CN103243143A - Method for extracting hydrolyzed collagen by using bone meal - Google Patents
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- CN103243143A CN103243143A CN2013101699076A CN201310169907A CN103243143A CN 103243143 A CN103243143 A CN 103243143A CN 2013101699076 A CN2013101699076 A CN 2013101699076A CN 201310169907 A CN201310169907 A CN 201310169907A CN 103243143 A CN103243143 A CN 103243143A
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Abstract
The invention provides a method for extracting hydrolyzed collagen by using bone meal. The target product, namely the hydrolyzed collagen, is formed by performing pickling treatment and high-pressure treatment on the bone meal, hydrolyzing the bone meal through papain and alkaline protease under the condition of a composite enzymolysis protection agent, and then sequentially performing high-temperature enzyme deactivation, filtration, decoloring, desalting, concentration and atomizing drying on the bone meal, wherein the bone meal is a byproduct produced in a bone grain preparation process; and the particle size of the bone meal is less than 5 mm. The hydrolyzed collagen prepared by the method has undivided molecule weight and meets requirements of foods, health protection products and cosmetics; furthermore, the technical difficulties that the bone meal is difficult to hydrolyze, hydrolyzed products are non-uniform in molecule weight, and the products have bad smell are solved; and the raw material cost of the hydrolyzed collagen is greatly reduced.
Description
Technical field
The invention belongs to technical field of bioengineering, relate to a kind of method of utilizing bone meal to extract Collagen Hydrolysate, be specifically related to a kind of method of from bovine bone powder, extracting Collagen Hydrolysate.
Background technology
Gelatin is the peptide molecule mixture made from the bone of animal or skin muscle tendon, in food, medicine and other fields extensive application.Since the eighties in last century, the technology that domestic and international manufacturing of gelatin producer has eliminated whole bone pickling is gradually used the skeletal grain pickling instead and is produced gelatin.Skeletal grain is the main raw material of producing edible Gelatinum oxhide and pharmagel, and ox bone has become 5 to 25mm skeletal grain through technologies such as graded crushing, oil removing, drying, sortings.During practice is produced, produced by product bone meal when making skeletal grain, this bone meal is can not be used for the particle diameter of manufacturing of gelatin less than the bone slag of 5mm.In the prior art, skeletal grain manufacturer can produce 4.5 tons of by product bone meal when utilizing per 100 tons of ox bones to produce 45 tons of skeletal grains.Because the particle diameter of bone meal is too small, can't be used for the production of gelatin, can only be used as protein fodder, greatly reduce the added value of manufacturing of gelatin raw material bone.See 6500 yuan of skeletal grain market prices per ton, and 2600 yuan of bone meal market prices per ton, 200 yuan/kg of the average market price of Collagen Hydrolysate with 2012 noons China domestic market situation.The present invention is processed into Collagen Hydrolysate with bone meal, and bone meal per ton can be produced Collagen Hydrolysate 130kg at least, is worth thereby greatly improve bone meal, improves the manufacturing of gelatin utilization ratio of raw materials.
Collagen Hydrolysate is the abundantest animal proteinum of Mammals in-vivo content; mainly be present in bone, tendon, flesh sheath, ligament, sarolemma, cartilage and the skin of animal; be extremely important a kind of structural protein in the reticular tissue; playing the function that supports organ and protection human body, also is the most important functions protein of forming intercellular substance.Collagen Hydrolysate can be applicable to industrial circles such as medical treatment, feed, food, makeup.Collagen protein is a kind of excellent performance, widely used biomaterial, can be made into hemostasis pulvis, burn dressing, pharmaceutical carrier, heart and distinguishes that the equivalent material of film, blood vessel, oesophagus and tracheae, surgery are with variforms such as sutures.The substratum that can be used as biological fermentation can effectively promote the growth of mycelia, substitutes that imported fish meal is used for mixing, mixed feed production and granulated feed binding agent.Can be used as collagen class functional foodstuff and healthcare products.Can also improve the quality of meat, as packaging material for food, beverage finings.Also can be used for producing superior cosmetics, can replenish the required nutrient of skin layer, replenish 17 kinds of amino acid to the human body beneficial; Repair tissue; Preserve moisture; Regulate and stable pH value stable foam, emulsifying collid.
