CN103333939A - Method for preparing gelatin from bone collagen fibers by alkali-enzyme composite degradation - Google Patents
Method for preparing gelatin from bone collagen fibers by alkali-enzyme composite degradation Download PDFInfo
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Abstract
The invention discloses a method for preparing gelatin from bone collagen fibers by alkali-enzyme composite degradation. The method comprises: (1) a step of pretreating the bone collagen fibers (bone extract particles) in an alkaline solution after the bone collagen fibers are degreased and demineralized; (2) a step of degrading the collagen fibers under the combined action of an alkali and a protease; (3) a step of filtering, washing with water, and performing enzyme deactivation by using boiling water; (4) a step of adding deionized water, extracting at a certain temperature, and filtering to obtain a gelatin crude extract; and (5) a step of performing ion exchanging, decoloring, condensing, drying, smashing and packaging on the gelatin crude extract. According to the method provided by the invention, alkali-enzyme composite degradation is adopted. The method has advantages, over traditional bone gelatin preparation technologies using an enzyme method, of improved degradation efficiency, short pretreatment period, and high gelatin purity. The method provides the basis for green preparation of gelatin.
Description
Technical field
The present invention relates to a kind of preparation method of gelatin, relate in particular to the method that a kind of alkali enzyme composite degradation collagenous fiber prepares gelatin.
Background technology
At present, China increases year by year to the bone gelatin(e) demand, the bone gelatin(e) industrial scale also enlarges year by year, but the bone gelatin(e) production technique of each manufacturing of gelatin enterprise still is difficult to obtain big breakthrough, and traditional alkaline process prepares bone gelatin(e) technology, because of its production cycle long (the liming operation is consuming time to reach 40～60 days), it is big to produce power consumption, and production efficiency is low, and water loss is big, therefore, be badly in need of the bone gelatin(e) new process of production of a kind of efficient energy-saving of exploitation.
From the fifties in last century, people are incorporated in the manufacturing of gelatin with regard to the Degradation that begins to attempt proteolytic enzyme catalysis collagen protein.Compare with traditional alkaline process adhesive-preparing technology, the Production by Enzymes cycle will shorten greatly.Therefore, domestic and international gelatin worker takes much count of the research to the enzyme process glue always.But up to the present, also still enzyme process is not prepared the gelatine process application in the big production of bone gelatin(e), this is relevant with the difficult control of enzyme liberating collagen reaction.
Patent publication No. CN 1473901A discloses a kind of method of preparing gelatin from enzyme degradation bone collagen, specifically may further comprise the steps: adopting the skeletal grain of not demineralization of degreasing is raw material, be milled to size at the skeletal grain of 4～8mm, add acid or slight alkaline protease, transferring pH is 1.5～4 or 7～8, the control collagen degradation, enzyme concentration is 2～8 ‰ of bone mud weight; Hot-water extraction again, separating and filtering gelatin extract obtains highly purified hard capsule gelatin solution.Being ground into from skeletal grain that glue produces only is 7～10 hours, and in addition, the process water amount is considerably less, has reversed fundamentally that the traditional technology water loss is big, the production cycle long, with serious pollution problem; Reduce production cost, improved economy, environmental benefit.But, select for use the direct enzymolysis of skeletal grain of not demineralization of degreasing to carry glue, cause residual a large amount of inorganic ions in the gelatin solution, cause difficulty for follow-up separation circuit.
Patent publication No. CN 101107974A discloses a kind of method of preparing edible gelatin with combined zyme, concrete steps are as follows: the aspartic protease that adds dried ossein quality 1～3 ‰, carried out degrading the first time 2～5 hours in 55～60 ℃, the high temperature enzyme that goes out is carried glue, add 2～3 ‰ neutrality or slight alkaline protease in the remaining ossein, in 55～60 ℃ of following enzymolysis for the second time, the enzyme that goes out is carried glue.Enzymolysis gained gelatin viscosity is 2.3mPa.s for the first time, freezes power 139Bloom g; Enzymolysis gained gelatin viscosity is 3.4mPa.s for the second time, freezes power 101Bloom g.This method proposes to utilize the different proteolytic enzyme of specificity enzymolysis ossein successively, increased restriction enzyme site, ossein is degraded fully as far as possible, the enzymolysis process with two kinds of enzymes separates simultaneously, it is less that the gelatin peptide of small molecular weight is produced, but this will certainly cause the gelatine molecular weight that obtains low.So the Gel Strength of Gelatin of the proteolytic enzyme that this technology is selected for use preparation is lower.
