JP4344950B2 - Shrinking of animal hair fiber - Google Patents

Description

  The present invention relates to an improved shrink-proofing method for imparting shrink-proofing properties capable of washing and washing to a textile product containing animal hair fibers such as wool, cashmere, alpaca, and Angola.

Animal hair fibers such as wool, cashmere, alpaca, and angora have excellent moisture absorption and release characteristics, heat retention, texture (soft touch), etc., and are said to be ideal fibers. Therefore, textile products such as woven fabrics, knitted fabrics, and non-woven fabrics using the fibers are repeatedly washed (except for dry cleaning), so that the tip protrusions of the scales are entangled and shrunk between multiple fibers. Produce. This shrinkage is due to the anisotropy of the surface friction coefficient of the scale, which is called felt shrinkage, and is a drawback unique to animal hair.
Various methods have been developed to eliminate this drawback. One of them is the chlorinated resin method (oxidation / resin shrinkage processing method), which is currently the mainstream of the wool shrinkage treatment method. In this method, the tip of the hydrophobic scale on the surface of the wool is scraped by chlorination using a chlorinating agent to reduce the friction coefficient anisotropy, and the animal fiber body is relatively hydrophilic. Is partially covered, and then the surface is covered with a hydrophilic resin (shrink-proofing agent) such as urethane resin. According to this method, the shrink-resistant agent is firmly bonded to the fiber surface and does not easily peel off even when the number of washings is increased, and thus the product has excellent shrinkage resistance. However, since this method requires the use of a chlorinating agent, it has a serious adverse effect of polluting the environment by discharging harmful absorptive organic halogen compounds (AOX). In Western Europe, where wastewater regulations are strict, industrial implementation of the method is currently becoming difficult. Moreover, this chlorinated resin method has drawbacks such that the chlorinating agent used penetrates into the inside of the fiber and damages the fiber, or increases the weight loss. In addition, since the resin as a shrink-proof agent coats the fiber surface, the resulting fiber has a disadvantage that it impairs the original excellent texture (soft touch) of the animal hair fiber and makes it feel hard. Furthermore, there is an unavoidable disadvantage of the so-called “yellowing phenomenon” in which the chlorine bonded to the fiber generates double bonds over time and yellows the fiber.
There is also known a clean shrink-proofing method that does not use a chlorinating agent and therefore does not discharge harmful substances. The method includes a method using protease and a method using electron energy (plasma). The former is a method of modifying the fiber surface by degrading and removing the scale tip protrusions that cause shrinkage by protease, and the latter introduces active groups to the fiber surface by plasma irradiation to make the fiber hydrophilic. This is a method of reforming.
However, none of the proteases proposed to be used in this type of animal hair fiber shrink-proofing process can selectively decompose and remove the keratinous scale covering the fiber surface. In order to obtain sufficient shrinkage using such an enzyme, a long-time treatment using a large amount of enzyme is required. However, in such an enzyme treatment, cortex cells and cell membrane complexes (cell membrane plain complex, CMC) inside the fiber as well as the scale are decomposed by the enzyme, resulting in a large weight reduction and a significant strength reduction. This reduction in strength is a fatal defect. Thus, the conventional shrink-proofing method using protease cannot perform sufficient shrink-proofing without reducing the strength of the fiber.
Moreover, the shrink-proof processing method using a plasma is only what modifies the hydrophobic scale surface to hydrophilicity. It does not reduce the anisotropy of the coefficient of friction on the scale surface, which is the cause of shrinkage. It is impossible to impart sufficient shrinkage resistance only by this method. In general, the shrink-proofing treatment of animal hair fiber products using the plasma is recommended to be used in combination with the above-described coating treatment of the fiber surface with a shrink-proofing agent (hydrophilic resin), enzyme treatment or the like. More specifically, after low-temperature plasma treatment, a resin is processed, and a lost texture is further improved by using protease (see, for example, Japanese Patent No. 2905311), and a combination of low-temperature plasma treatment and protease treatment ( For example, see Japanese National Publication No. 10-511437).
However, even by these methods, a fiber product having satisfactory shrinkage resistance, strength and texture cannot be obtained. That is, the resin processing method using a shrink-preventing agent has the disadvantage of essentially masking the natural soft texture of animal hair fibers, and an enzyme that selectively acts on the scale is still being developed in the enzyme processing method. Therefore, there is a fatal disadvantage that causes a decrease in strength of the fiber itself. The method using these in combination only reduces the disadvantages of the respective treatment methods to an acceptable range and satisfies the shrinkage resistance, texture, and strength within the range where each can be folded. In addition, the low-temperature plasma treatment used in combination with these methods requires work under reduced pressure, which is disadvantageous in terms of practicality in terms of equipment, cost and time, and is difficult to perform continuous treatment. There are also certain drawbacks.
Recently, a method for preventing shrinkage of animal hair fiber products using a pulse discharge plasma treatment under normal pressure (pulse corona treatment) and a resin processing using a urethane resin has been proposed (Japanese Patent Laid-Open No. 11-131365). No. publication). However, this method also has the above-mentioned disadvantages based on the use of the shrink-resistant resin, that is, the disadvantage that the texture of the fiber is impaired in order to impart sufficient shrink-proof property.
As described above, a method for shrink-proofing animal hair fibers without causing environmental pollution problems due to the use of a chlorinating agent, etc. is a request for conflict between the shrinkage resistance and the texture, and a request for conflict between the shrinkage resistance and the strength. However, there is an urgent need in the art for a new animal hair fiber shrink-proofing technology that satisfies all these requirements.

