JPS6247214B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a composition comprising chitosan and collagen. More specifically, heat resistance, moisture absorption resistance,
The present invention relates to a novel composition consisting of chitosan and collagen that has excellent mechanical properties and is non-absorbable to living bodies. Collagen is the connective tissue of mammals, birds, etc.
It is a hard protein that exists in bones, teeth, skin, dermis, fascia, etc., and is widely used in the form of edible casing materials, surgical sutures, vascular grafts, artificial skin, etc. Although collagen is superior to other materials used in the above applications, it is not always satisfactory. For example, beef, beef,
The intestines of food animals such as pigs and sheep, that is, natural collagen molded bodies, are used. However, the natural collagen molded body has a limited production amount and is manufactured through complicated processing steps, resulting in high cost. Moreover, the natural collagen molded bodies are not uniform in shape and quality, and therefore are difficult to be applied to ordinary high-speed meat stuffing machines in the production of hams, sausages, and the like. To overcome the above-mentioned drawbacks, a tube-shaped collagen fiber, ie, an artificial collagen casing, obtained by a method as described below has been proposed. However, the artificial collagen fibers have poor film-forming properties. Furthermore, the artificial collagen casing has a thick film thickness due to mechanical strength problems, and therefore has disadvantages such as poor appearance and significantly impaired texture. Furthermore, hams, sausages, etc. made of the artificial collagen casings are damaged during cooking, and undesirable phenomena such as separation between the filled meat and the casing occur. Recently, a method of further crosslinking the artificial collagen casing has been proposed to solve the problem during cooking. However, the crosslinked collagen casing is difficult to satisfy in terms of texture. On the other hand, collagen materials used as surgical medical materials such as surgical sutures, artificial skin, and hemostatic materials mentioned above have not only strength problems but also biostability (non-absorbability).
I am dissatisfied with this point. Various methods have been studied to improve the biostability of the collagen material. For example, there is a method of crosslinking collagen. However, the molded material obtained by this method has significantly reduced mechanical properties such as tensile strength and elongation. In addition, a method of solving the problem by forming a polyion complex between collagen and a polymeric substance having a carboxyl group or a sulfate group has also been attempted, which is represented by the following reaction formula. H ₃ N−−COO+Y−COO or Y−
SO ₃ →Y−COOH ₃ N−−COO or Y−SO ₃ H ₃ N−−COO (in the formula, H ₃ N−−COO represents collagen,
Y-COO or Y-SO ₃ represents a polymer substance. ) The resulting polyionplex has excellent biostability and antithrombotic properties. However, the polyionic complexes are soluble in aqueous media, such as inorganic salt solutions such as sodium chloride solutions, and have poor mechanical properties. Therefore, it is necessary to further perform stabilization treatment such as crosslinking treatment on the polyion complex. In view of the above circumstances, the present inventors, in the process of conducting intensive research into the development of new compositions suitable for a wide range of uses, found that a chitosan-collagen composition made by bringing chitosan and collagen into contact was extremely suitable. This discovery led to the present invention. That is, the present invention, based on the above findings, provides a chitosan/collagen structure obtained by bringing chitosan and collagen into contact in an acidic region, then deoxidizing the contact reaction product, removing solvent, and further crosslinking if necessary. A composition having a ratio of 0.01/99.99 to 99/1 (weight ratio) is provided. The composition of the present invention does not dissolve in the aqueous salt solution,
In addition, mechanical strength, blood coagulation, heat resistance, bacterial resistance,
