CN116606897A - High-purity bone peptide based on ultrafiltration technology and enzymolysis preparation system thereof - Google Patents
High-purity bone peptide based on ultrafiltration technology and enzymolysis preparation system thereof Download PDFInfo
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- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 73
- 238000000108 ultra-filtration Methods 0.000 title claims abstract description 26
- 238000002360 preparation method Methods 0.000 title claims abstract description 22
- 238000005516 engineering process Methods 0.000 title claims abstract description 12
- 239000007788 liquid Substances 0.000 claims abstract description 96
- 239000000243 solution Substances 0.000 claims abstract description 41
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- 238000002156 mixing Methods 0.000 claims abstract description 30
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- 238000011221 initial treatment Methods 0.000 claims abstract description 25
- 238000012958 reprocessing Methods 0.000 claims abstract description 22
- 238000001914 filtration Methods 0.000 claims abstract description 21
- 239000011259 mixed solution Substances 0.000 claims abstract description 19
- 239000000843 powder Substances 0.000 claims abstract description 19
- 238000012544 monitoring process Methods 0.000 claims abstract description 9
- 230000009849 deactivation Effects 0.000 claims abstract description 7
- 238000001035 drying Methods 0.000 claims abstract description 7
- 238000001694 spray drying Methods 0.000 claims abstract description 5
- 238000005507 spraying Methods 0.000 claims abstract description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 34
- 238000011282 treatment Methods 0.000 claims description 30
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
- C12M21/18—Apparatus specially designed for the use of free, immobilized or carrier-bound enzymes
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- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/12—Purification
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Abstract
The invention provides a high-purity bone peptide and an enzymolysis preparation system thereof based on an ultrafiltration technology, and compared with the prior art, the enzymolysis preparation system also comprises a mixing module for receiving bone powder and dissolving the bone powder to obtain a mixed solution, an enzymolysis module for preparing the mixed solution into an enzymolysis solution, first centrifugal equipment for centrifuging the enzymolysis solution, a decoloring module for decoloring the primary treatment solution after enzyme deactivation, second centrifugal equipment for centrifuging the decolored primary treatment solution, an ultrafiltration module for filtering the reprocessing solution to obtain bone polypeptide solution, and a spray drying device for drying and spraying the bone polypeptide solution to obtain the bone peptide. The invention can monitor and regulate the preparation process of bone peptide in real time through the functions of temperature control, liquid level monitoring, automatic liquid adding and the like, thereby ensuring the controllability and the stability of the production process.
Description
Technical Field
The invention relates to the technical field of polypeptide extraction and preparation, in particular to high-purity bone peptide based on an ultrafiltration technology and an enzymolysis and preparation system thereof.
Background
The bone collagen polypeptide is an active polypeptide substance extracted from bones of limbs, and is a polypeptide molecule with bone related biological activity. They play an important role in the formation, growth, repair, metabolism, etc. of bone tissue. Bone polypeptides may be derived from protein degradation products in the bone matrix, or may be synthesized and released by bone cells. During bone metabolism, bone polypeptides trigger a series of cellular signal transduction pathways through binding to specific receptors, regulating bone cell proliferation, differentiation and function. Bone polypeptides can affect bone cell activity, synthesis and mineralization of bone matrix, and bone remodeling and repair processes. Bone polypeptides have a variety of biological activities, including promoting bone cell proliferation and differentiation, enhancing bone matrix synthesis, inhibiting bone resorption, promoting bone mineralization, and the like. These activities make bone polypeptides an important area of research and application in the treatment of bone related diseases. In clinical application, bone polypeptides are widely studied for treating bone-related diseases such as osteoporosis, fracture healing disorder, bone defect repair and the like. By applying proper bone polypeptide, the regeneration capacity of bone tissue can be enhanced, the bone healing process can be accelerated, and the increase of bone density can be promoted, so that the treatment effect of diseases such as osteoporosis and the like can be improved. In general, bone polypeptides play an important role in bone tissue formation, growth, repair and metabolism as a class of polypeptide molecules having bone-related biological activity. The research and application of the novel polypeptide provide a new path and strategy for the treatment and rehabilitation of bone related diseases, and have wide application prospect.
The experimental team carries out browsing and researching of a large number of relevant record data for a long time aiming at the relevant technology of bone peptide production, and simultaneously depends on relevant resources, and carries out a large number of relevant experiments, and the prior art that discovers existence through a large number of searches is like CN211746738U, CN217568102U, CN112501229B and CN112501229A that prior art discloses, and a production line for preparing small molecule peptide by using fresh ox bone as disclosed in the prior art includes a bottom plate, the bottom fixedly connected with support column of bottom plate, the last fixedly connected with rubbing crusher of bottom plate, the bottom fixedly connected with conveyer pipe of rubbing crusher, the front fixedly connected with fixed supporting seat of rubbing crusher, the front rotation of fixed supporting seat is connected with the transfer line, the surface of transfer line is connected with the conveyer belt through the drive wheel transmission, the one end overlap joint of conveyer belt has the case of cooking, the inner wall of case is provided with inlet tube and outlet pipe respectively, the interior bottom wall of case of cooking has seted up the recess. This utilize production line of fresh ox bone preparation small molecule peptide through setting up cooking box, hot plate, fixed plate, gyro wheel, chain, movable tray, scraper blade, drive gear and dwang, is convenient for reach the effect of independently clearing up the garrulous ox bone in the cooking box in the production line of utilizing fresh ox bone preparation small molecule peptide.
