CN102433015A - Method for preparing blue-green algae pigment - Google Patents
Method for preparing blue-green algae pigment Download PDFInfo
- Publication number
- CN102433015A CN102433015A CN2011103114922A CN201110311492A CN102433015A CN 102433015 A CN102433015 A CN 102433015A CN 2011103114922 A CN2011103114922 A CN 2011103114922A CN 201110311492 A CN201110311492 A CN 201110311492A CN 102433015 A CN102433015 A CN 102433015A
- Authority
- CN
- China
- Prior art keywords
- blue
- green algae
- phycochrome
- preparation
- grams
- 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 52
- 241000192700 Cyanobacteria Species 0.000 title claims abstract description 51
- 239000000049 pigment Substances 0.000 title abstract description 16
- 108090000790 Enzymes Proteins 0.000 claims abstract description 29
- 102000004190 Enzymes Human genes 0.000 claims abstract description 29
- 238000000265 homogenisation Methods 0.000 claims abstract description 25
- 238000002360 preparation method Methods 0.000 claims abstract description 24
- 108010064696 N,O-diacetylmuramidase Proteins 0.000 claims description 20
- 108010059820 Polygalacturonase Proteins 0.000 claims description 17
- 238000012545 processing Methods 0.000 claims description 17
- 239000000243 solution Substances 0.000 claims description 17
- 239000006228 supernatant Substances 0.000 claims description 13
- BIGPRXCJEDHCLP-UHFFFAOYSA-N ammonium bisulfate Chemical compound [NH4+].OS([O-])(=O)=O BIGPRXCJEDHCLP-UHFFFAOYSA-N 0.000 claims description 12
- 239000013078 crystal Substances 0.000 claims description 12
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims description 11
- 241000195493 Cryptophyta Species 0.000 claims description 10
- 238000001035 drying Methods 0.000 claims description 8
- 239000000843 powder Substances 0.000 claims description 7
- 239000007864 aqueous solution Substances 0.000 claims description 5
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 4
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 4
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 4
- 239000002994 raw material Substances 0.000 claims description 4
- 238000002791 soaking Methods 0.000 claims description 4
- 238000001291 vacuum drying Methods 0.000 claims description 4
- 239000002244 precipitate Substances 0.000 claims description 3
- 238000000926 separation method Methods 0.000 claims description 3
- 101710089297 Polygalacturonase 1 Proteins 0.000 claims description 2
- 230000008021 deposition Effects 0.000 claims description 2
- 238000009987 spinning Methods 0.000 claims description 2
- 108090000623 proteins and genes Proteins 0.000 abstract description 8
- 102000004169 proteins and genes Human genes 0.000 abstract description 8
- 210000002421 cell wall Anatomy 0.000 abstract description 5
- 210000004027 cell Anatomy 0.000 abstract description 4
- 235000016709 nutrition Nutrition 0.000 abstract description 3
- 230000035764 nutrition Effects 0.000 abstract description 3
- 238000012870 ammonium sulfate precipitation Methods 0.000 abstract description 2
- 235000013373 food additive Nutrition 0.000 abstract 1
- 239000002778 food additive Substances 0.000 abstract 1
- ANRHNWWPFJCPAZ-UHFFFAOYSA-M thionine Chemical compound [Cl-].C1=CC(N)=CC2=[S+]C3=CC(N)=CC=C3N=C21 ANRHNWWPFJCPAZ-UHFFFAOYSA-M 0.000 abstract 1
- 229940088598 enzyme Drugs 0.000 description 27
- 239000000047 product Substances 0.000 description 17
- 238000000605 extraction Methods 0.000 description 14
- 108060006184 phycobiliprotein Proteins 0.000 description 13
- 241001517315 Habenaria Species 0.000 description 12
- 240000002900 Arthrospira platensis Species 0.000 description 11
- 235000016425 Arthrospira platensis Nutrition 0.000 description 11
- 229940082787 spirulina Drugs 0.000 description 11
- 239000007788 liquid Substances 0.000 description 8
- 230000008569 process Effects 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- 239000001055 blue pigment Substances 0.000 description 6
- 235000013305 food Nutrition 0.000 description 6
- 239000000523 sample Substances 0.000 description 6
- 239000012530 fluid Substances 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 4
- 239000012043 crude product Substances 0.000 description 4
- 239000012153 distilled water Substances 0.000 description 4
- 238000007654 immersion Methods 0.000 description 4
- 239000002953 phosphate buffered saline Substances 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- 230000008961 swelling Effects 0.000 description 4
- 238000005303 weighing Methods 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 238000004040 coloring Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000010438 heat treatment Methods 0.000 description 3
- 239000002932 luster Substances 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- QCGCETFHYOEVAI-GDVGLLTNSA-N (2s)-2-[(3-amino-3-carboxypropanoyl)amino]-5-(diaminomethylideneamino)pentanoic acid Chemical compound OC(=O)C(N)CC(=O)N[C@H](C(O)=O)CCCN=C(N)N QCGCETFHYOEVAI-GDVGLLTNSA-N 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 108010031546 cyanophycin Proteins 0.000 description 2
- 229920000976 cyanophycin polymer Polymers 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- 239000000989 food dye Substances 0.000 description 2
- 238000013467 fragmentation Methods 0.000 description 2
- 238000006062 fragmentation reaction Methods 0.000 description 2
- 239000004519 grease Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 240000006439 Aspergillus oryzae Species 0.000 description 1
- 235000002247 Aspergillus oryzae Nutrition 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 108010004032 Bromelains Proteins 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108010053210 Phycocyanin Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 208000031320 Teratogenesis Diseases 0.000 description 1
- 102000001742 Tumor Suppressor Proteins Human genes 0.000 description 1
- 108010040002 Tumor Suppressor Proteins Proteins 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 231100000315 carcinogenic Toxicity 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000013016 damping Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 235000002864 food coloring agent Nutrition 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000011132 hemopoiesis Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 239000003317 industrial substance Substances 0.000 description 1
- 231100000567 intoxicating Toxicity 0.000 description 1
- 230000002673 intoxicating effect Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 238000010297 mechanical methods and process Methods 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- -1 metals ion Chemical class 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 150000002790 naphthalenes Chemical group 0.000 description 1
- 231100000957 no side effect Toxicity 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 235000012736 patent blue V Nutrition 0.000 description 1
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000012797 qualification Methods 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 235000014347 soups Nutrition 0.000 description 1
- 238000013112 stability test Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
Images
Abstract
The invention belongs to the technical field of food additives, and provides a method for preparing blue-green algae pigment. The method comprises the steps of performing enzymolysis on blue-green algae, and performing high-pressure homogenization. The method for preparing the blue-green algae pigment specifically comprises the following steps of: performing enzymolysis on blue-green algae cells by using an enzyme preparation, crushing the blue-green algae cell walls, circulating blue-green algae cytochylema which is obtained after enzymolysis treatment in a high-pressure homogenization mode, and finally centrifuging, and performing ammonium sulfate precipitation to obtain the blue-green algae pigment. By the method, edible, high-purity, and high-valeur natural cyanine which also keeps the primary nutrition of protein of blue-green algae pigment can be obtained.