Excellent water resistance, film formability, and adhesion to fillers such as meat. Therefore, the composition of the present invention is applicable to various uses. One such use is as an edible casing material. In addition, as reported by Kiriyama et al. (ANI No. 71), chitosan has the following physiological activities a) Normalization of plasma cholesterol b Suppression of blood sugar c Removal of growth-inhibiting substances d Prevention of colorectal cancer The composition of the present invention can be added to foods in the form of spherical or fibrous materials, and can exert preventive or therapeutic effects on the above-mentioned symptoms as an edible material. Furthermore, since the composition of the present invention has both the non-absorbability of chitosan to the living body and the blood coagulation effect of collagen, it can be used in medical and surgical applications in the form of a film, fiber, or other three-dimensional structure. . Although the chitosan-collagen composition of the present invention is thrombogenic in itself, it can be used as an antithrombotic material by treatment with heparin, and can be used as medical materials such as artificial blood vessels, artificial skin, and artificial kidneys. Furthermore, since the composition of the present invention has excellent protein adsorption ability, it can also be used as a substrate for immobilizing microorganisms such as enzymes and microorganisms, or as an adsorbent. The reason why the composition of the present invention has the above-mentioned excellent mechanical properties, non-absorbability, and blood coagulation effect is not yet clear, but it is due to the amino groups derived from chitosan and collagen produced by the following reaction. It is inferred. H ₃ N−−COO＋Z−NH ₃ 〓→ H ₃ N−
-COOH ₃ N-Z (In the formula, H ₃ N--COO represents collagen, and H ₃ N-Z represents chitosan.) The present invention will be described in detail below. The chitosan according to the present invention has a degree of de-N-acetylation of 50 to 100%, preferably 70 to 100%, and a viscosity of 20 to 1000 cp from the viewpoint of solubility and workability. The chitosan may be commercially available, or it may be obtained by heat-treating chitin obtained by separating and purifying it from the shells of arthropods such as crabs and shrimps by conventional methods in a highly concentrated alkaline aqueous solution. can also be obtained. In addition, the above viscosity is 0.5wt% solution (solvent: 1wt
% acetic acid aqueous solution) at 20°C. The collagen according to the present invention is obtained by separating non-collagenous tissue components from the fibrous components, which are the main proteins in connective tissues of vertebrates and invertebrates, by chemical and mechanical treatments. The collagen can be produced, for example, by the following method. That is, depilated cowhide and beef Achilles tendon are shredded using a meat grinder or the like, then swollen in an acidic or alkaline medium, and then defibrated using a grinder or the like to form an aqueous dispersion.
Further, over-separate as necessary. The collagen used in the present invention has a fiber diameter of 1 to 3.
μ, fiber length 0.1-15 mm, molecular weight 5,000-1,000,000, preferably 20,000-500,000, but are not necessarily limited to these. For example, the collagen may be made into fine fibers by mechanical treatment or the like, or part or all of the telopeptide may be cleaved by enzyme treatment and used as soluble collagen. Also, mixtures thereof can be used. In this case, from the viewpoint of mechanical strength, it is preferable that the solubilized collagen be 40 parts by weight or less per 100 parts by weight of collagen fibers. however,
Depending on the application, it is not necessarily limited to this. In addition, waste materials that cannot be used as they are, such as scraps of artificial or natural collagen casings, can be cited. In addition, after the shredding step in the above production, it is preferable to pre-treat the fragments in order to improve the strength and homogenize the resulting composition. The pretreatment includes crosslinking amino groups in lysine residues of collagen with reagents such as formaldehyde, glutaraldehyde, dialdehyde starch, cryoxal, liquid smoke, or epihalohydrin, succinylation with succinic anhydride, and organic acid anhydride. Examples include esterification of carboxylic acids in side chains such as aspartic acid and glutamic acid in collagen using an acylating or methylating agent. The composition of the present invention can be obtained by bringing the above-described chitosan and collagen into contact in an acidic region, then deacidifying the contact reaction product, removing the solvent, and further subjecting it to crosslinking treatment if necessary. Can be done. Note that the acidic region referred to here means that the chitosan-collagen contact system has a pH of 1 to 6, preferably 3 to 6.
