CN106636275A - Method for extracting proteins in sipunculus nudus - Google Patents
Method for extracting proteins in sipunculus nudus Download PDFInfo
- Publication number
- CN106636275A CN106636275A CN201611143364.0A CN201611143364A CN106636275A CN 106636275 A CN106636275 A CN 106636275A CN 201611143364 A CN201611143364 A CN 201611143364A CN 106636275 A CN106636275 A CN 106636275A
- Authority
- CN
- China
- Prior art keywords
- sipunculus nudus
- pulp
- protein
- extracting
- hours
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/16—Extraction; Separation; Purification by chromatography
- C07K1/18—Ion-exchange chromatography
Abstract
The invention discloses a method for extracting proteins in sipunculus nudus. The extraction method comprises the following steps: 1, adding the sipunculus nudus into water according to the weight ratio of 1:1, and mincing the sipunculus nudus into insect pulp by adopting a high-speed stirrer; 2, placing the insect pulp in ultrasonic waves for 1 to 2 hours, and then placing the pulp in an electromagnetic field for 20 to 30 minutes; 3, adding peptidase into the insect pulp according to the weight ratio of 100:1, placing the pulp in a high-pressure reaction kettle, adding acetic acid to regulate the PH, stirring for 3 to 6 hours at the temperature of 50 to 60 DEG C, placing the insect pulp in a water bath for enzyme deactivation, thus obtaining enzymolysis insect pulp, centrifugally extracting supernatant, and freezing-drying the supernatant to obtain the proteins. According to the method, cells of the insect pulp are crushed to facilitate enzyme deactivation of the proteins in the sipunculus nudus after the pulp is placed in the ultrasonic waves and the electromagnetic field, and trypsin and papain are compounded for use by high-pressure enzymatic extraction, so that the extraction rate of the proteins in the sipunculus nudus is remarkably increased.
Description
Technical field
The present invention relates to Protein Extraction technical field, and in particular to method for extracting protein in a kind of Sipunculus nudus.
Background technology
Sipunculus nudus, are also called Sipunculus nudus, are commonly called as " sandworm "., like an intestines, in long tubular, body is long for its shape
About 10～20 centimetres, and it is nude without hair from head to foot, body wall longitudinal muscle bunchy, every circular muscle is staggered, and forms square clathrate decorative pattern, side
Although lattice siphon-worm without sea cucumber, shark's fin, abalone it is famous and precious, delicious flavour is tender and crisp, is sea cucumber, less than shark's fin.It is grown in edge
Seabeach applies, because very sensitive to the quality of growing environment, can not survive if pollution, thus have " environmental mark biological " it
Claim.Method for extracting protein is typically using extraction with aqueous solution method and organic solvent extraction, water extraction in current marine product
Protein extracting ratio is low, the easy dissolvent residual of organic solvent extraction, and Sipunculus nudus contain rich in protein, and currently without relating to
And method for extracting protein in a kind of Sipunculus nudus.
The content of the invention
One object of the present invention is provided in the Sipunculus nudus that a kind of simple and convenient, recovery rate is high, extraction purity of protein is high
Method for extracting protein.
The present invention provide technical scheme be：
A kind of method for extracting protein in Sipunculus nudus, the extracting method is comprised the following steps：
Step one, the sand in Sipunculus nudus is cleaned up, water is added using by weight 1: 1, using high-speed stirred
Machine blends adult slurry；
Step 2, worm slurry is placed in ultrasonic wave after 1-2 hours, then is placed into 20-30 minutes in electromagnetic field；
Step 3, by weight 100: 1 add protease toward worm slurry, in being placed in autoclave, add acetic acid by PH
Adjust to 3-5, temperature is 50-60 DEG C, after stirring 3-6 hours, then it is the 100-120 DEG C of enzyme that goes out to place temperature in a water bath, is obtained
Enzymolysis worm slurry, centrifugation is extracted supernatant, supernatant freeze-drying is obtained final product into the protein.
Preferably, also include that Sipunculus nudus are adopted into concentration divides for 80% mass before high-speed stirred is carried out in step one
After several sodium chloride solution immersion 1-2 hours, rinsed 2-3 time using clear water.
Preferably, protease described in step 3 is 1: 1 mixing by volume of trypsase and papain, described
The addition of protease is 2000-6000U/g.
Preferably, the pressure of step 3 mesohigh reactor is 1-2Mpa, and mixing speed is 100rmp/min.
Preferably, the operating frequency of ultrasonic wave is 300-500 hertz in step 2, and the induction of electromagnetic field is
3000-4000Gs。
Preferably, in step 3 before supernatant freeze-drying through ion exchange column 2-3 time.