The invention is made to solve the problems that the production quality and efficiency of bone peptide products are affected due to the fact that the filtration efficiency in the process of preparing bone peptide is too low, the purity of the produced bone peptide is unqualified due to the fact that the purification treatment efficiency in the production process is low, and the like in the prior art.
Disclosure of Invention
The invention aims at providing a high-purity bone peptide and an enzymolysis preparation system thereof based on an ultrafiltration technology aiming at the defects existing in the prior art.
In order to overcome the defects in the prior art, the invention adopts the following technical scheme:
the high-purity bone peptide and the enzymolysis preparation system thereof based on the ultrafiltration technology are characterized in that the bone peptide is prepared by the following steps:
s101: cleaning animal bone, freezing, cutting to obtain bone particles with diameter of 4-10cm,
s102: steaming and boiling bone particles at 130-150deg.C for 70-110min to obtain pretreated bone particles,
s103: washing the pretreated bone particles with water at 100 ℃ for at least two times, airing, crushing the aired pretreated aggregate, sieving to 20-40 meshes to obtain bone powder,
s104: mixing bone powder and water according to the mass ratio of 1:5-1:12, preserving heat for 3-3.5h at 105-120 ℃, then cooling to 15-35 ℃ to obtain mixed solution,
s105: mixing neutral protease and pepsin according to a mass ratio of 1:2 to obtain a treatment enzyme, adding the mixed enzyme into the mixed solution, placing the mixed enzyme and the mixed solution in a ratio of 600-690U:1mL for enzymolysis for 7h at 30-55 ℃ to obtain an enzymolysis solution,
s106: after the enzymolysis liquid is kept at 120 ℃ for 20min for enzyme deactivation treatment, the supernatant is centrifugally taken as primary treatment liquid,
s107: adding active carbon into the primary treatment liquid, standing at pH4.0-7.0 and 35-50deg.C for 30-80min to complete decolorizing treatment, centrifuging to obtain supernatant as reprocessing liquid,
s108: filtering the reprocessing solution with a filter membrane to obtain bone polypeptide solution, drying the bone polypeptide solution to obtain bone peptide,
the enzymolysis preparation system comprises a mixing module for receiving the bone powder and dissolving the bone powder to obtain a mixed solution, an enzymolysis module for preparing the mixed solution into an enzymolysis solution, a first centrifugal device for centrifuging the enzymolysis solution, a decoloring module for decoloring the primary treatment solution after enzyme deactivation, a second centrifugal device for centrifuging the decolored primary treatment solution, an ultrafiltration module for filtering the reprocessing solution to obtain bone polypeptide solution, and a spray drying device for drying and spraying the bone polypeptide solution to obtain bone peptide,
the mixing module comprises a mixing vessel, a storage vessel for placing the bone powder, a spiral discharging machine, a stirring mechanism, a temperature control device and a transfer pipe, wherein the spiral discharging machine is used for placing the storage vessel, the feeding end of the spiral discharging machine is communicated with the storage vessel, the discharging end of the spiral discharging machine is communicated with the mixing vessel, the stirring mechanism is arranged inside the mixing vessel in a matched mode, the temperature control device is used for adjusting the temperature of mixed liquid in the storage vessel, and the transfer pipe is used for transferring the mixed liquid in the mixing vessel to the enzymolysis module.
Further, the enzymolysis module comprises an enzymolysis dish for receiving the mixed liquid, a liquid storage tank for storing the treatment enzyme, a liquid inlet pipe for communicating the liquid storage tank with the enzymolysis dish, a driving pump for quantitatively driving the treatment enzyme in the liquid storage tank into the enzymolysis dish, a heater for heating the enzymolysis dish, a liquid level sensor for monitoring the liquid level of the mixed liquid in the enzymolysis dish, and a calculating unit for further generating transfer quantity of the treatment enzyme by the driving pump based on a liquid level monitoring value of the liquid level sensor.
Further, the decoloring module comprises a decoloring dish, a plurality of matching channels which are communicated with the upper dish wall of the decoloring dish, a filter cylinder which can vertically penetrate into the decoloring dish from the matching channels, an inner ring sleeved on the inner channel wall of the matching channels, a clamping ring sleeved on the outer cylinder wall near the top of the filter cylinder, a shielding cover for clamping and fixing the upper channel opening of the matching channels, and a transmission pipe for transferring reprocessing liquid obtained after decoloring in the decoloring dish to the outside of the decoloring dish,
wherein the diameter of the cross section of the matching channel is the same, the inner ring is arranged close to the lower channel opening of the matching channel, the ring outer diameter of the clamping ring is larger than the inner diameter of the inner ring, the ring outer diameter of the clamping ring is smaller than the inner diameter of the matching channel, the region of the matching channel above the inner ring is an upper channel region, the region of the matching channel below the inner ring is a lower channel region, the clamping ring can be movably matched in the upper channel region, the clamping ring can not enter the lower channel region under the limitation of the inner ring, the filter cartridge can movably penetrate through the upper channel region and the lower channel region,
the filter cylinder comprises a cylinder body with a cylindrical structure, a feeding hole arranged at the top of the cylinder body, and filtering holes uniformly distributed on the outer cylinder wall of the cylinder body, wherein the inside of the cylinder body is used for placing activated carbon, the aperture of the filtering holes is smaller than the particle size of the activated carbon, the activated carbon is limited inside the filter cylinder, when the clamping ring is abutted with the inner ring, at least part of the cylinder body is immersed into primary treatment liquid, and the activated carbon in the filter cylinder is contacted with the primary treatment liquid, so that the decoloring treatment of the primary treatment liquid is realized, and the filter cylinder is moved out of the decoloring dish from the matching channel by opening the shielding cover.
