CN1064969C - Obtusatus arthrospira phycocyanin and its application - Google Patents

Abstract

The present invention discloses new phycocyanin extracted from blunt-top arthrospira. The formula weight of the phycocyanin is 14500 dalton, the isoelectric point of the phycocyanin is 4.8, and the maximum spectral absorption value of the phycocyanin is 620mm. The phycocyanin is an oligomer composed of an alpha subunit and a beta subunit. The phycocyanin has a good health-care effect. The present invention also discloses a series of health-food compositions containing the phycocyanin and an application of the phycocyanin in the field of fluorescence detection.

Description

Obtusatus arthrospira phycocyanin and application thereof

The present invention relates to a kind of Phycocyanins, C-of novelty, more specifically, relate to the Phycocyanins, C-of obtusatus arthrospira, contain the composition of this Phycocyanins, C-, and uses thereof.

The formal name used at school of obtusatus arthrospira is Arthrospira platensis, is commonly called as spirulina, is a kind of valuable plant protein resource.It contains nutritive substances such as rich in protein, β-Hu Luobusu.

It is raw material with the blue green alga-spiral alga that Chinese patent application 89100036.4 discloses a kind of, through levigate, and leaching, hydrolysis makes the protein amino acid solution, prepares the method for blue transparent beverage again with this solution.

Chinese patent application No.94112205.0 is entitled as " artrospira spirulina health food composition and method for making thereof ", discloses a kind of food compositions that contains the artrospira spirulina enzymatic hydrolyzed extract, and method for making.

1986, the Schwartz and the Shklar of U.S. Harvard Medical School found after deliberation, and the algae egg white mixture that Oscillariaceae extracts from the algae of a kind of unknown kind of Spirullina has restraining effect to some cancer cells, and function such as antifatigue is arranged.Also the someone is applied to anti-ageing and treatment AIDS disease aspects such as (AIDS).

In a word, do not see the relevant report that from the algae of Spirullina, extracts Phycocyanins, C-in the prior art, especially do not extract the report of Phycocyanins, C-from obtusatus arthrospira.

An object of the present invention is to provide a kind of protein of novelty, the Phycocyanins, C-that this protein gets for extraction separation from obtusatus arthrospira.

Another object of the present invention provides a kind of method for making of Phycocyanins, C-.

The 3rd purpose of the present invention provides a series of food compositions that contain Phycocyanins, C-.

The 4th purpose of the present invention provides the purposes of this cyanophycin aspect fluoroscopic examination.

To achieve these goals, the inventor studied through the several years, extracted a kind of Phycocyanins, C-of novelty from obtusatus arthrospira, it is characterized in that, the molecular weight of this Phycocyanins, C-is about 14500 dalton, iso-electric point is 4.8, its maximum spectral absorption value is 620mm, in addition, and the oligomer that this Phycocyanins, C-is made up of α and β subunit, wherein α and β subunit are the polypeptide chains that is made of 160-180 amino acid, and two subunits have homology.

The present invention also provides and has contained a series of food compositions of an amount of Phycocyanins, C-as main component.

The present invention also provides the application of Phycocyanins, C-aspect fluoroscopic examination.

Extraction of the present invention from the Phycocyanins, C-of artrospira spirulina be some alga cells important catch photopigment albumen, closely related because of its splendid health-care effect again with the human lives.Phycocyanins, C-can be joined in the foodstuffs compositions as effective constituent, make different protective foodss.Food compositions not only can be made various people form common and commonly used like this, and can play splendid health-care effect.

Artrospira spirulina originates in Africa, environment such as happiness high temperature, long day and high pH value.China did not have this kind of plant originally.1978, the inventor at first tamed and dociled from U.S.'s introducing algae kind and educates.By discovering of aspects such as ultrastructure, be " spirulina " by domestic and international misnomer, reality is obtusatus arthrospira (Arthrospira platensis).About the cultural method and the condition of artrospira spirulina, and the method for results artrospira spirulina, can be referring to the inventor on June 18th, 1994 application, application number No.94112205.0 is entitled as the patent application of " artrospira spirulina health food composition and method for making thereof ".

