JPS6247213B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to protein-chitosan compositions. More specifically, a novel composition comprising chitosan, a fibrous protein insoluble in an aqueous solvent, and a protein soluble in the solvent, which has excellent heat resistance, moisture absorption resistance, and mechanical properties, and is non-absorbable to living organisms. Pertains to. Collagen, a typical protein, is found in mammals,
It is a hard protein that exists in the connective tissue, bones, teeth, skin, dermis, fascia, etc. of birds, etc., and is widely used in the form of edible casing materials, surgical sutures, vascular grafts, artificial skin, etc. It is being 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. In addition, 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 suture threads, artificial skin, and hemostatic materials described above are unsatisfactory not only in terms of strength but also in terms of stability (non-absorbability) in living bodies. Various methods have been studied to increase the stability of collagen materials in living organisms. 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 (wherein H ₃ N--COO represents collagen,
Y-COO or Y-SO ₃ represents a polymer substance. ) The resulting polyion complex 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-mentioned circumstances, the present inventors, in the process of conducting intensive research on the development of a new composite material suitable for a wide range of uses, discovered a fibrous protein - soluble protein - made by bringing chitosan and protein into contact with each other.
The present invention was achieved based on the discovery that a composition consisting of three components of chitosan is extremely suitable for the purpose. That is, the present invention based on the above knowledge is based on chitosan/
[Fibrous protein + soluble protein] Composition ratio 0.01/
The present invention provides a protein-chitosan composition having a weight ratio of 99.99 to 99/1 and a fibrous protein/soluble protein composition ratio of 99/1 to 60/40 (weight ratio). Conventionally, protein-based compositions such as collagen fiber-soluble collagen or collagen fiber-gelatin have been reported. However, since the protein-based composition has weak binding of soluble components, its strength decreases significantly in hot water. On the other hand, the composition of the present invention does not dissolve in salt aqueous solutions, does not lose its mechanical strength, especially in hot water, and has non-absorbability, blood coagulation properties, bacterial resistance, and Excellent film forming properties and adhesion to fillers such as meat. 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 chitosan forms a strong polyion complex with protein through the following reaction. That is, soluble protein and fibrous protein are combined with chitosan using the formula: H ₃ N−−COO+Z−NH ₃ →H ₃ N−−COOH ₃ N−Z (where H ₃ N−−COO represents protein, _H3N
-Z indicates chitosan. ), it is presumed that this is due to the strong bonding. 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. Further, the composition of the present invention can be used in medical and surgical applications in the form of a film, fiber, or other three-dimensional structure, since chitosan has both non-absorbability to living bodies and blood coagulation effects. Although the composition of the present invention is thrombogenic in itself, it can be used as an antithrombotic material by treatment with heparin.
It 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 present invention will be explained 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 proteins according to the present invention are of two types: fibrous proteins that are insoluble in aqueous solvents and proteins that are soluble in the aqueous solvents (hereinafter collectively referred to as protein components). The protein component can be obtained by separating and purifying the living body of various animals or by chemically treating the separated and purified protein. In addition, the aqueous solvent as used in the present invention means an aqueous solution of acid or alkali. Examples of acids include inorganic acids such as hydrochloric acid, and organic acids such as acetic acid, adipic acid, and propionic acid, either singly or in a mixture of two or more. Further, the alkali is a common alkali such as sodium hydroxide or potassium hydroxide. A typical example of fibrous protein is collagen fiber. Collagen fibers are obtained by separating non-collagenous tissue components from fiber components, which are the main proteins in the connective tissues of vertebrates and invertebrates, by chemical and mechanical treatments. The collagen is
For example, it can be manufactured by the following method. That is, depilated cow hide and beef Achilles tendon are shredded using a meat grinder or the like, then swollen in an acidic or alkaline medium, then defibrated using a grinder or the like to form an aqueous dispersion, and further subjected to over-separation if necessary. . 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 composite material. The pretreatment includes crosslinking of 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 acid residues in side chains such as aspartic acid residues and glutamic acid residues in collagen using an acylating or methylating agent. Furthermore, the collagen fibers include waste materials that cannot be used as casings, such as scraps of natural or artificial collagen casings. Other examples of the fibrous protein according to the present invention include fibroin present in silkworm silk, keratin, a component of hair, and fibrinogen in blood. The fibrous protein used in the present invention has a fiber diameter of 1 to 3μ, a fiber length of 0.1 to 15mm, and a molecular weight of 5000 to
1,000,000, preferably 20,000 to 500,000, but is not necessarily limited to this. On the other hand, proteins that are soluble in aqueous solvents include gelatin, which is used in the food industry on an industrial scale, or those that have been made completely soluble by treating the above-mentioned fibrous proteins with proteolytic enzymes, acids, or alkalis. I can give an example. Furthermore, by solubilizing a portion during the above treatment, it can be used as a mixture of fibrous protein and soluble protein components. The composition of the present invention can be obtained by bringing the chitosan described above into contact with a protein component in an acidic region, and then deacidifying the contact reaction product and removing the solvent. The composition ratio of chitosan and protein components is chitosan/protein: 0.01/99.99-99/depending on the intended use.