Description
Technical field
The invention belongs to the technical field of foodstuff additive, relate to the preparation method of food dye, particularly the preparation method of blue phycochrome.
Background technology
Food dye comprises natural pigment and synthetic colour.Synthetic colour has characteristics such as lovely luster, stable in properties, strong coloring force; But it comes from industrial chemicals more; Chromonic material contains groups such as phenyl ring, naphthalene nucleus more, takes in a certain amount of meeting human body is produced harm, even understand intoxicating, carcinogenic and teratogenesis; So synthetic colour has strict restriction on applicable scope and consumption, some kind is disabled.Edible natural pigment is from the tissue of animal, plant or mikrobe, to extract the coloring matter that obtains, and color and luster nature, tone are accepted by people more near whole food easily, the more important thing is to have no side effect, thereby extremely human consumer's welcome.Therefore, the development of edible natural pigment more and more receives people's attention, replaces the synthetic colour trend that is inevitable with natural pigment.
Algae blue pigment is a WOT-Al Floc, and different name is a blue phycochrome, spirulina blue color, spirulina protein pigment; Its staple is WOT-Al Floc, Phycocyanins, C-and polysaccharide, and Phycocyanins, C-belongs to the protein bound pigment, with protein same nature is arranged; For blue particle or powder, water-soluble, be insoluble to pure and mild grease; Stable down at weak acid and neutrallty condition (pH=4.5 to 8), precipitate when acid (pH=4.2), highly basic can cause decolouring; Isoelectric pH=3.4 are to the poor stability of metals ion, to heat, light, acid instability.
Phycocyanins, C-is a kind of important physical active substance; Can promote immunity system to resist multiple disease; Have anti-ageingly, anti-cancer tumor suppressor promotes the utilization of animal to iron; Critical functions such as promotion hematopoiesis (see [1] Hu Hongjun. tenaculat Habenaria biology and biotechnology principle [M]. Beijing: Science Press, 2003:81-97; [2] Li Dingmei. the optimal food of tenaculat Habenaria-global human [M]. Beijing: Police Officer Education Press, 1997; [3] Zhang Chengwu, Liu Yufeng, Wang Xixia. Spirulina phycocyanin is to human blood JEG-3 HL-60, the growth effect of K562 and U937 [J]. ocean science, 2000,24 (1): 45-48).Phycobiliprotein has beautiful sky blue, be a kind of good natural pigment (see [4] He Jia, Zhao Qimei, Tian Juan is etc. tenaculat Habenaria WOT-Al Floc Study on Stability [J]. biology magazine, 1998,15 (5): 10; [5] Wang Shuling, Liu Huanyun. the preliminary study of tenaculat Habenaria pigment stability [J]. foodstuffs industry, 2000 (5): 35-37; [6] Zhang Kun, dialect is male, Liang Zhibiao. spirulina blue color research [J]. and food and machinery, 1995 (4): 21-23; [7] Zhang Shaobin, the swallow peace, Liu Hui waits the influence [J] of .pH value to tenaculat Habenaria mutant strain (SP-Dz) growth and phycobiliprotein content, Anhui agricultural sciences, 2005,33 (8): 1562-1563).Form and can know from its amino acid, it can be used as the humans and animals indispensable amino acid the key protein source (see [8] soup morning sunlight, Jiang Jialun. the extraction of spirulina plalensis phycobiliprotein and characteristic preliminary study [J] thereof. ocean, the East Sea, 1993,11 (4): 49-53; [9] Shen Beiying, He Yuheng. phycobiliprotein extraction, separation and the purifying [J] of tenaculat Habenaria. grain and grease, 1999 (4): 38-40).Phycobiliprotein emission intensive fluorescence; The resorcinolphthalein that its fluorescence intensity ratio is commonly used is strong 30 times, (sees [10] Wu Ping, Gu Ming because of its outstanding fluorescent characteristic makes the exploitation of phycobiliprotein fluorescent probe especially noticeable; Qi Yihua; Deng. phycobiliprotein fluorescent probe and mark thereof [J]. life science, 2001,5 (2): 109-113.).Simultaneously, Phycocyanins, C-be used for the food research aspect especially the research of functional foodstuff obtained bigger progress, existing more than 10 kind of spiral algae sheet of China and capsule have passed through Ministry of Health's approval according to different functions and have been healthcare products.