It means that. The deoxidizing treatment is to adjust the pH to a level higher than the isoelectric point of the chitosan-collagen contact reaction product. Therefore, the deacidification treatment is a method in which the inside of the contact reaction system is treated with an alkali such as sodium hydroxide or potassium hydroxide to raise the pH to 7 or higher, or a method in which the excess acid present in the contact reaction product is removed by evaporation. , we can present a method of electrolytic deoxidation using electrodeposition method. According to the manufacturing method of the present invention, by combining collagen with chitosan, which has been unsatisfactory in moldability in the past, it is possible to improve moldability (film formability, etc.), and also to improve yield during manufacturing and stabilize products. Problems such as gender and uniformity can be solved. Furthermore, since it also reacts with free aldehyde groups obtained when collagen is crosslinked as a pretreatment, this is preferable from a safety standpoint. Hereinafter, a molded article (hereinafter referred to as
An example of manufacturing a molded article (referred to as a molded article of the present invention) will be described. One example is a method of mixing and dispersing a chitosan-containing acidic medium and a collagen-containing medium. In this case, the chitosan concentration in the chitosan-containing acidic medium is 5wt%, preferably 1wt% or less (solvent: dilute aqueous solution of acetic acid, hydrochloric acid, etc.), and the pH of the chitosan-containing acidic medium is 1 to 6, preferably 3 to 6. be. The collagen-containing medium is prepared by dispersing collagen in an acidic aqueous solution such as hydrochloric acid, and adjusting the pH to 3 to 6, preferably 3.
It was adjusted to ~4. The collagen-containing medium does not necessarily have to be acidic; it can also be alkaline. However, in this case, when mixed and dispersed with the chitosan-containing acidic medium, the mixed dispersion system needs to be in an acidic region. The collagen content of the collagen-containing medium may be arbitrary, but it is usually 5wt% or less, preferably 1wt% due to viscosity etc.
% or less. The mixing ratio of the chitosan-containing acidic medium and the collagen-containing medium is such that the chitosan/collagen composition ratio of the target composition is 0.01/99.99-99/
It can be appropriately selected within the range of 1 (weight ratio). In the present invention, the film-like molded product is obtained by degassing the mixture at the predetermined ratio, pouring the mixture onto a glass plate, deoxidizing it using a hot air dryer, etc., and then removing the solvent. It can be manufactured in The film-like molded article according to the present invention contains 1 fibrous protein.
Although it is a constituent component, it is characterized by having no vertical and horizontal direction dependence in terms of strength. This characteristic can only be expressed in the presence of chitosan, and this fact can be said to be completely surprising. Moreover, a spherical molded body can be produced by dropping and dispersing the liquid mixture in a hydrophobic organic solvent such as toluene or xylene, followed by deacidification treatment with an alcohol-water solution, and then desolvation. Furthermore,
By extruding the above-mentioned liquid mixture through a hole or slit-shaped nozzle into a highly concentrated salt or alkaline aqueous solution such as ammonium hydroxide, it can be formed into a fibrous, film-like, or hollow film-like form. In addition, a protein dispersion aqueous solution containing the chitosan of the present invention is introduced into an electrode bath equipped with at least one cathode and at least one anode, and a DC voltage is applied between the two electrodes to cause it to accumulate on a predetermined electrode surface. In this case, the molded article of the present invention can be molded extremely quickly. The tube-shaped molded product of the present invention obtained by the electrodeposition method has an extremely thin wall, and is suitable as an edible casing material that has excellent strength and good texture during electrodeposition and in a wet state. When the composition of the present invention is used as a film-like hollow body for an edible casing material, the composition ratio of chitosan/collagen is 0.01/1 from the viewpoint of texture, strength during smoking and cooking, heat denaturation temperature, and heat sealability.
A range of 99.99 to 60/40 is preferred. Also, when used as an edible food additive, 0.01/99.99-99/
It may be within the range of 1, and as an edible material, it is preferable to have a large amount of chitosan component. Another example of a manufacturing method is a method in which a collagen molded object previously formed into a sheet, film, tube, fiber, etc. is immersed in an acidic medium containing chitosan, followed by deacidification treatment and solvent removal. in this case,
The resulting molded product exhibits extremely unique properties such as improved mechanical properties, heat resistance, and antibacterial properties due to the chitosan component on the surface. However, a molded article obtained from a mixed solution of collagen and chitosan is superior in strength to a molded article obtained by the dipping method. The molded article of the present invention obtained by the above method etc. can be further crosslinked, succinylated with anhydride, etc.