Summary of the invention
In order to overcome the defective of prior art, the present inventor has carried out further investigation repeatedly.
The object of the present invention is to provide a kind of alkali and enzyme composite degradation collagenous fiber to prepare the novel method of gelatin.
The present inventor is surprisingly found out that following technical scheme can realize above-mentioned purpose.
A kind of alkali enzyme composite degradation collagenous fiber prepares the method for gelatin, comprises the steps:
(1) collagenous fiber (ossein particle) after pulverizing and the screening degreasing demineralization, immersion treatment in basic solution;
(2) product of (1) is degraded under the acting in conjunction of alkali and proteolytic enzyme;
(3) filter after, the boiling water enzyme that goes out;
(4) be washed to certain pH;
(5) add deionized water, extracting is at a certain temperature filtered then and is obtained the gelatin crude extract;
(6) the gelatin crude extract carry out ion-exchange, decolouring, concentrate, dry, pulverize and packing.
According to method of the present invention, preferably, step (1) is: pulverize and screening degreasing demineralization after the ossein particle, make particle diameter at 5～12mm, in pH is 8.0～10.0 basic solution, soaked 4～12 hours then.
According to method of the present invention, preferably, in step (1), the ossein particle after the degreasing demineralization and the volume ratio of basic solution are 1: 2～1: 5; Basic solution is calcium hydroxide aqueous solution, aqueous sodium hydroxide solution or potassium hydroxide aqueous solution.
According to method of the present invention, preferably, in step (2), alkali content is 0.1～0.5% in the solution of alkali and proteolytic enzyme, and protease concentration is 1～10ppm in the solution of alkali and proteolytic enzyme.
According to method of the present invention, preferably, in step (2), be the ossein particle weight of unit with the gram with the ratio that is the liquor capacity of the alkali of unit and proteolytic enzyme with the cubic centimetre it is 1: 1～1: 6.
According to method of the present invention, preferably, in step (2), used proteolytic enzyme is Sumizyme MP, and alkali is calcium hydroxide, sodium hydroxide or potassium hydroxide, and degradation temperature is 20～50 ℃, and degradation time is 1～15 hour.
According to method of the present invention, preferably, in step (3), the go out temperature of enzyme of boiling water is 80～110 ℃; Go out time of enzyme of boiling water is 10～30 minutes.
According to method of the present invention, preferably, in step (4), be washed to pH7.0～7.5.
According to method of the present invention, preferably, in step (5), be that the ossein weight of unit is 1: 1～1: 6 with the ratio that is the volume of water of unit with the cubic centimetre with the gram, the temperature of hot-water extraction is 60～90 ℃.
According to method of the present invention, preferably, ossein after the degreasing demineralization of step (1) obtains as follows: it is 2～5cm that animal skeleton is crushed to particle diameter, adopts the degreasing of high-pressure hydraulic method then, and adopting concentration again is the hydrochloric acid soln decalcification of 0.2～1.0mol/L.
Produce bone gelatin(e) technology with traditional alkaline process and compare, the present invention adopts alkali and enzyme composite degradation collagenous fiber, has shortened the production time, has reduced the difficulty of realization of industrialization.For example, pre-treatment (liming) cycle foreshortened to about 15 hours by original 60～90 days.In addition, method of the present invention has reduced the discharging of a large amount of alkali lye, has reduced environmental pollution.Prepare bone gelatin(e) with existing enzyme process and compare, the present invention adopts Sumizyme MP and alkali compound, and used enzyme amount and alkali number are less.In addition, method of the present invention has reduced environmental pollution, has improved economic benefit.Directly the enzymolysis gelatin extraction technique is not compared with removing mineral substance, and method of the present invention adopts the decalcification ossein, and the gelatin crude extract greatly reduces inorganic salt content in the gelatin solution through filtration, ion exchange process, has improved the quality of gelatin.