Further, the ultrafiltration module comprises an upper receiving dish, a lower receiving dish positioned below the upper receiving dish, a fixed support above a preset distance for fixing the lower receiving dish to the ground, two fixed supports symmetrically fixed on the side wall of the upper receiving dish, a lifting mechanism with the top end fixed on the fixed support and the bottom end fixed to the ground, an opening frame horizontally fixed on the bottom wall of the receiving dish, a funnel with the top fixed with the bottom of the upper receiving dish and the bottom extending downwards, a clamping groove which is arranged on the lower receiving dish and can be clamped with the opening frame, a first rubber layer which is laid on the bottom wall of the opening frame, a second rubber layer which is laid on the opening edge of the lower opening of the funnel, a liquid outlet pump with one end communicated with the side dish wall of the upper receiving dish and the other end inclined downwards, a filter membrane mechanism for driving the residual liquid in the lower receiving dish to flow out of the liquid outlet pump from the liquid outlet pipe, an adsorption pipe with one end communicated with the bottom wall of the lower receiving dish, an adsorption pipe fixedly connected with the other end of the adsorption pipe, and a one-way valve embedded in the adsorption pipe,
the lower opening of the funnel and the frame bottom wall of the opening frame are positioned on the same horizontal plane, the opening frame is a frame part, the middle part formed by closing and surrounding four connecting rods is an opening for receiving, the top wall of the dish of the upper receiving dish is an upper dish top wall, the bottom wall of the dish of the upper receiving dish is an upper dish bottom wall, the top wall of the dish of the lower receiving dish is a lower dish top wall, the bottom wall of the dish of the lower receiving dish is a lower dish bottom wall, the middle part of the upper dish bottom wall is provided with a discharge hole, the middle part of the top wall of the lower dish is provided with a receiving port, wherein the discharge hole is communicated with the upper opening of the funnel in a sealing way,
the cross section diameter length of funnel reduces gradually from last to the setting down, receive the bore length and be greater than the lower opening diameter length setting of funnel, just receive the bore length and be less than the long setting of opening frame diameter, the block groove sets up in lower ware roof, and at least part bottom of opening frame can the block be fixed in the block groove to under the deformation extrusion of second rubber and the groove diapire of block groove, realize receiving the ware and receive the airtight connection of ware under with, discharge gate, receiving mouth and the coaxial setting of lower opening of funnel.
Further, two grooves are symmetrically formed in the lower receiving dish, the grooves are of an open groove structure which is formed by sinking the top wall of the lower dish relative to the bottom wall of the lower dish, one groove is used as a first groove, the other groove is used as a second groove, the top opening end of the first groove is used as a first opening, and the top opening end of the second groove is used as a second opening.
Further, the filter membrane mechanism comprises a first rotary rod rotationally matched in the first groove, a second rotary rod rotationally matched in the second groove, a driving motor embedded in the first groove and fixedly connected with one end of the power output shaft and one end of the first rotary rod, a recovery motor embedded in the second groove and fixedly connected with one end of the power output shaft and one end of the second rotary rod, a filter membrane at least partially tensioned and wound on the first rotary rod, an ultraviolet lamp bead embedded in the bottom wall of the first groove, a first matched cover covered on the first opening, a second matched cover covered on the second opening, a linear through hole arranged on the first matched cover and a linear through hole arranged on the second matched cover, wherein the first matched cover is detachably covered on the first opening through clamping, the second matched cover is detachably covered on the second opening through clamping, and the first rotary rod, the second rotary rod, the linear through hole and the linear through hole are mutually parallel.
The beneficial effects obtained by the invention are as follows:
1. the invention can monitor and regulate the preparation process of the bone peptide in real time through the functions of temperature control, liquid level monitoring, automatic liquid adding and the like, thereby ensuring the controllability and the stability of the production process and improving the purity and the production efficiency of the bone peptide.
2. According to the invention, the active carbon is arranged in the filter cartridge, so that pigments and impurities in the primary treatment liquid can be effectively removed, and a high-quality reprocessing liquid is obtained, and the purity of the bone peptide product produced by the method is effectively improved.
3. Through the cooperation passageway, filter cartridge and shelter from the cooperation between the lid inseparable, and can open or close in a flexible way and shelter from the lid, conveniently in time replace and maintain to filter cartridge, the active carbon is limited inside the filter cartridge, can reuse, and then effectively reduced the use amount and the waste of active carbon to the resource has been practiced thrift.
4. According to the invention, the reprocessing liquid is purified and concentrated through the ultrafiltration module, so that the bone polypeptide liquid with correspondingly improved purity and quality of bone peptide is further obtained, the design and technical measures of the ultrafiltration module improve the filtration and production efficiency of the bone peptide liquid, ensure the safety and stability, and are beneficial to improving the production efficiency and quality of high-purity bone peptide.
