CN103725740A - Method for extracting glycoprotein by using scollop blanching liquid - Google Patents
Method for extracting glycoprotein by using scollop blanching liquid Download PDFInfo
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- CN103725740A CN103725740A CN201310649382.6A CN201310649382A CN103725740A CN 103725740 A CN103725740 A CN 103725740A CN 201310649382 A CN201310649382 A CN 201310649382A CN 103725740 A CN103725740 A CN 103725740A
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Abstract
The invention relates to a method for extracting glycoprotein by using scollop blanching liquid. The method comprises the following steps: a fresh blanching liquid produced by processing scollop is selected as a raw material, and is subjected to concentration under reduced pressure, ethanol precipitation and centrifugation to obtain a precipitation, and the precipitation is made into powder after dried; light petroleum or acetone is added into the powder for ungrease treatment, the mixture is dried to obtain a solid powder, a proteolytic enzyme is added into the solid powder for enzymolysis, and enzyme deactivation is performed; scollop enzymatic hydrolysate is cooled to low temperature, centrifugation is carried out, and a supernatant fluid is collected and subjected to ultrafiltration to desalinate; a retentate fluid is subjected to refrigeration and drying to obtain a dry scollop glycoprotein product. The extraction method provided by the invention is reasonable in process, low in equipment investment, advanced in technology, practicable in operation and suitable for large-scale industrial production. A test show that the scollop glycoprotein extracted and separated by the method has the characteristics that the content of polyose is 10-20%, and the content of protein is 50-60%.
Description
Technical field
The present invention relates to extract food protein composition from other marine animals especially a kind of method of utilizing scallop blanching solution to extract glycoprotein.
Background technology
Research in recent years shows, in sea mollusk, glycoprotein has tumor inhibition effect, and wherein the aqueous extract of abalone, Patinopecten yessoensis, Turbo cornulus is to transplantability S
180ascitic type tumour all has strong restraining effect.Particularly in seashells, extract separated glycoprotein and there is stronger anti-tumor activity, cause extensive concern.
Scallop, is one of main cultivated shellfish of China coast, and the scallop of producing in the world has more than 60 kind, and China accounts for half.Common scallop culture kind has chlamys farreri, bay scallop and Patinopecten yessoensis.At present, the processing method that adopts scallop to produce scallop glycoprotein has been reported.For example, " preparation of Glycoprotein From Chlamys Farreri and the anti-tumor activity preliminary study thereof " that Gu Qianqun etc. deliver at < < ocean science > > (fifth phase in 1998), its processing step is as follows: get fresh scallops limit, add 10 times of calorimetric poach and carry, filter cleaner; After filtrate spraying is dry Powdered crude extract, preserve at 4 ℃; Crude extract dissolves by suitable quantity of water, injects dialysis tubing and dialyses, and dialyzate is separated by DEAE-cellulose column, collects elutriant, and after concentrating, then by Sephadex G-100 column chromatography purification, lyophilize obtains Glycoprotein From Chlamys Farreri.There is the deficiency that raw materials cost is high in this technique.
In actual production, scallop, except marketing fresh, is processed into dry products or tin product mostly.In processing scallop process, need to adopt boiling water to carry out blanching processing to scallop, therefore, can produce a large amount of scallop blanching solutions, if scallop blanching solution is not processed, cause the great wasting of resources and environmental pollution.According to surveying and determination: in every kilogram of scallop blanching solution, include protein 1.8～3.6g, polysaccharose substance 5.5～7.5g, ash content 1.5～3.5g and a small amount of VITAMIN etc.Therefore, how to strengthen the processing and utilizing to scallop blanching solution, become problem in the urgent need to address.
At present, yet there are no the report that utilizes scallop blanching solution to extract glycoprotein.
Summary of the invention
In order to overcome, in prior art, utilize fresh scallops limit to produce the high and scallop blanching solution of scallop glycoprotein cost not carry out the deficiency that processing and utilizing easily causes environmental pollution and the wasting of resources, the invention provides a kind of operation rationally, the scallop blanching solution that utilizes of advanced technology, operation feasible extracts the method for glycoprotein.