In addition to the above properties, high blood coagulability, high blood coagulability,
Improved acid resistance is exhibited. Crosslinking can be carried out, for example, by the following method. First, the molded article of the present invention is immersed in an aqueous solution in which a crosslinking agent is dissolved in a phosphate buffer solution having a pH of 7.4. The immersion is carried out at a temperature of 10 to 50°C for 0.1 to 1 hour. After the reaction is completed, a washing step is performed to obtain a water-insoluble crosslinked molded product. The crosslinking treatment can cause not only chitosan crosslinking but also chitosan-collagen crosslinking, thereby further increasing surface bonding strength. Crosslinking occurs, for example, between the same or different functional groups of amino and hydroxyl groups of chitosan and collagen.
Examples of the crosslinking agent include formaldehyde, glutaraldehyde, dialdehyde starch, glyoxal, liquid smoke, and epihalohydrin. Polyhydric alcohols and sugars such as ethylene glycol, glycerin, and sorbitol are also used. In the production of the film-like hollow body of the present invention by electrodeposition, an example of a method is to add and disperse the crosslinking agent in the aqueous dispersion solution and carry out crosslinking treatment during electrodeposition. Examples of the fibrous protein according to the present invention include fibroin in silkworm silk, keratin in hair, and fibrinogen in blood. As described above, the present invention has a characteristic that overcomes the drawbacks of chitin and collagen, which exist in large amounts in nature but whose fields of application are limited due to their high crystallinity and chemical stability. The present invention provides a novel composition having the following characteristics, and its contribution to industry can be said to be significant. The present invention will be explained below with reference to Examples. Example 1 100 kg of salted stearhide skin from North America was soaked in water for 15 hours, washed with water, and then the fat was scraped off using a lining machine. The lined skin was then subjected to a lime soak treatment. Lime immersion treatment uses slaked lime 2%, Na ₂ S 0.5%
Add 90 kg of the above skin to 450 kg of liquid containing 0.5% (C ₂ H ₅ ) _{2 NH} 2
was added and gently stirred at 25°C for 24 hours. After the treatment, the skin was depilated using a depilation roll and decomposed products produced by the lime immersion treatment were removed. The treated skin was then divided into pieces using a splitting machine, cut into 5 mm wide pieces using a meat slicer, and further cut into 5 cm long pieces using a meat grinder. 80 kg of the skin pieces were dispersed in 800 kg of water, and 3 kg of acetic acid was added thereto, and after gentle stirring for 12 hours, the skin pieces were separated from the dispersion, dehydrated using a centrifugal dehydrator, and washed with ion-exchanged water. The washing was terminated when the electrical conductivity of the washing water became 20 μ/cm or less. The ash content in the skin at this point is 0.1
% or less. 40 kg of this skin strip was dispersed in 400 kg of an aqueous solution containing glutaraldehyde at a concentration of 0.015% and adjusted to pH 3.0 with hydrochloric acid, and was gently stirred at 20° C. for 12 hours to undergo crosslinking treatment. Next, the treated skin was separated using a wire mesh, and the fiber bundle was dissolved in 113 kg of cold water using a pulp refiner.
Make a dispersion of collagen fibers and add hydrochloric acid to it.
Adjusted to PH3.0. This dispersion has a thickness of 1 to 3μ,
A mixture of collagen fibers with a length of 1 to 10 mm and further finely divided collagen was obtained, resulting in 125 kg of liquid containing 2.5% collagen. On the other hand, 1 kg of chitosan was dissolved in 10 kg of an aqueous hydrochloric acid solution with a pH of 3, and the pH was adjusted to 3.0 to obtain a chitosan solution with a total weight of 20 kg. 50 kg of ion-exchanged water was added to 1.5 kg of the crosslinked collagen dispersion and stirred rapidly. Then, 374 g of chitosan aqueous solution was added to this dispersion and stirred and mixed for 1 hour until the weight ratio of collagen/chitosan was 100/5.