5. The ultrafiltration module is used for filtering the reprocessing liquid through a filter membrane by a membrane separation technology of selectively separating substances with molecular size so as to effectively remove larger molecular proteins, macromolecular impurities and particles in the reprocessing liquid and obtain bone polypeptide liquid retaining target bone peptide molecules, thereby realizing concentration and purification of bone peptide and obtaining high-purity bone peptide products,
drawings
The invention will be further understood from the following description taken in conjunction with the accompanying drawings. The components in the figures are not necessarily to scale, emphasis instead being placed upon illustrating the principles of the embodiments. Like reference numerals designate corresponding parts throughout the different views.
FIG. 1 is a schematic diagram of the enzymatic hydrolysis preparation system of the present invention.
Fig. 2 is a schematic view of a part of the structure of an ultrafiltration module according to the present invention.
FIG. 3 is a schematic view of a part of the structure of the upper receiving dish of the present invention.
FIG. 4 is a schematic view of a part of the structure of the lower receiving dish of the present invention.
Fig. 5 is a schematic view of a further part of the structure of the lower receiving dish according to the present invention.
Fig. 6 is a schematic diagram of a part of the structure of the decoloring module according to the present invention.
Reference numerals illustrate: 1-upper receiving dish; 2-fixing a support; 3-a lifting mechanism; 4-fixing a bracket; 5-lower receiving dish; 6-filtering membrane; 7-removing the tube; 8-opening frame; 9-funnel; 10-a first groove; 11-bottom wall of the lower dish; 12-a linear through-hole; 13-a first mating cover; 14-a second groove; 15-a second mating cap; 16-a linear penetration port; 17-a first rotary stick; 18-bearing seats; 19-a clamping groove; 20-receiving opening; 21-a second roller; 22-a recovery motor; 23-driving a motor; 24-shielding cover; 25-decolorizing dishes; 26-a filter cartridge; 27-an inner ring; 28-a clamping ring; 29-mating channels.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in further detail with reference to the following examples thereof; it is noted that the specific embodiments described herein are for purposes of illustration only and are not intended to be limiting. Other systems, methods, and/or features of the present embodiments will be or become apparent to one with skill in the art upon examination of the following detailed description. And the terms describing the positional relationship in the drawings are merely for illustrative purposes and are not to be construed as limiting the present patent, and specific meanings of the terms can be understood by those of ordinary skill in the art according to specific circumstances.
Embodiment one: referring to fig. 1, fig. 2, fig. 3, fig. 4, fig. 5 and fig. 6, this embodiment constructs a high-purity bone peptide based on ultrafiltration technology and an enzymolysis preparation system thereof, wherein the bone peptide is prepared by the following steps:
s101: cleaning animal bone, freezing, cutting to obtain bone particles with diameter of 4-10cm,
s102: steaming and boiling bone particles at 130-150deg.C for 70-110min to obtain pretreated bone particles,
s103: washing the pretreated bone particles with water at 100 ℃ for at least two times, airing, crushing the aired pretreated aggregate, sieving to 20-40 meshes to obtain bone powder,
s104: mixing bone powder and water according to the mass ratio of 1:5-1:12, preserving heat for 3-3.5h at 105-120 ℃, then cooling to 15-35 ℃ to obtain mixed solution,
s105: mixing neutral protease and pepsin according to a mass ratio of 1:2 to obtain a treatment enzyme, adding the mixed enzyme into the mixed solution, placing the mixed enzyme and the mixed solution in a ratio of 600-690U:1mL for enzymolysis for 7h at 30-55 ℃ to obtain an enzymolysis solution,
s106: after the enzymolysis liquid is kept at 120 ℃ for 20min for enzyme deactivation treatment, the supernatant is centrifugally taken as primary treatment liquid,
s107: adding active carbon into the primary treatment liquid, standing at pH4.0-7.0 and 35-50deg.C for 30-80min to complete decolorizing treatment, centrifuging to obtain supernatant as reprocessing liquid,
s108: filtering the reprocessing solution by a filter membrane to obtain bone polypeptide solution, and drying the bone polypeptide solution to obtain bone peptide;
the enzymolysis preparation system comprises a mixing module for receiving the bone powder and dissolving the bone powder to obtain a mixed solution, an enzymolysis module for preparing the mixed solution into an enzymolysis solution, first centrifugal equipment for carrying out centrifugal treatment on the enzymolysis solution, a decoloring module for carrying out decoloring treatment after enzyme deactivation on the primary treatment solution, second centrifugal equipment for carrying out centrifugal treatment on the primary treatment solution after the decoloring treatment, an ultrafiltration module for carrying out filtration treatment on the reprocessing solution to obtain bone polypeptide solution, and a spray drying device for drying and powder spraying the bone polypeptide solution to obtain bone peptide;
the mixing module comprises a mixing vessel, a storage vessel for placing the bone powder, a spiral discharging machine, a stirring mechanism, a temperature control device and a transfer pipe, wherein the spiral discharging machine is used for placing the bone powder, the feeding end of the spiral discharging machine is communicated with the storage vessel, the discharging end of the spiral discharging machine is communicated with the mixing vessel, the stirring mechanism is matched with the mixing vessel, the temperature control device is used for adjusting the temperature of mixed liquid in the storage vessel, and the transfer pipe is used for transferring the mixed liquid in the mixing vessel to the enzymolysis module, wherein centrifugal equipment, the spray drying device, the temperature control device and the stirring mechanism are of the prior art and are not repeated herein;
