CN105039480A - Method for purifying collagen derived from pelodiscus sinensis - Google Patents
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Abstract
The invention relates to a method for purifying collagen derived from pelodiscus sinensis. The method is characterized by comprising the steps of soaking, fat removal and enzymolysis; in an enzymolysis process, carinas of the pelodiscus sinensis, of which impurities are removed, are put into acetic acid with the concentration of 0.5mol/L, wherein the ratio of the carinas to the acetic acid is 1:25, pepsase enzymatic hydrolysate is added for enzymolysis treatment, and extraction is carried out at 4-10 DEG C for 24-48 h, so that a pelodiscus sinensis carina collagen coarse extracting solution is obtained; in a salting-out purification process, the coarse extracting solution is dissolved into a Tris-HCL buffer solution containing NaCl, the mixed solution is centrifuged to obtain supernatant, NaCl is added into the supernatant, salting-out is carried out, centrifugation is carried out again, and precipitates and the supernatant are respectively collected; and precipitates are dissolved into acetic acid with the concentration of 0.5 mol/L, and freeze-drying is carried out after dialysis, so that the pelodiscus sinensis collagen freeze-dried product is obtained. Compared with the prior art, the method provided by the invention has the advantages that collagen extracted through enzymolysis is purified through a salting-out purification method, so that no environment pollution is realized, the extraction efficiency is high and the obtained product has good biological characteristics.
Description
Technical field
The present invention relates to a kind of purification process of Trionyx sinensis (Wiegmann) derived collagen albumen, belong to natural active matter field.
Background technology
Collagen protein is structural protein important in organism, is the chief component of sustentacular tissue and reticular tissue.The amino acid structure of triple helices specific to collagen protein makes it have many very useful characteristics, as the performance of high tensile, good biocompatibility, low antigenicity, low irritant and low cytotoxicity and Promote cell's growth.These characteristics had all become a kind of desirable bio-medical material.With collagen protein the bio-medical material that derives by raw material, become one of the fastest growth point of field of medical materials, world market annual sales amount has reached more than 40 hundred million dollars.
Tradition extracts the raw material that uses of collagen protein mainly from the reticular tissue such as skin of the animal such as pig and ox.But due to problems such as mad cow disease, bird flu, foot and mouth disease, the national collagen product to animal tissues source carries out strict control, and the Chinese government forbids all with the import collagen product being organized as raw material extraction of ox at present.The biological collagen finding new source is the important topic that current urgent need will solve.Compared with terrestrial animal, even if aquatic animal collagen also dissolves in neutral salt solution or acidic solution at low temperatures, than being easier to obtained soluble collagen solution, this provides favourable condition for its utilization.
Current people have extracted collagen protein from multiple fishery products, and relevant report also gets more and more.But for the nutritious tonifying good merchantable brand that China is traditional, being inherently rich in the aquatic animal of collagen protein---Trionyx sinensis (Wiegmann) but compares shortage as the research of new bio collagen sources.Trionyx sinensis (Wiegmann) (Pelodiscussinensis), is commonly called as soft-shelled turtle, is exactly the traditional excellent tonic product of China from ancient times, has very high nutritive value and health-care effect.Knot around turtle shell is formed soft tissue and is commonly called " shirt rim ", collagen content is very abundant, premenstruum, experiment showed, its dry weight content is more than 70%, by literature search, be only limitted to the report of its separation and Extraction and structural characterization about the research of collagen protein in Trionyx sinensis (Wiegmann), but then have no report about the purifying process of soft-shelled turtle derived collagen albumen and biology performance research.
Summary of the invention
Technical problem to be solved by this invention is the purification process providing a kind of purifying process simple Trionyx sinensis (Wiegmann) derived collagen albumen for above-mentioned prior art.
