CN104130319B - A kind of extracting method of phycoerythrin - Google Patents
A kind of extracting method of phycoerythrin Download PDFInfo
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- CN104130319B CN104130319B CN201410409502.XA CN201410409502A CN104130319B CN 104130319 B CN104130319 B CN 104130319B CN 201410409502 A CN201410409502 A CN 201410409502A CN 104130319 B CN104130319 B CN 104130319B
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- extracting method
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- 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 present invention relates to a kind of extracting method of phycoerythrin, belong to biomedicine technical field.In order to solve the problem of existing broken wall efficiency is low and big for environment pollution, a kind of extracting method of phycoerythrin is provided, this method includes adding the fresh algae containing phycoerythrin or the algae powder Jing Guo pre-soaking in 50 DEG C~10 DEG C of ice powder, mixing speed control is subjected to broken wall treatment under conditions of 1000r/min~3000r/min, the shell-broken liquid containing phycoerythrin is obtained；The obtained shell-broken liquid containing phycoerythrin is separated, purified and dried again, finished product phycoerythrin is obtained.The method of the present invention has short broken wall cycle, efficiency high and environment amenable advantage, and obtained product purity is high.
Description
Technical field
The present invention relates to a kind of extracting method of phycoerythrin, belong to biomedicine technical field.
Background technology
Phycoerythrin is that the important of many marine algas catches photochromic fibroin, the algae courage egg such as it and phycocyanin, allophycocyanin
Bar-shaped phycobilisome is constituted in vain, and phycobilisome can capture and transmit luminous energy.Phycoerythrin is mainly distributed in the red algae of marine alga, by
In the presence of pigment molecular, it has stronger absorption in visible region 480-570nm wave bands, and can produce strong fluorescence.Algae
Lactoferrin can be applied to the industries such as food, cosmetics, dyestuff as natural pigment, fluorometric reagent be may also be fabricated which, for clinic
The research field such as medical diagnosis and immunochemistry and bioengineering.Phycoerythrin or a kind of important physiological activator, can
Sensitising agent is made applied to photodynamic therapy for cancer.However, at present, isolating and purifying for phycoerythrin is main using ammonium sulfate point
Level precipitation, the method such as ion-exchange chromatography are carried out, and it is cumbersome to isolate and purify process, complex operation, and time-consuming, and cost height.
There is the process using the processing of multigelation method.Such as Chinese patent application (publication number：CN102241754A a kind of red hair) is disclosed
The isolation and purification method of phycoerythrin in algae, including by red hair algae with 1：20~25m/v ratios are dissolved in distilled water, by freezing
Tie-melt, stir, after tissue mashing, ultrasonic disruption, filtering, centrifugation, supernatant is crude extract；Add ammonium sulfate progress
Saltout, the foreign protein that molecular weight is less than 30KDa is removed with the milipore filter concentration of 30KDa molecular weight, then using adsorbing separation desorption,
Obtain phycoerythrin.But its use freeze-thaw method exist the cycle it is long the problem of, and albumen qualitative change is easily caused using supercritical ultrasonics technology
Property inactivation, product yield is low, and it uses pollution of the salting out method to environment also larger.
The content of the invention
The present invention for above problems of the prior art there is provided a kind of extracting method of phycoerythrin, with carrying
Take efficiency and sporoderm-broken rate height and the few effect of environmental pollution.
The purpose of the present invention technical scheme is that, a kind of extracting method of phycoerythrin, the party
Method comprises the following steps：
A, broken wall：In the ice powder that red algae is added to -50 DEG C~-10 DEG C, by mixing speed control 1000r/min~
Broken wall treatment is carried out under conditions of 3000r/min, the shell-broken liquid containing phycoerythrin is obtained；
B, the obtained shell-broken liquid containing phycoerythrin separated, purified and dried, obtain finished product phycoerythrin.