the enzymolysis module comprises an enzymolysis dish for receiving the mixed liquid, a liquid storage tank for storing treatment enzymes, a liquid inlet pipe which is communicated with the liquid storage tank and the enzymolysis dish, a driving pump for quantitatively driving the treatment enzymes in the liquid storage tank into the enzymolysis dish, a heater for heating the enzymolysis dish, a liquid level sensor for monitoring the liquid level of the mixed liquid in the enzymolysis dish, and a calculating unit for further generating transfer quantity of the treatment enzymes by the driving pump based on the liquid level monitoring value of the liquid level sensor;
the calculating unit comprises a liquid level sensor signal interface for receiving liquid level information detected by a liquid level sensor, a preprocessing subunit for filtering and denoising the received liquid level information, a processing subunit for calculating the amount of processing enzyme required to be added in enzymolysis reaction based on the preprocessed liquid level information and combining with a predesigned enzymolysis reaction dynamics model, a transmitting subunit for transmitting the calculated processing enzyme transfer amount of the processing subunit to a driving pump controller, and a driving pump controller for further controlling the quantitative transfer of the driving pump to the processing enzyme according to the processing enzyme transfer amount transmitted by the processing subunit;
the calculating unit can be realized by an embedded system or a computer program selected by a person skilled in the art according to actual requirements, and is not described herein, one end of the transfer pipe is communicated with the mixing vessel, the other end of the transfer pipe is communicated with the enzymolysis vessel, the spiral discharging machine is used for quantitatively conveying bone meal into the mixing vessel, the stirring mechanism is used for stirring the mixed liquid in the mixing vessel, and the mixed liquid obtained by mixing in the mixing vessel is transferred from the transfer pipe into the enzymolysis vessel;
the invention can monitor and regulate the preparation process of the bone peptide in real time through the functions of temperature control, liquid level monitoring, automatic liquid adding and the like, thereby ensuring the controllability and the stability of the production process and improving the purity and the production efficiency of the bone peptide.
Embodiment two: in addition to the content of the above embodiments, the decolorizing module includes a decolorizing dish, a plurality of matching channels penetrating through the upper dish wall of the decolorizing dish, a filter cartridge capable of vertically penetrating into the decolorizing dish from the matching channels, an inner ring sleeved on the inner channel wall of the matching channels, a clamping ring sleeved on the outer cylinder wall near the top of the filter cartridge, a shielding cover for clamping and fixing the upper channel opening of the matching channels, and a transmission pipe for transferring reprocessing liquid obtained after decolorizing in the decolorizing dish to the outside of the decolorizing dish, wherein the filtering cartridge is combined with the accompanying fig. 1, fig. 2, fig. 3, fig. 4, fig. 5 and fig. 6;
the cross section of the fit channel is the same in diameter length, the inner ring is arranged close to the lower channel opening of the fit channel, the outer diameter of the clamping ring is larger than the inner diameter of the inner ring, the outer diameter of the clamping ring is smaller than the inner diameter of the fit channel, the region of the fit channel, which is positioned above the inner ring, is an upper channel region, the region of the fit channel, which is positioned below the inner ring, is a lower channel region, the clamping ring can be movably matched in the upper channel region, and the clamping ring cannot enter the lower channel region under the limitation of the inner ring, and the filter cartridge can movably penetrate through the upper channel region and the lower channel region;
the filter cylinder comprises a cylinder body with a cylindrical structure, a feed inlet arranged at the top of the cylinder body, and filter holes uniformly distributed on the outer cylinder wall of the cylinder body, wherein the inside of the cylinder body is used for placing activated carbon, the aperture of the filter holes is smaller than the particle size of the activated carbon, the activated carbon is limited in the filter cylinder, when the clamping ring is abutted against the inner ring, at least part of the cylinder body is immersed in primary treatment liquid, the activated carbon in the filter cylinder is contacted with the primary treatment liquid, so that the decoloring treatment of the primary treatment liquid is realized, and the filter cylinder is moved out of the decoloring dish from the matching channel by opening the shielding cover;
according to the invention, the active carbon is arranged in the filter cartridge, so that pigments and impurities in the primary treatment liquid can be effectively removed, and high-quality reprocessing liquid is obtained, meanwhile, the shielding cover can be flexibly opened or closed through the matching channel, the filter cartridge and the shielding cover, the filter cartridge is convenient to replace and maintain in time, the active carbon is limited in the filter cartridge, and the active carbon can be reused, so that the use amount and waste of the active carbon are effectively reduced, and resources are saved.