The present invention solves the problems of the technologies described above adopted technical scheme: the purification process of this Trionyx sinensis (Wiegmann) derived collagen albumen, is characterized in that: comprise the following steps:
(1) soak: get methods of soft-shell turtle skirt homogenate and smash, soak in saline solution by alkalescence and at 4 DEG C ~ 10 DEG C, place 24h ~ 48h, distilled water repetitive scrubbing, fully drain with gauze rear for subsequent use;
(2) degrease: after step (1) is soaked, his material has been collected and has been soaked 24h ~ 48h to my 10% ~ 15% aqueous isopropanol in 4 DEG C ~ 10 DEG C again, and to remove fat, distilled water repetitive scrubbing, fully drains rear for subsequent use with gauze;
(3) enzymolysis: the methods of soft-shell turtle skirt of impurity elimination is placed in 0.5mol/L acetic acid, solid-liquid ratio 1:25, adds in stomach en-enzymolysis solution and carries out enzymolysis processing, extracts 24h ~ 48h at 4 DEG C ~ 10 DEG C, obtains methods of soft-shell turtle skirt collagen protein crude extract;
(4) to saltout purifying: the crude extract getting step (3) is dissolved in the Tris-HCl damping fluid containing NaCl, and centrifuging and taking supernatant liquor, adds NaCl in supernatant liquor, saltout, recentrifuge, respectively collecting precipitation and supernatant liquor; To be precipitated and dissolved in the acetic acid of 0.5mol/L, after dialysis, lyophilize, obtains Trionyx sinensis (Wiegmann) collagen protein dried frozen aquatic products.
Further, have studied time, NaCl concentration, collagen concentration three factors respectively to the impact of the collagen protein rate of recovery in the purge process of saltouing of described step (4), wherein, research method for time factor is: the described crude extract getting 10mL, all dilute crude extract collagen concentration to 10g/L, and add 3mol/LNaCl all wherein, saltout 1h, 12h, 24h, 36h, 48h at 4 DEG C; And for the research method of NaCl concentration factor be: the described crude extract getting 10mL, adds the NaCl of 1mol/L, 2mol/L, 3mol/L, 4mol/L, 5mol/L respectively, and saltout 24h at 4 DEG C; And for the research method of collagen concentration factor be: the described crude extract getting 10mL, dilute crude extract collagen concentration respectively to 2g/L, 4g/L, 6g/L, 8g/L, 10g/L, add 3mol/LNaCl all wherein, saltout 24h at 4 DEG C.Research emphasis of the present invention is to have studied collagen concentration first for separation and purification role, for the factor that other purification experiment is of little use, reason is: collagen concentration is too high will cause coagulation, need a best to saltout scope, during neutral salt precipitating proteins, the actual concentrations of Proteins In Aqueous Solutions has larger impact to the effect be separated, also combine time and NaCl concentration two factors simultaneously, with by the research to time, NaCl concentration, collagen concentration three factors, to determine best separation and purification effect.
As preferably, described in the best of breed of saltouing be: the time is 24h, NaCl concentration is 3mol/L, and collagen concentration is 8g/L.
Further, the dialysis procedure of described step (4) is specially: by described be precipitated and dissolved in acetic acid after, the dialysis tubing being respectively 25KD, 50KD, 100KD with molecular weight cut-off is dialysed, dialyzate is pure water, dialyse respectively 24h and 48h at 4 DEG C ~ 10 DEG C, change water between dialysis period 4 ~ 5 times, the collagen solution getting each dialysis time carries out SDS-PAGE and detects purity, and the collagen solution lyophilize after dialysis is obtained collagen protein dried frozen aquatic products.
Further, in described step (1), the consumption that described alkalescence soaks saline solution is 2 times ~ 4 times of described methods of soft-shell turtle skirt quality.
Further, described alkalescence is soaked containing NaCl and NaOH in saline solution, and the massfraction that described alkalescence soaks NaCl in saline solution is 2% ~ 4%, and the concentration that described alkalescence soaks NaOH in saline solution is 0.5% ~ 1.5%.