In the extracting method of above-mentioned phycoerythrin, described separation can be using the conventional method in this area, such as using heavy
Shallow lake method separation or centrifugal separation method etc..Preferably, separation is specially described in step B：
The cotton post being made of medical absorbent cotton, which is inserted in the above-mentioned shell-broken liquid containing phycoerythrin, inhales phycoerythrin
It is attached on cotton post, the RED sector on separation cotton post is rinsed using extruding and water, obtains the aqueous solution containing phycoerythrin.
The purity of product can not only be improved using the separation method of the present invention, and it is environmentally friendly, without using chemistry such as ammonium sulfate
Reagent.Certainly, using the separation method of the present invention, and this area conventional wall-breaking method such as multigelation method, ultrasonic wave are used
Broken wall method, can equally realize the effect for improving product purity.
In the extracting method of above-mentioned phycoerythrin, preferably, purifying and drying are specially described in step B：
The obtained aqueous solution containing phycoerythrin is filtered and concentrated using milipore filter, is obtained containing the dense of phycoerythrin
Contracting liquid, then the concentrate containing phycoerythrin is dried, obtain finished product phycoerythrin.
The extracting method of the phycoerythrin of the present invention, by the way that under the conditions of high-speed stirred, the algae by ice powder and red algae is thin
The frictional force and shearing force produced between born of the same parents, makes the effect for reaching breaking-wall cell, so that realizing makes phycoerythrin from intracellular stream
The effect gone out.And be that can be achieved for separation, purifying and the dry method conventional using this area.But in order to preferably realize this
Invention, is inserted in the above-mentioned shell-broken liquid containing phycoerythrin by the cotton post that will be made of medical absorbent cotton, makes cotton pillar height degree
It is most of more than liquid level, and the fraction of cotton pillar height degree is inserted below liquid level, allow to realize more effective separation.
The 2/3~3/4 of the height of cotton post is such as set to be maintained at more than liquid level, and the height and width of cotton post can be carried out according to actual needs
Adjustment, preferably makes the height of cotton post be more than the width of cotton post.Again because phycoerythrin has highly hydrophilic, and water can
Constantly it is drawn onto more than liquid level by cotton post, constantly diffusion and the evaporation on cotton post with dampening, phycoerythrin is also constantly carried
Get on cotton post, and adsorb on cotton fiber (absorbent cotton).After the absorbent cotton on cotton post fully absorbs water, take away more than liquid level
The red part of absorbent cotton, and new absorbent cotton can be filled, until shell-broken liquid is blotted；On the other hand, due to shell-broken liquid
In substantial amounts of undissolved cell fragment can not be trapped in container bottom with water diffusion, and chlorophyll and carrotene etc.
Pigment is fat-soluble, can not be trapped in container bottom, the intracellular thing such as polysaccharides and ribonucleic acid with water diffusion
Matter is not so good as phycoerythrin due to hydrophily and water solubility, then the diffusion on cotton fiber is relatively slow, is trapped under cotton post
Portion, so as to reach the effect of separation, the separation method using the present invention has preferable effect, after testing the purity of the extract solution
A565nm/A280nm can be reached>1.0, the A565nm/A280nm generally used considerably beyond food service industry>0.7 requirement.
In the extracting method of above-mentioned phycoerythrin, red algae described in step A is selected from Porphyra yezoensis, porphyra haitanensis, multitube
Algae, asparagus, haematococcus or red hair algae.Preferably, the red algae be Porphyra yezoensis or porphyra haitanensis, and the Porphyra yezoensis or
Porphyra haitanensis carries out immersion treatment using water in advance.By immersion treatment, the abundant water swelling of seaweed can be made, made in high-speed stirred
Under the conditions of, it is easier to make to produce friction and shear effect between ice powder and seaweed, shell-broken effect is more preferable.
In the extracting method of above-mentioned phycoerythrin, preferably, ice powder described in the step A using 0.5~
1.0mol/L sodium-chloride water solution is made.Corresponding ice powder is made again by adding sodium chloride in water, it is possible to increase ice powder
Hardness, improve the friction between the frustule of ice powder and red algae and shear effect, reach more preferable shell-broken effect.