A mixed solution was obtained. This mixed solution has a viscosity of
It had an electrical conductivity of 200 cp and an electrical conductivity of 250 μ/cm. Next, a film was formed from the mixed solution using an electrodeposition device.
The electrodeposition device has an inner diameter of 100m/mφ and a height of 700m/m.
A platinum wire mesh supported by a PVC cylinder with holes is used as an anode, and a wire with an outer diameter of 17.5 m/mφ and a length of 700 mm is inserted into the anode.
It had a stainless steel tube cathode measuring m/m, and a propylene fiber cloth was attached to a PVC diaphragm holder with an inner diameter of 56 m/m and a thickness of 1 m/m to partition the electrodes. In the anode tank, an aqueous hydrochloric acid solution with a pH of 3.0 was introduced from the bottom of the tank, overflowed from the top of the tank, fell into the anolyte holder, and was circulated again from the bottom of the tank by a pump. The membrane raw material is introduced from the bottom of the cathode tank,
The mixture was allowed to overflow from the top and returned to the mixed liquid storage tank. The overflow part of the anode tank should be 50mm lower than the overflow surface of the cathode tank.
The aqueous hydrochloric acid solution was prevented from flowing into the cathode tank from the diaphragm. The temperature was kept below 10℃. The chitosan/collagen mixture was placed in the electrodeposition device, circulated, and a DC voltage of 300 V was applied between the electrodes. The positively charged collagen-chitosan moves to the cathode by electrophoresis, is neutralized by OH ^- generated by water electrolysis, approaches the isoelectric point PH, releases hydration water, and aggregates. , a film was formed on the cathode surface. The membrane pulling speed was 10 m/min. This tube-like material immediately after electrodeposition has a water content of 2 g-
H ₂ O/g and tensile strength of 200 Kg/cm ² . The pulled membrane was immersed in a 7% glycerin aqueous solution and then heated with air while maintaining a water column pressure of 300 m/m.
It was dried with hot air at 75° C. for 2 minutes to obtain a tube-shaped molded article (sample A) of the present invention. For comparison, a comparative collagen membrane (comparative sample A) was obtained in the same manner using a collagen dispersion to which chitosan was not added. In addition, the tube moisture content of comparative sample A immediately after electrodeposition was 7 g-H ₂ O/g, and the tensile strength was 100.
It was Kg/ ^cm2 . Table 1 shows the results of measuring the tensile strength and tear strength of the molded product in a wet state according to JIS P-8113 and JIS P-8116. As is clear from this, the tensile strength was significantly increased with the addition of chitosan, and an extremely thin film was obtained. Table 2 shows the results of immersing these molded bodies in a 0.2N aqueous sodium chloride solution at 20°C for 24 hours, and then examining the detent strength using the above method. As a result, it was revealed that the molded article of the present invention has excellent strength and is not only bound to collagen through simple ionic bonds, but also contains a binder-like element.

【table】

[Table] Example 2 The ratio of the chitosan-collagen mixture in Example 1 to 100 parts by weight of collagen was
The film was formed under the same conditions, except that the volume was 20 parts and the film pulling speed was 25 m/min, to obtain a continuous film-like molded article of the present invention (sample B). For comparison, a dispersion of collagen alone (0.58 wt%) was also tested at the same membrane pulling speed, but it was not possible to pull it up continuously. The mechanical properties of the molded article of the present invention were a film thickness of 8 μm, a tensile strength of 510 Kg/cm ² , a tear strength of 32 g·cm/cm, and an elongation of 32%. Example 3 After stirring and mixing 10 kg of the 2.5% collagen dispersion obtained in Example 1 and 1.5 kg of the 5% chitosan aqueous solution, the mixture was subjected to vacuum defoaming treatment. This was placed in a coagulation solution containing 0.1N ammonium hydroxide aqueous solution with a width of 10cm and a spacing of 0.3mm.
