CN106701874B - Preparation method of phycocyanin polypeptide - Google Patents
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- 108010053210 Phycocyanin Proteins 0.000 title claims abstract description 119
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 37
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 37
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 35
- 238000002360 preparation method Methods 0.000 title abstract description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 26
- 102000004190 Enzymes Human genes 0.000 claims abstract description 22
- 108090000790 Enzymes Proteins 0.000 claims abstract description 22
- 229940088598 enzyme Drugs 0.000 claims abstract description 22
- 239000007788 liquid Substances 0.000 claims abstract description 19
- 238000003756 stirring Methods 0.000 claims abstract description 19
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 17
- 239000000047 product Substances 0.000 claims abstract description 16
- 108090000526 Papain Proteins 0.000 claims abstract description 14
- 239000004365 Protease Substances 0.000 claims abstract description 14
- 229940055729 papain Drugs 0.000 claims abstract description 14
- 235000019834 papain Nutrition 0.000 claims abstract description 14
- 238000009210 therapy by ultrasound Methods 0.000 claims abstract description 14
- 230000000694 effects Effects 0.000 claims abstract description 13
- 238000001816 cooling Methods 0.000 claims abstract description 10
- 230000009849 deactivation Effects 0.000 claims abstract description 10
- 238000010438 heat treatment Methods 0.000 claims abstract description 10
- 238000000034 method Methods 0.000 claims abstract description 10
- 238000001694 spray drying Methods 0.000 claims abstract description 9
- 239000006228 supernatant Substances 0.000 claims abstract description 9
- 238000001291 vacuum drying Methods 0.000 claims abstract description 7
- 238000006243 chemical reaction Methods 0.000 claims description 10
- 239000012153 distilled water Substances 0.000 claims description 9
- 239000012528 membrane Substances 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 7
- 239000007795 chemical reaction product Substances 0.000 claims description 6
- 108090000145 Bacillolysin Proteins 0.000 abstract description 12
- 102000035092 Neutral proteases Human genes 0.000 abstract description 12
- 108091005507 Neutral proteases Proteins 0.000 abstract description 12
- 102000004169 proteins and genes Human genes 0.000 abstract description 11
- 108090000623 proteins and genes Proteins 0.000 abstract description 11
- 238000010008 shearing Methods 0.000 abstract description 3
- 239000000758 substrate Substances 0.000 abstract description 3
- -1 DPPH free radical Chemical class 0.000 description 10
- 235000016425 Arthrospira platensis Nutrition 0.000 description 5
- 240000002900 Arthrospira platensis Species 0.000 description 5
- 230000007062 hydrolysis Effects 0.000 description 5
- 238000006460 hydrolysis reaction Methods 0.000 description 5
- 229940082787 spirulina Drugs 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 3
- 150000003254 radicals Chemical class 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 102000003952 Caspase 3 Human genes 0.000 description 2
- 108090000397 Caspase 3 Proteins 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 230000007071 enzymatic hydrolysis Effects 0.000 description 2
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 2
- 235000013376 functional food Nutrition 0.000 description 2
- 238000003760 magnetic stirring Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000010979 pH adjustment Methods 0.000 description 2
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- 230000002797 proteolythic effect Effects 0.000 description 2
- 230000002000 scavenging effect Effects 0.000 description 2
- 241000195493 Cryptophyta Species 0.000 description 1
- 102000015636 Oligopeptides Human genes 0.000 description 1
- 108010038807 Oligopeptides Proteins 0.000 description 1
- 230000002292 Radical scavenging effect Effects 0.000 description 1
- 241000206572 Rhodophyta Species 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
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- 230000004071 biological effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- MGJZITXUQXWAKY-UHFFFAOYSA-N diphenyl-(2,4,6-trinitrophenyl)iminoazanium Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1N=[N+](C=1C=CC=CC=1)C1=CC=CC=C1 MGJZITXUQXWAKY-UHFFFAOYSA-N 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
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- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000004112 neuroprotection Effects 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 230000000243 photosynthetic effect Effects 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000000527 sonication Methods 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