The whole process that the present invention extracts Phycocyanins, C-is as follows: cultivate artrospira spirulina, make the exsiccant artrospira spirulina after the results, through broken wall, the centrifugal crude extract that makes, crude extract is again through saltouing, dialysis, and steps such as ion exchange chromatography make purified Phycocyanins, C-.

More specifically, it is an amount of that the artrospira spirulina dry algae powder is added water, and (for 15～20 times of dry powder weight) → put-4 ℃ is freezing, and → 25 °～35 ℃ melt, and → freezing again melting repeated 3～5 times; (or carry out enzymolysis with enzyme) → centrifugal → abandon throw out gets supernatant liquor, and add ammonium sulfate and divide gold-plating, → abandon the lower sediment thing, get topper, through dissolving and dialysis → carry out ion exchange chromatography with the DEAE fibre columns, lyophilize obtains purified Phycocyanins, C-.

Wherein, breaking cell wall can adopt methods such as freeze-thaw method or enzymolysis process to carry out.

Wherein, the enzyme of selecting for use during enzymolysis can be one or more of (but being not limited to) following enzyme: cellulase, papain, Traumanase, aspergillus oryzae, Monascus ruber, amylase, saccharifying enzyme, helicase etc.Enzymolysis is generally at pH3-8, and temperature 20-50 ℃ was carried out 18-24 hour, and enzymatic hydrolysis condition also can be by the condition of manufacturer's suggestion.

Frozen-thaw process can carry out one or many.Generally with repeatedly, especially 3-5 time for well.

The Phycocyanins, C-that makes finds that after deliberation its spectral response curve is as follows: obtained the maximum absorption is 620mm, and the fluorescence maximum value is 640mm.

This shows that Phycocyanins, C-absorption spectrum and green plants photosynthesis spectrum are complementary just, and this is wide to distributing, and originally productivity lacks the algae of chlorophyll b greatly, and it is important more to seem.

The absorption spectrum of Phycocyanins, C-in the aqueous solution is more hypsochromic slightly than peak value (λ max) in live body, its spectral shape and intensity depend primarily on the character of chromophore, quantity and state of aggregation, in the general chromophore 4 pyrroles systems conjugated double bond the more, λ max is more to red shift; (α, absorption spectrum β) is α, the superposition that beta subunit absorbs respectively, but further polymerization then can cause the red shift of long wave absorption peak, and increase with uptake factor (ε M).

Phycocyanins, C-is rare fluorescent in life cell, and they in a single day are released in the solution and when giving an amount of illumination, PE just sends brown fluorescent, and PC then sends red fluorescent.Relatively absorption and the emmission spectrum of PE, PC, AP and Chla between the PE-PC, all have stronger absorption to overlap between the PC-Ap and between the Ap-Chla, and the pass order of this hint energy between these photosynthetic pigments is: PE-PC-Ap-Chla.

Structure to Phycocyanins, C-is furtherd investigate, and finds that Phycocyanins, C-generally forms oligomer by α and beta monomers, wherein the polypeptide chain be made up of about 160-180 amino acid of α and β subunit.

The homology of Phycocyanins, C-subunit also showing on the higher level on similar polymerization behavior and the polymerizable molecular conformation, its step by step polymerization process established architecture basics for its transmission ofenergy function of in cell, undertaking.

Phycocyanins, C-behind the purifying all obtains single colour band with polyacrylamide gel (PAGE) electrophoresis and isoelectric focusing, its iso-electric point (IEP) value is respectively 4.8, Phycocyanins, C-by the visible obtusatus arthrospira of 12%SDS polyacrylamide gel (SDS-PAGE) electrophoresis is made up of α and two subunits of β, and its molecular weight is 14500.

The amino acid of Phycocyanins, C-is formed and content, and after measured, data are listed in table 1.

Table 1.

The amino acid of Obtusatus arthrospira phycocyanin is formed and content (%)

The amino acid Phycocyanins, C-

Aspartic acid Asp 6.15

Threonine Thr 6.56

Serine Ser 6.29

L-glutamic acid Glu 5.45

Glycine Gly 11.48

L-Ala Ala 10.13

Gelucystine Cys 5.37

Xie Ansuan Val 6.91

Methionine(Met) Met-

Isoleucine Ile 6.29

Leucine Leu 5.48

Tyrosine Tyr 5.70

Phenylalanine Phe 5.46

Methionin Lys 5.58

Proline(Pro) Pro-

Arginine Arg 4.10

Histidine His-

Tryptophane Try ND ^*

* undetermined.