It was extruded through a slit, washed with water, and then subjected to the same post-treatment as in Example 1 to obtain a film-shaped molded article of the present invention (sample C). The film-like material was measured in the same manner as in Example 1 and found to have a thickness of 30μ, a tensile strength of 520Kg/cm ² , and a tear strength.
It had a weight of 75 g·cm/cm and an elongation of 30%. In addition, the film (comparative sample B) molded under the same conditions without adding chitosan had a film thickness of 30 μ, a tensile strength of 200 Kg/cm ² , a tear strength of 48 g cm/cm, and an elongation of 26%.
It was hot. From the above results, it is clear that the molded article of the present invention has excellent strength. Example 4 Mechanical filling suitability test Tube-shaped sample B obtained in Example 2 and Example 1
A practical test was conducted using Comparative Sample A obtained in Example 1 as a sausage casing. The meat emulsion was a frankfurter or wiener type mixture whose composition was as follows. Salted pork 1300g Pork back fat 750g Egg white 90g Ice water 750g Starch 120g Sodium glutamate 9g Allspice 12g Sugar 9g Phosphoric acid 6g Koshiyo 6g The emulsion is manufactured using the following process using commonly used meat equipment. did. 1 Salting pork Cut the pork into 3cm cubes, coat with salt, mix well, and salt for 12 hours at 15℃. 2 Grind the pork fat twice in a meat grinder (2φ hole),
Cooled at ℃ for 12 hours. 3 Grind the salted pork twice in a meat grinder and bring it to 4℃.
Cool to 4 Put 1300g of ground meat into a silent cutter and add water.
Added 400g and added 90g of egg white while mixing at low speed. 5 Next, add 350g of water to 750g of pork fat that has been ground in a meat grinder and cooled to 4℃, and add starch while mixing.
Added 120g. 6 Finally, add sodium glutamate 9 as a seasoning.
g, 6 g of koshiyo, 12 g of allspice, 9 g of sugar, and 6 g of phosphate were added and mixed at high speed. Table 3 shows the results of a filling test when the mixture thus obtained was packed into two types of casings using a semi-automatic stuffer worker 760. As is clear from Table 3, the chitosan additive can withstand severe filling tests. The casing of the present invention had a thin sausage knot and was excellent in appearance.

[Table] Example 5 Sausage samples No. 2 and No. 8 obtained in Example 4
was smoked. In this process, the wood chips are hung in a chain with 8 rings on a stick, placed in a smoking chamber, initially controlled at 55-60℃, dried for 30 minutes, raised to 75℃, and smoke generated from wood chips is introduced. Then, steam was introduced to keep the humidity in the smoking room constant, and the process was continued for 30 minutes. After the treatment was completed, steam was introduced and the temperature in the smoking chamber was maintained at 75 to 80° C. for 50 minutes, after which the chamber was taken out and cooled down by spraying with cold water. Assuming that the processed sausage would be cooked by a housewife at home, a test for bursting property during cooking was conducted.・Fry test Pour oil (product name: Nissin Tempura Oil) into a frying pan with a temperature controller, maintain it at 160℃, and test 10 samples of each.
Prepare 2 sausages, add 2 sausages at a time to prevent the oil temperature from dropping, and make sure the casing is
The amount that bursts in 30 seconds was measured.・ Frying pan test To keep the temperature of the surface of the frying pan uniform, dip the bottom of the frying pan into an oil bath, 10mm below the oil surface.
Fix it so that the surface of the frying pan is 175
The temperature of the oil bath was controlled to ~180°C. A small amount of the above-mentioned tempura oil was placed in a frying pan and spread over the entire surface, 10 sausages were prepared for each sample, and the sausages were placed in groups of 5 and gently moved, and the amount of casing bursting in 2 minutes was measured. This was repeated twice for the same sample.・ Boil test Prepare a hot water pot (depth 50mm) and keep it in boiling state.
Prepare 10 sausages for each sample,
They were divided into 5 pieces and placed in a hot water pot and boiled for 5 minutes, and the amount that burst was measured. This was repeated twice for the same sample. These results are shown in Table 4. As is clear from Table 4, it was also extremely excellent in practical cooking tests.

