JPH08296177A - Treatment of fiber - Google Patents

Abstract

PURPOSE: To extremely improve handle and touch without reducing tenacity and to recover an immobilized enzyme by treating a fiber with an immobilized enzyme prepared by bonding an enzyme to an amino acid (co)polymer. CONSTITUTION: An enzyme such as cellulase is bonded to an amino acid (co)polymer such as a poly-L-glutamic acid having 5-5,000, preferably 10-1,000 degree of polymerization, etc., by using a cross-linking agent such as glutaraldehyde to form an immobilized enzyme. A fiber to be subjected to enzyme hydrolysis such as cotton, etc., is immersed in an alcohol, de-fatted, dried, treated by the enzyme by immersing the fiber in an aqueous solution of the immobilized enzyme adjusted to an optimum pH for a given time, washed and dried. The aqueous solution of the immobilized enzyme is mixed with an acid or an alkali to change pH and the immobilized enzyme is precipitated and readily recovered.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to fiber treatment using immobilized enzymes. More specifically, the present invention relates to a method for treating a fiber, which can improve the texture and feel of the fiber without lowering the strength of the fiber and can easily recover the immobilized enzyme.

[0002]

2. Description of the Related Art In recent years, enzymatic reactions have come to be used in various industrial fields, and in particular, enzymatic treatment is carried out to improve the texture and feel of fibers. Conventionally, most of the enzymes used when treating fibers have been enzyme soluble in solution. Collecting and reusing the enzymes used for fiber treatment is economical because it can use relatively expensive enzymes efficiently, and removing the enzyme components from the waste liquid can solve environmental pollution problems such as water pollution. Is important from. However, in order to recover the enzyme used for the fiber treatment from the reaction solution, it was necessary to make full use of a separation / purification technique, which was technically very difficult. What was considered to solve such a drawback is to immobilize the enzyme on a carrier insoluble in the reaction system to form an immobilized enzyme. By immobilizing the enzyme on the carrier, the enzyme can be easily recovered from the reaction system or the product. Further, by encapsulating such an immobilized enzyme in a column or the like, continuous reaction becomes possible, and improvement in production efficiency can be expected.
Furthermore, in the past, the enzyme reaction was controlled by inactivating the enzyme by heat or chemical treatment, but by using an immobilized enzyme, the enzyme can be easily separated from the reaction system, and it can be efficiently Many advantages are possible, including the ability to control the reaction. The technology for producing immobilized enzymes has been devised for a long time.For example, immobilized enzymes include activated carbon, silica gel, inorganic carriers such as alumina, natural polymer carriers such as cellulose, starch, chitin, alginic acid and collagen, and polyacrylamide. , Synthetic polymer carriers such as polyaminopolystyrene and ethylene-maleic acid derivatives are used by physical adsorption, ionic bond, covalent bond, lattice, microencapsulation, etc., and some of these immobilized enzymes have already been put to practical use. Has been done.

However, in such a conventional fiber treatment using an immobilized enzyme, the reaction system becomes heterogeneous because the enzyme is insoluble in water, resulting in inconvenience such as reduction in reaction efficiency. That is, by binding the enzyme to the water-insoluble carrier, the conformation of the enzyme may be changed, or the interaction between the substrate and the enzyme may be changed by the steric effect with the carrier or the electrostatic effect. High and consequently requires long reaction times. Therefore, it is desired to use an immobilized enzyme which becomes water-soluble during the reaction and provides a place for a homogeneous reaction system, and the enzyme itself has a property as an immobilized enzyme. An immobilized enzyme is disclosed in Japanese Patent Application Laid-Open No. 62-14782 as a means for achieving such an object. However, in order to achieve the purpose of the immobilized enzyme, it is necessary to add a metal salt such as calcium or magnesium or to use a chelating agent such as ethylenediaminetetraacetic acid or citric acid to remove the metal salt. Many enzymes require metal ions to express their activity, which limits the enzymes that can be used with the above-described immobilized enzymes using chelating agents. Further, the necessity of using a metal salt or an expensive chelating agent causes an economical problem.

[0004]

The present invention solves the above-mentioned drawbacks of conventional immobilized enzymes, has both the advantages of water solubility and water insolubility, and can be used in a simple method with excellent enzyme reaction efficiency. It is an object of the present invention to provide a method for treating a fiber using the immobilized enzyme.