これらの表に示される結果から、本酵素は市販酵素である製品ＡおよびＢと比較して、ケラチン分解活性が２倍以上高く、コラーゲン分解活性およびエラスチン分解活性は低く、ケラチンに対する特異性の高いことがわかる。従って、本酵素は市販酵素に比して、コラーゲンおよびエラスチンへの作用が少なく、獣毛繊維製品の処理に際して繊維内部のコルテックス細胞や細胞質複合体の分解が小さく、重量減少や強度低下を招くおそれが非常に少ないことが明らかである。An object of the present invention is to provide an animal hair fiber shrink-proofing technique that meets the above-described requirements, that is, without using a chemical with a problem of environmental pollution such as a chlorine-based chemical, and imparts a shrink-proof property that does not shrink even by washing with water. In addition, an object of the present invention is to provide an improved animal hair fiber shrink-proofing technique that does not impair the natural texture of animal hair fibers and does not substantially reduce strength.
As a result of earnest research, the present inventor selectively acts on the scale portion of the surface of the animal hair fiber to decompose it, but the cell membrane complex (CMC, cell membrane complex) layer, which is the fiber main body, is substantially We found an alkaline protease that should be called an ideal anti-shrinking enzyme that does not have an adverse effect, and met the above purpose when performing pulse corona treatment or oxidation treatment with a non-chlorine oxidant prior to shrink-proofing treatment with this specific enzyme. It was found that new animal hair fiber shrink-proof processing technology can be provided. The present invention has been completed based on this finding. The present invention provides the inventions according to items 1 to 12 below.
Item 1. A method for preventing shrinkage of animal hair fibers, wherein the animal hair fibers are subjected to pulse corona treatment or oxidation treatment using a non-chlorine oxidant, and then have a keratin degradation activity of 70 AKU or more and a collagen degradation activity with respect to the keratin degradation activity A method for preventing shrinkage of animal hair, comprising performing an enzyme treatment using an alkaline protease having a ratio of 2 or less and an elastin degradation activity ratio with respect to keratin degradation activity of 4 or less.
Item 2. The animal hair fiber shrink-proofing method according to claim 1, wherein the animal hair fiber is subjected to pulse corona treatment and then subjected to an enzyme treatment using an alkaline protease.
Item 3. Item 3. The shrink-proofing method according to Item 1 or 2, wherein the pulse corona treatment is performed at room temperature under conditions of an average electrolytic strength of 6 to 100 kv / cm, a pulse frequency of 10 pps or more, and a pulse width of 0.1 μs or more.
Item 4. The animal hair fiber shrink-proofing method according to claim 1, wherein the animal hair fiber is oxidized with a non-chlorine oxidant and then subjected to an enzyme treatment using an alkaline protease.
Item 5. The method according to claim 1 or 3, wherein the non-chlorine oxidant is at least one selected from the group consisting of hydrogen peroxide, persulfuric acid and salts thereof, and dichloroisocyanurate.
Item 6. Oxidation treatment with a non-chlorine oxidant is carried out at a bath ratio of 1:15 to 25, a temperature of 30 to 60 ° C., a pH of 4.0 to 4.5, and a time of 1 to 8 hours using 2 to 4 g of oxidant per 1 g of fiber. The method according to claim 5, which is performed under conditions.
Item 7. Item 7. The shrink-proofing method according to any one of Items 1 to 6, wherein the alkaline protease is derived from actinomycetes.
Item 8. Item 7. The shrink-proofing method according to any one of Items 1 to 6, wherein the alkaline protease is produced by Nocardiopsis sp. TOA-1 strain (FERM BP-08603).
Item 9. The contact with the alkaline protease was performed using an alkaline protease having an enzyme titer of 300 to 1000 APU per gram of fiber, a bath ratio of 1:15 to 25, a temperature of 30 to 60 ° C., a pH of 7 to 9.5, and a time of 2 to 8 hours. Item 9. The shrink-proofing method according to any one of Items 1 to 8, wherein
Item 10. Item 4. An alkaline protease used in the animal hair fiber shrink-proofing method according to any one of Items 1 to 3, wherein the keratin degradation activity is 70 AKU or more, the collagen degradation activity ratio to keratin degradation activity is 2 or less, and keratin degradation An alkaline protease having an elastin degrading activity ratio of 4 or less.
Item 11. Item 11. The alkaline protease according to Item 10, which is produced by Nocardiopsis sp. TOA-1 strain (FERM P-18676).
Item 12. Item 10. An animal hair fiber that has been shrink-proofed obtained by the method according to any one of Items 1 to 9.
Hereinafter, the shrink-proofing method of the animal hair fiber of the present invention will be described in detail.
(1) Animal hair fibers The animal hair fibers to which the method of the present invention is applied include various conventionally known animal hair fibers such as wool, cashmere, alpaca, and angora. The animal hair fibers to which the method of the present invention is applied are not only the animal hair fibers themselves, but also the animal hair yarns obtained by spinning the animal hair fibers, the animal hair knitted fabrics obtained by knitting and weaving the animal hair yarns, It may be in the form of an animal hair fiber product such as an animal hair nonwoven fabric obtained by accumulating animal hair fibers. In the method of the present invention, by applying this to an animal hair fiber product, the animal hair fiber in the product can be shrunk, and thus a shrunk animal fiber product can be obtained. Furthermore, these fiber products such as animal hair yarns, animal hair knitted fabrics, and animal hair nonwoven fabrics do not need to be composed of animal hair fibers alone, but are made of a mixture of animal hair fibers and other types of fibers. May be. This other kind of fiber is not limited at all. Typical examples include cellulose fiber, acrylic fiber, polyester fiber, polyamide fiber and the like. The mixing ratio of the other kinds of fibers is not particularly limited, and the animal hair fibers in the mixture can be shrunk by the method of the present invention at any ratio. The mixing ratio of the animal hair fibers that are required to be shrink-proofed may be at least 5% by weight of the total product weight. Specifically, for example, a baseball cap composed of 15% by weight of wool fiber, 83% by weight of acrylic fiber and 2% by weight of spandex, and a knitted product composed of 5% by weight of wool fiber, 15% by weight of nylon fiber and 80% by weight of acrylic fiber. Can be the subject of the shrink-proofing method according to the present invention.