The technical solution adopted for the present invention to solve the technical problems is: a kind of method of utilizing scallop blanching solution to extract glycoprotein, is characterized in that: process following process steps:
(1) raw material is chosen and is processed that to choose the fresh blanching solution that scallop is processed to form be raw material, utilizes concentrating under reduced pressure equipment, is concentrated into solid content 35%～40%, obtains concentrated solution;
(2) alcohol precipitation is that 90～95% ethanolic solns precipitate by adding concentration in the concentrated solution of step (1) gained, to its final concentration be 55%～65%;
(3) centrifugal solution after step (2) ethanol precipitation is sent in whizzer, centrifugal, collecting precipitation;
(4) dry the precipitation of taking in step (3) is dry after, recycling pulverizer is ground into powder;
(5) degreasing adds sherwood oil or acetone to carry out skimming treatment 6～10h in the resulting powder of step (4); Wherein, the consumption of described sherwood oil or acetone is powder quality 4～8 times; After degreasing, remove organic solvent, collect solid materials, dry, obtain the pressed powder after degreasing;
(6) enzymolysis is placed in enzymatic vessel by the pressed powder after resulting degreasing in step (5), adds the pure water of 20～45 times of pressed powder quality, and fully swelling, is heated to 100 ℃ and keeps 20～30min, killing microorganisms; Be cooled to 40～55 ℃, add the proteolytic ferment of pressed powder quality 3～8 ‰, carry out enzymolysis 2～5h; Wherein, described proteolytic ferment is one or more the combination in compound protease, food flavor enzyme, papoid, Sumizyme MP; After enzymolysis, be heated to 100 ℃ of maintenances enzyme 20～30min that goes out, obtain scallop enzymolysis solution;
(7) centrifugal, ultrafiltration is cooled to 2～5 ℃ by (6) resulting scallop enzymolysis solution, sends in whizzer, centrifugal, collects supernatant liquor; The filter membrane that is 3000Da by molecular weight cut-off by supernatant liquor carries out ultrafiltration, collects trapped fluid; Trapped fluid is added to pure water, then carry out ultrafiltration 3～4 times, remove salinity in trapped fluid;
(8) dry by the trapped fluid lyophilize after the desalination obtaining in step (7), obtain scallop glycoprotein dry product.
The present invention is that to choose the fresh blanching solution that scallop is processed to form be raw material, through concentrating under reduced pressure, and alcohol precipitation, centrifugal, after drying precipitate, make powder; Powder adds sherwood oil or acetone to carry out skimming treatment, dry, obtains pressed powder and adds proteolytic ferment to carry out enzymolysis, and enzyme goes out; Scallop enzymolysis solution is cooled to low temperature, centrifugal, collects supernatant liquor ultrafiltration desalination; Trapped fluid lyophilize, obtains scallop glycoprotein dry product.The present invention utilizes the blanching solution that scallop is processed to form to extract glycoprotein, and its raw materials cost is low; After alcohol precipitation, adopt sherwood oil or acetone to carry out skimming treatment, it is high that it obtains thing purity; And through proteolytic ferment, carry out enzymolysis, the one, increase extract resolvability and improve bioavailability; The 2nd, it is higher that the protein matter in removal extract makes to extract glycoprotein analog material purity; Ultrafiltration adopts the filter membrane of 3000Da, and it removes the material such as the oligopeptide in solution and amino acids after enzymolysis in desalination.Adopt method of the present invention to extract separated scallop glycoprotein, after measured: polysaccharide content is between 10-20%, and protein content is between 50-60%.Through experimentation on animals, confirm, it has good auxiliary anti-tumor activity.Extracting method of the present invention, its technique is reasonable, and equipment investment is low, advanced technology, operation feasible, is applicable to large-scale industrialized production.
