CN101418328A - Method for producing fish scale collagen protein - Google Patents
Method for producing fish scale collagen protein Download PDFInfo
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Abstract
The invention relates to a method for preparing fish scale collagen. The method comprises the following steps: fresh fish scales are taken as a material, undergo cleaning, drying and crushing, are soaked in NaOH solution for degreasing, and soaked in hydrochloric acid solution for deashing; the fresh fish scale is added with water of which the weight is 5 to 10 times of that of the fish scales, the pH value is adjusted to between 2 and 4 by acid solution, pepsin of which the weight accounting for the weight of the mixing solution is between 1 and 5 percent is added, the mixing solution undergoes enzymolysis at a low temperature for 4 to 10 hours, and enzyme is killed; enzymatic liquid is filtered twice to obtain extracting solutions, the extracting solutions are mixed, the mixing solution is decolored, sodium chloride of which the weight accounting for the weight of the mixing solution is between 5 and 15 percent is added into the mixing solution for overnight salting out, deposit is separated and collected, and acetic acid of which the weight accounting for the weight of the mixing solution is between 5 and 20 percent is added into the deposit for dissolution to obtain protein solution; the protein solution is dialyzed and desalted to obtain purified collagen liquid; and the collagen liquid is subjected to freeze-drying or spray-drying and micro-crushing to obtain solid particle. The method has the advantages of reasonable process, simple operation, safety, low cost, little loss of nutrient matters of the product, low ash content and high yield and purity of the collagen.
Description
Technical field
The present invention relates to peptide or proteinic preparation, especially a kind of method that from fish scale, prepares collagen protein.
Background technology
Collagen protein, be a kind of important function protein, have very strong biological activity and biological function, be widely used in industrial circles such as medicine, food, daily-use chemical industry, biosynthesizing, as capsule for medicine, surgical material, edible additive, makeup etc.At present, the many collagen protein tissue extracts of applied collagen protein from Mammals such as ox, pig and chicken or poultry animal.In recent years, the mad cow disease of ox, the foot and mouth disease of pig and the domestic animals such as bird flu of poultry, poultry eqpidemic disease take place in succession, and the security of the collagen protein of domestic animal, poultry is under suspicion, and adds that some human consumers use the livestock collagen protein that tendency hypersensitive is arranged.Therefore, use the land to produce the collagen protein that animal is extracted, have the secret worry on the safety.
Fish scale is the sclerotin that the collagen of fish skin corium generates, and formal name used at school is an ichthylepidin, and it accounts for 2%～3% of fish body weight.Scale sees it is transparent in appearance, and as petal, and the edge is small and curls, and white gloss slightly has a fish like smell, and quality is hard and soft, and water content is 16.4%～17.8%, and contains abundant collagen protein.Contain multiple amino acids in such collagen protein, can improve human body cell nutrition, promote the metabolism of skin and bone tissue, can make cell become plentiful, complete, keep skin elasticity and moistening, make skin lubricated more fine and smooth, prevented that it is aging.In addition, to preventing osteoporosis disease and rheumatic arthritis, raising human body exempt from service power, strengthen vitality and have good effect.Fish scale collagen and other collagen protein ratios, its protein easily decomposes, easily by digestion, absorption.Can increase the amino acid in the blood after people take, can promote collagen protein to generate simultaneously again.
The year amount of finish of China aquatic products fish is bigger, and the fish scale annual production behind the processing fish has reached many ten thousand tons of 30-50.Become the new research topic of people one as what tankage after with the processing of these fish being become useless for calmly, further improving its added value." a kind of method of extracting collagen protein from fish scale " disclosed in openly illustrating as disclosed CN1888075A Chinese invention patent on the 3rd application January in 2007, " a kind of Fishscale collagen production process " disclosed in the CN100370033C Chinese invention patent specification sheets that on February 20th, 2008 was announced, disclosed in the disclosed CN1986828A Chinese patent application prospectus on June 27th, 2007 a kind of " fish scale collagen and manufacture method thereof ", a kind of " utilizing enzyme engineering technology to extract the method for micromolecular collagen from fish scale " disclosed in the disclosed CN1814782A Chinese invention patent ublic specification of application on August 9th, 2006, disclosed processing method all is to adopt biological enzyme technology with the fish scale hydrolysis in the above-mentioned open file, again after filtration, concentrate, dry fish scale collagen, adopt that aforesaid method makes fish scale collagen exist yield and the low deficiency of purity.