It is several that the preparation method of Collagen Hydrolysate mainly is divided into pyrohydrolysis, acid hydrolysis, basic hydrolysis and enzymic hydrolysis etc., at present both at home and abroad in producing the Collagen Hydrolysate process, the most range of molecular weight distributions of Collagen Hydrolysate that the tissues such as skin, bone and fish scale by the ordinary method hydrolysis of animal obtain extensively, the molecular weight homogeneous is poor; And a large amount of total free aminoacidss that produce in the hydrolytic process make the biological activity of product reduce greatly, have both influenced the product taste, and justice influences effect.The raw material that Collagen Hydrolysate production is adopted usually has the tissues such as skin, bone and fish scale of animal.Because the restriction of raw materials cost, the price of Collagen Hydrolysate is always far above protein products such as whey-protein, soybean proteins.
Summary of the invention
The present invention overcomes the above-mentioned defective of prior art, propose a kind of by product bone meal that utilizes skeletal grain to produce and extract Collagen Hydrolysate, at first the by product bone meal with skeletal grain production carries out the pickling processing, clean up the back and adopt autoclaving, adopt two kinds of proteolytic enzyme directional enzymatics then, add the complex enzyme hydrolysis protective material simultaneously, effectively guarantee the biological activity of product; The high temperature enzyme after-filtration that goes out is removed and to be failed behind the impurity of hydrolysis, decolours, desalination and concentration and spraying drying again, has finally obtained the Collagen Hydrolysate product that molecular weight is single-minded, meet food and healthcare products, makeup requirement.The present invention is basic material with the by product bone meal of skeletal grain production, has solved bone meal and has been difficult to the difficult problem that hydrolysis, hydrolysate molecular weight heterogeneity, product have peculiar smell, also greatly reduces the raw materials cost of Collagen Hydrolysate simultaneously.
The present invention proposes a kind of method of utilizing bone meal to extract Collagen Hydrolysate; described bone meal is after pickling processing, autoclaving; under complex enzyme hydrolysis protective material condition through the hydrolysis of papoid, Sumizyme MP; then; through high temperature go out enzyme, filtration, decolouring, desalination and concentration, spraying drying, obtain purpose product Collagen Hydrolysate successively.
Among the present invention, described bone meal is the by product in the skeletal grain preparation process---particle diameter is less than the bone meal of 5mm.That is, described bone meal refers to that the by product that produces is that particle diameter is less than the bone bits of 5mm in the process of the main raw material skeletal grain for preparing bone gelatin(e).Preferably, refer to bovine bone powder.
Among the present invention, it is that by product bone meal with skeletal grain production is raw material that described pickling is handled, and the hydrochloric acid that increases concentration by certain gradient gradually is that solvent is handled in pickling, stops up to avoid directly handling the bone meal that causes with high concentrated acid.Preferably, adopt the hydrochloric acid of suitable any gradient concentration to be used for the pickling processing.More preferably, described pickling processing is that described bone meal was respectively soaked 6～8 hours with the dilute hydrochloric acid of 0.9%, 1.8%, 2.7%, 3.6%, 4.5% concentration respectively.Further, regularly stir bone meal during immersion, make its immersion more even.Preferably, leach bone meal after pickling finishes and clean up.Among the present invention, the employing pickling handles to remove the calcareous inorganic salt such as grade in the bone meal as far as possible, and reduces the loss of collagen as far as possible.
The present invention adopts the dilute hydrochloric acid with 0.9%, 1.8%, 2.7%, 3.6%, 4.5% 5 concentration to carry out pickling and handles, and its purpose is to remove inorganic salt in the bone and calcareous.The present invention adopts low-concentration hcl to press certain gradient and handles, and acidolysis reaction is relatively gentleer, can not produce a large amount of products the short period of time, stirs in the process in stirring at regular intervals, allows acid hydrolysate be dissolved in the acid solution, is unlikely to result in blockage.After acid solution thickens gradually, then can be deep into the inside of bone meal, can more thoroughly carry out pickling and handle.If directly handle with high concentrated acid, the bone meal surface is very fast to be soaked into, and the acid hydrolysate of generation can be distributed in the bone meal that the surface of bone meal causes and stop up, and causes acid solution can not enter bone meal inside.