Embodiment
Below in conjunction with embodiment, the present invention will be further described.
embodiment 1
Utilize a method for scallop blanching solution extraction glycoprotein, process following process steps:
(1) raw material is chosen and is processed that to choose the fresh blanching solution that scallop is processed to form be raw material, utilizes concentrating under reduced pressure equipment, is concentrated into solid content 37%, obtains concentrated solution;
(2) alcohol precipitation is that 93% ethanolic soln precipitates by adding concentration in the concentrated solution of step (1) gained, to its final concentration be 60%;
(3) centrifugal solution after step (2) ethanol precipitation is sent into link-suspended basket centrifuge, control rotating speed 2000rpm, centrifugal, collecting precipitation;
(4) dry the precipitation of taking in step (3) is dry after, recycling pulverizer is ground into powder;
(5) degreasing adds sherwood oil to carry out skimming treatment 8h in the resulting powder of step (4); Wherein, the consumption of described sherwood oil is powder quality 6 times; Centrifugal removal sherwood oil after degreasing, collects solid materials, and vacuum-drying, obtains the pressed powder after degreasing;
(6) enzymolysis is placed in enzymatic vessel by the pressed powder after resulting degreasing in step (5), adds the pure water of 30 times of pressed powder quality, and fully swelling, is heated to 100 ℃ and keeps 25min, killing microorganisms; Be cooled to 45 ℃, add the conjugated protein lytic enzyme of pressed powder quality 6 ‰, enzymolysis 1h, then adds the food flavor enzyme enzymolysis 2.5h of pressed powder quality 4 ‰; After enzymolysis, be heated to 100 ℃ of maintenances enzyme 25min that goes out, obtain scallop enzymolysis solution; Wherein, conjugated protein lytic enzyme, food flavor enzyme enzymolysis are commercially available prod;
(7) centrifugal, ultrafiltration is cooled to 4 ℃ by (6) resulting scallop enzymolysis solution, sends into link-suspended basket centrifuge, controls rotating speed 2000rpm, centrifugal, collects supernatant liquor; The filter membrane that is 3000Da by molecular weight cut-off by supernatant liquor carries out ultrafiltration, collects trapped fluid; Trapped fluid is added to pure water, then carry out ultrafiltration 3 times, remove salinity in trapped fluid;
(8) dry by the trapped fluid lyophilize after the desalination obtaining in step (7); Wherein, material temperature is controlled at-22 ℃, and plate temperature control is at 40 ℃, and vacuum degree control is at 100Pa; Obtain scallop glycoprotein dry product.
The scallop blanching solution that utilizes that the present embodiment is given extracts the method for glycoprotein, and its technique is reasonable, and equipment investment is low, advanced technology, operation feasible.It extracts separated scallop glycoprotein, after measured: polysaccharide content is 19.6%, and protein content is 57.9%.Through experimentation on animals, confirm, it has good auxiliary anti-tumor activity.