Further, in described step (3), the temperature of enzymolysis processing is 30 DEG C ~ 50 DEG C, and the pH value of enzymolysis processing is 2 ~ 4, and the time of enzymolysis processing is 8h ~ 12h.
Compared with prior art, the invention has the advantages that:
1, the present invention take methods of soft-shell turtle skirt as the collagen protein that raw material production can be widely used in the fields such as food, medicine, makeup, biomaterial, both extend industrial chains, improves added value of product, increases economic and social benefit;
2, collagen protein exists with the form of collegen filament in body, has higher stability.The extraction of tradition collagen protein adopts the methods such as salt, acid, alkali, hot water mostly, there is extraction efficiency lower, and environmental pollution is large, the problems such as unstable product quality.The present invention utilizes the extraction collagen protein of enzymolysis and carries out purifying by the method for purifying of saltouing, not only non-environmental-pollution, extraction efficiency is high, and products obtained therefrom has good biological characteristics, and the dialysis tubing comparing different size molecular retention amount is to the purification effect of collagen protein crude extract, be intended to for identifying the physico-chemical property of collagen protein sterling further from now on, and and the collagen proteins in other sources compare, simultaneously for comprehensive development and utilization soft-shelled turtle derived collagen albumen prepares the theoretical foundation that biological medicine material provides certain.
Accompanying drawing explanation
Fig. 1 be in the embodiment of the present invention NaCl concentration on the impact of the collagen protein rate of recovery;
Fig. 2 be in the embodiment of the present invention collagen concentration on the impact of the collagen protein rate of recovery;
Fig. 3 be in the embodiment of the present invention time on the impact of the collagen protein rate of recovery;
Fig. 4 is the electrophoretogram that in the embodiment of the present invention, methods of soft-shell turtle skirt collagen protein is saltoutd;
Fig. 5 is that (A, a swimming lane is respectively the purification effect of dialysis tubing dialysis 24h, 48h with molecular weight cut-off 25KD to the electrophoretogram that in the embodiment of the present invention, methods of soft-shell turtle skirt collagen protein is dialysed; B, b swimming lane is respectively the purification effect of dialysis tubing dialysis 24h, 48h with molecular weight cut-off 50KD; C, c swimming lane is respectively the purification effect of dialysis tubing dialysis 24h, 48h with molecular weight cut-off 100KD).
Embodiment
Below in conjunction with accompanying drawing embodiment, the present invention is described in further detail.
Embodiment 1
The purification process of Trionyx sinensis (Wiegmann) derived collagen albumen, comprises the following steps:
(1) soak: get methods of soft-shell turtle skirt homogenate and smash, soak in saline solution by alkalescence and at 4 DEG C, place 24h, distilled water repetitive scrubbing, fully drain with gauze rear for subsequent use;
(2) degrease: the methods of soft-shell turtle skirt after step (1) being soaked cleans, add 10% aqueous isopropanol and soak 24h in 4 DEG C, to remove fat, distilled water repetitive scrubbing, fully drains rear for subsequent use with gauze;
(3) enzymolysis: the methods of soft-shell turtle skirt of impurity elimination is placed in 0.5mol/L acetic acid, solid-liquid ratio 1:25, adds in stomach en-enzymolysis solution and carries out enzymolysis processing, extracts 24h at 4 DEG C, obtains methods of soft-shell turtle skirt collagen protein crude extract;
(4) to saltout purifying: the crude extract getting step (3) is dissolved in the Tris-HCl damping fluid containing NaCl, and centrifuging and taking supernatant liquor, adds NaCl in supernatant liquor, saltout, recentrifuge, respectively collecting precipitation and supernatant liquor; To be precipitated and dissolved in the acetic acid of 0.5mol/L, after dialysis, lyophilize, obtains Trionyx sinensis (Wiegmann) collagen protein dried frozen aquatic products.