Description of drawings
Fig. 1 is the production process route of a kind of embodiment of the present invention.
Embodiment
The present invention is further illustrated below in conjunction with embodiment, but protection scope of the present invention is not limited to this.
In the present invention, described " particle diameter ", no specified otherwise is all represented passing-screen size.
＜collagenous fiber 〉
The invention provides the method that a kind of alkali enzyme composite degradation collagenous fiber prepares gelatin.The animal skeleton that is used for the inventive method can be preferably sheep bone or ox bone, more preferably the sheep bone for sheep bone, ox bone, pig bone or chicken bone etc.
Collagenous fiber (ossein) after the degreasing demineralization of step of the present invention (1) can obtain as follows: animal skeleton is pulverized, adopted high-pressure hydraulic method degreasing and acid solution decalcification.For example, it is 2～5cm that animal skeleton is crushed to particle diameter, is preferably 3～4cm.The acid solution that decalcification is used can be hydrochloric acid, and preferably concentration is 0.2～1.0mol/L hydrochloric acid, and being more preferably concentration is 0.5～0.8mol/L hydrochloric acid, and preferably concentration is 0.6～0.7mol/L hydrochloric acid again.
＜bone gelatin(e) production technique 〉
Fig. 1 is the bone gelatin(e) production process route of a kind of embodiment of the present invention.After the processing of alkali enzyme, carry out hot water again and carry glue.Can shorten the production time like this, and reduce the consumption of enzyme and the consumption of alkali.
The invention provides the method that a kind of alkali enzyme composite degradation collagenous fiber prepares gelatin and comprise the steps: (1) basic solution soaking step; (2) alkali and enzyme liberating step; (3) the boiling water enzyme step of going out; (4) hot-water extraction step; (5) ion-exchange step; (6) steps such as decolouring, drying, pulverizing.To be described in detail below.
＜basic solution soaking step 〉
Prepare the method for gelatin at alkali enzyme composite degradation collagenous fiber of the present invention, step (1) is: the ossein after the degreasing demineralization is soaked in basic solution.Ossein after the degreasing demineralization can be crushed to particle diameter 5～12mm, be preferably 7～10mm.Being conducive to osso-albumin like this degrades fast.The pH that is used for the basic solution of the ossein particle after the soak degreasing demineralization is 8.0～10.0, is preferably 8.5～9.5.Ossein after degreasing demineralization soaking temperature in basic solution can be room temperature.Ossein particle soak time in basic solution after the degreasing demineralization is preferably 4～12 hours, more preferably 6～11 hours, is preferably 8～10 hours again.Can guarantee that like this osso-albumin degraded is abundant and efficient is higher.
In the step (1) of the inventive method, the ossein after the degreasing demineralization and the volume ratio of basic solution can be 1: 2～1: 5, are preferably 1: 3～1: 4.The basic solution of the ossein after the soak degreasing demineralization can be aqueous sodium hydroxide solution, calcium hydroxide aqueous solution or potassium hydroxide aqueous solution; Be preferably aqueous sodium hydroxide solution or potassium hydroxide aqueous solution; Aqueous sodium hydroxide solution more preferably.The carrying out that is conducive to the subsequent disposal operation like this.
＜alkali and enzyme liberating step 〉
Prepare the method for gelatin at alkali enzyme composite degradation collagenous fiber of the present invention, step (2) is: degrade in the solution of alkali and proteolytic enzyme.In order to guarantee the vigor of enzyme, in step (2), degradation temperature can be 20～50 ℃, is preferably 25～45 ℃, more preferably 30～40 ℃, most preferably is 38 ℃.Degradation time can be 1～15 hour, is preferably 5～12 hours, more preferably 8～10 hours.Can guarantee that like this enzymolysis process of collagenous fiber is abundant and efficient is higher.