As can be seen from Table 1, dicarboxylic amino acid (aspartic acid, L-glutamic acid) contained in the Phycocyanins, C-is higher with the ratio of basic aminoacids (Methionin, arginine), so its iso-electric point (IEP) is lower, is 4.8.In addition, Phycocyanins, C-lacks methionine(Met), proline(Pro) and Histidine.

Surprisingly, the inventor finds that Phycocyanins, C-has extremely important health-care effect: it has higher erythropoietin activity, the CFU-E colony is formed to have stronger hormesis (table 2) simultaneously.

Table 2.

Obtusatus arthrospira phycocyanin is to the influence of tire mouse liver cell CFU-E in late period

Sample	Dosage	The CFU-E colony is counted Counts of CFU-E colony (/ 2 * 10 4FLC)	Suitable EPO unit	Than Specific activity alive (μ/mg)
Blank Control (1640)		69 ±28．5
Standard EPO	62．5mu 125．0mu 250．0mu 500．0mu 1000．0mu	97±29．4 128±53．2 165±67．3 223±54．7 283±77．9
Phycocyanins, C-C-Phycocyanin	12．5ng	271±60．4	850	68000

As seen from Table 2, Spirulina phycocyanin has higher erythropoietin activity, the CFU-E colony is formed to have stronger hormesis simultaneously.

Phycocyanins, C-is also influential to the formation of normal mouse CFU-GM.Data are listed in table 3.

The influence that the external adding Obtusatus arthrospira phycocyanin of table 3. forms normal mouse CFU-GM

Rank Group	Dosage (μ/ml)	The CFU-GM colony is counted CFU-GM (Colony/10 5BMC)
Control group Control (MCM)	0	119．67±6．1
C-PC+MCM	10 20 50 100 200	120．33±3．06 127．33±4．73 143．67±9．02* 175．33±5．14* * * 140．33±3．45*
SPP C-PC	100 100	0 0

Annotate: MCM mouse flesh conditioned medium (Conditioned Medium of micemusal)

C-PC: the Phycocyanins, C-that derives from artrospira spirulina.

X ± SD n=5*P＜0.05**P＜0.01***P＜0.001 (compared with the control).

Observed ⁶⁰Recovery situation behind the Co gamma-rays 5Gy irradiation small white mouse grain monosystem progenitor cell in 30 days, find, 1-5 days small white mouse grain list progenitor cell colonies after being subjected to photograph form minimum, began in the 10th day to recover, and giving abdominal injection Phycocyanins, C-test group mouse, its CFU-GM productive rate is significantly higher than control group.Illustrate that Obtusatus arthrospira phycocyanin can significantly promote to be subjected to shine the proliferation and differentiation of mouse grain monosystem progenitor cell.

In addition, Phycocyanins, C-forms also influential to extracorporeal irradiation medullary cell CFU-GM.

Normal mouse marrow is made behind the single cell suspension after the Phycocyanins, C-of different concns is hatched warp ⁶⁰Carry out CFU-GM behind the Co gammairradiation again and cultivate, it the results are shown in Table 4.

Table 4.

The influence of Obtusatus arthrospira phycocyanin to being subjected to form according to bone marrow cells in mice CFU-GM

Group Group	Dosage Dose (μ g/ml)	The CFU-GM colony is counted CFU-GM (Colony/10 5BMC)	P
Contrast Control	0	51±4.48
Phycocyanins, C-C-PC	10 50 100 200	60.6±2.93 72±5.83 88.4±5.37 69±2.24	<0.01 <0.001 <0.001 <0.001

As can be seen from Table 4, the medullary cell of hatching with the Phycocyanins, C-of different concns forms the CFU-GM productive rate apparently higher than control group, this shows that Phycocyanins, C-can improve the capability of resistance to radiation of medullary cell.

Also studied the influence of Phycocyanins, C-to the mice serum colony stimulating activity.