Among the present invention, described autoclaving is the bone meal after the pickling processing to be placed handled under 0.14～0.15MPa high pressure 100～120 minutes.Preferably, bone meal is put into 126 ℃ of autoclaves, autoclaving 120 minutes.
Among the present invention; the described papain hydrolysis of carrying out under complex enzyme hydrolysis protective material condition is reacted; refer in the isothermal reaction jar, put into water and the bone meal after autoclaving; add papoid from plant extract in the ratio of 16000～18000U/g substrate; add the complex enzyme hydrolysis protective material again; hydrolysis temperature is 48～50 ℃, stirs enzymolysis 4～6 hours.Particularly, add the pure water of 2.6 times of bone meal weight in the isothermal reaction jar, put into the bone meal after autoclaving, intensification also keeps the retort feed temperature between 48～50 ℃.
Among the present invention, the described hydrolysis by novo that carries out under complex enzyme hydrolysis protective material condition reacts, and is to regulate pH to 9.0～9.5 with sodium hydroxide; ratio in the 9000U/g substrate adds the Sumizyme MP for preparing through microbial fermentation; hydrolysis temperature is 48～50 ℃, stirs enzymolysis 4～6 hours.
Among the present invention, papain hydrolysis reaction, hydrolysis by novo reaction are to carry out under 48～50 ℃ medium temperature condition.Under this temperature, the peptide chain in the bone meal can resolve into the polypeptide of molecular weight homogeneous slowly under the effect of proteolytic enzyme, kept higher biological activity.
Among the present invention, the described high temperature enzyme that goes out is to be warming up to 80～85 ℃, and the enzyme 15～20 minutes of going out is cooled to 30～40 ℃, is adjusted to pH6.5～7.0.
Among the present invention, it is to add gac that described decolouring is handled, and is incubated 30～40 ℃ and stirs 2～3 hours.
Among the present invention, described desalination and concentration are that the feed liquid after the decolouring is filtered through the clarification filter plate, carry out desalination and concentration through nanofiltration membrane again, collect trapped fluid.Further, the trapped fluid of collecting is carried out spraying drying, obtain described Collagen Hydrolysate.Preferably, the specification of described nanofiltration membrane is 200Da.
The product that the present invention prepares be molecular weight single-minded, meet the Collagen Hydrolysate that food and healthcare products, makeup require, the collagen content that comprises in this product Collagen Hydrolysate reaches more than 95%, and the molecular weight homogeneous is distributed in 1000Da to 2200Da scope.
Prior art is to utilize skeletal grain to be processed into gelatin.In the skeletal grain production process, be raw material with bone, adopt technique known processing and preparing such as graded crushing, boiling, centrifugal, dried, classified to obtain skeletal grain, obtain by product bone meal simultaneously.Must handle (be about to make the collagen peptide chain in the gelatine open slowly in the skeletal grain immersion aqua calcis, become the gelatin that the higher power of freezing is arranged) in the prior art in the subsequent technique of manufacturing of gelatin through the liming more than month.But, because the particle diameter of bone meal is too small, cause the area that bone meal contacts with aqua calcis in above-mentioned liming is handled excessive, thereby caused the situation of undue alkaline hydrolysis, cause obtaining the gelatin of the very low power of freezing.And because the particle diameter heterogeneity of bone meal, therefore particle size range, even the independent liming of bone meal is handled, also is difficult to time and the concentration of control liming from less than 0.5mm to 4～5mm.Therefore, the producer for preparing gelatin or Collagen Hydrolysate at present all can't utilize bone meal as raw materials for production.Can not be done cheap the processing as protein fodder usually for the production of gelatin because of bone meal in the prior art, its utilization ratio and added value are all very low.