In the step (2) of the inventive method, being the ossein weight of unit with the gram can be 1: 1～1: 6 with the ratio that with the cubic centimetre is the liquor capacity of the alkali of unit and proteolytic enzyme, is preferably 1: 2～1: 5, more preferably 1: 3～1: 4.It is the ossein weight of unit with the ratio that is the liquor capacity of the alkali of unit and proteolytic enzyme with the cubic centimetre is 1: 2,1: 4,1: 6 that concrete example comprises restraining.
In the step (2) of the inventive method, alkali content is 0.1～0.5% in the solution of alkali and proteolytic enzyme, is preferably 0.3～0.4%, is conducive to improve the vigor of enzyme like this.In the solution of alkali and proteolytic enzyme, protease concentration is 1～10ppm, is preferably 2～8ppm, more preferably 3～6ppm.Can guarantee that like this enzyme gives full play to enzymolysis and render a service.In the present invention, used proteolytic enzyme can be for being papoid (70,000 U/g), trypsin 2,500,000 U/g) and Sumizyme MP (100,000 U/mL), be preferably Sumizyme MP; Alkali is calcium hydroxide, sodium hydroxide or potassium hydroxide, is preferably sodium hydroxide.The water of preparing described aqueous solution use is preferably deionized water, distilled water or MILLIPORE pure water.
Sumizyme MP is biological catalyst, has the height specificity, it is the action site difference of different proteolytic enzyme peptide chain, but the protein restriction endonuclease is very difficult to the degraded of collagenous fiber, in industrial production, usually the liming that adopts is strong and the time spent is longer to the degraded randomness of collegen filament, makes the controlled relatively poor of its technology.Present method mainly realizes the controlled degradation of collegen filament by alkali and Sumizyme MP synergy, shortened the treatment time, and controlling of degraded is strong, has improved efficient and the quality of manufacturing of gelatin simultaneously, and industrialized feasibility.In the present invention, Sumizyme MP is known in the art, repeats no more here.
＜boiling water enzyme the step of going out 〉
Prepare the method for gelatin at alkali enzyme composite degradation collagenous fiber of the present invention, step (3) is: degraded after-filtration, the boiling water enzyme that goes out then.The method of filtering be known in the art those, repeat no more here.Preferably, described boiling water is the deionized water of boiling, the distilled water of boiling or the MILLIPORE pure water of boiling.The go out temperature of enzyme of boiling water can be 80～110 ℃, is preferably 85～105 ℃, more preferably 90～100 ℃.Go out time of enzyme of boiling water can be 10～30 minutes, is preferably 12～25 minutes, more preferably 15～20 minutes.
＜hot-water extraction step 〉
The method for preparing gelatin at alkali enzyme composite degradation collagenous fiber of the present invention, step (4) is: be washed to certain pH, step (5) is: adding water, is that the ossein weight of unit is 1: 1～1: 6 with the ratio that is the volume of water of unit with the cubic centimetre with the gram, is preferably 1: 2～1: 5; Extracting is at a certain temperature filtered then and is obtained the gelatin crude extract.Be washed to pH7.0～7.5; The temperature of hot-water extraction can be 60～90 ℃, is preferably 65～85 ℃, more preferably 70～80 ℃.If temperature is higher than 90 ℃, will destroy the viscosity of glue; If temperature is lower than 60 ℃, will reduce productive rate.The time of hot-water extraction can be 1～5 hour, is preferably 2～4 hours.Can guarantee the gelatin extracting abundant like this.The solution that extracting obtains filters to remove impurity.The method of filtering be known in the art those, repeat no more here.The water that hot-water extraction is used is preferably deionized water, distilled water or MILLIPORE pure water.
＜ion-exchange step 〉
Prepare the method for gelatin at alkali enzyme composite degradation collagenous fiber of the present invention, step (6) is: the gelatin crude extract carries out ion-exchange.In ion-exchange step of the present invention, after the filtration of gelatin crude extract, by ion-exchange resin purification, obtain gelatin solution.Preferably, after the filtration of gelatin crude extract, by the Zeo-karb exchange, by the anionite-exchange resin exchange, obtain gelatin solution then.Can guarantee purification effect like this.