Get normal mouse, the abdominal injection Phycocyanins, C-, administration was got blood from eyeball after 1～10 day, and (Colony stimulating factor, CSF) CFU-GM is cultivated in the source to gained serum as G CFS.Data are listed in table 5.

Table 5.

Obtusatus arthrospira phycocyanin is right

The influence of mice serum colony-stimulating vigor

Group Group	Administration time (my god)	The CFU-GM colony is counted CFu-GM (Colony/10 5BMC) aSerum bSerum+MCM	P
				Control group

Control	-	70±1 113±9.17
				Phycocyanins, C-C-PC	1 5 10	86.33±3.05 125±2 98±3.61 137±5.29 109.33±2.54 138.67±1.54	＞0.05 ＜0.05 ＜0.05

Annotate: a. final concentration 10%

B. final concentration 5%

From mouse boosting cell generation the influence that G CFS caused of Phycocyanins, C-, also can find out the health-care effect of Phycocyanins, C-to mitogenstimulated.

Normal mice by intraperitoneal injection Phycocyanins, C-is after 5 days, and extracting spleen cell was by 5 * 10 in the 6th day ⁶Ml concentration is incubated in the serum-free medium that contains 5 μ g/ml ConA, cultivate back centrifuging and taking supernatant, detect the colony stimulating activity of supernatant liquor, the same Phycocyanins, C-that adds different concns in the spleen cell cultures system, gained supernatant liquor carry out colony stimulating activity and measure.The detected result of the spleen cell cultures supernatant liquor of two kinds of methods preparation sees Table 6 and table 7.

Table 6.

Inducing mouse spleen in the Obtusatus arthrospira phycocyanin body

Produce the influence of G CFS

Group Group	The CFU-GM colony is counted CFU-GM (Colony/10 5BMC)
		aSF-SCCM bF-SCCM+MCM
	Control group Control		71.73±3.49 106.47±12.68
Phycocyanins, C-C-PC		101.87±9.98*** 126.2±7.98**

A. final concentration 10%

B. final concentration 5%

SF-SCCM serum-free splenocyte conditioned medium

* P＜0.01, * * * P＜0.001 (compared with the control)

By table 6 as seen, the abdominal injection Phycocyanins, C-, the inducing mouse splenocyte produces G CFS more significantly.

Table 7.

The external evoked mouse spleen cell of Obtusatus arthrospira phycocyanin

Produce the influence of G CFS

Group Group	Dosage Dose (μ g/ml)	The CFU-GM colony is counted CFU-GM (Colony/10 5BMC) aF-SCCM bSF-SCCM+MCM
				Control group Control	-	81.67±5.03	115±6
Phycocyanins, C-C-PC	10 50 100	85.67±3.05 106.67±3.5l** 117±4**	97.67±3.05 118.33±4.17 142.33±3.05**

A. final concentration 10%

B. final concentration 5%

* P＜0.05, * * P＜0.01, * * * P＜0.001 (compared with the control)

As can be seen from Table 7, external adding Phycocyanins, C-more significantly the inducing mouse splenocyte produce G CFS.

In addition, by Phycocyanins, C-to mouse CFU-E (CFU-E, BFU-E) influence of proliferation and differentiation.

(a) influence that normal mouse and anemia mice CFU-E and BFU-E are formed.

Normal mice by intraperitoneal injection Phycocyanins, C-5 days, carried out the cultivation of CFU-E and BFU-E on the 6th day, experimental result shows that Obtusatus arthrospira phycocyanin can improve CFU-E of mouse and the productive rate of BFU-E by the utmost point significantly, illustrates that Obtusatus arthrospira phycocyanin can promote the small white mouse erythroid hematopoiesis.

Normal small white mouse cervical vertebra dislocation is put to death, take out femur and make single cell suspension, do the cultivation of CFU-E, culture system replaces the Phycocyanins, C-that directly adds different concns when EPO or EPO exist with the Phycocyanins, C-of different concns and cultivates CFU-E, make blank with 1640, only adding EPO is normal control, listed in result such as the table 8.Hence one can see that, and Phycocyanins, C-has direct hormesis to CFU-E, promotes the formation of CFU-E colony.Phycocyanins, C-can significantly improve the productive rate (comparing with the normal control group) of CFU-E when existing simultaneously with EPO.