The present invention proposes to utilize the by product bone meal of skeletal grain production process to be raw materials for production first with innovating.Adopt the pickling treatment step among the present invention, be the dilute hydrochloric acid with 0.9%, 1.8%, 2.7%, 3.6%, 4.5% 5 concentration, increase concentration of hydrochloric acid gradually according to certain gradient and handle bone meal, thereby avoid directly handling the bone meal stopping state that causes with high concentrated acid.The hydrolysis temperature that adopts among the present invention is 48～50 ℃ medium temperature condition, has guaranteed that the peptide chain in the bone meal can slowly resolve into the polypeptide of molecular weight homogeneous under the proteolytic enzyme effect, thereby has kept higher biological activity.
The complex enzyme hydrolysis protective material that the present invention adopts refers to adopting papoid and Sumizyme MP to carry out the complex enzyme hydrolysis protective material that adds in the enzyme digestion reaction process.Specifically refer to " M0111-9 enzymolysis protective material ", " M0111-9 " is the model of this reagent, and it mainly acts on is to protect the specific peptide chain in the enzymolysis process not to be destroyed, thereby obtains the product of molecular weight homogeneous.People's Republic of China's light industry standard code, Collagen Hydrolysate molecular weight ranges are between 500 dalton to 20000 dalton, and molecular weight is 1000 to 2200 daltonian Collagen Hydrolysates, is fit to more be absorbed by the body.
The Sumizyme MP that the present invention adopts, refer to " 2709 Sumizyme MP ", it namely is a kind of Bacillus licheniformis 2709 that is gone out by bacterium protoplastis mutagenic and breeding, through submerged fermentation, extraction and the refining a kind of proteolytic ferment that forms, its main enzyme component is Bacillus licheniformis proteolytic enzyme, it is a kind of endo-protease of serine-type, it can generate polypeptide or amino acid by protein hydrolysate molecule peptide chain, ability with stronger decomposing protein is widely used in food, medical treatment, brewages, industry such as washing, silk, process hides.
The object of the invention is that the by product bone meal that fully efficiently utilizes skeletal grain to produce extracts Collagen Hydrolysate.On Technology, improved that routine techniques raw materials cost height, biological activity are destroyed, range of molecular weight distributions is wide in the product, active constituent content is low, product has defectives such as peculiar smell.The by product bone meal that the present invention utilizes skeletal grain to produce is raw material; after carrying out pre-treatment such as pickling and high pressure; under complex enzyme hydrolysis protective material condition through the hydrolysis of papoid, Sumizyme MP; again successively through the enzyme that goes out, filtration; decolouring; desalination and concentration and spraying drying obtain the Collagen Hydrolysate that molecular weight is single-minded, meet food and healthcare products, makeup requirement.Collagen Hydrolysate is the abundantest animal proteinum of Mammals in-vivo content, can be applied in medical treatment, food, the cosmetic industry, can be made into hemostasis pulvis, burn Ao material, pharmaceutical carrier, heart and distinguish that the equivalent material of film, blood vessel, oesophagus and tracheae, surgery are with variforms such as sutures, as the substratum of biological fermentation, mixed feed production and granulated feed binding agent; Collagen class functional foodstuff and healthcare products; Can also improve the quality of meat, as packaging material for food, beverage finings with produce superior cosmetics.
Advantage of the present invention comprises that adopting the by product bone meal that derives from skeletal grain production is main raw material, greatly reduces raw materials cost.Be raw material with the bovine bone powder, adopt the papoid of plant extract, adopt the Sumizyme MP of microbial fermentation preparation, do not add other animal derived zymins, do not adopt other non-Islamic raw materials, can guarantee the Islamic of product, meet to the requirement of Muslim country's supplying food and healthcare products, daily use chemicals raw material.Adopt the technology of two step directional enzymatics of papoid and Sumizyme MP; adopt the complex enzyme hydrolysis protective material simultaneously; guaranteed that proteolytic enzyme does not destroy the biological activity of hydrolysate when carrying out the directional enzymatic reaction; guarantee all one's life of molecular weight of product, effectively ensured biological activity and the fine quality of product.The hydrolysis temperature that adopts is 48～50 ℃ medium temperature condition, and under this temperature, the peptide chain in the bone meal can resolve into the polypeptide of molecular weight homogeneous slowly under the effect of proteolytic enzyme, kept higher biological activity.Adopt the nanofiltration membrane of 200Da to carry out membrane sepn, effectively removed small molecular weight impurity, also feed liquid is concentrated simultaneously, significantly reduced the energy consumption of spray-drying process.The present invention adopts the technology of papoid and 2709 Sumizyme MPs, two steps directional enzymatic; add MO111-9 complex enzyme hydrolysis protective material simultaneously; biochemical extraction and separation method such as comprehensive utilization membrane sepn and hig h-speed centrifugal spray drying; to the full extent raw material is hydrolyzed to the protein product of 1000Da to 2200Da; effectively guarantee its biological activity; and effectively remove small molecular weight impurity in the product; improve active constituent content in the feed liquid; save the energy that drying process consumes, it is single-minded to obtain molecular weight; meet the Islamic quality requirements; the Collagen Hydrolysate product that meets food and healthcare products and makeup requirement.The present invention is suitable for suitability for industrialized production.