Steps such as＜decolouring, drying, pulverizing 〉
Prepare the method for gelatin at alkali enzyme composite degradation collagenous fiber of the present invention, preferably include following steps: the gelatin after (6) ion-exchange carries out charcoal absorption decolouring, dry and pulverize.Can obtain the higher gelatin of quality like this.
In drying step of the present invention, gelatin solution is concentrated, then warm air drying.Preferably, gelatin solution is concentrated into 1/2 of raw material volume, is preferably 1/3, more preferably 1/4.Can shorten time of drying like this, improve the quality of products.Thickening equipment is known in the art those, for example the economic benefits and social benefits thickener.Preferably, the gelatin solution after concentrating below 70 ℃, below 60 ℃, below 50 ℃, more preferably dry in 40～50 ℃ the hot blast; Perhaps dry down at 20 ℃～40 ℃.Be dried to that moisture content is lower than 20wt% in the gelatin, preferred 14wt%, more preferably 12wt% gets final product.Gelatin can be preserved well like this.
Among the embodiment hereinafter, ossein: the solution of alkali and proteolytic enzyme (W/V) is quality and volume ratio, the g/mL of unit; Ossein: water (W/V) is quality and volume ratio, the g/mL of unit, unless specifically stated otherwise; Used basic protein enzyme activity is 100,000 U/ml; Described alkali lye is aqua calcis.
In the present invention, the group method of following examples is as follows:
With fresh sheep bone, reject muscle, meat and the periosteum on surface, to pound and be broken to particle diameter 2～5cm, the hydrochloric acid decalcification of 0.2mol/L～1.0mol/L is used in the waterpower degreasing again; Neutralization, washing obtain sheep collagenous fiber (ossein particle).
Pulverizing and screening ossein particle, make particle diameter at 10～15mm, is 8.0～10.0 dipping by lye again with 2 times of volume pH, is stabilized in about 8.0～8.5 to pH, continues immersion 4～12 hours.
Preparation pH is 8.0～10.0, the basic protein enzyme concn is the alkali of 1～10ppm and the solution of proteolytic enzyme, presses ossein: the solution of alkali and proteolytic enzyme (W/V)=1: 1,1: 2,1: 4 or 1: 6, under 30～45 ℃, degrade; The DeR time is 2～15 hours.
After degraded is finished, filter, add boiling water and stir the enzyme 10～30 minutes of going out fast, being washed to pH is 7.0～7.5.
Press ossein: water (W/V)=1: 2,1: 4 or 1: 5 are with the two mixing; Under 60～90 ℃, carry glue.
The muddy gelatin solution that obtains is carried out centrifugation, remove the solid filter residue, obtain the gelatin crude extract.
The thick liquid of isolated gelatin filters with filter cotton, removes inorganic ion through the ion exchange resin exchange.Concrete steps are: the gelatin solution after the filtration exchanges by Zeo-karb with the flow velocity of 8 times of column volume/h earlier, and the flow velocity with 8 times of column volume/h exchanges by anionite-exchange resin then; Collect the gelatin solution after exchanging.
Gelatin solution after the ion-exchange obtains gelatin through charcoal absorption decolouring, drying, pulverizing.Described drying step is specific as follows: be concentrated into 1/2 of original liquid concentration with the economic benefits and social benefits thickener; 40 ℃ of warm air drying gelatin are to moisture content＜14wt%.
The testing method of various details gelatin.
＜strength test the method for congealing 〉
Used instrument is the fabric analysis instrument, with reference to GB6783-1994.
The testing method of＜molecular weight 〉
Institute's use instrument is U.S. Waters HPLC, is equipped with the Waters2414 refraction detector, three chromatographic column series connection, and column temperature is 30 ℃, leacheate is mixed phosphate salts solution (potassium primary phosphate, Sodium phosphate dibasic, sodium azide), flow velocity 0.8mL/min.