Table 8.

The influence that external adding Obtusatus arthrospira phycocyanin forms CFU-E

Group Group	Dosage Dose (μ g/ml)	The CFU-E colony is counted CFU-E clone counting (Colony/5 * 10 4BMC)
Control group I Control (I) EPO (+) control group I Control (I) EPO (-)		152±11．3 51±17．2
(EPO) C-PC (EPO) for Phycocyanins, C-	0．015 1．000 10．00 50．00 100．00	147±17．9 172．2±14．8* 261．3±15．7* * * 227．7±19．6* * * 156．9±20．4
Phycocyanins, C-(+EPO) C-PC (+EPO)	0．015 1．000 10．00 50．00 100．0	244．5±13．7* * 273．3±18．2* * * 297．8±20．7* * * 301．3±22．7* * * 279．9±27．7* * *

EPO erythropoietin (Erythropoietin) * P＜0.05, * * P＜0.01, * * * P＜0.001, ((I) compares with control group)

In the BFU-E culture system, be that the Phycocyanins, C-of 0.015 μ g/ml replaces EPO or BPA with final concentration, or replace EPO and BPA simultaneously, and EPO and BPA add Phycocyanins, C-when existing simultaneously and cultivate BFU-E.Other establishes 4 groups of contrasts, the normal control that EPO and BPA add simultaneously, and one of them adding of EPO and BPA and EPO and BPA do not add.Experimental result such as table 9.

The influence that Obtusatus arthrospira phycocyanin forms BFU-E under table 9. different condition

Group Group	Control group Control				Experimental group
											Ⅰ	Ⅱ	Ⅲ	Ⅳ	1	2	3	4	5	6
	EPO BPA * C-PC HS L-glutamine mercapto alcohol methylcellulose RPMI-1640	+ + - + + + + +	- + - + + + + +	+ - - + + + + +	- - - + + + + +	- + - + + + + +	- + + + + + + +	- - - + + + + +	- - + + + + + +	+ + - + + + + +											+ + + + + + + +
BFU-E clone counting (clone/5 10 4BMC)											30.7 ±5.4	13.4 ±3.7	0	0	21.3** ±5.1	31.8** ±7.4	0	5.1 ±4.3	(**) 43.3 ±5.5	(**) 51.2 ±7.7

A. final concentration 0.015 μ g/ml (* *) P 0.01, I compares with control group, and * * P 0.01 compares with control Group II.

The BPA burst promoting factor

The HS horse serum

As can be seen from Table 9, adding Phycocyanins, C-BFU-E productive rate does not descend identical with normal control, and when no BPA existed, BFU-E can not form, but after having added Phycocyanins, C-, can form few colony.Therefore can know the activity that Phycocyanins, C-has EPO by inference, and the effect of certain BPA sample is arranged.

The mouse warp ⁶⁰Form Anemia behind Co gammairradiation and the abdominal injection phenylhydrazine hydrochloride, its CFU-E and BFU-E productive rate significantly descend.But no matter be according to preceding or according to after give mouse peritoneal injection Phycocyanins, C-, CFU-E of mouse and BFU-E productive rate are significantly higher than control group.

The health-care effect that the Phycocyanins, C-of purifying from obtusatus arthrospira in sum, has mainly shows:

Have higher erythropoietin activity, the CFU-E colony is formed have stronger hormesis simultaneously: can promote and acceleration is subjected to ⁶⁰The leukocytic recovery of Co gamma-rays (5Gy) irradiation back small white mouse; Can improve small white mouse bone marrow nucleated cell number more significantly, can resist or significantly opposing irradiation and make its recovery reach normal value to the damage of small white mouse bone marrow nucleated cell; Promote the proliferation and differentiation of mouse grain monosystem progenitor cell; The inducing mouse splenocyte produces G CFS significantly; To the promoter action that is formed with of mouse bone marrow cells matrix progenitor cell, that is to marrow microcirculation has certain influence.

So, thisly can promote immunity system in order to develop, the natural protein of opposing various diseases, on experiment basis, the inventor further provides a series of health food compositions that contain Phycocyanins, C-as active princlple.

Health food composition generally contains 95% Phycocyanins, C-and 5% water and/or auxiliary material.