embodiment 2
Utilize a method for scallop blanching solution extraction glycoprotein, process following process steps:
(1) raw material is chosen and is processed that to choose the fresh blanching solution that scallop is processed to form be raw material, utilizes concentrating under reduced pressure equipment, is concentrated into solid content 40%, obtains concentrated solution;
(2) alcohol precipitation is that 95% ethanolic soln precipitates by adding concentration in the concentrated solution of step (1) gained, to its final concentration be 65%;
(3) centrifugal solution after step (2) ethanol precipitation is sent into link-suspended basket centrifuge, control rotating speed 1500rpm, centrifugal, collecting precipitation;
(4) dry the precipitation of taking in step (3) is dry after, recycling pulverizer is ground into powder;
(5) degreasing adds acetone to carry out skimming treatment 10h in the resulting powder of step (4); Wherein, the consumption of described acetone is powder quality 4 times; After degreasing, suction filtration is removed acetone, collects solid materials, and vacuum-drying, obtains the pressed powder after degreasing;
(6) enzymolysis is placed in enzymatic vessel by the pressed powder after resulting degreasing in step (5), adds the pure water of 45 times of pressed powder quality, and fully swelling, is heated to 100 ℃ and keeps 30min, killing microorganisms; Be cooled to 55 ℃, add Sumizyme MP 40g hydrolysis 2h, add the Sumizyme MP of pressed powder quality 8 ‰, enzymolysis 2h; Then add pressed powder quality 3 ‰ papoids, enzymolysis 3h; After enzymolysis, be heated to 100 ℃ of maintenances enzyme 30min that goes out, obtain scallop enzymolysis solution; Wherein, Sumizyme MP, papoid are commercially available prod;
(7) centrifugal, ultrafiltration is cooled to 2 ℃ by (6) resulting scallop enzymolysis solution, sends in link-suspended basket centrifuge, controls rotating speed 1800rpm, centrifugal, collects supernatant liquor; The filter membrane that is 3000Da by molecular weight cut-off by supernatant liquor carries out ultrafiltration, collects trapped fluid; Trapped fluid is added to pure water, then carry out ultrafiltration 4 times, remove salinity in trapped fluid;
(8) dry by the trapped fluid lyophilize after the desalination obtaining in step (7); Wherein, control material temperature-25 ℃, plate temperature control is at 50 ℃, and vacuum degree control is at 120Pa; Obtain scallop glycoprotein dry product.
The scallop blanching solution that utilizes that the present embodiment is given extracts the method for glycoprotein, and its technique is reasonable, and equipment investment is low, advanced technology, operation feasible.It extracts separated scallop glycoprotein, after measured: polysaccharide content is 16.9%, and protein content is 55.4%.Through experimentation on animals, confirm, it has good auxiliary anti-tumor activity.
embodiment 3
Utilize a method for scallop blanching solution extraction glycoprotein, process following process steps:
(1) raw material is chosen and is processed that to choose the fresh blanching solution that scallop is processed to form be raw material, utilizes concentrating under reduced pressure equipment, is concentrated into solid content 35%, obtains concentrated solution;
(2) alcohol precipitation is that 90% ethanolic soln precipitates by adding concentration in the concentrated solution of step (1) gained, to its final concentration be 55%;
(3) centrifugal solution after step (2) ethanol precipitation is sent into link-suspended basket centrifuge, control rotating speed 1500rpm, centrifugal, collecting precipitation;
(4) dry the precipitation of taking in step (3) is dry after, recycling pulverizer is ground into powder;
(5) degreasing adds sherwood oil to carry out skimming treatment 6h in the resulting powder of step (4); Wherein, the consumption of described sherwood oil is powder quality 8 times; Centrifugal removal sherwood oil after degreasing, collects solid materials, and vacuum-drying, obtains the pressed powder after degreasing;
(6) enzymolysis is placed in enzymatic vessel by the pressed powder after resulting degreasing in step (5), adds the pure water of 20 times of pressed powder quality, and fully swelling, is heated to 100 ℃ and keeps 20min, killing microorganisms; Be cooled to 40 ℃, add the food flavor enzyme of pressed powder quality 3 ‰, enzymolysis 1.5h; Then the papoid that adds pressed powder quality 8 ‰, enzymolysis 1h; After enzymolysis, be heated to 100 ℃ of maintenances enzyme 20min that goes out, obtain scallop enzymolysis solution; Wherein, food flavor enzyme, papoid are commercially available prod;
(7) centrifugal, ultrafiltration is cooled to 5 ℃ by (6) resulting scallop enzymolysis solution, then sends in link-suspended basket centrifuge, controls rotating speed 2000rpm, centrifugal, collects supernatant liquor; The filter membrane that is 3000Da by molecular weight cut-off by supernatant liquor carries out ultrafiltration, collects trapped fluid; Trapped fluid is added to pure water, then carry out ultrafiltration 3 times, remove salinity in trapped fluid;
(8) dry by the trapped fluid lyophilize after the desalination obtaining in step (7); Wherein, material temperature-20 ℃, plate temperature control is at 30 ℃, and vacuum degree control is 90; Control to obtain scallop glycoprotein dry product.