Embodiment 2
The purification process of Trionyx sinensis (Wiegmann) derived collagen albumen, comprises the following steps:
(1) soak: get methods of soft-shell turtle skirt homogenate and smash, soak in saline solution by alkalescence and at 10 DEG C, place 48h, distilled water repetitive scrubbing, fully drain with gauze rear for subsequent use;
(2) degrease: the methods of soft-shell turtle skirt after step (1) being soaked cleans, add 15% aqueous isopropanol and soak 48h in 10 DEG C, to remove fat, distilled water repetitive scrubbing, fully drains rear for subsequent use with gauze;
(3) enzymolysis: the methods of soft-shell turtle skirt of impurity elimination is placed in 0.5mol/L acetic acid, solid-liquid ratio 1:25, adds in stomach en-enzymolysis solution and carries out enzymolysis processing, extracts 48h at 10 DEG C, obtains methods of soft-shell turtle skirt collagen protein crude extract;
(4) to saltout purifying: the crude extract getting step (3) is dissolved in the Tris-HCl damping fluid containing NaCl, and centrifuging and taking supernatant liquor, adds NaCl in supernatant liquor, saltout, recentrifuge, respectively collecting precipitation and supernatant liquor; To be precipitated and dissolved in the acetic acid of 0.5mol/L, after dialysis, lyophilize, obtains Trionyx sinensis (Wiegmann) collagen protein dried frozen aquatic products.
Embodiment 3
The purification process of Trionyx sinensis (Wiegmann) derived collagen albumen, comprises the following steps:
(1) soak: get methods of soft-shell turtle skirt homogenate and smash, soak in saline solution by alkalescence and at 8 DEG C, place 30h, distilled water repetitive scrubbing, fully drain with gauze rear for subsequent use;
(2) degrease: the methods of soft-shell turtle skirt after step (1) being soaked cleans, add 12% aqueous isopropanol and soak 30h in 8 DEG C, to remove fat, distilled water repetitive scrubbing, fully drains rear for subsequent use with gauze;
(3) enzymolysis: the methods of soft-shell turtle skirt of impurity elimination is placed in 0.5mol/L acetic acid, solid-liquid ratio 1:25, adds in stomach en-enzymolysis solution and carries out enzymolysis processing, extracts 30h at 8 DEG C, obtains methods of soft-shell turtle skirt collagen protein crude extract;
(4) to saltout purifying: the crude extract getting step (3) is dissolved in the Tris-HCl damping fluid containing NaCl, and centrifuging and taking supernatant liquor, adds NaCl in supernatant liquor, saltout, recentrifuge, respectively collecting precipitation and supernatant liquor; To be precipitated and dissolved in the acetic acid of 0.5mol/L, after dialysis, lyophilize, obtains Trionyx sinensis (Wiegmann) collagen protein dried frozen aquatic products.
Embodiment 4 is saltoutd single factor experiment-NaCl concentration
NaCl concentration is on the experimental technique of the impact of the collagen protein rate of recovery: get 10mL crude extract, add 1mol/L, 2mol/L, 3mol/L, 4mol/L, 5mol/LNaCl respectively, and saltout 24h at 4 DEG C;
As shown in Figure 1, NaCl concentration is on the impact of the collagen protein rate of recovery: when NaCl concentration is 1mol/L-3mol/L, the collagen protein rate of recovery increases with the increase of NaCl concentration, increases slowly after 2mol/L, maximum value is reached at 3mol/L, there is slow downtrending after 3mol/L, significantly decline after 4mol/L.This may be due to NaCl concentration lower time, along with the increase of NaCl concentration, the increase of salt ionic concentration in solution, therefore, the rate of recovery of collagen protein also can increase; Will cause the coagulation of collagen protein when salt ionic concentration is excessive in solution, then increase NaCl concentration, the collagen protein rate of recovery will decline, and NaCl excessive concentration, may affect the activity of collagen protein.According to the collagen protein rate of recovery in figure with the variation tendency of NaCl concentration, NaCl concentration 3mol/L is selected to be optimum point.
