continuous illumination and hungry nutritive salt Combined Stress under cultivate, regularly sample every day and observe frustule colour-change, after after 7 days, frond cell reddens completely, collect the astaxanthin that frustule utilizes organic solvent extraction frustule, now the accumulation volume of its astaxanthin has reached the highest.
Embodiment 3
(1) Haematocoocus Pluvialls adopts Haematocoocus Pluvialls bacterial strain Haematococcus pluvialisLUGU, utilize BG-11 substratum, at 26 DEG C, light intensity 2950lx, cultivate under continuous light condition, be cultured to Haematocoocus Pluvialls and arrive logarithmic phase, now frustule concentration reaches 10
6cells mL
-1, utilize the BG-11 substratum of nitrogen stress to be diluted to 2 × 10
5cells mL
-1as induction algae liquid.
(2) 10g L is made into pure water
-1xanthohumic acid mother liquor, xanthohumic acid mother liquor is added in the induction algae liquid diluted and makes xanthohumic acid concentration reach 5.05mg L
-1; Then by above-mentioned algae liquid 28 DEG C, light intensity 10000lx, cold light lamp continuous illumination and hungry nutritive salt Combined Stress under cultivate, regularly sample every day and observe frustule colour-change, after after 6 days, frond cell reddens completely, collect the astaxanthin that frustule utilizes organic solvent extraction frustule, now the accumulation volume of its astaxanthin has reached the highest.
Xanthohumic acid can promote the g and D of plant effectively as a plant growth regulators, utilize certain density xanthohumic acid as inductor, induction Haematocoocus Pluvialls accumulation astaxanthin, extraction astaxanthin is carried out to each group by adopting the extracting method of above-mentioned astaxanthin, result shows: it is 10 days that blank group (not adding xanthohumic acid) accumulates the time that astaxanthin completes, and when the 10th day, the content of astaxanthin is 11.1375mg L
-1, the ratio control group after xanthohumic acid process accumulates the cycle time of astaxanthin 20-30%, and the yield increased group of astaxanthin improves 82.3-86.9%, reaches 20.3-20.817mg L
-1.
Claims (5)
1. utilize xanthohumic acid to promote a method for haematococcus pluvialis to produce astaxanthin, it is characterized in that, comprise the following steps:
The preparation of step 1, algae liquid: cultivate Haematocoocus Pluvialls to the logarithmic phase later stage, also obtains the algae liquid of induction with the BG-11 substratum dilution algae liquid of nitrogen stress;
Step 2, induction Haematocoocus Pluvialls accumulation astaxanthin: be made into 10g L with pure water
-1xanthohumic acid mother liquor, xanthohumic acid mother liquor is added in the algae liquid of the induction of having diluted and makes xanthohumic acid concentration reach 4.95-5.05mg L
-1; Then above-mentioned algae liquid is cultivated, after frond cell reddens completely, collect frustule;
Step 3, utilize the astaxanthin of organic solvent extraction frustule.
2. the method utilizing xanthohumic acid to promote haematococcus pluvialis to produce astaxanthin according to claim 1, it is characterized in that, described Haematocoocus Pluvialls is Haematocoocus Pluvialls bacterial strain Haematococcus pluvialisLUGU.
3. the method utilizing xanthohumic acid to promote haematococcus pluvialis to produce astaxanthin according to claim 1, it is characterized in that, in described step 1, the preparation of algae liquid is specially: at 24-26 DEG C, light intensity 2900-3000lx, cultivate under continuous light condition, cultivate Haematocoocus Pluvialls and arrive the logarithmic phase later stage, now cell concn reaches 10
6cells mL
-1, utilize the BG-11 substratum of nitrogen stress to be diluted to 2 × 10
5cells mL
-1as induction algae liquid.
4. the method utilizing xanthohumic acid to promote haematococcus pluvialis to produce astaxanthin according to claim 1, it is characterized in that, the culture condition of inducing Haematocoocus Pluvialls to accumulate in astaxanthin in described step 2 is: 26-28 DEG C, light intensity 9500-10000lx, cold light lamp continuous illumination and hungry nutritive salt Combined Stress.
5. the method utilizing xanthohumic acid to promote haematococcus pluvialis to produce astaxanthin according to claim 1, it is characterized in that, utilize the astaxanthin of organic solvent extraction frustule to be specially in described step 3: to get the above-mentioned algae liquid of 5ml, the centrifugal 5min of 4000r/min, abandon supernatant and obtain frustule precipitation; Add 2mL 30% methyl alcohol and 5% KOH mixed solution, the centrifugal 10min of 60 DEG C of water-bath 15min, 4000r/min respectively in frustule precipitation, except supernatant, precipitation adds pure water 2 times, except alkali lye remains; Then 5mL DMSO is added, 200w ultrasonication 15min, wherein, ultrasonic 1min, 10s interval, in 45 DEG C of water-bath 20min, extracting is turned white to frond, and collected by centrifugation supernatant surveys OD490; And according to formula
calculate the content of astaxanthin, wherein C is content astaxanthin, and A is OD value under 490nm, and Va is DMSO volume, and Vb is that algae liquid amasss.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106566775A (en) * | 2016-10-21 | 2017-04-19 | 青岛科海生物有限公司 | Preparation method of high-activity haematococcus pluvialis cells |
| CN107354122A (en) * | 2017-09-18 | 2017-11-17 | 深圳市德和生物科技有限公司 | A kind of method for promoting haematococcus pluvialis growing multiplication and redden |
| CN109679853A (en) * | 2019-01-18 | 2019-04-26 | 昆明理工大学 | The method for improving haematococcus pluvialis biomass and astaxanthin yield using fulvic acid |
| CN110804639A (en) * | 2019-11-27 | 2020-02-18 | 中国科学院天津工业生物技术研究所 | Method for promoting esterification of microalgae astaxanthin |
| CN112998111A (en) * | 2021-04-16 | 2021-06-22 | 馨辰生物(广东)有限公司 | Haematococcus pluvialis gel candy and preparation method thereof |
| CN114717288A (en) * | 2022-05-20 | 2022-07-08 | 云南中科雨虹生物科技有限公司 | Method for increasing yield of haematococcus pluvialis astaxanthin by adding organic acid |
| CN115044527A (en) * | 2022-02-15 | 2022-09-13 | 昆明理工大学 | Application of inositol in promoting haematococcus pluvialis to produce astaxanthin |
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| CN109679853A (en) * | 2019-01-18 | 2019-04-26 | 昆明理工大学 | The method for improving haematococcus pluvialis biomass and astaxanthin yield using fulvic acid |
| CN110804639A (en) * | 2019-11-27 | 2020-02-18 | 中国科学院天津工业生物技术研究所 | Method for promoting esterification of microalgae astaxanthin |
| CN110804639B (en) * | 2019-11-27 | 2022-04-08 | 中国科学院天津工业生物技术研究所 | Method for promoting esterification of microalgae astaxanthin |
| CN112998111A (en) * | 2021-04-16 | 2021-06-22 | 馨辰生物(广东)有限公司 | Haematococcus pluvialis gel candy and preparation method thereof |
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