Because of phycobiliprotein with very big using value arranged; Many people attempt phycobiliprotein gene is changed in the mikrobe to realize the scale operation of phycobiliprotein; But because a variety of causes; Produce phycobiliprotein with transgenic method and also have very long stretch to walk, separate phycobiliprotein from extracting algae at present and be still the main path that obtains phycobiliprotein.
The extraction of Phycocyanins, C-often is divided into albumen stripping and two processes of albumen precipitation.Phycocyanins, C-is to belong to intracellular protein, make its stripping at first want the cell walls of smudge cells, cytolemma that it is dissolved in the extracting solution with the dissolved state, adopts suitable method that it is precipitated again, should keep proteic activity at leaching process.
Before the present invention, the extraction of blue phycochrome also is in the way of single employing Mechanical Crushing cell walls, disclosed like the CN1210122 patented claim; Be the phosphoric acid buffer that adopts 5-20 times of volume, multigelation, centrifugal; Collect supernatant, lyophilize gets the algae blue pigment dry product.But because the mode of Mechanical Crushing is simple, efficient is low, therefore have the shortcoming that operational path is long, extraction is thorough, yield is low excessively, final pigment product purity is low, and the look valency is also lower.
CN1035425A also discloses a kind of process for extracting of blue phycochrome; This method be earlier with blue-green algae levigate after; Use flooding; Regulating the scope of pH 4 to 9, carry out enzymolysis processing, employed enzyme is papain, trypsinase, aspergillus oryzae, Traumanase or proteolytic enzyme that from Bacillus subtilus, extracts and glycase.But, still can't reach activity to blue phycochrome and proteic effective extraction and maintenance cyanophycin owing to receive the restriction of employed enzyme.
Also have chemical reagent facture, swelling method, supersonic method and tissue to smash process for extracting such as method to pieces in addition, these methods are all more loaded down with trivial details, and last handling process is also cumbersome, has not only increased production cost, has also caused environmental pollution simultaneously.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of blue phycochrome, this method comprises enzymolysis step of blue-green algae and high pressure homogenization step.
The zymin that enzymolysis step among the above-mentioned preparation method is used is the arbitrary combination of N,O-Diacetylmuramidase, polygalacturonase or these two kinds of enzymes.
The preparation method of blue phycochrome of the present invention comprises the steps:
A) use the EDTA aqueous solution soaking blue-green algae of concentration as 0.01M-0.03M;
B) with the blue-green algae solution of zymin enzymolysis processing step a);
The blue-green algae solution that c) will pass through the step b) enzymolysis processing carries out the high pressure homogenization processing;
D) will pass through the step c) high pressure homogenization and handle the blue-green algae solution centrifugal separation that obtains, get supernatant; In this supernatant, add ammonium sulfate, the deposition blue phycochrome carries out spinning again, gets the throw out blue phycochrome.
The step c) of aforesaid method is to carry out in the pressure range of 20kPa to 40kPa, and circulation is carried out 2 times to 3 times.High-pressure homogeneous speed is 100L/h to 400L/h (rise/hour), and temperature is 10 ℃ to 25 ℃.
Aforesaid method also comprises the step of the blue phycochrome drying precipitate powdered that step d) is obtained.Said drying is in vacuum-drying below 60 ℃.
In the step a) of aforesaid method, the amount ratio of said dried blue algae powder and EDTA solution is 100 grams to 300 grams: 1000 milliliters (ml); Soaking temperature is 10 ℃ to 25 ℃, and soak time 1 hour (h) was to 10 hours.
In the step b) of aforesaid method, said enzymolysis processing temperature is 30 ℃ to 50 ℃, and the constant temperature treatment time is 2-6 hour, and with respect to 1 kilogram of blue-green algae, the add-on of N,O-Diacetylmuramidase is that 1 gram is to 2.5 grams; The add-on of polygalacturonase is that 2 grams are to 6 grams; Perhaps, N,O-Diacetylmuramidase and polygalacturonase combination are used, and with respect to 1 kilogram of blue-green algae, N,O-Diacetylmuramidase 0.5 gram is to 1.25 grams, and polygalacturonase 1 gram is to 3 grams.
The ammonium sulfate that adds in the step d) of aforesaid method is ammonia sulfate crystal, and the weight ratio of said dried blue-green algae raw material and ammonia sulfate crystal is 1: 0.05-0.5 is preferably 1: 0.1.
The preparation method of blue phycochrome of the present invention comprises and uses zymin that the blue-green algae cell is carried out enzymolysis; Broken blue-green algae cell walls; Blue-green algae enchylema after the mode that adopts high pressure homogenization is again handled enzyme carries out circular treatment, and is centrifugal at last, use ammonium sulfate precipitation, obtains blue phycochrome.
Can obtain edible, high purity, high luminance relay valency and keep the natural blue pigment of the intrinsic nutrition of blue phycochrome albumen through method of the present invention.
Description of drawings
Fig. 1 is a blue phycochrome preparing method's of the present invention process flow sheet.
Embodiment
Method of the present invention can realize through following steps:
A) with concentration be the EDTA distilled water solution immersion blue-green algae of 0.01M to 0.03M; According to dried blue-green algae: EDTA distilled water solution=100 grams to 300 grams (g): the ratio stirring and evenly mixing of 1000 milliliters (ml); Soak in 10 ℃ to 25 ℃ temperature; Soak time 1 hour to 10 hours, preferred 2 hours to 4 hours.This treatment step makes the abundant swelling of blue-green algae, and can guarantee that with this understanding cyanophycin does not produce degraded, for the enzymolysis processing step has been created condition.