Embodiment 5 is saltoutd single factor experiment-NaCl concentration
NaCl concentration is on the experimental technique of the impact of the collagen protein rate of recovery: get 10mL crude extract, add 1mol/L, 2mol/L, 3mol/L, 4mol/L, 5mol/LNaCl respectively, and saltout 24h at 4 DEG C;
Result as shown in Figure 1, NaCl concentration is on the impact of the collagen protein rate of recovery: when NaCl concentration is 1mol/L-3mol/L, the collagen protein rate of recovery increases with the increase of NaCl concentration, increases slowly after 2mol/L, maximum value is reached at 3mol/L, there is slow downtrending after 3mol/L, significantly decline after 4mol/L.This may be due to NaCl concentration lower time, along with the increase of NaCl concentration, the increase of salt ionic concentration in solution, therefore, the rate of recovery of collagen protein also can increase; Will cause the coagulation of collagen protein when salt ionic concentration is excessive in solution, then increase NaCl concentration, the collagen protein rate of recovery will decline, and NaCl excessive concentration, may affect the activity of collagen protein.According to the collagen protein rate of recovery in figure with the variation tendency of NaCl concentration, NaCl concentration 3mol/L is selected to be optimum point.
Embodiment 6 is saltoutd single factor experiment-collagen concentration
Collagen concentration is on the experimental technique of the impact of the collagen protein rate of recovery: get 10mL crude extract, and dilute crude extract collagen concentration respectively to 2g/L, 4g/L, 6g/L, 8g/L, 10g/L, add 3mol/LNaCl all wherein, saltout 24h at 4 DEG C;
Result is as shown in Figure 2: when collagen concentration is 2g/L-8g/L, the collagen protein rate of recovery increases with the increase of collagen concentration, and rising tendency is remarkable, reaches maximum value when 8g/L; Significantly decline after 8g/L, this may be because collagen concentration is higher, and caused by salt concn is enough to collagen protein to saltout completely not.According to the collagen protein rate of recovery in figure with the variation tendency of collagen concentration, selection collagen concentration 8g/L is optimum point.
Embodiment 7 is saltoutd single factor experiment-time
Time is on the experimental technique of the impact of the collagen protein rate of recovery: get 10mL crude extract, and all dilute crude extract collagen concentration to 10g/L, and add 3mol/LNaCl all wherein, saltout 1h, 12h, 24h, 36h, 48h at 4 DEG C;
Result is as shown in Figure 3: at 1h-24h, collagen protein rate of recovery prolongation in time and increasing, this is because collagen protein needs time enough to saltout; Significantly decline after 24h, this may be because collagen protein is under long salting-out condition, can cause the sex change of collagen protein.According to collagen protein rate of recovery trend over time in figure, select time 24h is optimum point.
Embodiment 8 is saltoutd orthogonal test
On single factor experiment basis, investigate NaCl concentration, collagen concentration, time has been research object, devise the orthogonal test of 3 factor 3 levels, with the collagen protein rate of recovery for index (see table 1), purification effect of saltouing is detected with SDS-PAGE, resolving gel concentration is 8%, and concentrated gum concentration is 5%.
Table 1 orthogonal test designs table
Tab.1Orthogonaldesignofexperiment
Table 2 orthogonal experiments
Tab.2Resultoforthogonalexperiment
As shown in Table 2, three kinds of factor extreme difference sizes are followed successively by RB > RC > RA, namely NaCl concentration has the greatest impact to the collagen protein rate of recovery, secondly be collagen concentration, time effects effect is minimum, optimum extraction conditions is: A2B1C2, namely the time is 24h, NaCl concentration is 2mol/L, and when collagen concentration is 8g/L, the collagen protein rate of recovery is maximum.Under this final condition, done three groups of parallel laboratory tests, obtaining the collagen protein rate of recovery is 81.22%, 80.65%, 82.31%, and visible optimal conditions is feasible.