In the extracting method of above-mentioned phycoerythrin, preferably make system temperature during broken wall treatment described in step A
Insulation is carried out below 0 DEG C.By the temperature of control system below 0 DEG C, the ice powder of addition can be made to be not easy to melt, more had
Beneficial to reaching shell-broken effect.It can certainly carry out at ambient temperature, but because ice powder at room temperature easily melts, so,
During broken wall treatment as fruit ice powder melts excessive, then it can be stirred for reaching shell-broken effect by adding ice powder.As excellent
Choosing, system temperature control is below -10 DEG C during broken wall treatment described in step A, it is further preferred that during the broken wall treatment
System temperature is controlled between 0 DEG C~-20 DEG C.
In the extracting method of above-mentioned phycoerythrin, preferably, 2/3 height of cotton post described in step B is maintained at
It is more than liquid level.Make 2/3 height of cotton post more than liquid level, can more effectively isolate phycoerythrin, and make polysaccharides and core
The hydrophilies such as the intracellular matters such as ribosomal ribonucleic acid and the relatively weak impurity of water solubility remain in bottom, can further improve
The purity of phycoerythrin.
In the extracting method of above-mentioned phycoerythrin, preferably, purifying is specially described in step C：
Molecular weight is first used to be carried out for the milipore filter of 600,000~1,500,000 dalton the obtained aqueous solution containing phycoerythrin
Filtering, then, then uses molecular weight to be filtered and concentrated for the milipore filter of 300,000~500,000 dalton, obtains containing phycoerythrin
Concentrate.
Combined because phycoerythrin is intermolecular easily by hydrogen bond crosslinks, actually it tends to be formed in aqueous
Polymer high molecular weight protein, its polymer molecular weight can reach more than 400,000 dalton, therefore, be 600,000 by using molecular weight
The ultrafiltration membrance filter of~150 ten thousand dalton, in order to which can preferably remove other big molecular impurities again can be fully efficient
Phycoerythrin is reclaimed, the yield of phycoerythrin is improved, simultaneously, additionally it is possible to microorganism is waited except degerming, realized without using high temperature
The advantage of sterilizing.And use molecular weight more to remove small molecular weight impurity for the milipore filter of 300,000~500,000 dalton and reach
To the effect of concentration.
In the extracting method of above-mentioned phycoerythrin, preferably, red algae described in step A and the mass ratio 1 of ice powder：
10~20.
In the extracting method of above-mentioned phycoerythrin, preferably, the temperature of the ice powder described in step A is -30 DEG C
~-20 DEG C.
In the extracting method of above-mentioned phycoerythrin, preferably, the mean particle size of the ice powder be 20 mesh~
60 mesh.
In summary, the present invention has advantages below compared with prior art：
1. the extracting method of phycoerythrin of the present invention, by under conditions of high-speed stirred, making shape between ice powder and red algae
Into frictional force and shearing force, without using multigelation method, it becomes possible to realize shell-broken effect, and the cycle is short, greatly improves
Production efficiency.
2. the extracting method of the phycoerythrin of the present invention, is separated by using cotton post, can efficiently separate out algae
Lactoferrin, and with the high effect of purity, A565nm/ can be just reached by the purity of the isolated phycoerythrin of the step
A280nm>Substantially handled in 1.0 effect, whole processing procedure using water, with environment amenable advantage.
3. the extracting method of the phycoerythrin of the present invention, by first using the ultrafiltration membrance filter of 600,000~1,500,000 dalton,
Realize removing macromolecule foreign protein and cell fragment and play degerming effect.
Embodiment
Below by specific embodiment, technical scheme is described in further detail, but the present invention is simultaneously
It is not limited to these embodiments.