Summary of the invention
For yield and the low deficiency of purity that overcomes fish scale collagen in the prior art, provide a kind of and fish scale can be turned waste into wealth, technology is easy to operate, the product ash oontent is few, the preparation method of yield and purity height, fish scale collagen that nutrition leak is few.
The technical solution adopted for the present invention to solve the technical problems is: a kind of preparation method of fish scale collagen is characterized in that: through following technology
Fresh fish scale flushing with clean water is selected in the processing of a, fish scale for use, and oven dry under 40-50 ℃ of temperature condition, goes into then and pulverizes in the high-speed tissue mashing machine;
B, degreasing, the deliming fish scale after with above-mentioned pulverizing adds NaOH solution soaking 24-56 hour of fish scale weight 4%-20%, removes the lipid material in the fish scale, is washed till neutrality with distilled water; The hydrochloric acid soln that adds fish scale weight 5%-20% then soaked 12-36 hour, sloughed the ash content in the fish scale, was washed till neutrality with distilled water;
C, the enzymolysis fish scale after with above-mentioned degreasing, deliming adds its weight 5-10 water doubly, regulating the pH value with acid solution is 2-4, the stomach en-that adds fish scale weight 1%-5% then, under the 4-8 ℃ of low temperature during continuously stirring enzymolysis 4-10, enzymolysis is warming up to more than 80 ℃ after finishing, and keeps 15-20 minutes enzymes that go out;
The above-mentioned fish scale enzymolysis solution that makes is filtered in d, extraction, preserves extract just; Residue after extracting is repeated the said extracted process as raw material, with twice extract mixing just, standby;
E, decolouring add the activated carbon of its weight 1%-2% with the first extracting solution of above-mentioned collagen protein, stir 15-20 minutes at 40-60 ℃, decolour;
F, the sodium-chlor that the extracting solution after the above-mentioned decolouring is added its weight 5-15% of saltouing spend the night to saltouing, and adopt centrifugal method separated and collected precipitation, and the gained precipitation adds the acetate dissolving of its weight 5%-20%, obtains protein liquid;
G, the protein liquid immigration dialysis tubing of purifying and making above-mentioned are used distill water dialysis 3-5 days, and change dialysis 2-5 times every day, carries out desalination, obtains the collagen liquid of purifying;
H, the finished product collagen liquid after with above-mentioned purifying adopts lyophilize or spraying drying, carries out micronizing after the drying, obtains the collagen protein solid particulate.
Fish scale behind described degreasing, the deliming is regulated the used acid solution of pH value in enzymolysis step be citric acid solution or acetic acid solution.
After the present invention carried out degreasing, deliming processing with fish scale, ash oontent was few in its product; The fish scale enzymolysis solution that obtains behind the enzymolysis is carried out twice filtration extract, the nutritive substance in its fish scale runs off few; Adopt salting-out process precipitate and separate protein, this salt analysis method can not cause protein denaturation, fully keeps its biological activity, and simple to operate, safety, and cost is low, does not need expensive especially equipment; Saltout protein that the back obtains in solution because of its colloidal quality spot diameter is bigger, can not see through semi-permeable membranes, and inorganic salt and other lower-molecular substances can see through, so utilize the dialysis method can be further purifying through the protein of salting-out process gained.Technology of the present invention is reasonable, simple to operate, safety, and cost is low, the yield of its collagen protein and purity height.
Embodiment
The present invention will be further described below in conjunction with embodiment.
