CN103554250A - Method for extracting phycocyanin - Google Patents
Method for extracting phycocyanin Download PDFInfo
- Publication number
- CN103554250A CN103554250A CN201310592174.7A CN201310592174A CN103554250A CN 103554250 A CN103554250 A CN 103554250A CN 201310592174 A CN201310592174 A CN 201310592174A CN 103554250 A CN103554250 A CN 103554250A
- Authority
- CN
- China
- Prior art keywords
- phycocyanins
- resin
- spirulina
- phycocyanin
- powder
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000000034 method Methods 0.000 title claims abstract description 13
- 108010053210 Phycocyanin Proteins 0.000 title abstract 7
- 240000002900 Arthrospira platensis Species 0.000 claims abstract description 35
- 235000016425 Arthrospira platensis Nutrition 0.000 claims abstract description 35
- 229940082787 spirulina Drugs 0.000 claims abstract description 32
- 239000000843 powder Substances 0.000 claims abstract description 28
- 239000011347 resin Substances 0.000 claims abstract description 28
- 229920005989 resin Polymers 0.000 claims abstract description 28
- 238000000605 extraction Methods 0.000 claims abstract description 12
- 239000000287 crude extract Substances 0.000 claims abstract description 11
- 239000006228 supernatant Substances 0.000 claims abstract description 11
- 238000001694 spray drying Methods 0.000 claims abstract description 8
- 238000000265 homogenisation Methods 0.000 claims abstract description 7
- 238000001914 filtration Methods 0.000 claims abstract description 6
- 239000012528 membrane Substances 0.000 claims abstract description 6
- 238000001471 micro-filtration Methods 0.000 claims abstract description 6
- 238000003756 stirring Methods 0.000 claims abstract description 6
- 239000000284 extract Substances 0.000 claims abstract description 3
- 239000007788 liquid Substances 0.000 claims description 15
- 238000005119 centrifugation Methods 0.000 claims description 9
- 238000001035 drying Methods 0.000 claims description 6
- 238000001179 sorption measurement Methods 0.000 claims description 6
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 claims description 4
- 239000003456 ion exchange resin Substances 0.000 claims description 4
- 229920003303 ion-exchange polymer Polymers 0.000 claims description 4
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims description 2
- 229910019142 PO4 Inorganic materials 0.000 claims description 2
- 239000010452 phosphate Substances 0.000 claims description 2
- 229910052708 sodium Inorganic materials 0.000 claims description 2
- 239000011734 sodium Substances 0.000 claims description 2
- 239000000243 solution Substances 0.000 abstract description 11
- 239000000047 product Substances 0.000 abstract description 5
- 239000007853 buffer solution Substances 0.000 abstract description 4
- 239000003480 eluent Substances 0.000 abstract 1
- 229940011019 arthrospira platensis Drugs 0.000 description 3
- 229910000162 sodium phosphate Inorganic materials 0.000 description 3
- 239000001488 sodium phosphate Substances 0.000 description 3
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 241000195493 Cryptophyta Species 0.000 description 1
- 125000003275 alpha amino acid group Chemical group 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 239000003797 essential amino acid Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000002864 food coloring agent Nutrition 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 235000008935 nutritious Nutrition 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 229910001753 sapphirine Inorganic materials 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/795—Porphyrin- or corrin-ring-containing peptides
Abstract
The invention provides a method for extracting phycocyanin. The method comprises the following steps: adding buffer solution into spirulina powder, uniformly stirring and then performing high-pressure homogenization and wall breaking to obtain spirulina wall-breaking solution; centrifugally separating and then taking a supernatant; filtering by using microfiltration membranes of 2 Mu m, 0.45 Mu m and 0.2 Mu m respectively to obtain phycocyanin crude extract; decolorizing and purifying with over-combined resin at a flow rate of 2-5mL/min; collecting eluent to obtain phycocyanin extract; finally performing high-speed centrifugal spray drying, and collecting the powder to obtain the phycocyanin. According to the method, the phycocyanin can be extracted to the greatest extent, no secondary pollution is caused, the extraction rate of the phycocyanin in the spirulina is over 90.0 percent, and the purity of the product reaches A620/A280 which is greater than 3.2.
Description
Technical field
The present invention relates to a kind of method of extracting Phycocyanins, C-, belong to biological product technical field.
Background technology
Phycocyanins, C-is to take the algae such as spirulina as the isolated a kind of mazarine powder of main raw material.It is a kind of protein, is again a kind of fabulous natural food colour, is again good protective foods simultaneously.Phycocyanins, C-is one of rare chromoprotein of occurring in nature, not only bright-colored, and itself is a kind of nutritious protein, and its amino acid forms complete, and essential amino acids content is high.