[Table] As a method to support these facts, the membranes before being filled were immersed in hot water at 75℃ for 1 minute, and the shrinkage rate and heat denaturation temperature were examined.
Shown in the table.

[Table] In addition, the sausage using the chitosan-added casing had a mellow flavor and sweetness that spread throughout the mouth, and had an unparalleled texture. This phenomenon did not occur in sausages using a casing made only of collagen fibers. Example 6 20 g of chitosan was dissolved in 5% acetic acid aqueous solution 1 to make a homogeneous solution, and this was designated as Solution A. Take 50 g of the 2.5% collagen dispersion without crosslinking treatment in Example 1, add 1 g of acidic protease (manufactured by Nagase Sangyo), treat it at 37°C for 5 hours, and then perform operations such as neutralization centrifugation and washing with water to obtain 1N- Adjust the pH to 3.5 with an aqueous hydrochloric acid solution, bring the total amount to 1 kg, and further subdivide using a homogenizer while keeping the temperature below 10°C.
A solution B was obtained. Solution A was added to solution B, and a homogenized solution was obtained using a homogenizer. Separately, a 3-size flask equipped with a stirrer was prepared in which 2 decalin and 1 g of polyoxyethylene sorbitol (trade name Twin #80) were placed. Add 100 c.c. of the A-B mixture to the flask,
Stir and disperse at 1000 rpm. After dispersing for 1 hour, the dispersions were redispersed in 10 ml of ethanol to obtain insoluble matter. After repeating filtration and ethanol washing to remove the organic solvent, it was redispersed in 2% ammonium hydroxide aqueous solution 1, neutralized, and washed with ethanol.
Molded article of the present invention that becomes spherical when freeze-dried (Sample D)
was gotten. In addition, the size of sample D is 0.1 to 1.0 in diameter.
It was m/m. 1 g of sample D was taken, poured into physiological saline, filtered, and excess saline was removed. This has an inner diameter of 8m/
Fill a mφ glass column, then add 50 ml of rabbit blood to which 250 units of heparin was added at 20 ml/min.
perfusion for 15 minutes at 37°C. After stopping the circulation, the blood was immediately removed, the spherical molded body was taken out after washing with physiological saline, and the degree of adhesion of platelets and blood cells to the surface of the molded body was measured. As a result, it was found that neither platelets nor blood cells were clearly attached. Admitted. Example 7 5 g of chitosan was added to 1% aqueous acetic acid solution to create a homogeneous solution. (The solution viscosity at this time is
It was hot at 50cps. ) A molded article consisting only of collagen fibers of Comparative Sample A was immersed in this solution at 20°C for 1 hour, excess aqueous solution was removed, and the product was immersed in 0.1N ammonia water in a wet state, then washed with water to remove ammonium acetate. , ammonia was removed and the solvent was removed to obtain a molded article (sample E) of the present invention. The mechanical strength of this molded body is tensile strength 395Kg/
cm ² , tear strength 50 g·cm/cm, and elongation 33%.
The amount of impregnation was 2 parts by weight of chitosan per 100 parts by weight of collagen fibers. Example 8 50 kg of skin pieces dehydrated using a centrifugal dehydrator in Example 1 were immersed in 100% acetic anhydride at a liquid temperature of 15° C. for 8 hours to cause an acetylation reaction. The treated skin was washed under running ion-exchanged water.