Embodiment III: in addition to the content of the above embodiments, the ultrafiltration module further includes an upper receiving dish, a lower receiving dish located below the upper receiving dish, a fixed support above a preset distance for fixing the lower receiving dish to the ground, two fixed supports symmetrically fixed on the side walls of the upper receiving dish, a lifting mechanism with the top end fixed on the fixed support and the bottom end fixed to the ground, an opening frame with the top fixed on the bottom wall of the dish of the receiving dish in a horizontal manner, a funnel with the top communicated with the bottom of the upper receiving dish and the bottom extending downwards, a clamping groove arranged on the bottom of the lower receiving dish and capable of being clamped with the opening frame, a first rubber layer arranged on the bottom wall of the frame of the opening frame, a second rubber layer arranged on the opening edge of the lower opening, a moving-out pipe with one end communicated with the side dish wall of the upper receiving dish and the other end inclined downwards, a liquid driving pump for driving residual liquid in the lower receiving dish to flow out from the moving-out pipe, a motor mechanism at least partially transversely arranged on the top wall of the dish of the lower receiving dish, an adsorption valve arranged at least partially, wherein the top wall of the motor is connected with the lower receiving dish, an adsorption valve is connected with the adsorption valve arranged in the adsorption pipe, and the adsorption valve is connected with the adsorption pipe;
the lower opening of the funnel and the frame bottom wall of the opening frame are positioned on the same horizontal plane, the opening frame is a frame part, the middle part of which is formed by closing and surrounding four connecting rods, is an opening for receiving, the top wall of the dish of the upper receiving dish is an upper dish top wall, the bottom wall of the dish of the upper receiving dish is an upper dish bottom wall, the top wall of the dish of the lower receiving dish is a lower dish top wall, the bottom wall of the dish of the lower receiving dish is a lower dish bottom wall, the middle part of the upper dish bottom wall is provided with a discharge hole, the middle part of the top wall of the lower dish is provided with a receiving port, and the discharge hole is communicated with the upper opening of the funnel in a sealing way;
the top wall of the opening frame is fixedly connected with the bottom wall of the upper dish, the discharge hole is arranged in the middle of the bottom wall of the upper dish, the funnel is vertically arranged in the opening area of the opening frame, the cross section diameter of the funnel is gradually reduced from top to bottom, the receiving caliber is longer than the lower opening diameter of the funnel, the receiving caliber is shorter than the opening frame diameter, the clamping groove is formed in the top wall of the lower dish, at least part of the bottom of the opening frame can be clamped and fixed in the clamping groove, and under the deformation extrusion of the bottom wall of the second rubber and the clamping groove, the airtight connection of the upper receiving dish and the lower receiving dish is realized;
the lifting mechanisms at least comprise two, and each fixed support is at least fixedly provided with one lifting mechanism, and the lifting mechanism is used for driving the upper receiving dish to lift and move relative to the ground through telescopic cooperation, wherein the discharge port, the receiving port and the lower opening of the funnel are coaxially arranged;
two grooves are symmetrically formed in the lower receiving dish, the grooves are of an open groove structure which is concavely arranged from the top wall of the lower dish to the bottom wall of the lower dish, one of the grooves is used as a first groove, the other groove is used as a second groove, the top open end of the first groove is used as a first opening, the top open end of the second groove is used as a second opening, and the adsorption tube is communicated with a non-groove area of the top wall of the lower dish;
the filter membrane mechanism comprises a first rotary rod, a second rotary rod, a driving motor, a recovery motor, a filter membrane, an ultraviolet lamp bead, a first matching cover, a second matching cover, a linear through hole and a linear through hole, wherein the first rotary rod is rotatably matched in a first groove, the second rotary rod is rotatably matched in a second groove, the driving motor is embedded in the first groove, a power output shaft is fixedly connected with one end of the first rotary rod, the recovery motor is embedded in the second groove, the power output shaft is fixedly connected with one end of the second rotary rod, the filter membrane is at least partially tensioned and wound on the first rotary rod, the ultraviolet lamp bead is embedded in the bottom wall of the first groove, the first matching cover is covered on the first opening, the second matching cover is covered on the second opening, the linear through hole is formed in the first matching cover, and the linear through hole is formed in the second matching cover, and the first matching cover is detachably covered on the first opening and the second matching cover is detachably covered on the second opening through the clamping;
the first rotary rod, the second rotary rod, the linear through hole and the linear through hole are arranged in parallel, and two ends of the first rotary rod and the second rotary rod are respectively in rotary fit in the first groove and the second groove through bearing seats;
one end of the filter membrane wound on the first rotary rod in a cylindrical shape penetrates out of the first groove from the linear penetrating hole and penetrates into the second groove from the linear penetrating hole, one end of the filter membrane is fixed on the second rotary rod in advance, the driving motor is used for driving the first rotary rod to rotate in an axial mode so that the first rotary rod rotates the wound filter membrane and the unused filter membrane is released, the recycling motor is used for driving the second rotary rod to rotate in an axial mode so that the first rotary rod recycles the used filter membrane, the filter membrane located in the area between the linear penetrating hole and the linear penetrating hole is used as a working membrane, and the working membrane can be fully covered to the receiving hole;
the ultrafiltration module is arranged in such a way that when the lifting mechanism descends to a preset descending height, the first rubber is abutted to the operation film, part of the bottom of the opening frame is clamped into the clamping groove, the second rubber is deformed and extruded into the clamping groove, and meanwhile, gaps between the bottom wall of the groove and the bottom wall of the frame are filled, so that airtight abutting joint between the upper receiving dish and the lower receiving dish is realized, and under the suction operation of the negative pressure motor, reprocessing liquid positioned on the upper receiving dish is sequentially reserved from the lower opening of the funnel under the pressure difference and filtered into the lower receiving dish after passing through the operation film;
after the ultrafiltration module finishes the filtering operation of the reprocessing liquid with a preset volume, the residual liquid remained on the upper receiving dish flows out of the removal pipe through the liquid driving pump, the upper receiving dish is further driven to be far away from the lower receiving dish through the extending operation of the lifting mechanism, and the used operation film is recovered into the second groove through the rotating operation of the first rotating rod and the second rotating rod, so that the continuous and efficient filtering operation of the ultrafiltration module is ensured;
the ultrafiltration module is used for filtering the reprocessing liquid through a filter membrane by a membrane separation technology of selectively separating substances with molecular size so as to effectively remove larger molecular proteins, macromolecular impurities and particles in the reprocessing liquid and obtain bone polypeptide liquid retaining target bone peptide molecules, thereby realizing concentration and purification of bone peptide and obtaining a high-purity bone peptide product;
the ultrafiltration module provided by the invention realizes timely replacement of an operation membrane through the cooperation operation of the lifting mechanism and the filter membrane mechanism, is coaxially arranged through the discharge hole, the receiving port and the lower opening of the funnel so as to further ensure the orderliness of the ultrafiltration module for discharging and collecting bone polypeptide liquid, effectively improves the filtering and generating efficiency of the bone polypeptide liquid by adopting the negative pressure motor, realizes the airtight connection of the first receiving dish and the second receiving dish by adopting the second rubber point rubber sealing piece, and can effectively prevent the air leakage phenomenon of the clamping area of the first receiving dish and the second receiving dish, thereby effectively comprising the safe operation of the ultrafiltration module.