The extracting method of the phycoerythrin of the present invention, by the way that (fresh red algae is preferably first passed through into pulverization process in advance into phase
The algae powder answered) add in -50 DEG C~-10 DEG C of ice powder, it is preferably -30 DEG C~-20 DEG C, by mixing speed control in 1000r/
Simultaneously control system temperature is incubated progress broken wall treatment below 0 DEG C under conditions of min~3000r/min, after broken wall treatment terminates,
The ice powder in system is melted naturally, obtain the shell-broken liquid containing phycoerythrin, by detection, the sporoderm-broken rate of frustule reaches
More than 95%；The obtained shell-broken liquid containing phycoerythrin is separated, purified and dried again, finished product phycoerythrin is obtained.
Preferably, described ice powder is made of the sodium-chloride water solution using 0.5~1.0mol/L, preferably, the red algae selects
From Porphyra yezoensis, porphyra haitanensis, Marine Red Alga Polysiphonia Urceolata, asparagus, haematococcus or red hair algae.It is made again accordingly by adding sodium chloride in water
Ice powder, it is possible to increase the hardness of ice powder, improve the friction between ice powder and frustule containing phycoerythrin and shear effect,
Preferably realize shell-broken effect；The average grain diameter of the ice ice powder is preferably 20~60 mesh.Separation described above, purifying and dry
The conventional method in dry use this area, such as carries out isolated supernatant using centrifugal process, then using aqueous two-phase system,
Ammonium sulfate precipitation method is purified and chromatographic purification etc., then after being freeze-dried, obtains corresponding finished product phycoerythrin.
As another embodiment, using the conventional method (such as multigelation method) in this area by fresh red algae (most
It is good to first pass through pulverization process in advance into corresponding algae powder) broken wall treatment is carried out, the corresponding shell-broken liquid containing phycoerythrin is obtained, so
Afterwards, the cotton post being made of medical absorbent cotton is inserted makes phycoerythrin adsorb in cotton in the above-mentioned shell-broken liquid containing phycoerythrin
On post, the RED sector on separation cotton post is rinsed using extruding and water, obtains the aqueous solution containing phycoerythrin, after testing
The purity of the aqueous solution containing phycoerythrin can reach A620nm/A280nm>1.0, it will obtain containing the water-soluble of phycoerythrin
Liquid is filtered and concentrated using milipore filter, obtains the concentrate containing phycoerythrin, then the concentrate containing phycoerythrin is carried out
Dry, obtain finished product phycoerythrin.
Embodiment 1
Fresh Porphyra yezoensis is taken, is weighed, the weight ratio according still further to Porphyra yezoensis and ice powder is 1：20 ratio adds -50
DEG C ice powder, the average grain diameter of ice powder is 40~20 mesh, and above-mentioned Porphyra yezoensis and ice powder are added in agitator tank together, described
Porphyra yezoensis preferably first passes through immersion treatment in advance, then, and mixing speed is controlled under conditions of 3000r/min and control system
Temperature is incubated progress broken wall treatment 90min below 0 DEG C, after broken wall treatment terminates, the ice powder in system is melted naturally, obtains
Shell-broken liquid containing phycoerythrin, by detection, the sporoderm-broken rate of frustule reaches more than 95%；
The cotton post being made of medical absorbent cotton, which is inserted in the above-mentioned shell-broken liquid containing phycoerythrin, inhales phycoerythrin
It is attached on cotton post, the medical absorbent cotton can be fixed using stainless steel filtering net, the height of cotton post can be according to actual need
It is adjusted, meanwhile, make the 2/3 of cotton pillar height degree to be maintained at more than liquid level, until shell-broken liquid is blotted by cotton post, take out cotton post
The absorbent cotton (part of absorption phycoerythrin) of RED sector, because phycoerythrin is red material, it is easy to difference separation, leads to
Cross extruding and be rinsed using water, be rinsed until red absorbent cotton untill white, isolate the algae red egg on cotton post
In vain, the aqueous solution containing phycoerythrin is obtained；The purity A565nm/A280nm of this step phycoerythrin after testing>1.0；
The obtained aqueous solution containing phycoerythrin is filtered and concentrated using milipore filter, is obtained containing the dense of phycoerythrin
Contracting liquid；
Concentrate containing phycoerythrin is freeze-dried, the freeze-drying is obtained using conventional method
Finished product phycoerythrin, after testing, product purity A565nm/A280nm>2.0, it is fully able to reach medical rank.