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Abstract
A method for preparing phycocyanin polypeptide, comprising the steps of: adding water into phycocyanin, stirring to dissolve completely to obtain phycocyanin solution; adding papain or neutral protease into the phycocyanin solution, stirring uniformly, and performing ultrasonic treatment for 30-50 min to obtain an enzymolysis product; heating the enzymolysis product to 85-95 ℃ and preserving heat for 5-15 min, cooling after enzyme deactivation, centrifuging, and taking the supernatant as enzymolysis liquid; performing ultrafiltration treatment on the enzymolysis liquid to obtain ultrafiltrate; concentrating the ultrafiltrate in vacuum, and spray drying to obtain phycocyanin polypeptide. According to the preparation method of the phycocyanin polypeptide, ultrasonic-assisted enzymolysis is adopted, ultrasonic waves have cavitation effect, generated bubbles are extruded and broken, extremely strong mechanical shearing force can be generated instantaneously, protein is degraded or protein conformation is changed, hydrophilic groups of the protein are promoted to be exposed more, the solubility of phycocyanin in water is improved, the combination of enzymes and phycocyanin substrates is facilitated, and the enzymolysis efficiency of phycocyanin can be remarkably improved.
Description
Technical Field
The invention relates to the technical field of proteolysis, in particular to a preparation method of phycocyanin polypeptide.
Background
Phycocyanin (also called PC) is a photosynthetic auxiliary pigment existing in red algae and blue algae cells, and can capture light energy efficiently. Recent researches find that phycocyanin has various biological activities such as antioxidation, anticancer, anti-inflammatory and neuroprotection, and the research on phycocyanin has become a hotspot for researching natural ocean medicines. The research on phycocyanin in China starts from the 70 th century of 20, and is developed over 30 years, and most of the phycocyanin is in laboratory level although the phycocyanin has greatly progressed. Meanwhile, the functional foods mainly containing phycocyanin are not more, and the research and development of medicines are still blank. Enzymatic hydrolysis can improve the functional properties of proteins, and the resulting peptides have physicochemical properties that are not comparable to those of some proteins. If the enzymolysis method is adopted to degrade the phycocyanin into soluble oligopeptide, the physicochemical property of the phycocyanin in food can be greatly improved, and the physiological activity which is not available in the precursor of the phycocyanin can be possibly obtained. There are patents related to the enzymatic hydrolysis of phycocyanin. The enzymolysis product of the spirulina phycocyanin and the application thereof (application number 200910178865.6) provide the enzymolysis product of the spirulina phycocyanin, functional food comprising the enzymolysis product of the spirulina phycocyanin and a pharmaceutical composition comprising the enzymolysis product of the spirulina phycocyanin and a pharmaceutically acceptable carrier. Patent application number 201110377690.6 discloses a method for preparing Caspase-3 activation by using phycocyanin, which is used for extracting phycocyanin from spirulina dry powder, hydrolyzing the phycocyanin by using trypsin, separating to obtain peptides which are more effective than the phycocyanin itself in activating the enzyme activity of Caspase-3 and have no toxic activity on normal cells. However, the conventional phycocyanin enzymolysis method generally adopts a single enzymolysis technology to carry out enzymolysis on the phycocyanin, and has the defect of low proteolytic efficiency.
Disclosure of Invention
In view of the above, it is necessary to provide a method for producing phycocyanin polypeptide with high proteolytic efficiency.
A method for preparing phycocyanin polypeptide, comprising the steps of:
adding water into phycocyanin, stirring to dissolve completely to obtain phycocyanin solution;
adding papain or neutral protease into the phycocyanin solution, stirring uniformly, and performing ultrasonic treatment for 30-50 min to obtain an enzymolysis product;
heating the enzymolysis product to 85-95 ℃ and preserving heat for 5-15 min, cooling to 30-40 ℃ after enzyme deactivation, centrifuging, and taking the supernatant as enzymolysis liquid;
carrying out ultrafiltration treatment on the enzymolysis liquid to obtain ultrafiltrate;
and carrying out vacuum concentration on the ultrafiltrate, and carrying out spray drying treatment to obtain the phycocyanin polypeptide.