Embodiment 9 crude extract dialysis purifying
Get 10mL crude extract and be placed in the dialysis tubing that molecular weight cut-off is 25KD, 50KD, 100KD respectively, dialyzate is pure water, and dialyse 24h, 48h at 4 DEG C, and the collagen solution getting each time carries out SDS-PAGE detection purity, gum concentration is: resolving gel concentration is 8%, and concentrated gum concentration is 5%.
By the collagen protein of Fig. 4 SDS-PAGE collection of illustrative plates of saltouing, result can find out that methods of soft-shell turtle skirt collagen protein crude extract can obtain obvious purifying after oversalting, and methods of soft-shell turtle skirt collagen protein crude extract mix band neither be a lot, more demonstrate collagen content in methods of soft-shell turtle skirt very high.
Can find out that the dialysis tubing of 100KD is best relative to the purification effect of the dialysis tubing of other two kinds of specifications by Fig. 5 collagen protein dialysis SDS-PAGE collection of illustrative plates, and the time also has a certain impact to purification effect, the purification effect of 48h is better than 24h, and compared to saltouing, its purification effect is better.Visible, directly purification process carried out to crude extract with dialysis tubing simple and convenient, economical; Collagen protein of saltouing, this step of dialysis desalination can also be saved, decrease workload and shorten test period.
Claims (7)
1. a purification process for Trionyx sinensis (Wiegmann) derived collagen albumen, is characterized in that: comprise the following steps:
(1) soak: get methods of soft-shell turtle skirt homogenate and smash, soak in saline solution by alkalescence and at 4 DEG C ~ 10 DEG C, place 24h ~ 48h, distilled water repetitive scrubbing, fully drain with gauze rear for subsequent use;
(2) degrease: the methods of soft-shell turtle skirt after step (1) being soaked cleans, add 10% ~ 15% aqueous isopropanol and soak 24h ~ 48h in 4 DEG C ~ 10 DEG C, to remove fat, distilled water repetitive scrubbing, fully drains rear for subsequent use with gauze;
(3) enzymolysis: the methods of soft-shell turtle skirt of impurity elimination is placed in 0.5mol/L acetic acid, solid-liquid ratio 1:25, adds in stomach en-enzymolysis solution and carries out enzymolysis processing, extracts 24h ~ 48h at 4 DEG C ~ 10 DEG C, obtains methods of soft-shell turtle skirt collagen protein crude extract;
(4) to saltout purifying: the crude extract getting step (3) is dissolved in the Tris-HCl damping fluid containing NaCl, and centrifuging and taking supernatant liquor, adds NaCl in supernatant liquor, saltout, recentrifuge, respectively collecting precipitation and supernatant liquor; To be precipitated and dissolved in the acetic acid of 0.5mol/L, after dialysis, lyophilize, obtains Trionyx sinensis (Wiegmann) collagen protein dried frozen aquatic products.
2. the purification process of Trionyx sinensis (Wiegmann) derived collagen albumen according to claim 1, it is characterized in that: have studied time, NaCl concentration, collagen concentration three factors in the purge process of saltouing of described step (4) respectively to the impact of the collagen protein rate of recovery, wherein, research method for time factor is: the described crude extract getting 10mL, all dilute crude extract collagen concentration to 10g/L, and add 3mol/LNaCl all wherein, saltout 1h, 12h, 24h, 36h, 48h at 4 DEG C; And for the research method of NaCl concentration factor be: the described crude extract getting 10mL, adds the NaCl of 1mol/L, 2mol/L, 3mol/L, 4mol/L, 5mol/L respectively, and saltout 24h at 4 DEG C; And for the research method of collagen concentration factor be: the described crude extract getting 10mL, dilute crude extract collagen concentration respectively to 2g/L, 4g/L, 6g/L, 8g/L, 10g/L, add 3mol/LNaCl all wherein, saltout 24h at 4 DEG C.