Embodiment 2
Fresh porphyra haitanensis is taken, is weighed, the weight ratio according still further to porphyra haitanensis and ice powder is 1：15 ratio adds -40 DEG C
Ice powder, the average grain diameter of ice powder is 40~20 mesh, above-mentioned porphyra haitanensis and ice powder is added in agitator tank together, the porphyra haitanensis
Preferably first pass through immersion treatment in advance, then, by mixing speed control under conditions of 2000r/min and control system temperature-
Less than 10 DEG C insulations carry out broken wall treatment 80min, after broken wall treatment terminates, the ice powder in system is melted naturally, are contained
The shell-broken liquid of phycoerythrin, by detection, the sporoderm-broken rate of frustule reaches more than 95%；
The cotton post being made of medical absorbent cotton, which is inserted in the above-mentioned shell-broken liquid containing phycoerythrin, inhales phycoerythrin
It is attached on cotton post, the medical absorbent cotton can be fixed using stainless steel filtering net, the height of cotton post can be according to actual need
It is adjusted, meanwhile, make the 2/3 of cotton pillar height degree to be maintained at more than liquid level, until shell-broken liquid is blotted by cotton post, take out cotton post
The absorbent cotton (part of absorption phycoerythrin) of RED sector, because phycoerythrin is red material, it is easy to difference separation, leads to
Cross extruding and be rinsed using water, be rinsed until red absorbent cotton untill white, isolate the algae red egg on cotton post
In vain, the aqueous solution containing phycoerythrin is obtained；The purity A565nm/A280nm of this step phycoerythrin after testing>1.0；
Molecular weight is first used to be filtered for the milipore filter of 1,000,000 dalton the obtained aqueous solution containing phycoerythrin,
Then, then use molecular weight to be filtered and concentrated for the milipore filter of 300,000 dalton, obtain the concentrate containing phycoerythrin；
Concentrate containing phycoerythrin is freeze-dried, the freeze-drying is obtained using conventional method
Finished product phycoerythrin, after testing, purity A565nm/A280nm>2.0, it is fully able to reach medical rank.
Embodiment 3
Fresh porphyra haitanensis is taken, is weighed, the weight ratio according still further to porphyra haitanensis and ice powder is 1：15 ratio adds -40 DEG C
Ice powder, the average grain diameter of ice powder is 40~20 mesh, and described ice powder is the sodium-chloride water solution system using 0.5~1.0mol/L
Into, above-mentioned porphyra haitanensis and ice powder are added in agitator tank together, the porphyra haitanensis preferably first passes through immersion treatment in advance, then,
By mixing speed control, simultaneously control system temperature is incubated between 0 DEG C~-20 DEG C at progress broken wall under conditions of 2000r/min
70min is managed, after broken wall treatment terminates, the ice powder in system is melted naturally, obtains the shell-broken liquid containing phycoerythrin, by inspection
Survey, the sporoderm-broken rate of frustule reaches more than 95%；
The cotton post being made of medical absorbent cotton, which is inserted in the above-mentioned shell-broken liquid containing phycoerythrin, inhales phycoerythrin
It is attached on cotton post, the medical absorbent cotton can be fixed using stainless steel filtering net, the height of cotton post can be according to actual need
It is adjusted, meanwhile, make the 2/3 of cotton pillar height degree to be maintained at more than liquid level, until shell-broken liquid is blotted by cotton post, take out cotton post
The absorbent cotton (part of absorption phycoerythrin) of RED sector, because phycoerythrin is red material, it is easy to difference separation, leads to
Cross extruding and be rinsed using water, be rinsed until red absorbent cotton untill white, isolate the algae red egg on cotton post
In vain, the aqueous solution containing phycoerythrin is obtained；The purity A565nm/A280nm of this step phycoerythrin after testing>1.0；
Molecular weight is first used to be filtered for the milipore filter of 1,500,000 dalton the obtained aqueous solution containing phycoerythrin,
Then, then use molecular weight to be filtered and concentrated for the milipore filter of 400,000 dalton, obtain the concentrate containing phycoerythrin；
Concentrate containing phycoerythrin is freeze-dried, the freeze-drying is obtained using conventional method
Finished product phycoerythrin, after testing, purity A565nm/A280nm>2.0, it is fully able to reach medical rank.