In one embodiment, water is added to the phycocyanin and stirred to be fully dissolved, and in the operation of obtaining the phycocyanin solution, the feed liquid ratio of the phycocyanin to the water is 1:10-1:50.
In one embodiment, water is added to the phycocyanin and stirred to be fully dissolved, and constant-temperature magnetic stirring is adopted in the operation of obtaining the phycocyanin solution, wherein the temperature is 50-55 ℃.
In one embodiment, the mass ratio of papain to phycocyanin is 1:100 to 1:20.
In one embodiment, the mass ratio of neutral protease to phycocyanin is 1:30-1:20.
In one embodiment, the method further comprises the step of adjusting the pH of the phycocyanin solution to 5.0-7.0 prior to the addition of papain.
In one embodiment, the method further comprises the step of adjusting the pH of the phycocyanin solution to 6.0-7.0 prior to adding the neutral protease.
In one embodiment, the reaction temperature is 50-60 ℃ during the ultrasonic treatment by adding papain.
In one embodiment, the reaction temperature is 45℃to 55℃during the addition of neutral protease for the sonication.
In one embodiment, the ultrasonic power is 300-800W during the operation of ultrasonic treatment.
According to the preparation method of the phycocyanin polypeptide, ultrasonic-assisted enzymolysis is adopted, ultrasonic waves have cavitation effect, generated bubbles are extruded and broken, extremely strong mechanical shearing force can be generated instantaneously, protein is degraded or protein conformation is changed, hydrophilic groups of the protein are promoted to be exposed more, the solubility of phycocyanin in water is improved, the combination of enzymes and phycocyanin substrates is facilitated, and the enzymolysis efficiency of phycocyanin can be remarkably improved in a short time.
Drawings
FIG. 1 is a flow chart of a method for producing an phycocyanin polypeptide according to one embodiment;
FIG. 2 is a graph showing the effect of phycocyanin polypeptide concentration on DPPH radical scavenging obtained in example 1.
Detailed Description
The present invention will be further described in detail with reference to the following examples, for the purpose of making the objects, technical solutions and advantages of the present invention more apparent, and it should be understood that the specific examples described herein are only illustrative of the present invention and are not intended to limit the present invention.
Referring to fig. 1, a preparation method of an phycocyanin polypeptide according to an embodiment includes the following steps:
s10, adding water into the phycocyanin, and stirring to fully dissolve the phycocyanin to obtain the phycocyanin solution.
Wherein, the purity A of phycocyanin 620 /A 280 >2. The feed liquid ratio of phycocyanin to water can be 1:10-1:50.
In the operation of S10, the phycocyanin is dissolved by constant-temperature magnetic stirring, and the temperature can be 50-55 ℃. The stirring time can be 20-40 min.
S20, adding papain or neutral protease into the phycocyanin solution, stirring uniformly, and performing ultrasonic treatment for 30-50 min to obtain an enzymolysis product.
Wherein the ultrasonic treatment can be performed in a water bath ultrasonic device.
Wherein the papain has an enzyme activity of 5×10 5 ～10×10 5 U/g. The mass ratio of papain to phycocyanin can be 1:100-1:20. In the process of adding papain for ultrasonic treatment, the reaction temperature can be 50-60 ℃. The ultrasonic power may be 300-800W. The step of adjusting the pH of the phycocyanin solution to 5.0-7.0 may also be included before the addition of papain. Further, the pH adjustment of the phycocyanin solution can be performed with 1M hydrochloric acid.
Wherein the enzyme activity of the neutral protease is 1×10 5 ～3×10 5 U/g. The mass ratio of neutral protease to phycocyanin can be 1:30-1:20. In the process of adding neutral protease for ultrasonic treatment, the reaction temperature can be 45-55 ℃. The ultrasonic power may be 300-800W. The step of adjusting the pH of the phycocyanin solution to 6.0-7.0 may also be included before adding the neutral protease. Further, the pH adjustment of the phycocyanin solution can be performed with 1M hydrochloric acid.