Embodiment
Further set forth the present invention below in conjunction with specific embodiment.Below implement process of the present invention and method, except the following content of mentioning specially, be universal knowledege and the common practise of this area, the present invention is not particularly limited content.Protection content of the present invention is not limited to following examples.Under the spirit and scope that do not deviate from inventive concept, variation and advantage that those skilled in the art can expect all are included in the present invention, and are protection domain with the appending claims.Following examples only are used for explanation the present invention, and are not used for limiting the scope of the invention.
The present invention utilizes bone meal to extract the operational path of the method for Collagen Hydrolysate:
(by product that skeletal grain is produced) → pickling → autoclaving → one time directional protection enzymolysis → secondary directional protection enzymolysis → bone meal goes out enzyme → decolouring → filtration → membrane sepn → spraying drying → Collagen Hydrolysate
The technological process of the inventive method (carrying out in the following order):
(1) the by product bone meal with skeletal grain production is raw material, respectively soaks 6～8 hours with the dilute hydrochloric acid of 0.9%, 1.8%, 2.7%, 3.6%, 4.5% 5 concentration, stirs in per 1 hour during immersion once to make immersion more even; Leach bone meal after pickling finishes and clean up;
(2) the above-mentioned bone meal of handling well is put into 126 ℃ of autoclaves, autoclaving 120 minutes;
(3) add the pure water of 2.6 times of bone meal weight in the isothermal reaction jar, put into the good bone meal of autoclaving, heating up also keeps the retort feed temperature between 48～50 ℃; Add papoid from plant extract in the ratio of 16000～18000U/g substrate, add 0.03%MO111-9 complex enzyme hydrolysis protective material again, be incubated 48～50 ℃ of stirrings, enzymolysis 4 hours;
(4) after the primary enzymolysis, regulate pH to 9.0～9.5 with sodium hydroxide, add 2709 Sumizyme MPs in the ratio of 9000U/g substrate, be incubated 48～50 ℃ of stirrings, enzymolysis 4 hours;
(5) after enzymolysis finishes, be warming up to 80～85 ℃, the enzyme 15～20 minutes of going out is cooled to 30～40 ℃, is adjusted to pH6.5～7.0;
(6) add the gac of reduction of feed volume 3.2% again in the above-mentioned feed liquid, be incubated 30～40 ℃ of stirrings 3 hours;
(7) feed liquid after the above-mentioned decolouring is filtered through the clarification filter plate, the filtrate that obtains is carried out desalination and concentration through nanofiltration membrane again, collects trapped fluid;
(8) trapped fluid of collecting is carried out spraying drying under ℃ air outlet temperature condition of 180 ℃ of inlet temperature/85, obtain described Collagen Hydrolysate.
Step of the present invention (1) is that the by product bone meal with skeletal grain production is raw material, is that solvent is handled in pickling with the dilute hydrochloric acid that increases gradually by certain gradient.Described step (3), (4) are to carry out under 48～50 ℃ the medium temperature condition.The specification of nanofiltration membrane is 200Da described in the described step (7).In the Collagen Hydrolysate that step of the present invention (8) obtains, protein content wherein reaches more than 95%, and the molecular weight homogeneous is distributed in 1000Da to the 2200Da scope.The product Collagen Hydrolysate that the inventive method is extracted is used for experimentation on animals and clinical experiment, and experimental result shows that this product Collagen Hydrolysate can make skin recover elasticity, reappear solid compact, reduce sagging phenomenon, level and smooth microgroove and wrinkle; Make tired out, aging skin regenerate vigor and recovery anger; Can accelerate the more living process of skin, shine gloss.