(2) Pulse corona treatment The pulse corona treatment of animal hair fibers according to the present invention is carried out according to a conventionally known method, for example, the method described in JP-A-11-131365 described above. This is done by applying a voltage. That is, it is carried out by applying a constant voltage in a pulsed manner between both electrodes while passing animal hair fibers between the discharge electrode and the counter electrode. The distance between both electrodes should just be the range which an animal hair fiber can pass, and is specifically about 1-40 cm. The voltage applied between the two electrodes has an average electrolytic strength (the average value of the voltage applied between the two electrodes divided by the distance between both electrodes) of about 6 to 100 kv / cm, more preferably 7 to 50 kv / cm. It is preferable to be in a range of about cm, and by adopting such a voltage, a desired pulse corona treatment can be performed.
The pulse high voltage is assumed to be a non-continuous application of a predetermined voltage at a predetermined time interval. Here, the pulse frequency of the pulse high voltage to be applied (number of pulses generated per second) is generally 10 pps or more, and particularly preferably in the range of 50 to 100 pps. The pulse width (the time during which one pulse is generated) is generally about 0.1 μs or more, preferably about 0.5 to 10 μs.
It is important that the pulse corona treatment is performed under normal pressure, whereby a pulse high voltage can be applied efficiently and continuously to the animal hair fibers. Further, by applying a pulse high voltage under normal pressure, oxygen in the atmosphere is activated, and a plasma state in which activated oxygen and electrons exist can be generated. These oxygen and electrons collide with the animal hair fiber to introduce an active group such as a carboxyl group or a hydroxyl group on the surface of the animal hair fiber.
By the pulse corona treatment, the animal hair fiber surface is uniformly hydrophilic.
(3) Oxidant treatment The oxidation treatment of animal hair fibers according to the present invention is carried out using a non-chlorine oxidant. Examples of the non-chlorine oxidant include hydrogen peroxide, persulfuric acid, persulfates, for example, monopersulfate alkali metal salts such as potassium persulfate and potassium hydrogensulfate, and dipersulfate alkali metal salts, sodium dichloroisocyanurate, etc. Dichloroisocyanurate, monoperoxyphthalic acid, and the like. These can be used alone or in combination of two or more. Of these, particularly preferred oxidizing agents are persulfuric acid and its salts. Specific examples of the oxidizing agent include “potassium monohydrogen persulfate” commercially available from DuPont. This is composed mainly of potassium hydrogen persulfate and is a mixture of this and potassium persulfate.
Unlike chlorine-based oxidants, non-chlorine-based oxidants have the advantage that they are not accompanied by the production of AOX that causes environmental pollution. In addition, the treatment with the oxidizing agent can be carried out according to the wet method in the same manner as the subsequent enzyme treatment with the alkaline protease, and thus there is an advantage that both treatment steps can be combined more easily.
The oxidation treatment of animal hair fibers is carried out, for example, by immersing non-treated fibers in a liquid containing an oxidizing agent, or by applying or spraying a liquid containing an oxidizing agent to a fiber to be treated.
The purpose of the oxidation treatment is to make the fiber, especially the scale surface thereof hydrophilic, suitable for the enzyme treatment when the fiber obtained by this oxidation treatment is subsequently subjected to the enzyme treatment. Conditions for the oxidation treatment (bath ratio, temperature, time, solution pH, etc.) can be appropriately determined so that a desired shrink-proof fiber can be obtained by subsequent enzyme treatment. This condition can be appropriately determined according to the type of oxidizing agent used, the type of fiber to be treated, and the like. Preferably, an oxidizing agent amount of 2 to 4 g / L, a bath ratio of 1:15 to 25, a temperature of 30 to 60 ° C., a time of 1 to 8 hours, and a liquid pH of about 4.0 to 4.5 can be employed.
After the oxidation treatment, the obtained animal hair fiber can be subjected to an enzyme treatment directly or appropriately after washing with water. Moreover, the reduction process using a normal reducing agent can also be performed as needed.
(4) Enzyme treatment In the method of the present invention, the animal hair fiber treated in the above (2) or (3) is then subjected to an enzyme treatment. The enzyme treatment is performed by contacting and acting a specific protease. It is important that the protease is an alkaline protease having a keratin degradation activity of 70 AKU or more, a collagen degradation activity ratio to keratin degradation activity of 2 or less, and an elastinolysis activity ratio to keratin degradation activity of 4 or less. In particular, those having a keratin degradation activity of 120 AKU or more are preferred. Typical examples of the alkaline protease include alkaline proteases shown in (5) below.
Here, keratin degradation activity (AKU) is measured by the following measurement method. That is, 1 mL of enzyme solution prepared to 25 APU / mL of casein decomposition activity was mixed with 3 mL of 100 mmol / L borax-sodium carbonate buffer solution (pH 10.5) containing 0.04 g of Keratin Azure made by Sigma. Then, the mixture is allowed to react with stirring at 35 ° C. for 60 minutes, and then the dye released with decomposition is measured at an absorbance of 595 nm. 1 AKU is the amount of enzyme that increases the absorbance at 595 nm by 0.001 per hour under the above reaction conditions.
In the above, the casein degradation activity (APU / mL) of the enzyme solution is measured by the following measurement method. That is, 1 mL of 100 mmol / L borax-sodium carbonate buffer (pH 10.5) containing 1% hamal-sten dairy casein was mixed with 1 mL of enzyme solution, reacted at 35 ° C. for 10 minutes, and then 7.2% trichloro. The reaction is stopped by adding 2 mL of an acetic acid solution, allowed to stand at 35 ° C. for 20 minutes, then filtered through a filter paper (ADVANTEC, No. 6, manufactured by TOYO), and the proteolysate in the filtrate is measured by the forin method. 1 APU is the amount of enzyme that liberates 1 μg of tyrosine per minute in the above measurement method.