S30, heating the enzymolysis product to 85-95 ℃ and preserving heat for 5-15 min, cooling after enzyme deactivation, centrifuging, and taking the supernatant to obtain the enzymolysis liquid.
Wherein the heating rate can be 20-30 ℃/min.
In S30, after enzyme deactivation, cooling to 30-40 ℃ and centrifuging.
In the centrifugation operation, the centrifugation may be performed at 10000r/min for 10min.
And S40, performing ultrafiltration treatment on the enzymolysis liquid to obtain ultrafiltrate.
Wherein, ultrafiltration treatment can be carried out by adopting an ultrafiltration membrane with the molecular weight cutoff of 1 kD-10 kD.
S50, concentrating the ultrafiltrate in vacuum, and performing spray drying treatment to obtain the phycocyanin polypeptide.
According to the preparation method of the phycocyanin polypeptide, ultrasonic-assisted enzymolysis is adopted, ultrasonic waves have cavitation effect, generated bubbles are extruded and broken, extremely strong mechanical shearing force can be generated instantaneously, protein is degraded or protein conformation is changed, hydrophilic groups of the protein are promoted to be exposed more, the solubility of phycocyanin in water is improved, the combination of enzymes and phycocyanin substrates is facilitated, the enzymolysis efficiency of the phycocyanin can be remarkably improved in a short time, and the obtained polypeptide has relatively strong oxidation resistance.
The following is a detailed description of embodiments.
Example 1
(1) 5g phycocyanin (A) was weighed out 620 /A 280 >2) Placing into a container, adding 200mL of distilled water, stirring on a constant-temperature magnetic stirrer for 30min to fully dissolve the distilled water to obtain phycocyanin solution;
(2) The phycocyanin solution obtained in step (1) was adjusted to pH 6.0, followed by the addition of 1.5g papain (enzyme activity 8X 10) 5 U/g), uniformly stirring, and then performing ultrasonic treatment in a water bath ultrasonic device with the ultrasonic power of 600W and the reaction temperature of 55 ℃ for 50min to obtain an enzymolysis product;
(3) Rapidly heating the enzymolysis reaction product to 90 ℃, preserving heat for 10min, cooling to 30 ℃ after enzyme deactivation, centrifuging for 10min at 10000r/min, taking the supernatant as an enzymolysis liquid, and measuring that the hydrolysis degree of phycocyanin is 13.68%;
(4) Carrying out ultrafiltration treatment on the enzymolysis liquid obtained in the step (3) by adopting an ultrafiltration membrane with molecular weight cutoff of 5kD to obtain ultrafiltrate;
(5) Concentrating the ultrafiltrate obtained in the step (4) in vacuum, and spray drying to obtain phycocyanin polypeptide. The ability of phycocyanin polypeptide with different concentrations to scavenge DPPH free radicals is shown in figure 1, wherein one curve is a relation curve of the concentration of phycocyanin polypeptide and the scavenging rate of DPPH free radicals, and the other curve is a relation curve of the concentration of VC and the scavenging rate of DPPH free radicals. Therefore, the phycocyanin polypeptide has a certain relation to the clearance rate and concentration of DPPH free radical, and when the concentration of the phycocyanin polypeptide reaches 5mg/mL, the clearance rate of DPPH free radical reaches 87.31 percent.