Embodiment 1
A kind of preparation method of fish scale collagen:
After the processing of a, fish scale selects for use fresh fish scale with flushing with clean water, under 40 ℃ of temperature condition, dry, send into high-speed tissue mashing machine then and pulverize;
B, degreasing, the deliming fish scale after with above-mentioned pulverizing adds the NaOH solution soaking 56 hours of fish scale weight 5%, removes the lipid material in the fish scale, is washed till neutrality with distilled water; The hydrochloric acid soln that adds fish scale weight 10% then soaked 24 hours, sloughed the ash content in the fish scale, was washed till neutrality with distilled water again;
C, the enzymolysis fish scale after with above-mentioned degreasing, deliming adds the water of 5 times of its weight, with the lemon acid for adjusting pH value is 3, the stomach en-that adds fish scale weight 2% then, employing magnetic stirring apparatus continuously stirring enzymolysis is 10 hours under 8 ℃ of low temperature, enzymolysis is warming up to 95 ℃ after finishing, and keeps 15 minutes enzymes that go out;
The above-mentioned fish scale enzymolysis solution that makes is filtered in d, extraction, preserves extract just; Residue after extracting is repeated the said extracted process as raw material, with twice extract mixing just, standby;
E, decolouring with above-mentioned collagen protein through extracted twice just extracting solution add the activated carbon of its weight 1%, stirred 20 minutes at 50 ℃, decolour;
F, the sodium-chlor that the extracting solution after the above-mentioned decolouring is added its weight 10% of saltouing spend the night to saltouing, and adopt centrifugal method separated and collected precipitation, and the gained precipitation adds the acetate dissolving of its weight 5%, obtains protein liquid;
G, the protein liquid immigration dialysis tubing of purifying and making above-mentioned are used distill water dialysis 5 days, and change dialysis 4 times every day, carries out desalination, obtains the collagen liquid of purifying;
H, the finished product collagen liquid after with above-mentioned purifying adopts cryodesiccated method to carry out drying, and dry back collagen protein carries out micronizing, obtains the collagen protein solid particulate.
Embodiment 2
A kind of preparation method of fish scale collagen:
After the processing of a, fish scale selects for use fresh fish scale with flushing with clean water, under 45 ℃ of temperature condition, dry, send into high-speed tissue mashing machine then and pulverize;
B, degreasing, the deliming fish scale after with above-mentioned pulverizing adds the NaOH solution soaking 48 hours of fish scale weight 15%, removes the lipid material in the fish scale, is washed till neutrality with distilled water; The hydrochloric acid soln that adds fish scale weight 20% then soaked 12 hours, sloughed the ash content in the fish scale, was washed till neutrality with distilled water again;
C, the enzymolysis fish scale after with above-mentioned degreasing, deliming adds the water of 10 times of its weight, with the second acid for adjusting pH value is 4, adds the stomach en-of fish scale weight 5% then, and the continuously stirring enzymolysis is 7 hours under 4 ℃ of low temperature, enzymolysis is warming up to 90 ℃ after finishing, and keeps 18 minutes enzymes that go out;
The above-mentioned fish scale enzymolysis solution that makes is filtered in d, extraction, preserves extract just; Residue after extracting is repeated the said extracted process as raw material, with twice extract mixing just, standby;
E, decolouring with above-mentioned collagen protein through extracted twice just extracting solution add the activated carbon of its weight 2%, stirred 15 minutes at 60 ℃, decolour;
F, the sodium-chlor that the extracting solution after the above-mentioned decolouring is added its weight 5% of saltouing spend the night to saltouing, and adopt centrifugal method separated and collected precipitation, and the gained precipitation adds the acetate dissolving of its weight 10%, obtains protein liquid;
G, the protein liquid immigration dialysis tubing of purifying and making above-mentioned are used distill water dialysis 4 days, and change dialysis 3 times every day, carries out desalination, obtains the collagen liquid of purifying;
H, the finished product collagen liquid after with above-mentioned purifying adopts spray-dired method to carry out drying, and dry back collagen protein carries out micronizing, obtains the collagen protein solid particulate.