The isoelectric point of the raw material, cowhide, was PH6.5, but it changed to PH3.8 after this treatment. The treated skin was made into a dispersion of collagen fibers in the same manner as in Example 1, adjusted to pH 3.0 with hydrochloric acid, and swollen to 2.5
% collagen dispersion was prepared. As in Example 1, an aqueous chitosan solution was added to adjust the weight ratio of collagen/chitosan to 100/10, and the film was formed using the electrodeposition apparatus of Example 1 and then subjected to post-treatment. When its mechanical properties were examined based on Example 1, the film thickness was 9μ, tensile strength was 485Kg/cm ² , tear strength was 35g·cm/cm, and elongation was 33%. Example 9 The collagen-chitosan mixed solution of Example 3
200m/m x 200m/m plate with a thickness of 5m/m
After pouring it in and leaving it for one hour, it was immersed in a 0.1N ammonium hydroxide aqueous solution to neutralize the acid, and then washed with ion-exchanged water to remove salts. This was immersed in a 0.1 molar dibasic sodium phosphate aqueous solution containing 0.5% formaldehyde at 30°C for 1 hour, and then washed with water. The treated molded product was freeze-dried to obtain a sponge-like molded product of the present invention. It was found that the molded product had an absorption capacity of 20 g/g for physiological saline and had strong coagulability against blood, so it could be used as a sanitary product and an emergency tourniquet. Example 10 Glucose isomerase (2000 U/g manufactured by Nagase Sangyo) in 500 c.c. of 0.1 molar dibasic sodium phosphate aqueous solution
1 g of collagen-chitosan was dissolved, and 30 g of the collagen-chitosan spherical molded article in Example 6 was added thereto. After stirring at 5° C. for 2 hours, 2 c.c. of a 25% glutaraldehyde aqueous solution was further added and treated for 5 hours. After the treatment, the treated molded body was washed with a phosphate buffer of PH 7.0, and the treated molded body was added to 500 c.c. of a phosphate buffer of PH 7.0 containing 2 g of glucose.
When the amounts of glucose and fructose were measured after being treated at 70℃ for 60 minutes, they were 600 mg and 600 mg, respectively.
1400 mg, and the immobilized enzyme was 70%. Further, after adjusting the collagen-chitosan aqueous solution before dispersion in Example 6 to pH 6.0, 1 g of the above glucose isomerase was added at 5°C, and after dispersion in the same manner as in Example 6, it was washed with ethanol, and the pH was 7.0 phosphate buffered. After washing with a solution, the activity of the immobilized enzyme was examined by the above method and found to be 85%. These results indicate that it can be used as a carrier for immobilized enzymes and bacterial cells. Example 11 2 g of epichlorohydrin and 100 c.c. of water were added to 10 g of sample B obtained in Example 2, and after heating under reflux for 1 hour, 8N
- 100 g of aqueous sodium hydroxide solution was further added and heat treated on an oil bath at 120°C for 2 hours. The crosslinked product was washed with water to obtain a molded article of the present invention. The treated product was insoluble in acids, alkalis and organic solvents. still,
Mechanical properties were measured in the same manner as in Example 1 and found to be a tensile strength of 630 Kg/cm ² and an elongation of 15%. Example 12 Tubular shaped bodies (Samples F and G) were produced under the same conditions as in Example 1, except that the amount of chitosan added was changed so that the collagen/chitosan weight ratio was 100/1100/0.1. Obtained. This was filled under the conditions of Sample No. 2 of Example 4 and subjected to the smoking treatment of Example 5. Incidentally, no rupture was observed in any of the samples during filling. Next, a cooking test similar to that in Example 5 was conducted, and the results are shown in Table 6. As is clear from Table 6, it was also extremely excellent in practical cooking tests.

【table】

Claims (1)

[Claims] 1. Chitosan 0.01-99% by weight and collagen
Chitosan-collagen composition consisting of 99.99-1% by weight. 2. A method for producing a chitosan-collagen composition, which comprises bringing chitosan and collagen into contact in an acidic region, and then deacidifying and removing the solvent from the contact reaction product. 3 The pH of the acidic region is 1 to 6, preferably 3 to 6.
6. The manufacturing method according to claim 2. 4 Chitosan and collagen weight ratio: 0.01:
The manufacturing method according to claim 2, characterized in that the contact is carried out at a ratio of 99.99 to 99:1. 5. Claims 2 to 4, characterized in that the contact is carried out by mixing and dispersing the chitosan-containing acidic medium and the collagen-containing medium.
Manufacturing method described in Section. 6. The manufacturing method according to claim 2 or 4, wherein the contact is performed by immersing the collagen molded body in an acidic medium containing chitosan.