The gelatin sample concentration that detects is 4mg/mL, measures weight-average molecular weight.
＜ash content testing method 〉
Adopt the calcination method, with reference to GB 5009.4-2010.
Embodiment 1
Select the sheep ossein 30g of degreasing demineralization for use, particle diameter is at 7～10mm, and the pH that adds 2 times of volumes is 9.0 dipping by lye, is stabilized in about 8.5 to pH, continues to soak 8 hours.
The preparation sodium hydrate content is 0.3%, the basic protein enzyme concn is the sodium hydroxide of 6ppm and the solution of proteolytic enzyme, presses ossein: the solution of sodium hydroxide and proteolytic enzyme (W/V)=1: 2, under 38 ℃, carry out enzymolysis, and the DeR time is 8 hours.
The degraded after-filtration places boiling water, stirs fast enzyme to be lost activity in 20 minutes, and being washed to pH is 7.0～7.5.
Press ossein: water (W/V)=1: 4 in 70 ℃ of following extractings 3 hours, obtains the gelatin crude extract.
Adopt filter cotton filtration, anion-cation exchange resin purifying to obtain the gelatin settled solution, obtain the solid gelatin through charcoal absorption decolouring, drying, pulverizing again.
The gained gelatin congeal intensity more than 225Bloom g, weight-average molecular weight is greater than 60000, ash content is less than 0.1wt%.
Embodiment 2
Select the sheep bone ossein 30g of degreasing demineralization for use, particle diameter is at 7～10mm, and the pH that adds 2 times of volumes is 9.0 dipping by lye, is stabilized in about 8.5 to pH.
The preparation sodium hydrate content is 0.3%, the basic protein enzyme concn is the sodium hydroxide of 6ppm and the solution of proteolytic enzyme, presses ossein: the solution of sodium hydroxide and proteolytic enzyme (W/V)=1: 2, under 30 ℃, carry out enzymolysis, and the DeR time is 8 hours.
The degraded after-filtration places boiling water, stirs fast enzyme to be lost activity in 20 minutes, and being washed to pH is 7.0～7.5.
Press ossein: water (W/V)=1: 4, in 60 ℃, extracting 3 hours obtains the gelatin crude extract.
Adopt filter cotton filtration, anion-cation exchange resin purifying to obtain the gelatin settled solution, obtain the solid gelatin through charcoal absorption decolouring, drying, pulverizing again.
The gained gelatin congeal intensity more than 225Bloom g, weight-average molecular weight is greater than 60000, ash content is less than 0.1wt%.
Embodiment 3
Select the sheep bone ossein 30g of degreasing demineralization for use, particle diameter is at 7～10mm, and the pH that adds 2 times of volumes is 9.0 dipping by lye, is stabilized in about 8.5 to pH.
The preparation sodium hydrate content is 0.3%, the basic protein enzyme concn is the sodium hydroxide of 6ppm and the solution of proteolytic enzyme, presses ossein: the solution of sodium hydroxide and proteolytic enzyme (W/V)=1: 2, under 40 ℃, to degrade, and the DeR time is 8 hours.
The degraded after-filtration places boiling water, stirs fast enzyme to be lost activity in 20 minutes, and being washed to pH is 7.0～7.5.
Press ossein: water (W/V)=1: 2, in 65 ℃, extracting 3 hours obtains the gelatin crude extract.
Adopt filter cotton filtration, anion-cation exchange resin purifying to obtain the gelatin settled solution, obtain the solid gelatin through charcoal absorption decolouring, drying, pulverizing again.
The gained gelatin congeal intensity more than 230Bloom g, weight-average molecular weight is greater than 60000, ash content is less than 0.1%.
Embodiment 4
Select the sheep bone ossein 30g of degreasing demineralization for use, particle diameter is at 7～10mm, and the pH that adds 2 times of volumes is 9.0 dipping by lye, is stabilized in about 8.5 to pH.