At present, in spirulina, the extracting method of Phycocyanins, C-is a lot, but too many to the loss of Phycocyanins, C-, extraction yield is lower, is difficult to extract in a large number, and easily causes product secondary pollution, should not apply.Therefore, be necessary to research and develop high, the easy to operate Phycocyanins, C-extracting method of a kind of extraction yield.
Summary of the invention
For Phycocyanins, C-extraction yield in solution prior art is low, be difficult to the problems such as popularization, the invention provides a kind of method of extracting Phycocyanins, C-, by following technical proposal, realize.
The present invention is to provide a kind of like this technical scheme: a kind of method of extracting Phycocyanins, C-, is characterized in that through following each step:
(1) frond of fresh or dry spirulina plalensis is made to spirulina powder;
(2) press spirulina powder: the mass ratio of buffered soln=1:0.8～1:1.2, in the spirulina powder of step (1), add buffered soln, stir evenly, at pressure, be under 40～70MPa, to carry out high pressure homogenization broken wall 2～5 times, obtain breaking wall of spirullina princeps liquid;
(3) step (2) gained breaking wall of spirullina princeps liquid is carried out, after centrifugation, getting supernatant liquor, then supernatant liquor is filtered by the microfiltration membrane of 2 μ m, 0.45 μ m and 0.2 μ m successively, after filtration, obtain Phycocyanins, C-crude extract;
(4) by step (3) gained Phycocyanins, C-crude extract, with the flow velocity of 2～5mL/min, by the combined resin purifying that decolours, collect elutriant, be Phycocyanins, C-extracting solution; The consumption of described combined resin is 0.8～2.0 times of spirulina powder volume;
(5) step (4) gained Phycocyanins, C-extracting solution is carried out to hig h-speed centrifugal spray drying, collect powder, obtain Phycocyanins, C-.
The buffered soln of described step (2) is the sodium radio-phosphate,P-32 solution that pH is 6.5, concentration is 0.1M.
The high pressure homogenization broken wall of described step (2) is to complete on conventional high pressure homogenizer.
Centrifugation 20～30min under the condition that the centrifugation of described step (3) is is 3500～5000r/min at rotating speed.
The combined resin of described step (4) is two or more the combination in D900 type ion exchange resin, HPD-300L type resin, the large mesh nonpolar adsorption resin of BEHP-3 type, the large mesh nonpolar adsorption resin of BEHP-4 type, D112 sun resin, and portfolio ratio is arbitrarily.
The hig h-speed centrifugal spray drying of described step (5) is to be that 2～8L/h, inlet temperature are that 180～320 ℃, air outlet temperature are that 80～90 ℃, drying temperature are to carry out under the condition of 160～220 ℃ in liquid feeding speed.
The advantage that the present invention possesses and effect: by the present invention, can extract to greatest extent Phycocyanins, C-; and product can not be secondary polluted; the operation of the present invention from broken wall, separation and purification to drying and other steps all convenient and easy, feasibility is strong; can accomplish scale production; in production process, not discharging pollutants, is a kind of environment amenable Phycocyanins, C-extracting method.Phycocyanins, C-content in spirulina is generally 3～10%, and the extraction yield that adopts method of the present invention to extract Phycocyanins, C-in spirulina is that more than 90.0% product purity reaches A620/A280>3.2.
Embodiment
Below by embodiment, the present invention will be further described.
Embodiment 1
(1) frond of fresh high-quality spirulina plalensis (Arthrospira platensis) is removed after impurity, made routinely spirulina powder;
(2) press spirulina powder: the mass ratio of buffered soln=1:1, in the spirulina powder of step (1), add the buffer solution of sodium phosphate that pH is 6.5, concentration is 0.1M, after stirring evenly, sending in high pressure homogenizer, is under 60MPa, to carry out high pressure homogenization broken wall 4 times at pressure, obtains breaking wall of spirullina princeps liquid;
(3) under the condition that is 3500r/min by step (2) gained breaking wall of spirullina princeps liquid at rotating speed, carry out centrifugation 30min, get supernatant liquor, then supernatant liquor is filtered by the microfiltration membrane of 2 μ m, 0.45 μ m and 0.2 μ m respectively, after filtration, obtain Phycocyanins, C-crude extract;
(4) by step (3) gained Phycocyanins, C-crude extract with the flow velocity of 3mL/min by combined resin, the purifying that decolours, collects elutriant, is Phycocyanins, C-extracting solution; The consumption of described combined resin is 1.5 times of spirulina powder volume, and this combined resin is: D900 type ion exchange resin: the volume ratio of the large mesh nonpolar adsorption resin=2:1 of BEHP-3 type;
(5) by step (4) gained Phycocyanins, C-extracting solution, in liquid feeding speed, be that 5L/h, inlet temperature are that 220 ℃, air outlet temperature are that 80 ℃, drying temperature are to carry out hig h-speed centrifugal spray drying under the condition of 180 ℃, collect sapphirine powder, obtain Phycocyanins, C-.Extraction yield is more than 90.0%, and purity is A620/A280=3.4.