It can be appropriately selected within the range of 1 (weight ratio). The ratio of fibrous protein and soluble protein in the protein component is soluble protein/fibrous protein: 1/99 ~
The weight ratio is 40/60. When the composition ratio of the protein component is 40/60 or more, the amount of water absorbed by the molded product made from the composition of the present invention increases, making it difficult to maintain the shape of the molded product and causing a decrease in tensile strength and the like. Gelatin is one of the most preferred soluble proteins, and in this case, the amount is 0.01 to 0.4 parts by weight, preferably 0.05 to 0.3 parts by weight, per 1 part by weight of fibrous protein. The soluble protein component contributes to the flexibility of the composition, lowering the heat distortion temperature, improving mechanical strength, and further improving the texture and heat sealability of the casing material as described below. Note that the above-mentioned partially solubilized fibrous protein is used as a protein component. In the present invention, the acidic region refers to a chitosan-protein component contact system with a pH of 1 to 1.
6 preferably means 3 to 6. The deacidification treatment is to adjust the pH to a level higher than the isoelectric point of the chitosan-protein component contact reaction product. Therefore, the deacidification treatment can be carried out by treating the inside of the contact reaction system with an alkali such as sodium hydroxide or potassium hydroxide to raise the pH to 7 or higher, by treating it with an alkaline salt aqueous solution, or by treating the contact reaction system with an aqueous alkaline salt solution. A method of removing existing acid by evaporation or a method of electrolytic deoxidation by electrodeposition can be proposed. Gelatin is made by denaturing insoluble collagen with hot water or the like to make it soluble in aqueous solvents, and its isoelectric point, jelly strength, ash content, etc. vary depending on the treatment method. Therefore, when producing the above-mentioned protein-forming form using gelatin, it is necessary to change the production conditions depending on the type of gelatin. For example, when using gelatin obtained by acid treatment when producing a composition by the electrodeposition method described below, it is necessary to set the pH of the adjustment solution to 7 or less and to form a composite molded body on the cathode side. Further, in the case of gelatin obtained by alkaline treatment, the pH of the adjusting solution must be set to 7 or higher to form a molded body on the anode side. On the other hand, in the present invention, due to the action of chitosan, which is one of the constituent components of the composition, a molded article can be produced without the above-mentioned constraints in terms of production. In addition, according to the manufacturing method of the present invention, by combining the protein component, which has conventionally been unsatisfactory in moldability, with chitosan, it is possible to improve moldability (film formability, etc.), and at the same time, it is possible to improve the yield rate during manufacturing. and can solve problems such as product stability and uniformity. 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. The film-shaped molded article of the present invention is produced by defoaming a mixture of a chitosan-containing acidic medium and a protein component-containing medium at a predetermined ratio, pouring the mixture onto a glass plate, and then drying the mixture in a hot air dryer or the like. Deoxidized,
It can be produced by removing the solvent. Chitosan concentration in chitosan-containing acidic medium is 5wt% preferably 1wt%
The pH of the chitosan-containing acidic medium is 1 to 6, preferably 3 to 6. For the protein component-containing medium, the protein component is dispersed in an acidic aqueous solution such as hydrochloric acid, and the pH is adjusted to 3 to 3.