The preparation sodium hydrate content is 0.35%, the basic protein enzyme concn is the sodium hydroxide of 10ppm and the solution of proteolytic enzyme, presses ossein: the solution of sodium hydroxide and proteolytic enzyme (W/V)=1: 2, under 30 ℃, to degrade, and the DeR time is 8 hours.
The degraded after-filtration places boiling water, stirs fast enzyme to be lost activity in 20 minutes, and being washed to pH is 7.0～7.5.
Press ossein: water (W/V)=1: 4, in 80 ℃, extracting 3 hours obtains the gelatin crude extract.
Adopt filter cotton filtration, anion-cation exchange resin purifying to obtain the gelatin settled solution, obtain the solid gelatin through charcoal absorption decolouring, drying, pulverizing again.
The gained gelatin congeal intensity more than 225Bloom g, weight-average molecular weight is greater than 60000, ash content is less than 0.1wt%.
Embodiment 5
Select the sheep bone ossein 30g of degreasing demineralization for use, particle diameter is at 7～10mm, and the pH that adds 2 times of volumes is 9.0 dipping by lye, is stabilized in about 8.5 to pH.
The preparation sodium hydrate content is 0.4%, the basic protein enzyme concn is the sodium hydroxide of 6ppm and the solution of proteolytic enzyme, presses ossein: the solution of sodium hydroxide and proteolytic enzyme (W/V)=1: 2, under 30 ℃, to degrade, and the DeR time is 8 hours.
The degraded after-filtration places boiling water, stirs fast enzyme to be lost activity in 20 minutes, and being washed to pH is 7.0～7.5.
Press ossein: water (W/V)=1: 5, in 75 ℃, extracting 3 hours obtains the gelatin crude extract.
Adopt filter cotton filtration, anion-cation exchange resin purifying to obtain the gelatin settled solution, obtain the solid gelatin through charcoal absorption decolouring, drying, pulverizing again.
The gained gelatin congeal intensity more than 235Bloom g, weight-average molecular weight is greater than 60000, ash content is less than 0.1wt%.
Embodiment 6
Select the sheep bone ossein 30g of degreasing demineralization for use, particle diameter is at 7～10mm, and the pH that adds 2 times of volumes is 9.0 dipping by lye, is stabilized in about 8.5 to pH.
The preparation sodium hydrate content is 0.35%, the basic protein enzyme concn is the sodium hydroxide of 6ppm and the solution of proteolytic enzyme, presses ossein: the solution of sodium hydroxide and proteolytic enzyme (W/V)=1: 2, under 30 ℃, to degrade, and the DeR time is 8 hours.
The degraded after-filtration places boiling water, stirs fast enzyme to be lost activity in 20 minutes, and being washed to pH is 7.0～7.5.
Press ossein: water (W/V)=1: 4, in 85 ℃, extracting 3 hours obtains the gelatin crude extract.
Adopt filter cotton filtration, anion-cation exchange resin purifying to obtain the gelatin settled solution, obtain the solid gelatin through charcoal absorption decolouring, drying, pulverizing again.
The gained gelatin congeal intensity more than 225Bloom g, weight-average molecular weight is greater than 60000, ash content is less than 0.1wt%.
The present invention is not limited to above-mentioned embodiment, and under the situation that does not deviate from flesh and blood of the present invention, any distortion that it may occur to persons skilled in the art that, improvement, replacement all fall into scope of the present invention.
Claims (10)
1. an alkali enzyme composite degradation collagenous fiber prepares the method for gelatin, comprises the steps:
(1) collagenous fiber (ossein particle) after pulverizing and the screening degreasing demineralization, immersion treatment in basic solution;
(2) product of (1) is degraded under the acting in conjunction of alkali and proteolytic enzyme;
(3) filter after, the boiling water enzyme that goes out;
(4) be washed to certain pH;
(5) add deionized water, extracting is at a certain temperature filtered then and is obtained the gelatin crude extract;
(6) the gelatin crude extract carry out ion-exchange, decolouring, concentrate, dry, pulverize and packing.