Moreover, the collagen degradation activity (ACU / mL) and the elastin degradation activity (AEU / mL) shown by the relative ratio with respect to the said keratin degradation activity are calculated | required with the following measuring methods, respectively. That is, 1 mL of an enzyme solution prepared to 50 APU / mL of casein decomposition activity in 1 mL of 100 mmol / L borax-sodium carbonate buffer solution (pH 10.5) containing 2% of collagen Type I (manufactured by Sigma) or elastin (manufactured by Sigma) And the mixture is allowed to react with slow stirring for 2 hours at 35 ° C., then 2 mL of a 7.2% trichloroacetic acid solution is added to stop the reaction, and the mixture is allowed to stand at 35 ° C. for 20 minutes. Next, it is filtered through a filter paper (ADVANTEC, No. 6, manufactured by TOYO), and the proteolysate in the filtrate is measured by the forin method to determine the amount of increase in absorbance at 660 nm (based on the measured value of 660 nm at reaction time 0). As an increase for that). 1ACU and 1AEU are the amounts of enzyme that increase the absorbance at 660 nm by 0.001 per hour in the above measurement method.
Collagen-degrading activity and elastin-degrading activity indicated by the relative ratios based on the keratin-degrading activity indicate the specificity or selectivity of the enzyme for keratin. That is, the smaller the relative ratio of these collagenolytic activity and elastinolytic activity, the smaller the effect of the enzyme on collagen and elastin, thus indicating that it is specific for keratin.
An enzyme having such characteristics, that is, an enzyme that satisfies a specific keratin degradation activity and a specific collagen degradation activity ratio and an elastin degradation activity ratio with respect to the keratin degradation activity can be used to achieve excellent animal hair shrinkage. A processing effect can be produced. The reason is considered as follows. That is, the surface of the animal hair fiber is covered with keratin and is hydrophobic, but is polarized (hydrophilized) by pretreatment according to the method of the present invention, that is, pulse corona treatment or oxidizing agent treatment. When the animal hair fiber is then treated with the enzyme according to the present invention, the enzyme can uniformly act on the entire surface of the fiber that has been hydrophilized as described above, and the enzyme has high keratin specificity and keratin degradation activity. Thus, it selectively acts on the scale layer on the fiber surface and decomposes and disappears, but hardly acts on the CMC layer, and thus has excellent shrinkage resistance without substantially causing a decrease in strength or a decrease in texture. It is thought that it can be granted.
The enzyme treatment (contact) with the alkaline protease according to the present invention is generally performed by immersing the fiber to be treated in the enzyme solution in the form of an aqueous liquid, or by applying the enzyme solution to the fiber to be treated, spraying, or the like. Implemented by enforcing. The concentration of the enzyme in the enzyme solution is not particularly limited, and can be appropriately determined according to the type of fiber to be treated, the employed pulse corona treatment conditions, the enzyme treatment method, conditions, and the like. Usually, the enzyme is in the form of a solution having a concentration of 10-80 APU / mL, preferably 20-40 APU / mL, and has a titer of about 100-2400 APU, preferably about 300-1000 APU, per 1 g of animal hair fiber weight. Should be used in quantity.
In addition, calcium hydroxide as a pH adjuster, boric acid as a buffering agent, etc., as well as surfactants that are known to be added to and mixed with this type of enzyme treatment liquid as necessary. Etc. can be appropriately added and blended. Examples of the surfactant include surfactants having a chemical penetration promoting effect and surfactants having a degreasing effect, such as “Spralane UF” (manufactured by Zschimmer & Schwartz). The surfactant also includes a surface activity that can also exhibit an antiseptic effect, specifically, “cismoran BH” (manufactured by Bayer). The amount of these surfactants added is not particularly different from the amount in which they are usually used. In the present invention, the use of a surfactant as a penetrating agent is unnecessary, but, of course, does not prevent the use.
The enzyme treatment conditions are, for example, in the case of an immersion method, a bath ratio of about 1: 10-30, preferably about 1: 15-25, a temperature of about 30-60 ° C., preferably about 40-50 ° C. The reaction can be performed under conditions of about pH 7-9.5, preferably about 8.5-9, and time 0.5-8 hours, preferably 2-8 hours, more preferably 1-2 hours. The enzyme reaction can be stopped by heating at about 90 ° C. for about 10 minutes according to a conventional method. After completion of the reaction, it is possible to obtain animal hair fibers subjected to the desired shrink-proofing treatment of the present invention by washing thoroughly with running water and drying.
(5) Alkaline protease Examples of the alkaline protease suitable for the shrinkage-proofing method of the present invention include those derived from actinomycetes. A typical example thereof is one produced by the alkaliphilic Nocardiopsis SP TOA-1 strain found by the present inventors (Japanese Patent Application No. 2002-161099). Hereinafter, the alkaline protease produced by this TOA-1 strain is also referred to as “the present enzyme”.
The TOA-1 strain was obtained from Nocardiopsis sp. On January 16, 2002 at the National Institute of Advanced Industrial Science and Technology, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi 1-chome, Tsukuba, Ibaraki, Japan. It has been deposited with the designation TOA-1, and the deposit number is FERM P-18676. This was deposited internationally on January 29, 2004, and the deposit number is FERM BP-08603.
Cultivation of the above-mentioned bacterium and collection of this enzyme can be performed according to conventional methods. For example, since the bacterium is an alkalophilic actinomycete, the culture is carried out in an alkaline region in which a suitable alkali is added to a normal medium. Nutrient sources such as carbon source, nitrogen source, and other inorganic salts used in the medium may be ordinary ones used for culturing this kind of enzyme-producing bacteria. For example, glucose, soluble starch, cellulose and the like can be exemplified as the carbon source. Examples of the nitrogen source include inorganic substances such as nitrates and ammonium salts, urea, peptone, dry yeast, yeast extract, skim milk, soy flour, corn steep liquor, casein, meat extract, amino acids, and the like. Examples of other inorganic salts include magnesium salts, potassium salts, sodium salts, and phosphates. One of these nutrient sources may be used alone, or two or more of these nutrient sources may be used in combination. Their combination is also arbitrary. Examples of the alkali added to the medium include aqueous solutions of carbonates such as sodium carbonate and sodium hydrogen carbonate having a concentration of about 0.5-2%, aqueous sodium hydroxide, and aqueous ammonia. The pH of the medium is usually preferably about 8-11. Culturing can be carried out at a temperature of about 20-40 ° C., preferably about 30-35 ° C., for 2-7 days, aerobically with stirring or shaking. The desired enzyme is secreted and accumulated mainly in the culture medium.