Wherein a kind of health food composition is capsule or tablet, and it consists of 95% Phycocyanins, C-, and 5% auxiliary material.

Another kind of health food composition is a beverage, and it consists of 7% Phycocyanins, C-, 91% water and 2% auxiliary material.

The third health food composition is an oral liquid, and it consists of 30% Phycocyanins, C-, 68% water and 2% auxiliary material.

In the present invention, " auxiliary material " refers to constitute the complementary raw material of health food composition, i.e. other secondary material except that Phycocyanins, C-and water specifically comprise:

1. cereal; Starch; Dextrin;

2. vegetable puree;

3. sweeting agent-stevioside or protein sugar, or another name for sugar etc.;

4. essence;

5. carboxymethyl cellulose;

6. gum arabic; Tragakanta etc.

Adopt Phycocyanins, C-as a kind of specific health-care components in the food compositions, when making health food composition, method, technology and the equipment that can adopt food-processing and making field to know, and can add other known compositions with health-care effect, more balancedly to bring into play health-care effect.

In addition, than protein, because Phycocyanins, C-effective high fluorescent productive rate in the pH of broad value scope as a kind of novelty; Their fluorescent does not disappear because of the existence of other biomolecules; Can be better molten between water; Therefore stabilizer pole and can preserve reason such as long period all when existing with the liquid or solid state can be used for immunodetection, the fluorescent microscopy, etc. the aspect.

Below in conjunction with embodiments of the invention, set forth the present invention in further detail.

Unless dated especially, otherwise all per-cent all is weight percent.Embodiment 1

The making embodiment 1A of Phycocyanins, C-crude extract

Get dry algae powder 100g and add 2000ml phosphoric acid buffer (pH7.3) and add amylase 1.5g, put in 35 ℃ of water-baths 30 minutes, fully stir 30 minutes after, put quick-frozen under-20 ℃ of environment, take out after waiting to form ice cube, put in 35 ℃ of water-baths and thaw, restir 30 minutes, quick-frozen again, three times so repeatedly, after the freeze thawing, centrifugal 30 minutes of 4 ℃ of 1000rpm, collect supernatant liquor, this is crude extract.Embodiment 1B

In the 100g dry algae powder, add 25 ℃ of distilled water pH7.3 of 2000ml and add amylase 1.2g, put in 37 ℃ of water-baths 45 minutes, stir that quick-frozen becomes ice cube after 30 minutes, thaw under 30 ℃ of conditions, stir quick-frozen again in 40 minutes.Triplicate, 1500rpm is centrifugal, gets supernatant liquor, (this is crude extract).Embodiment 1C

Get the 100g dry algae powder and add saccharifying enzyme 1g, add 20 ℃ of distilled water 2000ml pH7.3 and put in 27 ℃ of water-baths 30 minutes, stirred 30 minutes, quick-frozen is thawed under 20 ℃ of conditions, quick-frozen, and 2000rpm is centrifugal behind the multigelation three times, gets supernatant liquor, promptly gets crude extract.Embodiment 1D

Get dry algae powder 10g and add 200ml phosphoric acid buffer pH7.3 induction stirring 30 minutes, put in-20 ℃ of environment 12 hours, taking-up is put in 37 ℃ of water-baths and is thawed, restir 30 minutes freezes to take out after 12 hours again and thaws, and stirs, freezing, so behind the multigelation three times, centrifugal 30 minutes of 4 ℃ of 1000rpm collect supernatant liquor (this is a crude extract).Embodiment 2 Phycocyanins, C-s separate, purifying:

The crude extract that obtains among the embodiment 1 is diluted one times with distilled water, last DEAE52 cellulose column (3 * 18cm).This capital is used 10mM pH7.3 phosphoric acid buffer balance in advance, and 280nm detects, treat that crude extract all enters fibre columns after, wash post to baseline with same damping fluid, use the phosphoric acid buffer of 10mM pH8.0 again instead, and add the Na-Cl of 0.3N and 0.6N, branch's wash-out and collection.