Embodiment 2
(1) frond of dry high-quality spirulina plalensis (Arthrospira platensis) is made to spirulina powder routinely;
(2) press spirulina powder: the mass ratio of buffered soln=1:0.8, in the spirulina powder of step (1), add the buffer solution of sodium phosphate that pH is 6.5, concentration is 0.1M, after stirring evenly, sending in high pressure homogenizer, is under 70MPa, to carry out high pressure homogenization broken wall 2 times at pressure, obtains breaking wall of spirullina princeps liquid;
(3) under the condition that is 4000r/min by step (2) gained breaking wall of spirullina princeps liquid at rotating speed, carry out centrifugation 25min, get supernatant liquor, and this supernatant liquor is filtered by the microfiltration membrane of 2 μ m, 0.45 μ m and 0.2 μ m respectively, after filtration, obtain Phycocyanins, C-crude extract;
(4) by step (3) gained Phycocyanins, C-crude extract, with the flow velocity of 5mL/min, by the combined resin purifying that decolours, collect elutriant, be Phycocyanins, C-extracting solution; The volumetric usage of this combined resin is 2.0 times of spirulina powder volume, and this combined resin is comprised of HPD-300L type resin, the large mesh nonpolar adsorption resin of BEHP-4 type and D112 sun resin, and its volume ratio is 3:2:2;
(5) by step (4) gained Phycocyanins, C-extracting solution, in liquid feeding speed, be that 8L/h, inlet temperature are that 180 ℃, air outlet temperature are that 90 ℃, drying temperature are under the condition of 220 ℃, carry out routinely hig h-speed centrifugal spray drying, collect powder, obtain Phycocyanins, C-.Extraction yield is more than 90.0%, and purity is A620/A280=3.5.
Embodiment 3
(1) frond of fresh high-quality spirulina plalensis (Arthrospira platensis) is made to spirulina powder routinely;
(2) press spirulina powder: the mass ratio of buffered soln=1:1.2, in the spirulina powder of step (1), add the buffer solution of sodium phosphate that pH is 6.5, concentration is 0.1M, after stirring evenly, sending in high pressure homogenizer, is under 40MPa, to carry out high pressure homogenization broken wall 5 times at pressure, obtains breaking wall of spirullina princeps liquid;
(3) under the condition that is 5000r/min by step (2) gained breaking wall of spirullina princeps liquid at rotating speed, carry out getting supernatant liquor after centrifugation 20min, then supernatant liquor is filtered by the microfiltration membrane of 2 μ m, 0.45 μ m and 0.2 μ m respectively, after filtration, obtain Phycocyanins, C-crude extract;
(4), by step (3) gained Phycocyanins, C-crude extract, with the flow velocity of 2mL/min, by the combined resin being comprised of by the volume ratio of 1:1 D900 type ion exchange resin and the D112 sun resin purifying that decolours, collection elutriant, is Phycocyanins, C-extracting solution; The consumption of this combined resin is 0.8 times of spirulina powder volume;
(5) by step (4) gained Phycocyanins, C-extracting solution, in liquid feeding speed, be that 2L/h, inlet temperature are that 320 ℃, air outlet temperature are that 85 ℃, drying temperature are to carry out hig h-speed centrifugal spray drying under the condition of 160 ℃, collect powder, obtain Phycocyanins, C-.Extraction yield is more than 90.0%, and purity is A620/A280=3.4.
Claims (5)
1. extract a method for Phycocyanins, C-, it is characterized in that through following each step:
(1) frond of fresh or dry spirulina plalensis is made to spirulina powder;
(2) press spirulina powder: the mass ratio of buffered soln=1:0.8～1:1.2, in the spirulina powder of step (1), add buffered soln, stir evenly, at pressure, be under 40～70MPa, to carry out high pressure homogenization broken wall 2～5 times, obtain breaking wall of spirullina princeps liquid;
(3) step (2) gained breaking wall of spirullina princeps liquid is carried out, after centrifugation, getting supernatant liquor, then supernatant liquor is filtered by the microfiltration membrane of 2 μ m, 0.45 μ m and 0.2 μ m successively, after filtration, obtain Phycocyanins, C-crude extract;
(4) by step (3) gained Phycocyanins, C-crude extract, with the flow velocity of 2～5mL/min, by the combined resin purifying that decolours, collect elutriant, be Phycocyanins, C-extracting solution; The consumption of described combined resin is 0.8～2.0 times of spirulina powder volume;
(5) step (4) gained Phycocyanins, C-extracting solution is carried out to hig h-speed centrifugal spray drying, collect powder, obtain Phycocyanins, C-.