The enzyme can be collected and purified from the culture solution according to a known method using the physicochemical properties of the enzyme. Since the present enzyme is mainly secreted outside the microbial cells (in the culture solution), for example, the microbial cells can be removed by an operation such as filtration or centrifugation to obtain a crude enzyme solution. The crude enzyme solution is further subjected to a conventional method, for example, a method of salting out using ammonium sulfate, a method of precipitation using an organic solvent such as methanol, ethanol, acetone, a method of adsorbing using keratin, etc., an ultrafiltration method They can be purified by various chromatographic methods such as gel filtration chromatography, ion exchange chromatography and hydrophobic chromatography. These purification operations can be used alone or in combination.
As one of the particularly preferred purification methods, 80% saturated ammonium sulfate is first added to the culture filtrate, salting out is performed, and the resulting precipitate is dissolved in a buffer solution, and then CM-Toyopearl 650M (manufactured by Tosoh Corporation), for example, An example is a method of performing ion exchange chromatography using DEAE-Toyopearl 650M (manufactured by the same company). By this method, a purified enzyme that is uniform in SDS electrophoresis can be obtained.
The enzyme thus obtained has the following properties.
(1) Action and substrate specificity Acts on proteins and peptides, and cleaves peptide bonds by an endo-type mechanism to produce low molecular weight oligopeptides and amino acids. It also exhibits strong activity against insoluble proteins such as keratin.
(2) Optimum pH and stable pH
As buffers, HCl / KCl (pH 1.0-1.5), glycine / NaCl / HCl (pH 2.0-3.0), acetic acid (pH 4.0-5.0), phosphoric acid (pH 6.0-7) 0.0), Tris-HCl (pH 7.0-9.0), Glycine / NaCl / NaOH (pH 9.0-12.0) and KCl / NaOH (pH 12.0-13.0). From the activity measurement results obtained according to the activity measurement method, the optimum pH of the enzyme is 11.0-11.5 when casein is used as a substrate at 30 ° C., and 12.0 or more when keratin is used as a substrate. It is.
Similarly, the stable pH range of the present enzyme determined by measuring the residual protease activity after holding the enzyme in a buffer solution of each pH at 30 ° C. for 24 hours is a wide range of 1.5 to 12.0. It is confirmed that
(3) Optimal temperature and stable temperature The optimal temperature of this enzyme is 70-75 ° C. when casein is used as a substrate, and 65-70 ° C. when keratin is used as a substrate. In addition, the enzyme was added to 100 mmol / L Tris-HCl buffer (pH 7.0), held at a temperature range of 40-80 ° C. for 10 minutes, and the residual protease activity was measured. Is stable. In addition, regarding the temperature stability of this enzyme, the effect of calcium addition (10 mM) is not recognized.
(4) Molecular weight The molecular weight of the enzyme was measured by SDS electrophoresis. As a result, the molecular weight was about 20,000. In addition, the molecular weight calculated from the amino acid sequence described in SEQ ID NO: 2 described later is 19,150.
(5) Isoelectric point The isoelectric point of the enzyme was measured by isoelectric focusing. As a result, the isoelectric point was 10.0 or more.
(6) Inhibition Each of the general enzyme inhibitors PMSF (phenylmethanesulfonyl fluoride), EDTA (ethylenediaminetetraacetic acid), and SSI (Streptomyces subtilisin inhibitor) is adjusted to a predetermined concentration of 50 mmol / L Tris-HCl. The resultant was dissolved in a buffer solution (pH 9.0), treated with this enzyme for 30 minutes at 30 ° C., and then a fixed amount was taken from the treated solution to measure its residual activity. As a result, this enzyme was inhibited by PMSF and SSI, and not inhibited by EDTA. This revealed that this enzyme is a serine protease.
(7) Amino terminal sequence The sequence from the amino terminal to the 25th amino acid of this enzyme was determined using a gas phase protein sequencer (manufactured by Shimadzu Corporation, PPSQ-21). As a result, the 1-25th sequence of SEQ ID NO: 2 was confirmed.
(8) Nucleotide sequence and amino acid sequence The gene and amino acid sequence of this enzyme can be obtained by conventional methods (for example, J. Sambrook, EF Fritsch, T. Maniatis: Molecular Cloning. A Laboratory Manual, 2nd. Ed. Cold Spring). According to Harbor Laboratory Press, 1989, etc.), it was determined according to the protocol of the equipment used, reagent kit, etc. First, the purified enzyme was treated with urea and then decomposed with lysyl endopeptidase (Wako Pure Chemical Industries), and the amino acid sequence of the obtained fragment was determined with a gas phase protein sequencer. From the amino acid sequence information thus obtained, appropriate two types of oligonucleotide primers were synthesized by the phosphoramidite method, and using these primers, gene amplification was performed according to PCR (Biometra, T-Gradient Thermoblock 050-801). went. As a result, a specific amplified fragment was observed around 0.5 kbp. Using this fragment as a probe, a full-length gene encoding this enzyme was screened from the genomic library of this strain TOA-1. A DNA sequencer (manufactured by LICOR, LICOR-4000) based on the dideoxy method (F. Sanger et.al., Proc. Natl. Acad. Sci., 74 , 5463-5467, 1977) was used as the base sequence of the obtained clone. ). The base sequence (564 bp) is shown in SEQ ID NO: 1. The amino acid sequence (188 amino acids) determined based on the base sequence is shown in SEQ ID NO: 2.
(9) Keratin specificity This enzyme has a particularly excellent keratin specificity. For example, Table 1 shows the results of comparing the keratin degradation activity, collagen degradation activity, and elastin degradation activity of this enzyme and commercially available representative alkaline proteases (referred to as products A and B). In addition, Table 2 shows the results of determining the relative activity ratio of collagen degradation activity and elastin degradation activity based on keratin degradation activity.