While the invention has been described above with reference to various embodiments, it should be understood that many changes and modifications can be made without departing from the scope of the invention. That is, the methods, systems and devices discussed above are examples. Various configurations may omit, replace, or add various procedures or components as appropriate. For example, in alternative configurations, the methods may be performed in a different order than described, and/or various components may be added, omitted, and/or combined. Moreover, features described with respect to certain configurations may be combined in various other configurations, such as different aspects and elements of the configurations may be combined in a similar manner. Furthermore, as the technology evolves, elements therein may be updated, i.e., many of the elements are examples, and do not limit the scope of the disclosure or the claims. And it is understood that various changes and modifications may be made by those skilled in the art after reading the description of the invention, and such equivalent changes and modifications are intended to fall within the scope of the invention as defined in the appended claims.
Claims (6)
1. The high-purity bone peptide and the enzymolysis preparation system thereof based on the ultrafiltration technology are characterized in that the bone peptide is prepared by the following steps:
s101: cleaning animal bone, freezing, cutting to obtain bone particles with diameter of 4-10cm,
s102: steaming and boiling bone particles at 130-150deg.C for 70-110min to obtain pretreated bone particles,
s103: washing the pretreated bone particles with water at 100 ℃ for at least two times, airing, crushing the aired pretreated aggregate, sieving to 20-40 meshes to obtain bone powder,
s104: mixing bone powder and water according to the mass ratio of 1:5-1:12, preserving heat for 3-3.5h at 105-120 ℃, then cooling to 15-35 ℃ to obtain mixed solution,
s105: mixing neutral protease and pepsin according to a mass ratio of 1:2 to obtain a treatment enzyme, adding the mixed enzyme into the mixed solution, placing the mixed enzyme and the mixed solution in a ratio of 600-690U:1mL for enzymolysis for 7h at 30-55 ℃ to obtain an enzymolysis solution,
s106: after the enzymolysis liquid is kept at 120 ℃ for 20min for enzyme deactivation treatment, the supernatant is centrifugally taken as primary treatment liquid,
s107: adding active carbon into the primary treatment liquid, standing at pH4.0-7.0 and 35-50deg.C for 30-80min to complete decolorizing treatment, centrifuging to obtain supernatant as reprocessing liquid,
s108: filtering the reprocessing solution with a filter membrane to obtain bone polypeptide solution, drying the bone polypeptide solution to obtain bone peptide,
the enzymolysis preparation system comprises a mixing module for receiving the bone powder and dissolving the bone powder to obtain a mixed solution, an enzymolysis module for preparing the mixed solution into an enzymolysis solution, a first centrifugal device for centrifuging the enzymolysis solution, a decoloring module for decoloring the primary treatment solution after enzyme deactivation, a second centrifugal device for centrifuging the decolored primary treatment solution, an ultrafiltration module for filtering the reprocessing solution to obtain bone polypeptide solution, and a spray drying device for drying and spraying the bone polypeptide solution to obtain bone peptide,
the mixing module comprises a mixing vessel, a storage vessel for placing the bone powder, a spiral discharging machine, a stirring mechanism, a temperature control device and a transfer pipe, wherein the spiral discharging machine is used for placing the storage vessel, the feeding end of the spiral discharging machine is communicated with the storage vessel, the discharging end of the spiral discharging machine is communicated with the mixing vessel, the stirring mechanism is arranged inside the mixing vessel in a matched mode, the temperature control device is used for adjusting the temperature of mixed liquid in the storage vessel, and the transfer pipe is used for transferring the mixed liquid in the mixing vessel to the enzymolysis module.
2. The enzymatic hydrolysis preparation system according to claim 1, wherein the enzymatic hydrolysis module includes an enzymolysis vessel for receiving the mixed solution, a liquid storage tank for storing the treatment enzyme, a liquid inlet pipe for communicating the liquid storage tank with the enzymolysis vessel, a drive pump for quantitatively driving the treatment enzyme in the liquid storage tank into the enzymolysis vessel, a heater for heating the enzymolysis vessel, a liquid level sensor for monitoring the liquid level of the mixed solution in the enzymolysis vessel, and a calculation unit for further generating a transfer amount of the treatment enzyme by the drive pump based on a liquid level monitoring value of the liquid level sensor.