B) in the solution of above-mentioned immersion blue-green algae, add zymin, carry out the constant temperature enzymolysis processing, wherein said zymin is the arbitrary combination of N,O-Diacetylmuramidase, polygalacturonase or these two kinds of enzymes; Said constant temperature scope is 30 ℃ to 50 ℃, and the constant temperature enzymolysis processing time is 2-6 hour, to guarantee that the cell walls of blue-green algae cell is fully dissolved fragmentation.Wherein, the N,O-Diacetylmuramidase add-on is 1g to a 2.5g enzyme: 1 kilogram of (kg) blue-green algae; The polygalacturonase add-on is 2g to a 6g enzyme: the 1kg blue-green algae; Combination is used like two enzymes, and then ratio should be N,O-Diacetylmuramidase 0.5g to 1.25g: polygalacturonase 1g to 3g: blue-green algae 1kg.
C) will in high pressure homogenizer, carry out homogenization treatment in the pressure range of 20kPa to 40kPa through the blue-green algae liquid after the enzymolysis processing; Control homogeneous speed is that 100 liters/hour (L/h) are to 400L/h; Temperature remains in 10 ℃ to 25 ℃, with circulate homogenization treatment 2-3 time of homogenizing fluid.
D) the blue-green algae liquid after high-pressure homogeneous is carried out centrifugal treating, remove throw out, throw out is recyclable as feed; In supernatant, add ammonia sulfate crystal; Add-on is that the weight ratio of said dried blue-green algae raw material and ammonia sulfate crystal is 1: 0.05-0.5, be preferably 1: 0.1, and make it to dissolve, leave standstill, be settled out thick WOT-Al Floc; The centrifugal throw out that gets is desired blue phycochrome product.
The throw out that obtains through aforesaid method can directly use, and also can carry out drying or purifying as required.Said drying can be selected any feasible drying means for use, but drying temperature should not be higher than 60 ℃, is destroyed to avoid pigment composition, so preferably adopt vacuum-drying.
Used blue-green algae wide material sources among the preparation method of blue phycochrome of the present invention; Multiple blue-green algae such as can adopt tenaculat Habenaria, basketball algae, deliver vegetables; All the other protein can not be destroyed in the blue algae slurry behind the extraction pigment, can not influence protein content as feedstuff protein equally; Make the best use of everything, make the sub product increment.
The route of the traditional extraction technique of blue phycochrome is oversize, and most of process need be carried out at low temperatures, and extracting cycle is long, thereby cost is high, efficient is low, production application is worth not high.The preparation method of blue phycochrome of the present invention is to be main raw material with the blue-green algae; In conjunction with utilizing biotechnological means that blue-green algae is carried out deep processing; And then obtain edible highly purified natural blue pigment; Also kept the intrinsic nutrition of blue-green algae simultaneously, blue phycochrome crude product yield＞40% that preparation method of the present invention obtains is far above about 20% of general low temperature freeze thawing mechanical methods.
According to national standard, the ratio of A620/A280 can be used as food grade greater than 0.7, can be used as pharmaceutical grade greater than 2.5.The thick product purity that the inventive method obtains is better, and A620/A280 can reach 1.0-1.5, can directly use as food color.In addition, product also has characteristics such as color and luster nature, stable in properties, strong coloring force, and experiment shows that the blue phycochrome under the dry powder at room temperature preserved 1 year, and its 620nm characteristic light absorption value does not change basically.
Below through experimental example explain enzyme of the present invention dissolve method, high pressure homogenization method, enzyme dissolve-condition of homogeneous method orthogonal test selects.
Experimental example 1, high pressure homogenization method
(1) take by weighing 9 parts of (every part of 100g) tenaculat Habenarias respectively and contain the zero(ppm) water of 0.02M EDTA in 1000ml, 25 ℃ are stirred and soaked 4 hours;
(2) with spirulina liquor different pressure (20,30,40kPa) with under the condition of different cycle index (1,2,3 time) carry out homogeneous, control homogeneous speed is 200L/h, temperature remains in 25 ℃;
(3) with homogenizing fluid centrifugal 20 minutes (min) under the 12000r/min condition, remove sediment, get supernatant.
(4) in supernatant, add the 10g ammonia sulfate crystal, make it dissolving, leave standstill, be settled out thick WOT-Al Floc.
(5) centrifugal 15min under the 10000r/min condition gets throw out.
(6) result and analysis
The cycle index of high pressure homogenization method is bigger than the influence of homogenization pressure to the influence of title product (WOT-Al Floc) extraction yield.Can draw, along with the increase of pressure and increasing of cycle index, the tenaculat Habenaria fragmentation is good more, and the yield of title product (WOT-Al Floc) is also high more.
Table 1, high-pressure homogeneous experimental result
Experimental example 2, enzyme dissolve method
(1) take by weighing 9 parts of (every part of 100g) tenaculat Habenarias respectively, be dissolved in the zero(ppm) water that 1000ml contains 0.02M EDTA, 25 ℃ are stirred and soaked 4 hours;
(2) in 9 parts of samples, add respectively different total amounts (100,200,300mg) N,O-Diacetylmuramidase, polygalacturonase (N,O-Diacetylmuramidase: polygalacturonase=1: 2) and different temperature (30,40,50 ℃) down constant temperature different time (2,4,6h) handle:
(3), remove throw out with the centrifugal 15min under the condition of 10000r/min of the spirulina liquor behind the enzymolysis;
(4) in supernatant, add the 10g ammonia sulfate crystal respectively, make it dissolving, leave standstill, be settled out thick WOT-Al Floc;
(5) centrifugal 15min under the condition of 10000r/min gets throw out.
Visible by following table 2 range analysiss, the enzyme treatment temp has the greatest impact to title product (WOT-Al Floc), and the influence of molten time of enzyme dosage and enzyme is littler comparatively speaking.Polygalacturonase=1: 2)/to dissolve the time be 4h for 10g (tenaculat Habenaria), enzyme, when temperature is 40 ℃ can find out that from analytical results enzyme dosage is that (N,O-Diacetylmuramidase:, title product (WOT-Al Floc) extraction yield is the highest for 20mg.