From the results shown in these tables, this enzyme has a keratin-degrading activity more than twice as high as that of products A and B, which are commercially available enzymes, has low collagen-degrading activity and elastin-degrading activity, and has high specificity for keratin. I understand that. Therefore, this enzyme has less action on collagen and elastin than commercially available enzymes, and cortex cells and cytoplasmic complexes in the fiber are less decomposed during the treatment of animal fiber products, leading to weight loss and strength reduction. It is clear that there is very little fear.

The invention's effect

The present invention provides a halogen-free process that combines a pulse corona treatment by applying a pulse high voltage or an oxidation treatment with a non-chlorine oxidant and an enzyme treatment with a protease having high keratin specificity, and is extremely clean. Therefore, it has low environmental impact and is highly practical.
The product made of animal hair fibers obtained by the present invention has good quality that has been sufficiently shrink-proofed without losing properties such as mechanical strength, can be washed in a home washing machine, and is dry-cleaned. There is no need. In addition, this product possesses the soft touch inherent to animal hair fibers, and its texture is very excellent.
In addition, the method of the present invention is practically excellent in terms of work, equipment, cost, and production time because there is no risk of discharging compounds harmful to the environment, and no work under reduced pressure is required.

Hereinafter, production examples of alkaline protease used in the present invention will be described as reference examples, and then the present invention will be described in more detail with reference to examples of the shrinkage-proofing method of the present invention.
In addition, the area shrinkage rate, the strong elongation, and the weight reduction rate of the samples obtained in the examples are according to the following measurement method and calculation method.
(1) Area shrinkage rate of sample 3 hours using sodium phosphate buffer (pH 7) and a Cubex tester (FLOATAIRE) according to IWS (International Wool Secretariat) test standard TM185. The area shrinkage ratio was calculated by multiplying the average lengths of the sample before and after the shrinkage test.
(2) Strength and elongation test of sample The maximum tensile strength (N) of each of 20 points of warp and weft was measured using an autograph manufactured by Shimadzu Corporation.
(3) Weight reduction rate of sample The weight change after drying for 1.5 hours at 100 ° C. before and after the shrink-proofing treatment was measured.
Reference Example 1 Preparation of Crude Enzyme Sample Nocardiopsis SP TOA-1 strain (FERM BP-08603) precultured solution was added 0.5% skim milk, 0.1% yeast extract and sterilized sodium carbonate A small jar fermenter containing 4000 mL of a medium (pH 10.5) containing 1.0% was inoculated and cultured at 30 ° C., aeration rate 1 v / v / min, 200 rpm / min for 3 days. After completion of the culture, the culture was centrifuged at 8,000 rpm for 10 minutes to remove the cells.
As a result, about 3,800 mL of a 45 APU / mL crude enzyme solution was obtained. To this crude enzyme solution, ammonium sulfate powder was added until it was 80% saturated. After standing overnight in the dark at 5 ° C., the resulting precipitate was collected by centrifugation at 8,000 rpm, and lyophilized. As a result, 8.9 g of a crude enzyme preparation of 15,100 APU / g was obtained.
This product had a keratin degradation activity of 72,500 AKU / g, a collagen degradation activity of 77,900 ACU / g, and an elastin degradation activity of 106,000 AEU / g. From these, the collagen degradation activity ratio to the keratin degradation activity of this enzyme is calculated as 1.08, and the elastin degradation activity ratio is calculated as 1.46.

各表に示す結果から、本発明方法によれば、酵素の使用量および処理時間を適宜決定することにより、面積収縮率５％以下で強度低下が実質的になく、重量減少率も３％以下の、優れた防縮加工製品を収得できることが判った。しかも、本発明によれば、未処理布と差のない優れた風合いを有する防縮加工製品が得られることが明らかとなった。本発明防縮加工製品は、ソフトで柔らかく白くなっており、染色性の向上も認められた。
また、得られた本発明処理布、未処理布およびパルスコロナ処理のみを行った対照布における羊毛繊維の形状を電子顕微鏡にて観察した。その結果、未処理羊毛布では、羊毛繊維のスケールが明瞭に観察されたのに対して、パルスコロナ処理のみを行った処理布では、スケールが削られたような状態で丸みを帯びている様子が観察され、本発明処理布（収縮率０％）では、スケールがさらに丸みを帯びている様子が明らかとなった
比較例１
実施例１と同様にしてパルスコロナ処理した試料に、市販のアルカリプロテアーゼ酵素製品（製品ＡおよびＢ、前記表１および２に示すケラチン分解活性、コラーゲン分解活性比およびエラスチン分解活性を有するもの）を作用させて、比較処理布を得た。
市販酵素製品ＡおよびＢを用いた酵素処理は、浴比１：２０、５０℃、Ｂｒｉｔｔｏｎ−Ｒｏｂｉｎｓｏｎ広域緩衝液（ｐＨ８．０）を用い、非イオン性界面活性剤「スコアロール１００」（北広ケミカル株式会社製繊維加工用薬剤）２ｇ／Ｌ中で行った。酵素製品Ａの場合は、力価２３３５ＡＰＵ／ｍＬで１６時間処理した。また酵素製品Ｂの場合は、力価４２８ＡＰＵ／ｍＬで１６時間処理した。
得られた比較処理布の性質を、未処理布および本発明処理布と比較して表５に示す。
尚、市販酵素製品の力価および処理時間を実施例１と一致させて行った比較試験では、得られる処理布の収縮率は５％を遙かに越えるものとなり、所望の防縮加工はできなかった。