2. method according to claim 1, step (1) is: pulverize and screening degreasing demineralization after the ossein particle, make particle diameter at 5～12mm, in pH is 8.0～10.0 basic solution, soaked 4～12 hours then.
3. method according to claim 1, it is characterized in that: in step (1), the ossein particle after the degreasing demineralization and the volume ratio of basic solution are 1: 2～1: 5; Basic solution is calcium hydroxide aqueous solution, aqueous sodium hydroxide solution or potassium hydroxide aqueous solution.
4. method according to claim 1, it is characterized in that: in step (2), the alkali massfraction is 0.1～0.5% in the solution of alkali and proteolytic enzyme, and the concentration of proteolytic enzyme is 1～10ppm.
5. method according to claim 1 is characterized in that: in step (2), be the ossein weight of unit with the gram with the ratio that is the liquor capacity of the alkali of unit and proteolytic enzyme with the cubic centimetre be 1: 1～1: 6.
6. method according to claim 1, it is characterized in that: in step (2), used proteolytic enzyme is Sumizyme MP, and alkali is calcium hydroxide, sodium hydroxide or potassium hydroxide, and temperature of reaction is 10～60 ℃, and degradation time is 1～15 hour.
7. method according to claim 1 is characterized in that: in step (3), the go out temperature of enzyme of boiling water is 80～110 ℃; Time is 10～30 minutes.
8. method according to claim 1 is characterized in that: in step (4), be washed to pH7.0～7.5.
9. method according to claim 1 is characterized in that: in step (5), be that the ossein weight of unit is 1: 1～1: 6 with the ratio that is the volume of water of unit with the cubic centimetre with the gram.The temperature of hot-water extraction is 60～90 ℃.
10. method according to claim 1, it is characterized in that: the ossein particle after the degreasing demineralization of step (1) obtains as follows: it is 2～5cm that animal skeleton is crushed to particle diameter, adopt the degreasing of high-pressure hydraulic method then, adopting concentration again is the hydrochloric acid soln decalcification of 0.2～1.0mol/L.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104630319A (en) * | 2015-02-07 | 2015-05-20 | 济宁医学院 | Method for preparing medical bone gelatin by biological enzyme method |
| CN114891446A (en) * | 2022-05-07 | 2022-08-12 | 李华雨 | Method for preparing leather gelatin by compounding alkaline method/acid method and enzymatic method |
| CN115368454A (en) * | 2022-09-30 | 2022-11-22 | 斐缦(长春)医药生物科技有限责任公司 | Gelatin and preparation method thereof |
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| CN102051130A (en) * | 2010-11-24 | 2011-05-11 | 中国科学院理化技术研究所 | Method for preparing gelatin by degrading ossein with protease |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104630319A (en) * | 2015-02-07 | 2015-05-20 | 济宁医学院 | Method for preparing medical bone gelatin by biological enzyme method |
| CN104630319B (en) * | 2015-02-07 | 2018-06-05 | 济宁医学院 | The method that biological enzyme prepares medical bone gelatin |
| CN114891446A (en) * | 2022-05-07 | 2022-08-12 | 李华雨 | Method for preparing leather gelatin by compounding alkaline method/acid method and enzymatic method |
| CN115368454A (en) * | 2022-09-30 | 2022-11-22 | 斐缦(长春)医药生物科技有限责任公司 | Gelatin and preparation method thereof |
| CN115368454B (en) * | 2022-09-30 | 2023-05-12 | 斐缦(长春)医药生物科技有限责任公司 | Gelatin and preparation method thereof |
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Address after: 100029 Chaoyang District, North Third Ring Road,, No. 15, Beijing University of Chemical Technology Applicant after: Beijing University of Chemical Technology Applicant after: Xinjiang Azhar Biological Technology Co., Ltd. Address before: 831100, 05-05, Changji hi tech Industrial Development Zone, the Xinjiang Uygur Autonomous Region, Changji, Xinjiang Applicant before: Xinjiang Azhar Biological Technology Co., Ltd. Applicant before: Beijing University of Chemical Technology |
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Application publication date: 20131002 |