6, preferably adjusted to 3 to 4. The protein component-containing medium does not necessarily have to be acidic, and may be neutral or 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. In addition, in the electrodeposition method described below, the protein component-containing medium is used in an acidic region due to the relationship between electrical conductivity and required power. The content of the protein component in the protein component-containing medium may be arbitrary, but it is usually 5 wt% or less, preferably 1 wt% or less, in view of viscosity and the like. Although the film-like molded article according to the present invention contains fibrous protein as one of its constituent components, it is characterized by having no dependence in the longitudinal and lateral directions 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 article can be produced by dropping and dispersing the mixed liquid in a hydrophobic organic solvent such as toluene or xylene, and then deoxidizing the mixture with an alcohol-water solution. Furthermore, the mixture can be extruded into a highly concentrated salt or alkaline aqueous solution such as ammonium hydroxide through a hole or slit nozzle to form a fibrous, film, or hollow shape. A fibrous protein dispersion aqueous solution containing soluble protein and chitosan 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 the dispersion to accumulate on a predetermined electrode surface. The tube-shaped molded product of the present invention has an extremely thin wall and has excellent strength, so it is suitable as an edible casing material with good texture. In addition, when the composition of the present invention is used as an edible casing, the composition ratio of chitosan/protein component is 0.01/99.99-60/ from the viewpoint of texture, strength during smoking and cooking, heat denaturation temperature, and heat sealability. A range of 40 is preferred. The composition ratio of the protein component is soluble protein/fibrous protein: 5/95 to 40/60 (weight ratio). Further, when the composition of the present invention is used as an edible food additive, the composition ratio of chitosan/protein component is preferably 1/99 to 99/1, especially the one with a large amount of chitosan component. In addition, gelatin used for forming by electrodeposition method has a low ash content due to the power required and quality.
The amount used is 0.5wt% or less, preferably 0.2wt% or less. As another method for producing a molded body made of the composition of the present invention, a sheet is prepared in advance in an acidic medium containing chitosan,
Another example is a method in which a protein molded object formed into a film, tube, fiber, etc. is immersed, and then deacidified and solvent removed. In this case, the molded product obtained exhibits extremely unique properties such as improved mechanical properties, heat resistance, and antibacterial properties due to the chitosan component in the surface portion. The molded article of the present invention obtained by the above method etc. can be further crosslinked, succinylated with succinic anhydride, acylated with anhydrous organic acid, or asparagine residue in collagen using a methylating agent.
By esterifying the carboxyl residue in the side chain of the glutamine residue, in addition to the above properties, high blood coagulability and improved acid resistance are 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-protein component crosslinking, thereby further increasing the surface binding 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 or epihalohydrin, polyhydric alcohols such as ethylene glycol, glycerin, and sorbitol, and sugars. According to the present invention, soybean protein,
It is also possible to use soluble proteins such as casein,
It can be applied for various purposes. As described above, the present invention more than makes up for the drawbacks of chitin and protein components, which exist in large quantities in nature but whose fields of application are limited due to their high crystallinity and chemical stability. It provides a novel composition with unique 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 hides were then lime soaked. 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 these 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%
It was below. 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 128 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. On the other hand, acid-treated gelatin (Miyagi Chemical F-795) 1
After dissolving Kg in 10 Kg of an aqueous hydrochloric acid solution with a pH of 3, the pH was adjusted to 3.0 to obtain a gelatin solution with a total weight of 20 Kg. Add ion-exchanged water to 15kg of the cross-linked collagen dispersion.
50 kg was added and stirred rapidly. Next, 374 g of a chitosan aqueous solution and 374 g of a gelatin aqueous solution were added to this dispersion, and the mixture was stirred and mixed for 1 hour to obtain a mixed solution with a weight ratio of collagen/gelatin/chitosan of 100/5/5. Obtained. 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 in diameter, 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. Inside 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-gelatin mixture was put into the electrodeposition device, circulated, and a DC voltage was applied between the electrodes.
I applied 300V. Positively charged collagen
Chitosan-gelatin moves to the cathode by electrophoresis, is neutralized by OH ^- generated by water electrolysis, approaches the isoelectric point PH, releases hydration water, aggregates, and forms a film on the cathode surface. was formed. The membrane pulling speed is
The speed was 10m/min. 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 molded article (sample A) of the present invention. For comparison, a comparative gelatin-collagen film (comparative sample A) was obtained in the same manner using a gelatin-collagen dispersion to which chitosan was not added. 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 has increased. Table 2 shows the results of immersing these molded bodies in hot water at 40° C. for 24 hours, and then examining their tensile strength using the above method. As a result, it became clear that the molded article of the present invention was also excellent in strength.

【table】

[Table] Example 2 Same conditions as in Example 1 except that the ratio of the chitosan-collagen-gelatin mixture was 20 parts chitosan and 5 parts gelatin per 100 parts by weight collagen, and the membrane pulling speed was 25 m/min. A film of the present invention (sample B) was obtained as a continuous film. For comparison, a collagen-gelatin mixture (100 parts by weight - 5 parts by weight) was prepared at the same membrane pulling speed.
I also tried this, but I couldn't raise it continuously. The mechanical properties of the molded article of the present invention were a film thickness of 8 μm, a tensile strength of 490 Kg/cm ² , a tear strength of 33 g·cm/cm, and an elongation of 33%. Example 3 In Example 1, gelatin was changed from acid-treated gelatin to alkali-treated gelatin (Miyagi Chemical F-
800) and the ratio of collagen-gelatin-chitosan mixture to 100 parts by weight of collagen.