Preferably, also include adding volume ratio to carry out degreasing for 1: 1 ethyl acetate toward worm slurry before step 2, by subtracting
Pressure condensed skimmed, while reclaiming ethyl acetate.
Beneficial effects of the present invention are as follows：
The present invention starches worm after ultrasonic wave and electromagnetic field, and the clasmatosis that worm is starched is beneficial to protein in Sipunculus nudus
Enzymolysis, then by high pressure enzymolysis and extraction, trypsase and papain compounding use, significantly improve albumen in Sipunculus nudus
The recovery rate of matter, is preferably processed using high concentration salt solution other side lattice siphon-worm before extraction so that cell process dehydration shape
State, get rid of intestinal parasites by means of drugs slurry when be easily broken, improve protein enzymatic hydrolyzation, enzymolysis before also carry out ungrease treatment and be centrifuged after through
Ion-exchange treatment, can significantly improve the purity of protein.
Specific embodiment
With reference to specific embodiment, the present invention is described in further detail, to make those skilled in the art's reference say
Bright book word can be implemented according to this.
Embodiment 1
Method for extracting protein is comprised the following steps in the present embodiment Sipunculus nudus：
Step one, the sand in Sipunculus nudus is cleaned up, water is added using by weight 1: 1, using high-speed stirred
Machine blends adult slurry；
Step 2, worm slurry is placed in ultrasonic wave after 1 hour, then is placed into 20 minutes in electromagnetic field；
Step 3, by weight 100: 1 add protease toward worm slurry, in being placed in autoclave, add acetic acid by PH
Adjust to 3-5, temperature is 50 DEG C, after stirring 3 hours, then place adopt in a water bath water-bath carry out water bath heating temperature for
100 DEG C of enzymes that go out, obtain digesting worm slurry, and centrifugation is extracted supernatant, supernatant freeze-drying is obtained final product into the protein.
Embodiment 2
Method for extracting protein is comprised the following steps in the present embodiment Sipunculus nudus：
Step one, the sand in Sipunculus nudus is cleaned up, Sipunculus nudus are adopted into concentration for 80% mass fraction
After sodium chloride solution soaks 2 hours, rinsed 3 times using clear water, using by weight 1: 1 water is added, stirred using homogenizer
It is broken into worm slurry；
Step 2, worm slurry is placed in ultrasonic wave after 2 hours, then is placed into 30 minutes in electromagnetic field；
Step 3, by weight 100: 1 add protease toward worm slurry, in being placed in autoclave, autoclave
Pressure is 1Mpa, and mixing speed is 100rmp/min, adds acetic acid PH to be adjusted to 3-5, and temperature is 60 DEG C, after stirring 6 hours,
It is 120 DEG C of enzymes that go out to place temperature in a water bath again, obtains digesting worm slurry, and centrifugation is extracted supernatant, is by supernatant freeze-drying
The protein is obtained, the protease is 1: 1 mixing by volume of trypsase and papain, the addition of the protease
Measure as 2000U/g.
Wherein, the operating frequency of ultrasonic wave is 300 hertz in step 2, and the induction of electromagnetic field is 3000Gs.
Wherein, in step 3 before supernatant freeze-drying through ion exchange column 2 times.
Wherein, also include adding volume ratio to carry out degreasing for 1: 1 ethyl acetate toward worm slurry before step 2, it is dense by reducing pressure
Contracting degreasing, while reclaiming ethyl acetate.
Embodiment 3
Method for extracting protein is comprised the following steps in the present embodiment Sipunculus nudus：
Step one, the sand in Sipunculus nudus is cleaned up, water is added using by weight 1: 1, using high-speed stirred
Machine blends adult slurry；
Step 2, worm slurry is placed in ultrasonic wave after 2 hours, then is placed into 30 minutes in electromagnetic field；
Step 3, by weight 100: 1 add protease toward worm slurry, in being placed in autoclave, add acetic acid by PH
Adjust to 3-5, temperature is 60 DEG C, after stirring 6 hours, then it is 120 DEG C of enzymes that go out to place temperature in a water bath, obtains digesting worm slurry,
Supernatant is extracted in centrifugation, and supernatant freeze-drying is obtained final product into the protein.
Wherein, also include before high-speed stirred is carried out for Sipunculus nudus adopting concentration for 80% mass fraction in step one
After sodium chloride solution soaks 2 hours, rinsed 3 times using clear water.
Wherein, protease described in step 3 is 1: 1 mixing by volume of trypsase and papain, the albumen
The addition of enzyme is 6000U/g.
Wherein, the pressure of step 3 mesohigh reactor is 2Mpa, and mixing speed is 100rmp/min.