表５に示す結果から次のことが明らかである。本発明方法では、本酵素を２ｇ／Ｌ用いて６時間処理することにより、収縮率３．６６％を達成できたのに対し、製品Ａでは２，３３５ＡＰＵ／ｍＬで１６時間、製品Ｂでは４２８ＡＰＵ／ｍＬで１６時間の処理によって、収縮率５％が達成できるにすぎなかった。また、本酵素を利用した本発明方法では、強度低下は未処理とほぼ同様であったのに対し、製品ＡおよびＢを用いた場合は、それぞれ０．５Ｎまで強度が低下（約５８％の強度低下が認められる）し、重量減少率も各々１３％および１８％と著しく、生地はボロボロの状態となることが明らかとなった。
本発明方法に採用するパルスコロナ処理は、それ自体温度上昇しないために羊毛繊維の風合いを低下させるおそれのないものであり、しかも本発明方法では引き続く酵素処理によって風合いがより柔らかくなっていた。従って、本発明方法では、当然ながら柔軟剤などの利用は不必要である。また、本発明方法では従来法において採用される如き酸化剤処理をしていないので、処理布は耐黄変性があり、染色性も向上している。さらに、本発明方法では、促進剤としての非イオン性界面活性剤の使用も必要がなく、これは廃液処理の観点からも優れた方法であるということができる。As a sample, a wool-attached white cloth according to JIS L 0803 was prepared. This sample was subjected to a pulse corona treatment by applying a pulse high voltage in an atmosphere of a relative humidity of 30 to 40% and room temperature using an atmospheric pressure plasma treatment apparatus “Plasma Atom” manufactured by Nippon Paint Co., Ltd. Pulse high voltage application is performed while transporting the sample at 760 mm / min under the conditions of a distance between electrodes of 23.5 mm, an average electric field strength of 32.3 kv / cm, a power output of 76 kv, a pulse frequency of 120 pps and a pulse width of 2.5 μs, Repeated several times.
Next, a sample subjected to pulse corona treatment was prepared under the conditions of 0.1 mol / L Tris-HCl buffer (pH 9.0), bath ratio 1:20, 50 ° C. and pH 9.0. The enzyme treatment was performed by contact treatment with L or 4 g / L (corresponding to 300 AKU / mL, 450 AKU / mL or 600 AKU / mL) for 2, 4, 6 or 8 hours, respectively.
Comparison of the properties of the enzyme-treated cloth of the present invention compared with the untreated cloth (white wool-attached cloth not subjected to pulse corona treatment or silicon treatment) used as a sample and the wool-attached white cloth subjected to pulse corona treatment alone. The results are shown in Table 3 (result of area shrinkage rate) and Table 4 (result of strong elongation test).

From the results shown in each table, according to the method of the present invention, by appropriately determining the amount of enzyme used and the treatment time, there is substantially no decrease in strength at an area shrinkage rate of 5% or less, and the weight reduction rate is 3% or less. It has been found that an excellent shrink-proof processed product can be obtained. Moreover, according to the present invention, it has been clarified that a shrink-proof processed product having an excellent texture that is not different from that of an untreated fabric can be obtained. The shrink-proof processed product of the present invention was soft, soft and white, and improved dyeability was also observed.
Moreover, the shape of the wool fiber in the obtained control cloth, this untreated cloth, and the control cloth which performed only the pulse corona treatment was observed with an electron microscope. As a result, in the untreated wool cloth, the scale of the wool fiber was clearly observed, whereas in the treated cloth subjected only to the pulse corona treatment, the scale was rounded in a state where the scale was shaved. As a result, it was clarified that the treated cloth of the present invention (shrinkage rate 0%) was further rounded.
Comparative Example 1
A commercially available alkaline protease enzyme product (products A and B having keratin degradation activity, collagen degradation activity ratio and elastin degradation activity shown in Tables 1 and 2) was applied to a sample subjected to pulse corona treatment in the same manner as in Example 1. A comparatively treated cloth was obtained by the action.
Enzymatic treatment using commercially available enzyme products A and B uses a bath ratio of 1:20, 50 ° C., Britton-Robinson broad-area buffer (pH 8.0), and a nonionic surfactant “Scoreol 100” (Kitahiro Chemical) It was carried out in 2 g / L of a textile processing agent manufactured by the same company. In the case of enzyme product A, it was treated with a titer of 2335 APU / mL for 16 hours. In the case of enzyme product B, it was treated with a titer of 428 APU / mL for 16 hours.
The properties of the comparative treated cloth obtained are shown in Table 5 in comparison with the untreated cloth and the treated cloth of the present invention.
In addition, in the comparative test conducted by matching the titer and treatment time of the commercially available enzyme product with those in Example 1, the shrinkage rate of the obtained treated cloth exceeds 5%, and the desired shrink-proofing process cannot be performed. It was.

From the results shown in Table 5, the following is clear. In the method of the present invention, a contraction rate of 3.66% was achieved by treating the enzyme with 2 g / L for 6 hours, whereas in product A, 2,335 APU / mL was applied for 16 hours, and in product B, 428 APU. A shrinkage of 5% could only be achieved by treatment for 16 hours at / mL. Further, in the method of the present invention using the present enzyme, the strength decrease was almost the same as that of the untreated, whereas when products A and B were used, the strength decreased to 0.5 N (about 58%). Strength reduction was observed), and the weight loss rates were 13% and 18%, respectively, and it became clear that the dough became tattered.
The pulse corona treatment employed in the method of the present invention itself does not raise the temperature, so that the texture of the wool fiber is not lowered, and in the method of the present invention, the texture is softened by the subsequent enzyme treatment. Therefore, in the method of the present invention, it is naturally unnecessary to use a softening agent or the like. Further, in the method of the present invention, since the oxidizing agent treatment as employed in the conventional method is not performed, the treated cloth has yellowing resistance and dyeability is improved. Furthermore, in the method of the present invention, it is not necessary to use a nonionic surfactant as an accelerator, which can be said to be an excellent method from the viewpoint of waste liquid treatment.