20 parts of gelatin and 10 parts of chitosan with membrane pulling speed
The film was formed under the same conditions except that the speed was 15 m/min.
A molded article of the present invention (sample C) which is a continuous film-like article was obtained. For comparison, a collagen-gelatin mixture (100 parts by weight - 20 parts by weight) was prepared at the same membrane pulling speed.
However, the film was not formed on the electrode and 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 9 μm, a tensile strength of 440 Kg/cm ² , a tear strength of 30 g·cm/cm, and an elongation of 34%. Example 4 10 kg of 2.5% collagen dispersion obtained in Example 1, 1 kg of 5% gelatin aqueous solution, and 5% chitosan aqueous solution
After stirring and mixing 1.5 kg, this was subjected to vacuum defoaming treatment. This was extruded through slits with a width of 10 cm and an interval of 0.3 mm into a coagulation bath containing a 0.1N ammonium hydroxide aqueous solution, and after washing with water, the same post-treatment as in Example 1 was performed. ) was obtained. The film-like material was measured in the same manner as in Example 1 and found to have a film thickness of 30μ, a tensile strength of 495Kg/cm ² , and a tear strength.
It had an elongation of 73 g·cm/cm and an elongation of 31%. In addition, the film (comparative sample B) molded under the same conditions without adding chitosan had a film thickness of 30μ, a tensile strength of 320Kg/cm ² , a tear strength of 65gcm/cm, and an elongation of 28%.
It was hot. From the above results, it is clear that the molded article of the present invention has excellent strength. Example 5 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 backfat 750g Egg white 90g Ice water 750g Starch 120g Sodium glutamate 9g Allspice 12g Sugar 9g Phosphate 6g Koshiyo 6g The emulsion is produced in the following process using commonly used meat equipment. Manufactured. 1 Salting pork Cut the pork into 3cm cubes, coat with salt, mix well, and salt at 15℃ for 12 hours. 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 6 Sausage samples No. 2 and No. 3 obtained in Example 5
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 and keep it at 160℃, 10 samples for 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 be ~180℃. 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 boiling.
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.

[Table] As is clear from Table 4, it was extremely excellent in practical theory tests as well. In addition, the sausage using the casing to which chitosan was added 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. In a sensory test conducted by a panel of 30 people, the overall evaluation was that the sausage had a smoother texture and almost no artificial taste compared to sausages made from natural sheep intestines. Example 7 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. Chitosan and gelatin aqueous solution were added to this in the same manner as in Example 1, and the weight ratio of collagen/gelatin/chitosan was adjusted to 100/10/10.
After film formation, post-treatment was performed using an electrodeposition apparatus of 1 to obtain a molded article of the present invention (sample E). When its mechanical properties were examined based on Example 1, the film thickness was 9μ, tensile strength was 480Kg/cm ² , tear strength was 34g·cm/cm, and elongation was 33%. Example 8 1 kg of skin strips before the crosslinking treatment in Example 1 were taken and immersed in Atkins-Pantin buffer 20 of PH9 containing 0.2% by weight of pronase (manufactured by Kaken Chemical Co., Ltd.) for 24 hours, and reacted with gentle stirring. Ta. After the reaction was completed, the lumps were purified by centrifugation, washing with water, etc. This is mixed with 1N hydrochloric acid aqueous solution.
The pH was adjusted to 3.5 to obtain a homogeneous solution with a total amount of 100 kg.
A portion of this solution was taken and freeze-dried, and the concentration of soluble collagen was examined, and it was found to be 0.95%. Take 13.5 kg of the 2.5% crosslinked collagen dispersion in Example 1, add 50 kg of ion exchange water,
Stir rapidly, then add the above soluble collagen solution
3.95Kg was added and mixed with stirring for 30 minutes. Furthermore, 748 g of the chitosan aqueous solution in Example 1,
374 g of gelatin aqueous solution was added and mixed by stirring. This mixed solution has a composition ratio of collagen fibers: soluble collagen: gelatin: chitosan of 90:10:10:20. This was formed into a film using the electrodeposition apparatus of Example 1 to obtain a molded article (sample F) of the present invention. Example 1
When the mechanical properties were examined based on this, the film thickness was 9 μm, tensile strength was 500 Kg/cm ² , tear strength was 35 g·cm/cm, and elongation was 35%. Example 9 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 crosslinked 2.5% collagen dispersion in Example 1, add 7.5 g of the gelatin aqueous solution in Example 1, adjust the pH to 3.5 with 1N hydrochloric acid aqueous solution, make a total amount of 1 kg, and heat with a homogenizer below 10°C. The mixture was further subdivided while maintaining the temperature to obtain Solution B. Solution A was added to solution B, and a homogenized solution was obtained using a homogenizer. Separately, a 3-sized flask equipped with a stirrer was prepared in which 2 g of decalin and 1 g of polyoxyethylene sorbito (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 G)
I got it. In addition, the size of sample G is 0.1 to 1.0 in diameter.