Embodiment 4
Fresh asparagus is taken, is weighed, the weight ratio according still further to asparagus and ice powder is 1：10 ratio adds -30 DEG C
Ice powder, the average grain diameter of ice powder is 40~20 mesh, and described ice powder is the sodium-chloride water solution system using 0.5~1.0mol/L
Into, above-mentioned asparagus and ice powder are added in agitator tank together, the asparagus preferably first passes through immersion treatment in advance, then,
By mixing speed control, simultaneously control system temperature is incubated progress broken wall treatment below -5 DEG C under conditions of 1000r/min
90min, after broken wall treatment terminates, makes the ice powder in system melt naturally, obtains the shell-broken liquid containing phycoerythrin, by inspection
Survey, the sporoderm-broken rate of frustule reaches more than 95%；
The cotton post being made of medical absorbent cotton, which is inserted in the above-mentioned shell-broken liquid containing phycoerythrin, inhales phycoerythrin
It is attached on cotton post, the medical absorbent cotton can be fixed using stainless steel filtering net, the height of cotton post can be according to actual need
It is adjusted, meanwhile, make the 2/3 of the height of cotton post to be maintained at more than liquid level, until shell-broken liquid is blotted by cotton post, take out cotton post
RED sector the absorbent cotton part of phycoerythrin (absorption), because phycoerythrin is red material, it is easy to difference separation,
By extruding and being rinsed using water, it is rinsed until red absorbent cotton untill white, isolates the algae red on cotton post
Albumen, obtains the aqueous solution containing phycoerythrin；The purity A565nm/A280nm of this step phycoerythrin after testing>1.0；
Molecular weight is first used to be filtered for the milipore filter of 600,000 dalton the obtained aqueous solution containing phycoerythrin, so
Afterwards, then use molecular weight to be filtered and concentrated for the milipore filter of 300,000 dalton, obtain the concentrate containing phycoerythrin；
Concentrate containing phycoerythrin is freeze-dried, the freeze-drying is obtained using conventional method
Finished product phycoerythrin, after testing, purity A565nm/A280nm>2.0, it is fully able to reach medical rank.
Embodiment 5
The algae powder of haematococcus is taken, is weighed, is 1 according still further to the algae powder of haematococcus and the weight ratio of ice powder：20 ratio addition-
20 DEG C of ice powder, the average grain diameter of ice powder is 40~20 mesh, and described ice powder is the aqueous sodium chloride using 0.5~1.0mol/L
Liquid is made, and the algae powder and ice powder of above-mentioned haematococcus is added in agitator tank together, the algae powder of the haematococcus is preferably passed through in advance
Immersion treatment is crossed, then, by mixing speed control is under conditions of 1000r/min and control system temperature is at -5 DEG C~-10 DEG C
Between insulation carry out broken wall treatment 85min, after broken wall treatment terminates, the ice powder in system is melted naturally, obtain containing algae red
The shell-broken liquid of albumen, by detection, the sporoderm-broken rate of frustule reaches more than 95%；
The concrete operation method be the same as Example 1 of following steps, is repeated no more here.
The purity A565nm/A280nm of the finished product phycoerythrin finally given>2.0, it is fully able to reach medical rank.