Wherein, the operating frequency of ultrasonic wave is 500 hertz in step 2, and the induction of electromagnetic field is 4000Gs.
Wherein, in step 3 before supernatant freeze-drying through ion exchange column 3 times.
Wherein, also include adding volume ratio to carry out degreasing for 1: 1 ethyl acetate toward worm slurry before step 2, it is dense by reducing pressure
Contracting degreasing, while reclaiming ethyl acetate.
Comparative example 1
It is same as Example 1, lack step 2.
Comparative example 2
It is same as Example 2, lack by Sipunculus nudus adopt concentration for the sodium chloride solution immersion of 80% mass fraction it is 2 little
Shi Hou, using clear water 3 the step are rinsed.
Comparative example 3
It is same as Example 2, lack in step 3 before supernatant freeze-drying through ion exchange column 2 times and in step
Two go to worm slurry to add volume ratio to carry out degreasing for 1: 1 ethyl acetate, by reduced pressure concentration degreasing, while reclaiming ethyl acetate.
Using ultraviolet absorption method test protein content, weigh sandworm and go weight M after sand, then weigh embodiment 1-3 and right
Ratio 1-3 obtains product weight M1, and re-test embodiment 1-3 and comparative example 1-3 obtain the protein content M2 in product, calculate egg
White matter recovery rate=M1/M*100% and lipidated protein=M2/M1*100%, as a result such as following table：
| Embodiment 1 | Embodiment 2 | Embodiment 3 | Comparative example 1 | Comparative example 2 | Comparative example 3 | |
| Recovery rate (%) | 8.2 | 10.2 | 9.8 | 5.85 | 6.4 | 7.2 |
| Purity (%) | 78.2 | 86.9 | 88.2 | 70.2 | 71.1 | 69.5 |
It is therefore seen that the extraction rate of protein and purity of the present invention significantly increase.
Although embodiment of the present invention is disclosed as above, it is not restricted to listed in specification and embodiment
With, it can be applied to completely various suitable the field of the invention, for those skilled in the art, can be easily
Other modification is realized, therefore under the universal limited without departing substantially from claim and equivalency range, the present invention is not limited
In specific details and shown here as the embodiment with description.
Claims (7)
1. method for extracting protein in a kind of Sipunculus nudus, it is characterised in that the extracting method is comprised the following steps：
Step one, the sand in Sipunculus nudus is cleaned up, using by weight 1: 1 water is added, stirred using homogenizer
It is broken into worm slurry；
Step 2, worm slurry is placed in ultrasonic wave after 1-2 hours, then is placed into 20-30 minutes in electromagnetic field；
Step 3, by weight 100: 1 add protease toward worm slurry, in being placed in autoclave, add acetic acid to adjust PH
To 3-5, temperature is 50-60 DEG C, and after stirring 3-6 hours, then it is the 100-120 DEG C of enzyme that goes out to place temperature in a water bath, is digested
Worm is starched, and supernatant is extracted in centrifugation, and supernatant freeze-drying is obtained final product into the protein.
2. method for extracting protein in Sipunculus nudus as claimed in claim 1, it is characterised in that carrying out height in step one
After also including for Sipunculus nudus adopting concentration for the sodium chloride solution immersion 1-2 hours of 80% mass fraction before speed stirring, adopt
Clear water is rinsed 2-3 time.
3. method for extracting protein in Sipunculus nudus as claimed in claim 1, it is characterised in that albumen described in step 3
Enzyme is trypsase and papain 1: 1 mixing by volume, and the addition of the protease is 2000-6000U/g.
4. method for extracting protein in Sipunculus nudus as claimed in claim 1, it is characterised in that step 3 mesohigh reacts
The pressure of kettle is 1-2Mpa, and mixing speed is 100rmp/min.
5. method for extracting protein in Sipunculus nudus as claimed in claim 1, it is characterised in that ultrasonic wave in step 2
Operating frequency is 300-500 hertz, and the induction of electromagnetic field is 3000-4000Gs.
6. method for extracting protein in Sipunculus nudus as claimed in claim 1, it is characterised in that supernatant is cold in step 3
It is lyophilized it is dry before through ion exchange column 2-3 time.