It was m/m. 1 g of sample G was taken, poured into physiological saline, filtered, and excess saline was removed. This inner diameter is 8
Fill a m/mφ glass column, then add 20ml/50ml of rabbit blood to which 250 units of heparin has been added.
Perfusion was performed for 15 minutes at 37°C at a rate of min. 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 on the surface of the molded body was measured.
Adhesion of both platelets and blood cells was clearly observed. Example 10 The collagen-gelatin-chitosan mixed solution of Example 2 was poured into a 200 m/m x 200 m/m plate with a thickness of 5 m/m, left to stand for 1 hour, and then 0.1N-
The sample was immersed in an aqueous ammonium hydroxide solution to neutralize the acid, and then washed with ion-exchanged water to remove salts. this
It was immersed in a 0.1 molar dibasic sodium phosphate aqueous solution containing 0.5% formaldehyde at 30°C for 1 hour.
Then, it was washed with water. The treated molded product was freeze-dried to obtain a sponge-like molded product of the present invention (Sample H). The molded product had an absorption capacity of 21 g/g for physiological saline and had strong coagulability against blood, so it was found that it could be used as a sanitary product and an emergency tourniquet. Example 11 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 the collagen-gelatin-chitosan spherical molded body (sample G) in Example 9.
After stirring at 5° C. for 2 hours, 2 c.c. of a 25% aqueous glutaraldehyde 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, and treated at 70°C for 60 minutes. When the amounts of glucose and fructose were measured, they were 600 mg and 1400 mg, respectively, and the immobilized enzyme was 70%. These results demonstrate that the molded article of the present invention can be used as a carrier for immobilized enzymes and bacterial cells. Example 12 5 g of chitosan was added to 1% aqueous acetic acid solution to form a homogeneous solution. (The solution viscosity at this time is
It was hot at 50cps. ) The collagen fiber-gelatin molded body of Comparative Sample A was immersed in this solution at 20°C for 1 minute, the excess aqueous solution was removed, and the body was immersed in 0.1N ammonia water while still wet, then washed with water and acetic acid. Ammonium and ammonia were removed and the product was dried with hot air to obtain a molded product of the present invention (Sample I). The mechanical strength of this compact is tensile strength 400Kg/
cm ² , tear strength of 30 g·cm/cm, and elongation of 30%.
The amount of impregnation was 2 parts by weight of chitosan per 100 parts by weight of collagen fibers.

Claims (1)

[Claims] 1. Chitosan/[fibrous protein + soluble protein]
Composition ratio of 0.01/99.99 to 99/1 (weight ratio), and fibrous protein/soluble protein composition ratio of 99/1 to
A protein-chitosan composition that is 60/40 (weight ratio). 2. The composition according to claim 1, wherein the fibrous protein is collagen fiber. 3. The composition according to claim 1, wherein the soluble protein is gelatin, solubilized collagen, or a mixture thereof. 4. The composition according to claim 1, wherein the chitosan has a degree of de-N-acetylation of 50 to 100%. 5. A method for producing a fibrous protein-soluble protein-chitosan composition, which comprises bringing chitosan into contact with a fibrous protein and a soluble protein in an acidic region, and then deacidifying and removing the solvent from the contact reaction product. 6 Ratio of fibrous protein and soluble protein is 99:1
6. The production method according to claim 5, wherein the protein and chitosan are brought into contact at a weight ratio of 99.99:0.01 to 1:99 (weight ratio). 7. The manufacturing method according to claim 5, wherein the fibrous protein is collagen fiber. 8. The production method according to claim 5, wherein the soluble protein is gelatin, solubilized collagen, or a mixture thereof. 9. The production method according to claim 5, wherein the chitosan has a degree of de-N-acetylation of 50 to 100%. 10. The manufacturing method according to claim 5, wherein the pH of the acidic region is 1 to 6. 11. The production method according to claim 5, wherein the contact is performed by mixing and dispersing the chitosan-containing acidic medium, the fibrous protein-containing medium, and the soluble protein-containing medium. 12. The production method according to claim 5, wherein the contact is performed by immersing the protein molded body in an acidic medium containing chitosan.