Embodiment 3
A kind of preparation method of fish scale collagen:
After the processing of a, fish scale selects for use fresh fish scale with flushing with clean water, under 50 ℃ of temperature condition, dry, send into high-speed tissue mashing machine then and pulverize;
B, degreasing, the deliming fish scale after with above-mentioned pulverizing adds the NaOH solution soaking 24 hours of fish scale weight 20%, removes the lipid material in the fish scale, is washed till neutrality with distilled water; The hydrochloric acid soln that adds fish scale weight 5% then soaked 36 hours, sloughed the ash content in the fish scale, was washed till neutrality with distilled water again;
C, the enzymolysis fish scale after with above-mentioned degreasing, deliming adds the water of 8 times of its weight, with the lemon acid for adjusting pH value is 2, the stomach en-that adds fish scale weight 2.5% then, employing magnetic stirring apparatus continuously stirring enzymolysis is 5 hours under 6 ℃ of low temperature, enzymolysis is warming up to 85 ℃ after finishing, and keeps 20 minutes enzymes that go out;
The above-mentioned fish scale enzymolysis solution that makes is filtered in d, extraction, preserves extract just; Residue after extracting is repeated the said extracted process as raw material, with twice extract mixing just, standby;
E, decolouring with above-mentioned collagen protein through extracted twice just extracting solution add the activated carbon of its weight 1.5%, stirred 18 minutes at 40 ℃, decolour;
F, the sodium-chlor that the extracting solution after the above-mentioned decolouring is added its weight 15% of saltouing spend the night to saltouing, and adopt centrifugal method separated and collected precipitation, and the gained precipitation adds the acetate dissolving of its weight 18%, obtains protein liquid;
G, the protein liquid immigration dialysis tubing of purifying and making above-mentioned are used distill water dialysis 3 days, and change dialysis 5 times every day, carries out desalination, obtains the collagen liquid of purifying;
H, the finished product collagen liquid after with above-mentioned purifying adopts spray-dired method to carry out drying, and dry back collagen protein carries out micronizing, obtains the collagen protein solid particulate.
Claims (2)
1, a kind of preparation method of fish scale collagen is characterized in that: through following technology
Fresh fish scale flushing with clean water is selected in the processing of a, fish scale for use, and oven dry under 40-50 ℃ of temperature condition, goes into then and pulverizes in the high-speed tissue mashing machine;
B, degreasing, the deliming fish scale after with above-mentioned pulverizing adds NaOH solution soaking 24-56 hour of fish scale weight 4%-20%, removes the lipid material in the fish scale, is washed till neutrality with distilled water; The hydrochloric acid soln that adds fish scale weight 5%-20% then soaked 12-36 hour, sloughed the ash content in the fish scale, was washed till neutrality with distilled water;
C, the enzymolysis fish scale after with above-mentioned degreasing, deliming adds its weight 5-10 water doubly, regulating the pH value with acid solution is 2-4, the stomach en-that adds fish scale weight 1%-5% then, under the 4-8 ℃ of low temperature during continuously stirring enzymolysis 4-10, enzymolysis is warming up to more than 80 ℃ after finishing, and keeps 15-20 minutes enzymes that go out;
The above-mentioned fish scale enzymolysis solution that makes is filtered in d, extraction, preserves extract just; Residue after extracting is repeated the said extracted process as raw material, with twice extract mixing just, standby;
E, decolouring add the activated carbon of its weight 1%-2% with the first extracting solution of above-mentioned collagen protein, stir 15-20 minutes at 40-60 ℃, decolour;
F, the sodium-chlor that the extracting solution after the above-mentioned decolouring is added its weight 5-15% of saltouing spend the night to saltouing, and adopt centrifugal method separated and collected precipitation, and the gained precipitation adds the acetate dissolving of its weight 5%-20%, obtains protein liquid;
G, the protein liquid immigration dialysis tubing of purifying and making above-mentioned are used distill water dialysis 3-5 days, and change dialysis 2-5 times every day, carries out desalination, obtains the collagen liquid of purifying;
H, the finished product collagen liquid after with above-mentioned purifying adopts lyophilize or spraying drying, carries out micronizing after the drying, obtains the collagen protein solid particulate.
2, the preparation method of a kind of fish scale collagen according to claim 1 is characterized in that: the fish scale behind described degreasing, the deliming is regulated the used acid solution of pH value in enzymolysis step be citric acid solution or acetic acid solution.
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| CN102108098A (en) * | 2010-08-11 | 2011-06-29 | 北京洪源澳达生物技术发展有限公司 | Crocodile collagen, and preparation and use thereof |
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Open date: 20090429 |