Table 2, enzyme dissolve experimental result
Experimental example 3, enzyme dissolve the combination of method and high pressure homogenization method
(1) take by weighing 9 parts of (every part of 100g) tenaculat Habenarias respectively, be dissolved in the zero(ppm) water that 1000ml contains 0.02M EDTA, 25 ℃ of stirrings were soaked 4 hours;
(2) in 9 parts of samples, add respectively different total amounts (100,200,300mg) N,O-Diacetylmuramidase, polygalacturonase (N,O-Diacetylmuramidase: polygalacturonase=1: 2) and different temperature (30,40,50 ℃) down constant temperature different time (2,4,6h) handle.
(3) with spirulina liquor different pressure (20,30,40kPa) with under the condition of different cycle index (1,2,3 time) carry out homogeneous, control homogeneous speed is 200L/h, temperature remains in 25 ℃;
(4), remove throw out with the centrifugal 15min under the condition of 10000r/min of the spirulina liquor behind the enzymolysis;
(5) in supernatant, add the 10g ammonia sulfate crystal respectively, make it dissolving, leave standstill, be settled out thick WOT-Al Floc;
(6) centrifugal 15min under the condition of 10000r/min gets throw out.
With reference to the analytical results of above-mentioned treatment process, can know that homogenization pressure in the high pressure homogenization method, cycle index are bigger to the influence of title product (WOT-Al Floc) extraction yield, and in three factors that enzyme is handled, temperature has the greatest impact to the extraction yield of title product (WOT-Al Floc).Fixingly do not change (be definite value, being respectively 30mg/10g, enzyme, to dissolve the time be 4h) so dissolve-adopt in the high pressure homogenization method ratio of components and time at enzyme, the influence of temperature, homogenization pressure, cycle index is studied.
Table 3 enzyme dissolves high-pressure homogeneous Orthogonal experiment results
Visible by above range analysis, high-pressure homogeneous round-robin number of times is the principal element that influences the algae blue pigment extraction yield, the taking second place of enzyme treatment temp.Conform to the test-results of method 1 with method 2, but enzyme dissolve-high pressure homogenization method in the WOT-Al Floc extraction yield than molten method of single enzyme or high-pressure homogeneous combined techniques height.
Therefore, enzyme dissolve-optimal conditions that the homogeneous method is extracted blue phycochrome is:
(1) take by weighing the 100g tenaculat Habenaria, be dissolved in the zero(ppm) water that 1000ml contains 0.02M EDTA, 25 ℃ are stirred and soaked 4 hours;
(2) in sample, add the about 200mg N,O-Diacetylmuramidase of total amount, polygalacturonase mixed enzyme, wherein N,O-Diacetylmuramidase respectively: polygalacturonase=1: 2, and in the processing in 4 hours of 40 ℃ of following constant temperature of different temperature.
(3) spirulina liquor is carried out homogeneous under the condition of different pressure 40kPa and cycle index 3 times, control homogeneous speed is 200L/h, and temperature remains in 25 ℃;
(4), remove throw out with the centrifugal 15min under the condition of 10000r/min of the spirulina liquor behind the enzymolysis;
(5) in supernatant, add the 10g ammonia sulfate crystal respectively, make it dissolving, leave standstill, be settled out thick WOT-Al Floc;
(6) centrifugal 15min under the condition of 10000r/min gets throw out.
Below in conjunction with accompanying drawing, through embodiment further explain the present invention, but these embodiment do not constitute the qualification to the scope of the invention.
The preparation of embodiment 1 solid-state blue phycochrome
In 20 ℃, the 1000g blue-green algae is soaked in 10 liters of (L) distilled water solutions of 0.02M EDTA, and stirring and evenly mixing, immersion makes the abundant swelling of blue-green algae, under 40 ℃ of conditions, adds N,O-Diacetylmuramidase 1g, (keeps 40 ℃ of temperature, reacts 4 hours.To under 40kPa pressure, pass through the high pressure homogenizer homogenization treatment through the blue-green algae liquid of enzymolysis processing, homogenizing fluid circulation homogeneous 2 times, homogenization treatment speed 200L/h, temperature are 25 ℃.With 1000r/min rotating speed centrifugal treating liquid 30 minutes (min), filter and discard throw out, in supernatant, add ammonia sulfate crystal 100 grams, make it to dissolve, leave standstill, be settled out thick WOT-Al Floc.To get throw out after the 1000r/min rotating speed centrifugal treating.Under vacuum tightness 0.09MPa, bed thickness 5mm drains throw out in the Vacuumdrier, guarantees that simultaneously the temperature of heating plate scope is 45 ℃-55 ℃, gets blue phycochrome powder crude product 447.2g, and thick product yield is 44.72%, and measuring A620/A280 is 1.1.
The preparation of embodiment 2 solid-state blue phycochromes
In 20 ℃, the 1000g blue-green algae is soaked in the 10L distilled water solution of 0.02MEDTA, and stirring and evenly mixing, immersion makes the abundant swelling of blue-green algae, under 40 ℃ of conditions, adds N,O-Diacetylmuramidase 0.5g, and polygalacturonase 1.0g keeps 40 ℃ of temperature, reacts 4 hours.To under 40kPa pressure, pass through the high pressure homogenizer homogenization treatment through the blue-green algae liquid of enzymolysis processing, homogenizing fluid circulation homogeneous 3 times, homogenization treatment speed 200L/h, temperature are 25 ℃.With 1000r/min rotating speed centrifugal treating liquid 30 minutes (min), filter and discard throw out, in supernatant, add ammonia sulfate crystal 100 grams, make it to dissolve, leave standstill, be settled out thick WOT-Al Floc.To get throw out after the 1000r/min rotating speed centrifugal treating.Under vacuum tightness 0.09MPa, bed thickness 7mm drains throw out in the Vacuumdrier, guarantees that simultaneously the temperature of heating plate scope is 45 ℃-55 ℃, gets blue phycochrome powder crude product 682.3g, and thick product yield is 68.2%, and measuring A620/A280 is 1.3.