Embodiment 6
The algae powder of haematococcus is taken, is weighed, is 1 according still further to the algae powder of haematococcus and the weight ratio of ice powder：15 ratio addition-
20 DEG C of ice powder, the average grain diameter of ice powder is 40~20 mesh, and described ice powder is the aqueous sodium chloride using 0.5~1.0mol/L
Liquid is made, and the algae powder and ice powder of above-mentioned haematococcus is added in agitator tank together, the algae powder of the haematococcus is preferably passed through in advance
Immersion treatment is crossed, then, by mixing speed control is under conditions of 3000r/min and control system temperature is protected below -10 DEG C
Temperature carries out broken wall treatment 75min, after broken wall treatment terminates, the ice powder in system is melted naturally, obtains containing phycoerythrin
Shell-broken liquid, by detection, the sporoderm-broken rate of frustule reaches more than 95%；
The concrete operation method be the same as Example 1 of following steps, is repeated no more here.
The purity A620nm/A280nm of the finished product phycoerythrin finally given>2.0, it is fully able to reach medical rank.
Embodiment 7
The algae powder of red hair algae is taken, is weighed, the weight ratio according still further to algae powder and ice powder is 1：15 ratio adds -10 DEG C of ice
Powder, the average grain diameter of ice powder is 40~20 mesh, and described ice powder is made of the sodium-chloride water solution using 0.5~1.0mol/L,
Above-mentioned algae powder and ice powder are added in agitator tank together, the red hair algae preferably first passes through immersion treatment in advance, then, will stir
Speed control is under conditions of 1500r/min and control system temperature is incubated progress broken wall treatment 90min below -15 DEG C, breaks
After wall processing terminates, the ice powder in system is melted naturally, obtain the shell-broken liquid containing phycoerythrin, by detection, frustule
Sporoderm-broken rate reach more than 95%；
The cotton post being made of medical absorbent cotton, which is inserted in the above-mentioned shell-broken liquid containing phycoerythrin, inhales phycoerythrin
It is attached on cotton post, the medical absorbent cotton can be fixed using stainless steel filtering net, the height of cotton post can be according to actual need
It is adjusted, meanwhile, make the 3/4 of cotton pillar height degree to be maintained at more than liquid level, until shell-broken liquid is blotted by cotton post, take out cotton post
The absorbent cotton (part of absorption phycoerythrin) of RED sector, because phycoerythrin is red material, it is easy to difference separation, leads to
Cross extruding and be rinsed using water, be rinsed until red absorbent cotton untill white, isolate the algae red egg on cotton post
In vain, the aqueous solution containing phycoerythrin is obtained；The purity A565nm/A280nm of this step phycoerythrin after testing>1.0；
Molecular weight is first used to be filtered for the milipore filter of 1,000,000 dalton the obtained aqueous solution containing phycoerythrin,
Then, then use molecular weight to be filtered and concentrated for the milipore filter of 500,000 dalton, obtain the concentrate containing phycoerythrin；
Concentrate containing phycoerythrin is freeze-dried, the freeze-drying is obtained using conventional method
Finished product phycoerythrin, after testing, purity A565nm/A280nm>2.0, it is fully able to reach medical rank.
Embodiment 8
The algae powder of Marine Red Alga Polysiphonia Urceolata is taken, is weighed, is 1 according still further to the algae powder of Marine Red Alga Polysiphonia Urceolata and the weight ratio of ice powder：10 ratio addition-
30 DEG C of ice powder, the average grain diameter of ice powder is 40~20 mesh, and described ice powder is the aqueous sodium chloride using 0.5~1.0mol/L
Liquid is made, and the algae powder and ice powder of above-mentioned Marine Red Alga Polysiphonia Urceolata is added in agitator tank together, the algae powder of the Marine Red Alga Polysiphonia Urceolata is preferably passed through in advance
Immersion treatment is crossed, then, at room temperature, mixing speed control broken wall treatment is subjected under conditions of 3000r/min, if broken wall
During ice powder melt excessive, can continue to stir by adding ice powder and carry out broken wall treatment, after broken wall treatment terminates, make system
Interior ice powder melts naturally, obtains the shell-broken liquid containing phycoerythrin, by detection, the sporoderm-broken rate of frustule reach 95% with
On；
The concrete operation method be the same as Example 1 of following steps, is repeated no more here.