Embodiment 1
The by product bovine bone powder of weighing skeletal grain production is 500kg, soaks respectively 8 hours with each 750kg of dilute hydrochloric acid of 0.9%, 1.8%, 2.7%, 3.6%, 4.5% 5 concentration, stirs in per 1 hour during immersion once to make immersion more even; Leach bone meal after pickling finishes and clean up; Put into 126 ℃ of autoclaves again, autoclaving 120 minutes.Add the pure water of 1300kg in the 2000L isothermal reaction jar, put into the good bone meal of autoclaving, heating up also keeps the retort feed temperature between 48～50 ℃; The papoid that adds 1,320,g80 ten thousand U/g adds 150 gram M0111-9 complex enzyme hydrolysis protective materials again, is incubated 48～50 ℃ of stirrings, enzymolysis 4 hours; After the primary enzymolysis, regulate pH to 9.0～9.5 with sodium hydroxide, add 2709 Sumizyme MPs of 5,94g,100 ten thousand U/g, be incubated 48～50 ℃ of stirrings, enzymolysis 4 hours; After enzymolysis finishes, be warming up to 80～85 ℃, the enzyme 15～20 minutes of going out is cooled to 30～40 ℃, is adjusted to pH6.5～7.0; The gac that adds 42kg again in the above-mentioned feed liquid be incubated 30～40 ℃ of stirrings 3 hours; Feed liquid after the above-mentioned decolouring is filtered through the clarification filter plate, and the filtrate that obtains is carried out desalination and concentration through nanofiltration membrane again, collects trapped fluid; The trapped fluid of collecting is carried out spraying drying under ℃ air outlet temperature condition of 180 ℃ of inlet temperature/85, get Collagen Hydrolysate 66.5kg.
Method according to the QB2732 regulation is measured product, protein content is 96.1% in the Collagen Hydrolysate, its mean molecule all is distributed in 1000Da to the 2200Da scope, transmittance A450nm=80.2%, A620nm=93.0%, other every indexs are accord with Q B2732 People's Republic of China (PRC) light industry ministerial standard all.
Embodiment 2
The by product bovine bone powder of weighing skeletal grain production is 500kg, soaks respectively 6 hours with each 750kg of dilute hydrochloric acid of 0.9%, 1.8%, 2.7%, 3.6%, 4.5% 5 concentration, stirs in per 1 hour during immersion once to make immersion more even; Leach bone meal after pickling finishes and clean up; Put into 126 ℃ of autoclaves again, autoclaving 100 minutes.Add the pure water of 1300kg in the 2000L isothermal reaction jar, put into the good bone meal of autoclaving, heating up also keeps the retort feed temperature between 48～50 ℃; The papoid that adds 1,320,g80 ten thousand U/g adds 150 gram M0111-9 complex enzyme hydrolysis protective materials again, is incubated 48～50 ℃ of stirrings, enzymolysis 4 hours; After the primary enzymolysis, regulate pH to 9.0～9.5 with sodium hydroxide, add 2709 Sumizyme MPs of 5,94g,100 ten thousand U/g, be incubated 48～50 ℃ of stirrings, enzymolysis 4 hours; After enzymolysis finishes, be warming up to 80～85 ℃, the enzyme 15～20 minutes of going out is cooled to 30～40 ℃, is adjusted to pH6.5～7.0; The gac that adds 42kg again in the above-mentioned feed liquid be incubated 30～40 ℃ of stirrings 3 hours; Feed liquid after the above-mentioned decolouring is filtered through the clarification filter plate, and the filtrate that obtains is carried out desalination and concentration through nanofiltration membrane again, collects trapped fluid; The trapped fluid of collecting is carried out spraying drying under ℃ air outlet temperature condition of 180 ℃ of inlet temperature/85, get Collagen Hydrolysate 59.5kg.