The preparation of embodiment 3 refining blue phycochromes
With the blue phycochrome powdered product 400g of gained among the embodiment 1 with 5000ml; 0.01mol/L, leave standstill after the dissolving of the phosphate buffered saline buffer of pH=7.0, with the PBS of crude product solution through 10mmol/L and counter-balanced DEAE-Sophadex A-25 and Win 40350 absorption upper prop gradient elution (continuous increase PBS ionic strength in wash-out (and respectively with 10,20,40,100, the PBS damping fluid of 200mmol/L, pH=7.0; And the NaCL eluant solution of 0.3mol/L in addition); Charateristic avsorption band 620nm Phycocyanins, C-part elutriant is merged, concentrates, in vacuum drying oven under vacuum tightness 0.09MPa; Bed thickness 5-7mm drains throw out; Guarantee that simultaneously the temperature of heating plate scope is 45 ℃-55 ℃, get blue phycochrome powder refined products 237.2g, yield is 59%; Measuring A620/A280 is 2.986, obtains SILVER REAGENT standard algae blue pigment.
The stability test of blue phycochrome bullion
One, method
Get the blue phycochrome bullion 500g that the foregoing description 2 obtains; In 25 ℃ ± 2 ℃, the condition that relative humidity 60% ± 5%, lucifuge place preserve is carried out stability experiment, with the sampling of investigating in 1,3,6,9,12 month; The row ultra-violet analysis; Investigate sample A620/A280 changing conditions, cofabrication solution is investigated the liquid colour-changeable situation.
Two, result
1, sample A620/A280 changing conditions (seeing following table for details)
Sampling in 1-12 month, the A620/A280 value meets the food grade colorant requirement all greater than 1.0, and purity is higher.
Table 1, sample A620/A280 changing conditions
| 1 | 3 | 6 | 9 | 12 | |
| A620/A280 | 1.3 | 1.2 | 1.2 | 1.1 | 1.0 |
2, sample aqueous solution colour-change situation (seeing following table for details)
1,3, June the blue phycochrome aqueous solution be blue-greenish colour, transparent liquid, pH is stable at more than 6.0; Color is not seen considerable change in the time of 9 months, and pH descends to some extent; Produce a little muddy thing during December, pH drops to about 5.0.
Table 2, sample aqueous solution colour-change situation
Claims (10)
1. the preparation method of a blue phycochrome, this method comprises enzymolysis step of blue-green algae and high pressure homogenization step.
2. the preparation method of blue phycochrome as claimed in claim 1, the zymin that wherein said enzymolysis step is used is the arbitrary combination of N,O-Diacetylmuramidase, polygalacturonase or these two kinds of enzymes.
3. the preparation method of blue phycochrome as claimed in claim 2, this method comprises the steps:
A) use the EDTA aqueous solution soaking blue-green algae of concentration as 0.01M-0.03M;
B) with the blue-green algae solution of zymin enzymolysis processing step a);
The blue-green algae solution that c) will pass through the step b) enzymolysis processing carries out the high pressure homogenization processing;
D) will pass through the step c) high pressure homogenization and handle the blue-green algae solution centrifugal separation that obtains, get supernatant; In this supernatant, add ammonium sulfate, the deposition blue phycochrome carries out spinning again, gets the throw out blue phycochrome.
4. the preparation method of blue phycochrome as claimed in claim 3, wherein said step c) is to carry out in the pressure range of 20kPa to 40kPa, and circulation is carried out 2 times to 3 times.
5. the preparation method of blue phycochrome as claimed in claim 3, this method also comprise the step of the blue phycochrome drying precipitate powdered that step d) is obtained.
6. the preparation method of blue phycochrome as claimed in claim 5, wherein said drying is in vacuum-drying below 60 ℃.
7. the preparation method of blue phycochrome as claimed in claim 3, wherein in said step a), the amount ratio of said dried blue algae powder and EDTA solution is 100 grams to 300 grams: 1000 milliliters; Soaking temperature is 10 ℃ to 25 ℃, soak time 1 hour to 10 hours.
8. the preparation method of blue phycochrome as claimed in claim 3, wherein in said step b), said enzymolysis processing temperature is 30 ℃ to 50 ℃, and the constant temperature treatment time is 2-6 hour, and with respect to 1 kilogram of blue-green algae, the add-on of N,O-Diacetylmuramidase is that 1 gram is to 2.5 grams; The add-on of polygalacturonase is that 2 grams are to 6 grams; Perhaps, N,O-Diacetylmuramidase and polygalacturonase combination are used, and with respect to 1 kilogram to 2 kilograms blue-green algae, N,O-Diacetylmuramidase 0.5 gram is to 1.25 grams, and polygalacturonase 1 gram is to 3 grams.