7. method for extracting protein in Sipunculus nudus as claimed in claim 1, it is characterised in that also include before step 2
Volume ratio is added to carry out degreasing for 1: 1 ethyl acetate toward worm slurry, by reduced pressure concentration degreasing, while reclaiming ethyl acetate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611143364.0A CN106636275A (en) | 2016-12-12 | 2016-12-12 | Method for extracting proteins in sipunculus nudus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611143364.0A CN106636275A (en) | 2016-12-12 | 2016-12-12 | Method for extracting proteins in sipunculus nudus |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106636275A true CN106636275A (en) | 2017-05-10 |
Family
ID=58824390
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611143364.0A Pending CN106636275A (en) | 2016-12-12 | 2016-12-12 | Method for extracting proteins in sipunculus nudus |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106636275A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117362084A (en) * | 2023-10-27 | 2024-01-09 | 中山市承铭农业技术开发有限公司 | Amino acid water-soluble fertilizer prepared from soybeans and production process |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103012615A (en) * | 2013-01-08 | 2013-04-03 | 广东海洋大学 | Method for efficiently extracting sepia acidic polysaccharose |
| CN103169081A (en) * | 2013-03-22 | 2013-06-26 | 薛命雄 | Sandworm polypeptide enteric capsule |
| CN105087729A (en) * | 2015-08-14 | 2015-11-25 | 浙江省海洋开发研究院 | Tuna bone collagen peptide preparation method |
-
2016
- 2016-12-12 CN CN201611143364.0A patent/CN106636275A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103012615A (en) * | 2013-01-08 | 2013-04-03 | 广东海洋大学 | Method for efficiently extracting sepia acidic polysaccharose |
| CN103169081A (en) * | 2013-03-22 | 2013-06-26 | 薛命雄 | Sandworm polypeptide enteric capsule |
| CN105087729A (en) * | 2015-08-14 | 2015-11-25 | 浙江省海洋开发研究院 | Tuna bone collagen peptide preparation method |
Non-Patent Citations (3)
| Title |
|---|
| 孙秀发: "《食品营养学》", 31 July 2011, 郑州大学出版社 * |
| 孙雪萍: "方格星虫蛋白酶解工艺优化", 《食品与机械》 * |
| 李乃胜: "《中国海洋水产品现代加工技术与质量安全》", 31 May 2015, 海洋出版社 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117362084A (en) * | 2023-10-27 | 2024-01-09 | 中山市承铭农业技术开发有限公司 | Amino acid water-soluble fertilizer prepared from soybeans and production process |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104086667B (en) | A kind of method preparing citruss skin slag pectin using ultrasonic assistant extraction | |
| CN101836687B (en) | Preparation method of instant fresh water fish peptide powder | |
| CN105361153A (en) | Processing method of sea cucumber glycopeptides chelated calcium | |
| JP2007068454A (en) | Method for producing rice protein, rice protein produced by the method, and food | |
| CN104073540A (en) | Polypeptide production method for enhancing utilization ratio of protein | |
| CN104498568A (en) | Method for preparing fish-skin collagen powder by use of fresh fish skin | |
| US20130035474A1 (en) | Collagen mixture and method of making the same | |
| CN106690140A (en) | Preparation method of oyster powder | |
| CN105779542A (en) | Method for preparing organic rapeseed polypeptide through enzyme method | |
| CN106906199A (en) | A kind of fast purifying squid viscera albumen enzyme method and application | |
| CN107198252A (en) | A kind of pupa albumen, its compounding albumen and preparation method thereof | |
| CN105475881A (en) | Desalination method for salted egg white, salted-egg-white protein peptide powder and preparation method | |
| CN107227328A (en) | A kind of selenium-enriched tea leaf polypeptide and application | |
| CN106636275A (en) | Method for extracting proteins in sipunculus nudus | |
| JP4468144B2 (en) | Yeast extract high in 5'-ribonucleotide and method for producing the same | |
| CN105483195A (en) | Method for preparing marine protein peptide by multi-step enzymolysis | |
| CN106804873A (en) | A kind of preparation technology of gel soybean protein isolate high | |
| CN105349600A (en) | Deodorizing method of Gadus protein hydrolysate | |
| CN107089869A (en) | A kind of preparation method of the molten slurry Protein fertilizer of fish | |
| CN104304639A (en) | Preparation method of concentrated silkworm chrysalis protein powder without abnormal smell | |
| CN107114713A (en) | A kind of biologic fishy smell removing method of snapper seed | |
| CN110229859A (en) | A kind of efficient preparation process of the anti-oxidant calcium ion chelating peptide in Optimization of Low Value Fish visceral protein source | |
| CN112779308A (en) | Preparation method of micromolecular oyster peptide | |
| CN104498569A (en) | Method for preparing fish-skin collagen powder by use of dry fish skin | |
| CN107868133A (en) | A kind of immersion process in processing of farm products production |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB03 | Change of inventor or designer information |
Inventor after: Wang Weijian Inventor after: Li Hongcuan Inventor after: Xie Meijuan Inventor after: Zhan Hao Inventor before: Zhong Shuming |
|
| CB03 | Change of inventor or designer information | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20170510 |
|
| WD01 | Invention patent application deemed withdrawn after publication |