Method according to the QB2732 regulation is measured product, protein content is 97.2% in the Collagen Hydrolysate, its mean molecule all is distributed in 1000Da to the 2200Da scope, transmittance A450nm=82.0%, A620nm=94.7%, other every indexs are accord with Q B2732 People's Republic of China (PRC) light industry ministerial standard all.
Embodiment 3
The by product bovine bone powder of weighing skeletal grain production is 500kg, soaks respectively 24 hours with each 1500kg of dilute hydrochloric acid of 2%, 4% two concentration, stirs in per 1 hour during immersion once to make immersion more even; Leach bone meal after pickling finishes and clean up.Add the pure water of 1300kg in the 2000L isothermal reaction jar, put into the bone meal of handling well, heating up also keeps the retort feed temperature between 48～50 ℃; The papoid that adds 1,980,g80 ten thousand U/g adds 150 gram M0111-9 complex enzyme hydrolysis protective materials again, is incubated 48～50 ℃ of stirrings, enzymolysis 4 hours; After the primary enzymolysis, regulate pH to 9.0～9.5 with sodium hydroxide, add 2709 Sumizyme MPs of 8,91g,100 ten thousand U/g, be incubated 48～50 ℃ of stirrings, enzymolysis 4 hours; After enzymolysis finishes, be warming up to 80～85 ℃, the enzyme 15～20 minutes of going out is cooled to 30～40 ℃, is adjusted to pH6.5～7.0; The gac that adds 42kg again in the above-mentioned feed liquid be incubated 30～40 ℃ of stirrings 3 hours; Feed liquid after the above-mentioned decolouring is filtered through the clarification filter plate, and the filtrate that obtains is carried out desalination and concentration through nanofiltration membrane again, collects trapped fluid; The trapped fluid of collecting is carried out spraying drying under ℃ air outlet temperature condition of 180 ℃ of inlet temperature/85, get Collagen Hydrolysate 36.8kg.
Method according to the QB2732 regulation is measured product, protein content is 90.7% in the Collagen Hydrolysate, its mean molecule all is distributed in 1000Da to the 2200Da scope, transmittance A450nm=78.7%, A620nm=86.1%, other every indexs are accord with Q B2732 People's Republic of China (PRC) light industry ministerial standard all.
Among the embodiment 2, reduced pickling time and autoclaving time, yield is slightly on the low side, and other indexs of finished product are suitable with embodiment 1.In embodiment 3, adopt the method for two kinds of conventional concentration picklings to handle, and do not adopt autoclaving, and adopt the ordinary method that directly strengthens enzyme dosage to be hydrolyzed, yield has only 55% of embodiment 1, and a large amount of collagen proteins does not come out from bone meal heavy water solution; The detection index of product also descends to some extent.
The above is preferred embodiment of the present invention only, is not to limit protection scope of the present invention.Be that all equivalences of doing according to content of the present invention change and modification, all should belong to category of the present invention.
Claims (9)
1. method of utilizing bone meal to extract Collagen Hydrolysate, it is characterized in that, described bone meal is after pickling processing, autoclaving, under complex enzyme hydrolysis protective material condition through the hydrolysis of papoid, Sumizyme MP, successively through high temperature go out enzyme, filtration, decolouring, desalination and concentration and spraying drying, obtain purpose product Collagen Hydrolysate again; Wherein, described bone meal is the by product in the skeletal grain preparation process, and described bone meal particle diameter is less than 5mm.
2. the method for claim 1 is characterized in that, described pickling is handled and referred to described bone meal was respectively soaked 6～8 hours with the dilute hydrochloric acid of 0.9%, 1.8%, 2.7%, 3.6%, 4.5% concentration respectively.
3. the method for claim 1 is characterized in that, described autoclaving is that the bone meal after pickling is handled placed under 0.14～0.15Mpa high pressure boiling 100～120 minutes.
4. the method for claim 1; it is characterized in that; describedly under complex enzyme hydrolysis protective material condition, carry out papain hydrolysis and refer to; in the isothermal reaction jar, put into water and the bone meal after autoclaving; add papoid from plant extract in the ratio of 16000～18000U/g substrate; add the complex enzyme hydrolysis protective material again, hydrolysis temperature is 48～50 ℃, continues to stir enzymolysis 4～6 hours.