9. the preparation method of blue phycochrome as claimed in claim 4, wherein in said step c), homogeneous speed is 100L/h to 400L/h, temperature is 10 ℃ to 25 ℃,
10. the preparation method of blue phycochrome as claimed in claim 3, the ammonium sulfate that wherein in said step d), adds is ammonia sulfate crystal, the weight ratio of said dried blue-green algae raw material and ammonia sulfate crystal is 1: 0.05-0.5 is preferably 1: 0.1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110311492.2A CN102433015B (en) | 2011-10-14 | 2011-10-14 | Method for preparing blue-green algae pigment |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110311492.2A CN102433015B (en) | 2011-10-14 | 2011-10-14 | Method for preparing blue-green algae pigment |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102433015A true CN102433015A (en) | 2012-05-02 |
| CN102433015B CN102433015B (en) | 2014-11-12 |
Family
ID=45981369
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201110311492.2A Active CN102433015B (en) | 2011-10-14 | 2011-10-14 | Method for preparing blue-green algae pigment |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102433015B (en) |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015007997A1 (en) | 2013-07-19 | 2015-01-22 | Roquette Freres | Optimised method for breaking chlorella cell walls by means of very high pressure homogenisation |
| CN109219640A (en) * | 2016-06-28 | 2019-01-15 | Dic株式会社 | The manufacturing method of dye material and dye material |
| CN109627201A (en) * | 2019-01-14 | 2019-04-16 | 中国科学院烟台海岸带研究所 | A method of extracting phycocyanobilin |
| CN110205135A (en) * | 2019-07-08 | 2019-09-06 | 山东多芬农业有限公司 | A method of soil and fixed nitrogen are administered using cyanobacteria |
| US10465159B2 (en) | 2013-07-04 | 2019-11-05 | Corbion Biotech, Inc. | Optimised method for breaking chlorella walls by mechanical crushing |
| WO2020144330A1 (en) | 2019-01-11 | 2020-07-16 | Fermentalg | Process for purifying phycocyanins |
| CN113583111A (en) * | 2021-08-04 | 2021-11-02 | 中国农业科学院农业质量标准与检测技术研究所 | High-pressure carbon dioxide non-thermal extraction method of high-quality phycocyanin pigment |
| CN113698473A (en) * | 2021-08-13 | 2021-11-26 | 河南工业大学 | Phycocyanin stability and acidolysis modification research method thereof |
| CN113956926A (en) * | 2021-11-17 | 2022-01-21 | 陈勇 | Natural plant detergent and preparation method thereof |
| FR3130842A1 (en) | 2021-12-22 | 2023-06-23 | CarbonWorks | METHOD FOR CAPTURING PHYTOTOXINS IN A BIOLOGICAL REACTOR |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6427446A (en) * | 1987-07-23 | 1989-01-30 | Toa Nenryo Kogyo Kk | Method for extracting blue pigment from blue-green algae |
| CN1035425A (en) * | 1988-12-29 | 1989-09-13 | 江西师范大学 | The manufacture method of blue green alga-spiral alga drink |
| JPH07118555A (en) * | 1993-10-26 | 1995-05-09 | Dainippon Ink & Chem Inc | Production of natural red pigment |
| JPH1027446A (en) * | 1996-07-08 | 1998-01-27 | Nec Corp | Carriage mechanism and magnetic disk device provided therewith |
| CN1210122A (en) * | 1997-09-04 | 1999-03-10 | 中国科学院武汉植物研究所 | Extraction of blue pigment and polysaccharide from spirulina and use of residues |
| JP2003342489A (en) * | 2002-05-28 | 2003-12-03 | Dainippon Ink & Chem Inc | Method of extracting phycocyanin from blue-green algae |
| CN101899102A (en) * | 2010-07-12 | 2010-12-01 | 华南理工大学 | Method for separating high purity phycocyanin from spirulina |
-
2011
- 2011-10-14 CN CN201110311492.2A patent/CN102433015B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6427446A (en) * | 1987-07-23 | 1989-01-30 | Toa Nenryo Kogyo Kk | Method for extracting blue pigment from blue-green algae |
| CN1035425A (en) * | 1988-12-29 | 1989-09-13 | 江西师范大学 | The manufacture method of blue green alga-spiral alga drink |
| JPH07118555A (en) * | 1993-10-26 | 1995-05-09 | Dainippon Ink & Chem Inc | Production of natural red pigment |
| JPH1027446A (en) * | 1996-07-08 | 1998-01-27 | Nec Corp | Carriage mechanism and magnetic disk device provided therewith |
| CN1210122A (en) * | 1997-09-04 | 1999-03-10 | 中国科学院武汉植物研究所 | Extraction of blue pigment and polysaccharide from spirulina and use of residues |
| JP2003342489A (en) * | 2002-05-28 | 2003-12-03 | Dainippon Ink & Chem Inc | Method of extracting phycocyanin from blue-green algae |
| CN101899102A (en) * | 2010-07-12 | 2010-12-01 | 华南理工大学 | Method for separating high purity phycocyanin from spirulina |
Non-Patent Citations (1)
| Title |
|---|
| 叶海辉等: "螺旋藻的破碎及藻蓝色素的提取", 《华南热带农业大学学报》, vol. 7, no. 3, 30 September 2001 (2001-09-30) * |
Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US10465159B2 (en) | 2013-07-04 | 2019-11-05 | Corbion Biotech, Inc. | Optimised method for breaking chlorella walls by mechanical crushing |
| WO2015007997A1 (en) | 2013-07-19 | 2015-01-22 | Roquette Freres | Optimised method for breaking chlorella cell walls by means of very high pressure homogenisation |
| CN109219640A (en) * | 2016-06-28 | 2019-01-15 | Dic株式会社 | The manufacturing method of dye material and dye material |
| CN109219640B (en) * | 2016-06-28 | 2021-02-19 | Dic株式会社 | Pigment material and method for producing pigment material |
| CN113518558A (en) * | 2019-01-11 | 2021-10-19 | 费尔曼塔格公司 | Method for purifying phycocyanin |
| WO2020144330A1 (en) | 2019-01-11 | 2020-07-16 | Fermentalg | Process for purifying phycocyanins |
| FR3091640A1 (en) * | 2019-01-11 | 2020-07-17 | Fermentalg | PROCESS FOR THE PURIFICATION OF PHYCOCYANINS |
| US20220112235A1 (en) * | 2019-01-11 | 2022-04-14 | Fermentalg | Process for purifying phycocyanins |