The purity A565nm/A280nm of the finished product phycoerythrin finally given>2.0, it is fully able to reach medical rank.
Specific embodiment described in the present invention is only to spirit explanation for example of the invention.Technology neck belonging to of the invention
The technical staff in domain can be made various modifications or supplement to described specific embodiment or be replaced using similar mode
Generation, but without departing from the spiritual of the present invention or surmount scope defined in appended claims.
It is skilled to this area although having been made a detailed description to the present invention and being cited some specific embodiments
For technical staff, as long as it is obvious that can make various changes or correct without departing from the spirit and scope of the present invention.
Claims (9)
1. a kind of extracting method of phycoerythrin, it is characterised in that this method comprises the following steps：
A, broken wall：In the ice powder that red algae is added to -50 DEG C~-10 DEG C, by mixing speed control in 1000r/min~3000r/
Carry out making system temperature insulation carry out below 0 DEG C during broken wall treatment, and the broken wall treatment under conditions of min, obtain
Shell-broken liquid containing phycoerythrin；
B, the obtained shell-broken liquid containing phycoerythrin separated, purified and dried, obtain finished product phycoerythrin；Described point
From specially：
Make algae in the shell-broken liquid containing phycoerythrin that will be obtained in the cotton post insertion above-mentioned steps A being made of medical absorbent cotton
Lactoferrin is adsorbed on cotton post, the RED sector on separation cotton post, is rinsed, is obtained containing phycoerythrin using extruding and water
The aqueous solution.
2. the extracting method of phycoerythrin according to claim 1, it is characterised in that tool is purified and dried described in step B
Body is：
The obtained aqueous solution containing phycoerythrin is filtered and concentrated using milipore filter, the concentration containing phycoerythrin is obtained
Liquid；The concentrate containing phycoerythrin is dried again, finished product phycoerythrin is obtained.
3. the extracting method of phycoerythrin according to claim 1, it is characterised in that the red algae is selected from Porphyra yezoensis, altar
Seaweed, Marine Red Alga Polysiphonia Urceolata, asparagus, haematococcus or red hair algae.
4. the extracting method of the phycoerythrin according to claim 1-3 any one, it is characterised in that described in step A
Ice powder be made of 0.5~1.0mol/L sodium-chloride water solution.
5. the extracting method of phycoerythrin according to claim 1, it is characterised in that the height 2/3 of cotton post described in step B
It is maintained at more than liquid level.
6. the extracting method of phycoerythrin according to claim 1, it is characterised in that purifying described in step B is specially：
Molecular weight is first used to be carried out for the milipore filter of 600,000~1,500,000 dalton the obtained aqueous solution containing phycoerythrin
Filter, then, then uses molecular weight to be filtered and concentrated for the milipore filter of 300,000~500,000 dalton, obtains containing phycoerythrin
Concentrate.
7. the extracting method of phycoerythrin according to claim 1-3 any one, it is characterised in that red described in step A
Algae and the mass ratio 1 of ice powder：10~20.
8. the extracting method of phycoerythrin according to claim 1 or 5, it is characterised in that the temperature of the ice powder described in step A
Spend for -30 DEG C~-20 DEG C.
9. the extracting method of phycoerythrin according to claim 1, it is characterised in that the particle of ice powder described in step A
Size is the mesh of 20 mesh~60.
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| EP3684326A4 (en) * | 2017-09-18 | 2020-11-25 | Seiberg Consulting, LLC | Compositions containing natural products and use thereof for skin and hair |
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