Example 2
(1) 5g phycocyanin (A) was weighed out 620 /A 280 >2) Placing the mixture into a container, adding 50mL of distilled water, and stirring on a constant-temperature magnetic stirrer until the mixture is fully dissolved to obtain an phycocyanin solution;
(2) Adjusting pH of the phycocyanin solution obtained in step (1) to 5.0, and adding papain (enzyme activity 10×10) at 5% of the weight of phycocyanin 5 U/g), uniformly stirring, and then performing ultrasonic treatment in a water bath ultrasonic device with ultrasonic power of 300W and reaction temperature of 50 ℃ for 30min to obtain an enzymolysis product;
(3) Rapidly heating the enzymolysis reaction product to 85 ℃, preserving heat for 15min, cooling to 30 ℃ after enzyme deactivation, centrifuging for 10min at 10000r/min, taking the supernatant as an enzymolysis solution, and measuring that the hydrolysis degree of phycocyanin is 12.97%;
(4) Carrying out ultrafiltration treatment on the enzymolysis liquid obtained in the step (3) by adopting an ultrafiltration membrane with the molecular weight cutoff of 10kD to obtain ultrafiltrate;
(5) Concentrating the ultrafiltrate obtained in the step (4) in vacuum, and spray drying to obtain phycocyanin polypeptide. The obtained phycocyanin polypeptide has a certain relationship between the clearance rate and the concentration of DPPH free radical, and when the concentration of the phycocyanin polypeptide reaches 5mg/mL, the clearance rate of DPPH free radical reaches 80.98 percent.
Example 3
(1) 10g phycocyanin (A) was weighed out 620 /A 280 >2) Placing into a container, adding 500mL of distilled water, and stirring on a constant-temperature magnetic stirrer until the distilled water is fully dissolved to obtain an phycocyanin solution;
(2) Adjusting pH of the phycocyanin solution obtained in step (1) to 7.0, and adding papain (enzyme activity 10×10) at 1% of the weight of phycocyanin 5 U/g), uniformly stirring, and then performing ultrasonic treatment in a water bath ultrasonic device with the ultrasonic power of 800W and the reaction temperature of 60 ℃ for 50min to obtain an enzymolysis product;
(3) Rapidly heating the enzymolysis reaction product to 95 ℃, preserving heat for 5min, cooling to 30 ℃ after enzyme deactivation, centrifuging for 10min at 10000r/min, taking the supernatant as an enzymolysis liquid, and measuring that the hydrolysis degree of phycocyanin is 13.59%;
(4) Carrying out ultrafiltration treatment on the enzymolysis liquid obtained in the step (3) by adopting an ultrafiltration membrane with molecular weight cutoff of 1kD to obtain ultrafiltrate;
(5) Concentrating the ultrafiltrate obtained in the step (4) in vacuum, and spray drying to obtain phycocyanin polypeptide. The obtained phycocyanin polypeptide has a certain relationship between the clearance rate and the concentration of DPPH free radical, and when the concentration of the phycocyanin polypeptide reaches 5mg/mL, the clearance rate of DPPH free radical reaches 85.16 percent.
Example 4
(1) 5g phycocyanin (A) was weighed out 620 /A 280 >2) Placing into a container, adding 200mL of distilled water, stirring on a constant-temperature magnetic stirrer for 30min to fully dissolve the distilled water to obtain phycocyanin solution;
(2) Adjusting pH of the phycocyanin solution obtained in step (1) to 6.0, and adding neutral protease (enzyme activity 3×10) at 3% of phycocyanin weight 5 U/g), uniformly stirring, and then performing ultrasonic treatment in a water bath ultrasonic device with the ultrasonic power of 600W and the reaction temperature of 55 ℃ for 50min to obtain an enzymolysis product;
(3) Rapidly heating the enzymolysis reaction product to 95 ℃, preserving heat for 5min, cooling to 30 ℃ after enzyme deactivation, centrifuging for 10min at 10000r/min, taking the supernatant as enzymolysis liquid, and measuring that the hydrolysis degree of phycocyanin is 11.82%;
(4) Carrying out ultrafiltration treatment on the enzymolysis liquid obtained in the step (3) by adopting an ultrafiltration membrane with molecular weight cutoff of 1kD to obtain ultrafiltrate;
(5) Concentrating the ultrafiltrate obtained in the step (4) in vacuum, and spray drying to obtain phycocyanin polypeptide. The obtained phycocyanin polypeptide has a certain relationship between the clearance rate and the concentration of DPPH free radical, and when the concentration of the phycocyanin polypeptide reaches 6mg/mL, the clearance rate of DPPH free radical reaches 75.82 percent.