| CN109627201A (en) * | 2019-01-14 | 2019-04-16 | 中国科学院烟台海岸带研究所 | A method of extracting phycocyanobilin |
| CN110205135A (en) * | 2019-07-08 | 2019-09-06 | 山东多芬农业有限公司 | A method of soil and fixed nitrogen are administered using cyanobacteria |
| CN113583111A (en) * | 2021-08-04 | 2021-11-02 | 中国农业科学院农业质量标准与检测技术研究所 | High-pressure carbon dioxide non-thermal extraction method of high-quality phycocyanin pigment |
| CN113583111B (en) * | 2021-08-04 | 2022-05-31 | 中国农业科学院农业质量标准与检测技术研究所 | High-pressure carbon dioxide non-thermal extraction method of high-quality phycocyanin pigment |
| CN113698473A (en) * | 2021-08-13 | 2021-11-26 | 河南工业大学 | Phycocyanin stability and acidolysis modification research method thereof |
| CN113956926A (en) * | 2021-11-17 | 2022-01-21 | 陈勇 | Natural plant detergent and preparation method thereof |
| FR3130842A1 (en) | 2021-12-22 | 2023-06-23 | CarbonWorks | METHOD FOR CAPTURING PHYTOTOXINS IN A BIOLOGICAL REACTOR |
| WO2023118401A1 (en) | 2021-12-22 | 2023-06-29 | CarbonWorks | Method for capturing phytotoxins in a biological reactor |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102433015B (en) | 2014-11-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102433015B (en) | Method for preparing blue-green algae pigment | |
| MišurCoVá et al. | Nitrogen content, dietary fiber, and digestibility in algal food products | |
| CN105586377B (en) | A kind of enzymolysis processing method of seaweed | |
| CN101401821B (en) | Bee pollen peptide slice and preparation method thereof | |
| Affan et al. | Variation of Spirulina maxima biomass production in different depths of urea-used culture medium | |
| CN107751697A (en) | Chlorella extract phycocyanin drink with function preparation method | |
| CN106047749A (en) | Zaralenone degrading bacteria and application thereof | |
| CN104382951A (en) | Extracting method for algal polyphenols | |
| Gómez-Jacinto et al. | Iodine speciation in iodine-enriched microalgae Chlorella vulgaris | |
| CN108522900A (en) | A kind of aquatic products anti-stress feed addictive | |
| CN104643178A (en) | Technology for increasing content of protein of ginkgo nuts by performing solid fermentation on ginkgo nuts | |
| Chen et al. | Could Chlorella pyrenoidosa be exploited as an alternative nutrition source in aquaculture feed? A study on the nutritional values and anti-nutritional factors | |
| CN102524529B (en) | Clean production method for functional compound feed and product by clean production method | |
| CN116987178A (en) | Preparation method of fish bone collagen polypeptide chelated calcium powder | |
| CN104164467B (en) | Method for extracting antibacterial peptide from chicken blood | |
| CN102690322A (en) | Method for decolorizing silkworm chrysalis protein enzymatic hydrolysis peptide | |
| CN109354601A (en) | The extracting method of selenoprotein in a kind of pear fruit | |
| Cho et al. | Single-and repeated-dose toxicities of aloe fermentation products in rats | |
| CN109007232A (en) | Extraction and purification process based on the boron-rich high white nutrient of boron high-selenium eggs for planting plant of selenium-rich | |
| CN101856089B (en) | Lespedeza bicolor Turcz. bee pollen polysaccharide oral liquid and preparation method thereof | |
| CN101664107A (en) | High-efficiency shrimp immunomodifier | |
| CN107125679B (en) | Method for removing nitroimidazole and quinolone drug residues in honey | |
| CN108047307B (en) | Edible and medicinal fungus source selenium chelating peptide and preparation method thereof | |
| CN109160947A (en) | A kind of preparation method and applications of spirulina phycocyanin | |
| CN109897873B (en) | Enrichment method based on mixotrophic haematococcus pluvialis gamma-aminobutyric acid |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20170322 Address after: 231100 Hefei province high tech Zone Industrial Park, mechanical and Electrical Industrial Park, the 5 floor of the 13 Patentee after: Anhui Fengrun biotechnology Limited by Share Ltd Address before: 230088 Hefei science and technology zone, Anhui high tech Zone, No. four, layer 122 Patentee before: Chen Yong |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20171215 Address after: 231100 Hefei province high tech Zone Industrial Park, mechanical and Electrical Industrial Park, the 5 floor of the 13 Co-patentee after: Advanced technology research institute of China Science & Technology University Patentee after: Chen Yong Address before: 231100 Hefei province high tech Zone Industrial Park, mechanical and Electrical Industrial Park, the 5 floor of the 13 Patentee before: Anhui Fengrun biotechnology Limited by Share Ltd |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20190107 Address after: 231100 Electrical and Mechanical Industrial Park, Hefei High-tech Zone, Anhui Province, 13 buildings and 5 floors Patentee after: Chen Yong Address before: 231100 Electrical and Mechanical Industrial Park, Hefei High-tech Zone, Anhui Province, 13 buildings and 5 floors Co-patentee before: Advanced technology research institute of China Science & Technology University Patentee before: Chen Yong |
|
| EE01 | Entry into force of recordation of patent licensing contract | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20120502 Assignee: Anhui blue peptide Biotechnology Co.,Ltd. Assignor: Chen Yong Contract record no.: X2021980007295 Denomination of invention: Preparation method of cyanobacteria pigment Granted publication date: 20141112 License type: Exclusive License Record date: 20210805 |