Get the elution peak of the NaCl of 0.3N, add solid ammonium sulfate to saturation ratio, 4 ℃ to leave standstill after 4 hours 2500rpm centrifugal, collecting precipitation adds water 10ml dissolving, and water is dialysed, change the dialyzate secondary, dialysis equilibrium spends the night in the phosphoric acid buffer of the most rearmounted 500ml 10mM pH7.3.

Liquid after the dialysis, the centrifugal precipitation of going of 2500rpm, (1.5 * 95cm) gel columns are used the phosphoric acid buffer balance of 10mM pH7.3 to supernatant liquor in advance with Seohade * G75 gel filtration column, once go up sample, with same buffer elution, fraction collection, and do CFU-E and electrophoretic analysis, lyophilize gets finished product.One of embodiment 3 health food compositions: the making of capsule (or tablet)

(1) prescription:

A. phycocyanin extract 56%, starch 4%, β dextrin 20%, Icing Sugar 20%;

B. phycocyanin extract 66%, β dextrin 4%, starch 30%;

C. phycocyanin extract 85%, β dextrin 5%, carboxymethyl cellulose 1%, dressing 9%;

D. phycocyanin extract 95%, β dextrin and carboxymethyl cellulose 5%

(its ratio: 85: 15)

(2) method for making:

Purifying Phycocyanins, C-powder+auxiliary material (by prescription) → kneading is agglomerating, feel suitably → 18 mesh sieves, sieve series become 80 ℃ of following drying in oven of particle → put → respectively by hand or machine be filled in the capsule → check (the weighing error must not greater than ± 0.1g → encapsulation → ⁶⁰The Co width of cloth is penetrated sterilization → finished product;

Or the particle → compacting that will dry is in blocks, and → bag or not sugar coating → envelope (bag) dress → check → width of cloth are penetrated sterilization → finished product.(3) advantage

Phycocyanins, C-is comprised in the capsule; Can make its disintegration in stomach or intestines according to the requirement of using; It not only can effectively prevent its oxidation, and can better preserving property, effective ingredient, improve health-care effect.Embodiment 4

Two of health food composition: the making of beverage and oral liquid,

A. the dry Phycocyanins, C-50g that gets purification adds the 2.5g gum arabic, in dried mortar, grind well, once add the 25ml distilled water, wear into breast in the homogenizer rapidly, it is an amount of to add the stevioside aqueous solution then, slowly adds tragacanth mucilage (0.7g powder) again and adds water and make slurry in right amount, adds 0.1% potassium sorbate, add distilled water to 1000ml, with its filling bottle, the sterilization.

B Phycocyanins, C-70% Yelkin TTS 30g grinds to form breast in the homogenizer, it is an amount of to add the stevioside aqueous solution, continues emulsification and adds potassium sorbate 0.1%, adds distilled water to its filling bottle of 1000ml, sterilization.

C. Phycocyanins, C-30g, stevioside, citric acid is an amount of, and sterilization water makes into 100ml, the bottling disinfection.

The characteristics of oral liquid and beverage:

Fully natural green; Do not have any artificial color sugar-free and contain multiple amino acids and element and trace element.

Although, the present invention sets forth in conjunction with above specific embodiment, but, by reading specification sheets of the present invention and claims, those skilled in the art can fully understand feature of the present invention and advantage, and can under the prerequisite of purport of the present invention, make many modifications to specific embodiments of the invention, changes such as increase or deletion, but this falls within claims institute restricted portion of the application equally.

Claims (5)

1. Phycocyanins, C-matter, it is characterized in that, this protein extracts from obtusatus arthrospira, its molecular weight is 14500 dalton, iso-electric point is 4.8, and maximum spectral absorption value is 620nm, and the oligomer of being made up of α and β subunit, wherein α and β subunit are the polypeptide chains that is made of 160-180 amino acid, and two subunits have homology.

2. a health food composition is characterized in that, it contains Phycocyanins, C-as claimed in claim 1 as one of component.

3. a health food composition as claimed in claim 2 is characterized in that, this health food composition is capsule or tablet.

4. a health food composition as claimed in claim 2 is characterized in that, this health food composition is beverage or oral liquid.

5. the purposes of a Phycocyanins, C-as claimed in claim 1 is characterized in that, this Phycocyanins, C-is used for fluoroscopic examination, immunodetection and fluorescent microscopy.