The scallop blanching solution that utilizes that the present embodiment is given extracts the method for glycoprotein, and its technique is reasonable, and equipment investment is low, advanced technology, operation feasible.It extracts separated scallop glycoprotein, after measured: polysaccharide content is 18.3%, and protein content is 57.2%.Through experimentation on animals, confirm, it has good auxiliary anti-tumor activity.
Claims (1)
1. utilize scallop blanching solution to extract a method for glycoprotein, it is characterized in that: process following process steps:
(1) raw material is chosen and is processed that to choose the fresh blanching solution that scallop is processed to form be raw material, utilizes concentrating under reduced pressure equipment, is concentrated into solid content 35%～40%, obtains concentrated solution;
(2) alcohol precipitation is that 90～95% ethanolic solns precipitate by adding concentration in the concentrated solution of step (1) gained, to its final concentration be 55%～65%;
(3) centrifugal solution after step (2) ethanol precipitation is sent in whizzer, centrifugal, collecting precipitation;
(4) dry the precipitation of taking in step (3) is dry after, recycling pulverizer is ground into powder;
(5) degreasing adds sherwood oil or acetone to carry out skimming treatment 6～10h in the resulting powder of step (4); Wherein, the consumption of described sherwood oil or acetone is powder quality 4～8 times; After degreasing, remove organic solvent, collect solid materials, dry, obtain the pressed powder after degreasing;
(6) enzymolysis is placed in enzymatic vessel by the pressed powder after resulting degreasing in step (5), adds the pure water of 20～45 times of pressed powder quality, and fully swelling, is heated to 100 ℃ and keeps 20～30min, killing microorganisms; Be cooled to 40～55 ℃, add the proteolytic ferment of pressed powder quality 3～8 ‰, carry out enzymolysis 2～5h; Wherein, described proteolytic ferment is one or more the combination in compound protease, food flavor enzyme, papoid, Sumizyme MP; After enzymolysis, be heated to 100 ℃ of maintenances enzyme 20～30min that goes out, obtain scallop enzymolysis solution;
(7) centrifugal, ultrafiltration is cooled to 2～5 ℃ by (6) resulting scallop enzymolysis solution, sends in whizzer, centrifugal, collects supernatant liquor; The filter membrane that is 3000Da by molecular weight cut-off by supernatant liquor carries out ultrafiltration, collects trapped fluid; Trapped fluid is added to pure water, then carry out ultrafiltration 3～4 times, remove salinity in trapped fluid;
(8) dry by the trapped fluid lyophilize after the desalination obtaining in step (7), obtain scallop glycoprotein dry product.
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Cited By (3)
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| CN104497117A (en) * | 2014-11-27 | 2015-04-08 | 山东好当家海洋发展股份有限公司 | Method for extracting glycoprotein from oyster water boiled liquid |
| CN104610423A (en) * | 2015-02-06 | 2015-05-13 | 山东省科学院生物研究所 | Method for extracting glycoprotein by utilizing trepang processing blanching liquid |
| CN105087736A (en) * | 2015-09-28 | 2015-11-25 | 刘媛 | Argopecten irradias polypeptide and application thereof |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104497117A (en) * | 2014-11-27 | 2015-04-08 | 山东好当家海洋发展股份有限公司 | Method for extracting glycoprotein from oyster water boiled liquid |
| CN104610423A (en) * | 2015-02-06 | 2015-05-13 | 山东省科学院生物研究所 | Method for extracting glycoprotein by utilizing trepang processing blanching liquid |
| CN104610423B (en) * | 2015-02-06 | 2018-06-19 | 山东省科学院生物研究所 | Utilize the method for Holothurian machining blanching solution extraction glycoprotein |
| CN105087736A (en) * | 2015-09-28 | 2015-11-25 | 刘媛 | Argopecten irradias polypeptide and application thereof |
| CN105087736B (en) * | 2015-09-28 | 2018-12-21 | 刘媛 | A kind of bay scallop polypeptide and its application |
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Application publication date: 20140416 |