各表に示す結果から、次のことが明らかである。即ち、酸化剤処理のみでは十分な防縮性は得られないが、酸化剤処理と酵素処理とを組み合わせるときには、酵素の使用量および処理時間を適宜決定することにより、面積収縮率５％を遙かに下回りしかも強度低下も殆どなく、重量減少率も３％以下の、優れた防縮加工製品を収得できることが判る。しかも、本発明によれば、未処理布と差のない優れた風合いを有する防縮加工製品が得られることが明らかとなった。本発明防縮加工製品は、ソフトで柔らかく白くなっており、染色性の向上も認められた。
また、得られた本発明処理布、未処理布および酸化剤処理のみを行った対照布における羊毛繊維の形状（乾燥状態）を電子顕微鏡にて観察した。その結果、未処理羊毛布では、羊毛繊維のスケールが明瞭に観察され、酸化剤処理のみを行った処理布でも、ほぼ同様にスケールが観察されたのに対して、本発明処理布（収縮率０％）では、スケールが削られて丸みを帯びている様子が明らかとなった。尚、この例で得られた本発明処理布を、実施例１で得た本発明処理布と対比すると、この例で得られた処理布では羊毛繊維表面に若干の損傷が見られる場合があり、これが機械的強度を実施例１の場合に比して若干低下させるものと考えられる。As a sample, a wool-attached white cloth according to JIS L 0803 was prepared. This sample was immersed in a solution (pH 4.0) in which 2 g of potassium hydrogen persulfate (“Oxone ^™ ” manufactured by DuPont) was dissolved in 1 L of water at a bath ratio of 1:20 and 50 ° C. for 30 minutes. Oxidant treatment was performed.
Next, the oxidant-treated sample was subjected to 2 g / L or 3 g / L of this enzyme under the conditions of 0.1 mol / L Tris-HCl buffer (pH 9.0), bath ratio 1:20, 50 ° C. and pH 9.0. The enzyme treatment was performed by contact treatment with L (corresponding to 300 AKU / mL or 450 AKU / mL) for 2, 3, 4 or 6 hours, respectively.
The properties of the obtained enzyme-treated cloth of the present invention were compared with the untreated cloth (white cloth attached with wool which was not subjected to oxidizing agent treatment or enzyme treatment) and the white cloth attached with wool which was only treated with oxidizing agent. Table 6 (results of area shrinkage) and Table 7 (results of high elongation test).

From the results shown in each table, the following is clear. That is, sufficient shrinkage cannot be obtained only by the oxidant treatment, but when combining the oxidant treatment and the enzyme treatment, the area shrinkage rate can be increased by 5% by appropriately determining the amount of the enzyme used and the treatment time. In addition, it can be seen that an excellent shrink-proof processed product having a lower strength and almost no decrease in strength and a weight reduction rate of 3% or less can be obtained. Moreover, according to the present invention, it has been clarified that a shrink-proof processed product having an excellent texture that is not different from that of an untreated fabric can be obtained. The shrink-proof processed product of the present invention was soft, soft and white, and improved dyeability was also observed.
Moreover, the shape (dry state) of the wool fiber in the obtained treated cloth of the present invention, the untreated cloth, and the control cloth subjected only to the oxidizing agent treatment was observed with an electron microscope. As a result, in the untreated wool cloth, the scale of the wool fiber was clearly observed, and in the treated cloth subjected only to the oxidizing agent treatment, the scale was observed almost in the same manner, whereas the treated cloth (shrinkage rate) 0%), it became clear that the scale was cut and rounded. In addition, when the treated cloth of the present invention obtained in this example is compared with the treated cloth of the present invention obtained in Example 1, the treated cloth obtained in this example may have some damage on the wool fiber surface. This is considered to reduce the mechanical strength slightly as compared with the case of Example 1.

According to the present invention, it is possible to obtain a quality animal hair fiber product that has been sufficiently shrink-proofed without substantially losing properties such as mechanical strength. This animal hair fiber product can be washed in a home washing machine and does not require dry cleaning. Moreover, it has the soft touch inherent to animal hair fibers, and its texture is very excellent.
The method of the present invention is practically excellent in terms of work, equipment, cost, and production time because there is no risk of discharging compounds harmful to the environment, and there is no need for work under reduced pressure.

Claims (7)

A shrink proofing method of animal fiber, after pulsed corona treatment the animal fiber, elastin degradation collagenolytic activity ratio keratolytic activity against a is and keratolytic activity than 70AKU is for 2 or less and keratin hydrolyzing activity A method for preventing shrinkage of animal hair, comprising performing an enzyme treatment using an alkaline protease having an activity ratio of 4 or less. 2. The shrink-proofing method according to claim 1 , wherein the pulse corona treatment is performed at normal temperature under conditions of an average electrolytic strength of 6 to 100 kv / cm, a pulse frequency of 10 pps or more, and a pulse width of 0.1 μs or more. The shrink-proofing method according to claim 1 or 2 , wherein the alkaline protease is derived from actinomycetes. Alkaline protease, Nocardioides Opsys sp (Nocardiopsis sp.) Shrink proofing method according to claim 3 TOA-1 strain (FERM BP-08603) is intended to produce. The contact with the alkaline protease was performed using an alkaline protease having an enzyme titer of 300 to 1000 APU per gram of fiber, a bath ratio of 1:15 to 25, a temperature of 30 to 60 ° C., a pH of 7 to 9.5, and a time of 2 to 8 hours. The shrink-proof processing method as described in any one of Claims 1-4 performed by this. The animal hair fiber by which shrink-proof processing obtained by the method as described in any one of Claims 1-5 was carried out. A textile product comprising animal hair fibers subjected to shrink-proof processing obtained by the method according to any one of claims 1 to 5 .