3. the purification process of Trionyx sinensis (Wiegmann) derived collagen albumen according to claim 2, is characterized in that: described in the best of breed of saltouing be: the time is 24h, NaCl concentration is 3mol/L, and collagen concentration is 8g/L.
4. the purification process of Trionyx sinensis (Wiegmann) derived collagen albumen according to claim 1, it is characterized in that: the dialysis procedure of described step (4) is specially: by described be precipitated and dissolved in acetic acid after, the dialysis tubing being respectively 25KD, 50KD, 100KD with molecular weight cut-off is dialysed, dialyzate is pure water, dialyse respectively 24h and 48h at 4 DEG C ~ 10 DEG C, water is changed 4 ~ 5 times between dialysis period, the collagen solution getting each dialysis time carries out SDS-PAGE and detects purity, and the collagen solution lyophilize after dialysis is obtained collagen protein dried frozen aquatic products.
5. the purification process of Trionyx sinensis (Wiegmann) derived collagen albumen according to claim 1, is characterized in that: in described step (1), and the consumption that described alkalescence soaks saline solution is 2 times ~ 4 times of described methods of soft-shell turtle skirt quality.
6. the purification process of Trionyx sinensis (Wiegmann) derived collagen albumen according to claim 5, it is characterized in that: described alkalescence is soaked in saline solution containing NaCl and NaOH, the massfraction that described alkalescence soaks NaCl in saline solution is 2% ~ 4%, and the concentration that described alkalescence soaks NaOH in saline solution is 0.5% ~ 1.5%.
7. the purification process of Trionyx sinensis (Wiegmann) derived collagen albumen according to claim 1, is characterized in that: in described step (3), the temperature of enzymolysis processing is 30 DEG C ~ 50 DEG C, and the pH value of enzymolysis processing is 2 ~ 4, and the time of enzymolysis processing is 8h ~ 12h.
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| CN107034259A (en) * | 2017-06-21 | 2017-08-11 | 江苏省农业科学院 | A kind of preparation method of high thermal stability Shelled Turtle Trionyx Sinensis byproduct collagen albumen |
| CN109221391A (en) * | 2018-10-24 | 2019-01-18 | 浙江卫斯敦环境科技有限公司 | A kind of compound fruit and vegetable fresh-keeping film-coating and preparation method thereof |
| CN113234659A (en) * | 2021-05-28 | 2021-08-10 | 浙江万里学院 | Method for in vitro culture of pelodioblasts at embryo stage of Chinese softshell turtle and construction of damage model |
| CN117024571A (en) * | 2023-07-31 | 2023-11-10 | 中科国康(浙江)生命科学有限公司 | System and method for efficiently synthesizing recombinant humanized collagen |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106220727A (en) * | 2016-08-31 | 2016-12-14 | 江苏省农业科学院 | Response phase method is utilized to optimize the ultrasonic wave added acid extracting method of collagen protein |
| CN106674345A (en) * | 2016-12-12 | 2017-05-17 | 江苏省农业科学院 | Method for preparing collagen through ultrasonic enzyme extraction |
| CN107034259A (en) * | 2017-06-21 | 2017-08-11 | 江苏省农业科学院 | A kind of preparation method of high thermal stability Shelled Turtle Trionyx Sinensis byproduct collagen albumen |
| CN109221391A (en) * | 2018-10-24 | 2019-01-18 | 浙江卫斯敦环境科技有限公司 | A kind of compound fruit and vegetable fresh-keeping film-coating and preparation method thereof |
| CN113234659A (en) * | 2021-05-28 | 2021-08-10 | 浙江万里学院 | Method for in vitro culture of pelodioblasts at embryo stage of Chinese softshell turtle and construction of damage model |
| CN117024571A (en) * | 2023-07-31 | 2023-11-10 | 中科国康(浙江)生命科学有限公司 | System and method for efficiently synthesizing recombinant humanized collagen |
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Application publication date: 20151111 |