5. the method for claim 1; it is characterized in that; describedly under complex enzyme hydrolysis protective material condition, carry out hydrolysis by novo; be to regulate pH to 9.0～9.5 with sodium hydroxide; ratio in the 9000U/g substrate adds the Sumizyme MP for preparing through microbial fermentation; hydrolysis temperature is 48～50 ℃, continues to stir enzymolysis 4～6 hours.
6. the method for claim 1 is characterized in that, the described high temperature enzyme that goes out is to be warming up to 80～85 ℃, and the enzyme 15～20 minutes of going out is cooled to 30～40 ℃, is adjusted to pH6.5～7.0.
7. the method for claim 1 is characterized in that, it is to add gac that described filtration and decolouring are handled, and 30～40 ℃ were stirred 2～3 hours.
8. the method for claim 1 is characterized in that, described desalination and concentration are that the feed liquid after the decolouring is filtered through the clarification filter plate, carry out desalination and concentration through nanofiltration membrane again, collect trapped fluid.
9. the method for claim 1 is characterized in that, the collagen content in the described product Collagen Hydrolysate reaches more than 95%, and the molecular weight homogeneous is distributed in 1000Da to 2200Da scope.
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| CN105104754A (en) * | 2015-09-02 | 2015-12-02 | 湖南伍星生物科技有限公司 | Bone glue enzyme hydrolyzed protein feed processed by residues obtained by extracting chondroitin sulfate and processing method for bone glue enzyme hydrolyzed protein feed |
| CN105549520A (en) * | 2014-10-28 | 2016-05-04 | 兰州理工大学 | Automation control system of acid preparation process in production process of gelatin |
| CN106309372A (en) * | 2016-08-31 | 2017-01-11 | 湖北瑞邦生物科技有限公司 | Bovine bone polypeptide sustained-release preparation and preparation method thereof |
| EP3170393A1 (en) | 2015-11-18 | 2017-05-24 | Institutul National de Cercetare-Dezvoltare Pentru Textile si Pielarie (INCDTP) Sucursala Institul de Cercetare Pielarie Incaltaminte (ICPI) | Collagen polydispersions for the treatment of cereal seeds and process thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105549520A (en) * | 2014-10-28 | 2016-05-04 | 兰州理工大学 | Automation control system of acid preparation process in production process of gelatin |
| CN105104754A (en) * | 2015-09-02 | 2015-12-02 | 湖南伍星生物科技有限公司 | Bone glue enzyme hydrolyzed protein feed processed by residues obtained by extracting chondroitin sulfate and processing method for bone glue enzyme hydrolyzed protein feed |
| EP3170393A1 (en) | 2015-11-18 | 2017-05-24 | Institutul National de Cercetare-Dezvoltare Pentru Textile si Pielarie (INCDTP) Sucursala Institul de Cercetare Pielarie Incaltaminte (ICPI) | Collagen polydispersions for the treatment of cereal seeds and process thereof |
| CN106309372A (en) * | 2016-08-31 | 2017-01-11 | 湖北瑞邦生物科技有限公司 | Bovine bone polypeptide sustained-release preparation and preparation method thereof |
| CN106309372B (en) * | 2016-08-31 | 2019-12-24 | 湖北瑞邦生物科技有限公司 | Bovine bone polypeptide sustained release agent and preparation method thereof |
| CN108112990A (en) * | 2017-12-29 | 2018-06-05 | 银川中科元昊科技有限公司 | After a kind of ox bone acid treatment in sediment beneficiating ingredient extracting method |
| CN110269243A (en) * | 2018-03-17 | 2019-09-24 | 北京神飞航天应用技术研究院 | A kind of dedicated raising bone density nutritional preparation of spacefarer |
| CN110656142A (en) * | 2018-06-29 | 2020-01-07 | 台盐实业股份有限公司 | High-purity undenatured collagen and method for producing same |
| CN114365848A (en) * | 2020-10-14 | 2022-04-19 | 上海阿敏生物技术有限公司 | Collagen tablet and preparation method and application thereof |
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