Example 5
(1) 5g phycocyanin (A) was weighed out 620 /A 280 >2) Placing the mixture into a container, adding 50mL of distilled water, and stirring on a constant-temperature magnetic stirrer until the mixture is fully dissolved to obtain an phycocyanin solution;
(2) Adjusting pH of the phycocyanin solution obtained in step (1) to 7.0, and adding neutral protease (enzyme activity 1×10) at 5% of phycocyanin weight 5 U/g), uniformly stirring, and then performing ultrasonic treatment in a water bath ultrasonic device with the ultrasonic power of 800W and the reaction temperature of 45 ℃ for 30min to obtain an enzymolysis product;
(3) Rapidly heating the enzymolysis reaction product to 85 ℃, preserving heat for 15min, cooling to 40 ℃ after enzyme deactivation, centrifuging for 10min at 10000r/min, taking the supernatant as an enzymolysis liquid, and measuring that the hydrolysis degree of phycocyanin is 11.06%;
(4) Carrying out ultrafiltration treatment on the enzymolysis liquid obtained in the step (3) by adopting an ultrafiltration membrane with the molecular weight cutoff of 10kD to obtain ultrafiltrate;
(5) Concentrating the ultrafiltrate obtained in the step (4) in vacuum, and spray drying to obtain phycocyanin polypeptide. The obtained phycocyanin polypeptide has a certain relationship between the clearance rate and the concentration of DPPH free radical, and when the concentration of the phycocyanin polypeptide reaches 6mg/mL, the clearance rate of DPPH free radical reaches 70.03 percent.
The foregoing is merely a preferred embodiment of the invention, and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the invention, which are intended to be comprehended within the scope of the invention.
Claims (1)
1. A method for preparing phycocyanin polypeptide, comprising the steps of:
(1) 5g phycocyanin, A, was weighed out 620 /A 280 >2, placing the mixture into a container, adding 200mL of distilled water, and stirring the mixture on a constant-temperature magnetic stirrer for 30min to fully dissolve the mixture to obtain an phycocyanin solution;
(2) Adjusting the pH of the phycocyanin solution obtained in step (1) to 6.0, then adding 1.5g papain, and the enzyme activity is 8×10 5 U/g, after stirring uniformly, performing ultrasonic treatment in a water bath ultrasonic device with ultrasonic power of 600W and reaction temperature of 55 ℃ for 50min to obtain an enzymolysis product;
(3) Rapidly heating the enzymolysis reaction product to 90 ℃, preserving heat for 10min, cooling to 30 ℃ after enzyme deactivation, centrifuging for 10min at 10000r/min, and taking the supernatant as enzymolysis liquid;
(4) Carrying out ultrafiltration treatment on the enzymolysis liquid obtained in the step (3) by adopting an ultrafiltration membrane with molecular weight cutoff of 5kD to obtain ultrafiltrate;
(5) Concentrating the ultrafiltrate obtained in the step (4) in vacuum, and spray drying to obtain phycocyanin polypeptide.
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| CN110106218A (en) * | 2019-04-24 | 2019-08-09 | 北海生巴达生物科技有限公司 | A kind of algae small-molecular peptides extracting method |
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| CN112226476A (en) * | 2020-10-10 | 2021-01-15 | 暨南大学 | Enzymolysis method of phycocyanin and application thereof |
| CN113355383A (en) * | 2021-06-04 | 2021-09-07 | 中国水产科学研究院黄海水产研究所 | Comprehensive utilization and processing method of spirulina |
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| CN115716870B (en) * | 2022-11-21 | 2023-10-20 | 北京工商大学 | Phycocyanin anti-tumor peptide and preparation method and application thereof |
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Application publication date: 20170524 Assignee: Nantong Zhongke Binmei Biotechnology Co.,Ltd. Assignor: YANTAI INSTITUTE OF COASTAL ZONE RESEARCH, CHINESE ACADEMY OF SCIENCES Contract record no.: X2024210000063 Denomination of invention: Preparation method of phycocyanin peptide Granted publication date: 20230516 License type: Common License Record date: 20240925 |