2. the method for extraction Phycocyanins, C-according to claim 1, is characterized in that: the buffered soln of described step (2) is the sodium radio-phosphate,P-32 solution that pH is 6.5, concentration is 0.1M.
3. the method for extraction Phycocyanins, C-according to claim 1, is characterized in that: centrifugation 20～30min under the condition that the centrifugation of described step (3) is is 3500～5000r/min at rotating speed.
4. the method for extraction Phycocyanins, C-according to claim 1, it is characterized in that: the combined resin of described step (4) is two or more the combination in D900 type ion exchange resin, HPD-300L type resin, the large mesh nonpolar adsorption resin of BEHP-3 type, the large mesh nonpolar adsorption resin of BEHP-4 type, D112 sun resin, and portfolio ratio is arbitrarily.
5. the method for extraction Phycocyanins, C-according to claim 1, is characterized in that: the hig h-speed centrifugal spray drying of described step (5) is to be that 2～8L/h, inlet temperature are that 180～320 ℃, air outlet temperature are that 80～90 ℃, drying temperature are to carry out under the condition of 160～220 ℃ in liquid feeding speed.
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| CN201310592174.7A CN103554250B (en) | 2013-11-22 | 2013-11-22 | Method for extracting phycocyanin |
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| CN201310592174.7A CN103554250B (en) | 2013-11-22 | 2013-11-22 | Method for extracting phycocyanin |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105055251A (en) * | 2015-08-29 | 2015-11-18 | 云南蓝钻生物科技股份有限公司 | Anti-aging cosmetic and preparation method thereof |
| CN105055250A (en) * | 2015-08-29 | 2015-11-18 | 云南蓝钻生物科技股份有限公司 | Cosmetic containing phycocyanin and preparation method |
| CN108059669A (en) * | 2017-10-26 | 2018-05-22 | 东台市赐百年生物工程有限公司 | A kind of extracting method for spraying broken wall formula phycocyanin |
| CN108794565A (en) * | 2018-04-27 | 2018-11-13 | 胡梦丽 | A kind of preparation method of spirulina protein powder |
| CN110272486A (en) * | 2019-03-08 | 2019-09-24 | 北海生巴达生物科技有限公司 | The extracting method of phycocyanin in a kind of cyanobacteria |
| CN111116737A (en) * | 2020-01-13 | 2020-05-08 | 江西理工大学 | High-purity phycocyanin and preparation method thereof |
| CN111493204A (en) * | 2020-04-26 | 2020-08-07 | 武汉舜泽生物工程有限公司 | Production process and application of spirulina concentrated protein protoplasm |
| US10822373B2 (en) | 2019-01-09 | 2020-11-03 | Chenghui Zheng | Methods for synthesizing phycocyanin using a biological substance |
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105055251A (en) * | 2015-08-29 | 2015-11-18 | 云南蓝钻生物科技股份有限公司 | Anti-aging cosmetic and preparation method thereof |
| CN105055250A (en) * | 2015-08-29 | 2015-11-18 | 云南蓝钻生物科技股份有限公司 | Cosmetic containing phycocyanin and preparation method |
| CN105055250B (en) * | 2015-08-29 | 2018-06-15 | 云南蓝钻生物科技股份有限公司 | A kind of cosmetics containing phycocyanin and preparation method thereof |
| CN105055251B (en) * | 2015-08-29 | 2018-06-15 | 云南蓝钻生物科技股份有限公司 | A kind of anti-aging cosmetics and preparation method thereof |
| CN108059669A (en) * | 2017-10-26 | 2018-05-22 | 东台市赐百年生物工程有限公司 | A kind of extracting method for spraying broken wall formula phycocyanin |
| CN108059669B (en) * | 2017-10-26 | 2021-05-25 | 东台市赐百年生物工程有限公司 | Extraction method of phycocyanin capable of breaking wall by spraying |
| CN108794565A (en) * | 2018-04-27 | 2018-11-13 | 胡梦丽 | A kind of preparation method of spirulina protein powder |
| US10822373B2 (en) | 2019-01-09 | 2020-11-03 | Chenghui Zheng | Methods for synthesizing phycocyanin using a biological substance |
| CN110272486A (en) * | 2019-03-08 | 2019-09-24 | 北海生巴达生物科技有限公司 | The extracting method of phycocyanin in a kind of cyanobacteria |
| CN110272486B (en) * | 2019-03-08 | 2023-04-07 | 北海生巴达生物科技有限公司 | Method for extracting phycocyanin from blue algae |
| CN111116737A (en) * | 2020-01-13 | 2020-05-08 | 江西理工大学 | High-purity phycocyanin and preparation method thereof |
| CN111493204A (en) * | 2020-04-26 | 2020-08-07 | 武汉舜泽生物工程有限公司 | Production process and application of spirulina concentrated protein protoplasm |
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| CN103554250B (en) | 2015-06-24 |
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