3. The enzymolysis preparation system of claim 2, wherein the decolorizing module comprises a decolorizing dish, a plurality of matching channels penetrating through the upper dish wall of the decolorizing dish, a filter cartridge capable of vertically penetrating into the decolorizing dish from the matching channels, an inner ring sleeved on the inner channel wall of the matching channels, a clamping ring sleeved on the outer cylinder wall near the top of the filter cartridge, a shielding cover for clamping and fixing the upper channel opening of the matching channels, and a transmission pipe for transferring the reprocessing liquid obtained after the decolorizing in the decolorizing dish to the outside of the decolorizing dish,
wherein the diameter of the cross section of the matching channel is the same, the inner ring is arranged close to the lower channel opening of the matching channel, the ring outer diameter of the clamping ring is larger than the inner diameter of the inner ring, the ring outer diameter of the clamping ring is smaller than the inner diameter of the matching channel, the region of the matching channel above the inner ring is an upper channel region, the region of the matching channel below the inner ring is a lower channel region, the clamping ring can be movably matched in the upper channel region, the clamping ring can not enter the lower channel region under the limitation of the inner ring, the filter cartridge can movably penetrate through the upper channel region and the lower channel region,
the filter cylinder comprises a cylinder body with a cylindrical structure, a feeding hole arranged at the top of the cylinder body, and filtering holes uniformly distributed on the outer cylinder wall of the cylinder body, wherein the inside of the cylinder body is used for placing activated carbon, the aperture of the filtering holes is smaller than the particle size of the activated carbon, the activated carbon is limited inside the filter cylinder, when the clamping ring is abutted with the inner ring, at least part of the cylinder body is immersed into primary treatment liquid, and the activated carbon in the filter cylinder is contacted with the primary treatment liquid, so that the decoloring treatment of the primary treatment liquid is realized, and the filter cylinder is moved out of the decoloring dish from the matching channel by opening the shielding cover.
4. The enzymatic hydrolysis preparing system as set forth in claim 3, wherein the ultrafiltration module includes an upper receiving dish, a lower receiving dish located below the upper receiving dish, a fixing bracket above a predetermined distance for fixing the lower receiving dish to the ground, two fixing supports symmetrically fixed to side walls of the upper receiving dish, a lifting mechanism with a top end fixed to the fixing supports and a bottom end fixed to the ground, an opening frame horizontally fixed to a bottom wall of the receiving dish, a funnel with a top fixed to a bottom of the upper receiving dish and a bottom extending downward, an engaging groove provided on the lower receiving dish and capable of engaging with the opening frame, a first rubber layer laid on a bottom wall of the frame of the opening frame, a second rubber layer laid on an opening edge of a lower opening of the funnel, a liquid-driving pump with one end connected to a side dish wall of the upper receiving dish and the other end inclined downward, a top wall filter mechanism at least partially disposed across the lower receiving dish, an adsorption tube with one end connected to the lower receiving dish, a negative pressure adsorption tube with the other end fixedly connected to the adsorption tube, a one-way valve disposed in the adsorption tube,
the lower opening of the funnel and the frame bottom wall of the opening frame are positioned on the same horizontal plane, the opening frame is a frame part, the middle part formed by closing and surrounding four connecting rods is an opening for receiving, the top wall of the dish of the upper receiving dish is an upper dish top wall, the bottom wall of the dish of the upper receiving dish is an upper dish bottom wall, the top wall of the dish of the lower receiving dish is a lower dish top wall, the bottom wall of the dish of the lower receiving dish is a lower dish bottom wall, the middle part of the upper dish bottom wall is provided with a discharge hole, the middle part of the top wall of the lower dish is provided with a receiving port, wherein the discharge hole is communicated with the upper opening of the funnel in a sealing way,
the cross section diameter length of funnel reduces gradually from last to the setting down, receive the bore length and be greater than the lower opening diameter length setting of funnel, just receive the bore length and be less than the long setting of opening frame diameter, the block groove sets up in lower ware roof, and at least part bottom of opening frame can the block be fixed in the block groove to under the deformation extrusion of second rubber and the groove diapire of block groove, realize receiving the ware and receive the airtight connection of ware under with, discharge gate, receiving mouth and the coaxial setting of lower opening of funnel.
5. The enzymatic hydrolysis process system of claim 4, wherein two grooves are symmetrically disposed in the lower receiving vessel, wherein the grooves are open-ended groove structures recessed from the top wall of the lower vessel toward the bottom wall of the lower vessel, one of the grooves is a first groove, the other groove is a second groove, the top open end of the first groove is a first opening, and the top open end of the second groove is a second opening.
6. The enzymatic hydrolysis preparation system as claimed in claim 5 wherein the filter membrane mechanism includes a first rotatable rod rotatably fitted in the first recess, a second rotatable rod rotatably fitted in the second recess, a drive motor fitted in the first recess and having a power output shaft fixedly connected to one end of the first rotatable rod, a recovery motor fitted in the second recess and having a power output shaft fixedly connected to one end of the second rotatable rod, a filter membrane at least partially tensioned and wound around the first rotatable rod, an ultraviolet lamp bead fitted in a bottom wall of the first recess, a first fitting cap fitted to the first opening, a second fitting cap fitted to the second opening, a linear through-opening provided in the first fitting cap, and a linear through-opening provided in the second fitting cap, wherein the first fitting cap is detachably fitted to the first opening by snap-fit, the second fitting cap is detachably fitted to the second opening by snap-fit, and the first rotatable rod, the second fitting cap, and the linear through-opening are disposed in parallel to each other.
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