[0005]

The present invention provides a method for treating a fiber, which comprises contacting an immobilized enzyme (hereinafter, simply referred to as an immobilized enzyme) in which an amino acid (co) polymer and an enzyme are bound to the fiber. Is provided. The present invention will be described in detail below. The amino acid (co) polymer used in the present invention is obtained by homopolymerizing or copolymerizing amino acid monomers. As the amino acid monomer, in addition to naturally occurring amino acids, amino acids having a non-natural structure can be used. In the present invention, the amino acid monomer, the polymer is required to take a water-soluble-water-insoluble reversible morphological change with the change of the pH of the solution, for example glutamic acid,
Examples include aspartic acid and lysine. The amino acid copolymer is a block copolymer composed of a block component (A) composed of a polyamino acid and a block component (B) composed of another polymer, for example, a (A)-(B) block copolymer. , (A)-(B)-(A) block copolymers and the like. Examples of the block component (B) include polybutadiene, polyisoprene, styrene-butadiene copolymer, styrene-isoprene copolymer, butadiene-isoprene copolymer, and the like. A group such as an amino group or an alkoxy group is introduced into at least one of the terminals,
A block copolymer can be obtained by copolymerizing with an amino acid monomer. In the amino acid (co) polymer of the present invention, it is preferable that the amino acid monomer is (co) polymerized in an amount of 20 mol% or more. When the amino acid monomer content is less than 20 mol%, it becomes difficult to take a sufficient reversible water-insoluble morphological change due to the pH change of the solution. In the present invention, a known method may be used for the (co) polymerization of amino acid monomers, for example, N-carboxy-α-
A ring-opening polymerization method of an amino acid anhydride, a method using various active esters or a condensing agent, etc. can be used. The degree of polymerization of the amino acid (co) polymer is usually in the range of 5 to 5000,
It is preferably 10 to 1000. When the degree of polymerization of the amino acid (co) polymer is less than 5, it becomes difficult to separate when the immobilized enzyme is insolubilized, and when the degree of polymerization exceeds 5000, the viscosity of the reaction system becomes high and the handling becomes inconvenient.

As the enzyme used in the present invention, an enzyme effective for the intended fiber treatment may be selected. Specifically, aminopeptidase, chymotrypsin, papain,
Urokinase, urease, amidase, asparaginase, protease, glucose phosphatase, amylase, chitinase, lysozyme, cellulase, invertase, pectinase, lipase, ribonuclease, various restriction enzymes (endodeoxyribonuclease)
Hydrolases such as lysine racemase, UDP galactose epimerase, glucose isomerase, lactase racemase, isomerization enzymes such as lysine aminomutase, glycine methyltransferase, transketolase, lysine acetyltransferase, glutamyltransferase, phosphorylase, hexokinase, phosphase Foglyceromutase, glutamate aminotransferase, aspartate aminotransferase, leucine aminopeptidase, creatine phosphokinase and other transferases, alcohol dehydrogenase, aldehyde dehydrogenase, fumarate reductase, lactate dehydrogenase, glucose dehydrogenase Genase, alcohol oxidase, amino acid oxidase, nicotine Chromatography with Dinner hydrogenase kinase, hydroxylamine oxidase, glutathione dehydrogenase, cytochrome c oxidase, catalase,
Redox enzymes such as peroxidase, lipoxygenase, superoxide dismutase, etc., ornithine decarboxylase, lysine decarboxylase, enolase, tryptophan synthetase, lyase such as adenylate cyclase, t-RNA synthetase, acetyl CoA synthetase, glutamine synthetase. Ligase such as.

The bond between the enzyme and the amino acid (co) polymer is
It is achieved by a covalent bond method or an ionic bond method. Coupling by the covalent bond method includes a carboxyl group, an amino group, a hydroxyl group, a sulfhydryl group or the like of an amino acid (co) polymer, a terminal carboxyl group or an amino group, and a carboxyl group, an amino group, a hydroxyl group or a sulfhydryl group of an enzyme. However, any reaction method may be used. For example, if the peptide method is used, the side chain or terminal carboxyl group of the amino acid (co) polymer is made into a derivative such as azide, chloride, or isocyanate, and this is reacted with the amino group of the enzyme, or by carbodiimidization. Examples include a method of reacting a carboxyl group or amino group of an amino acid (co) polymer with a carboxyl group or amino group of an enzyme using a reagent or a condensation reagent such as Woodward reagent K. In addition, in the alkylation method, an amino acid (co) polymer may be activated with a halogenated acetyl derivative, a triazinyl derivative, a halogenated methacryl derivative, etc. and reacted with an amino group, a phenolic hydroxyl group or a thiol group of an enzyme. The (co) polymer and the amino group of the enzyme may be bonded with a crosslinking agent such as glutaraldehyde or hexamethylene diisocyanate. Among these binding methods, the method using glutaraldehyde is simple and particularly preferable. In the present invention, the ratio of the amino acid (co) polymer bound to one molecule of the enzyme is usually 20 molecules or less, preferably 1 to 10 molecules. If the binding ratio of the amino acid (co) polymer exceeds 20 molecules per 1 molecule of the enzyme, there is a risk of affecting the structure of the enzyme molecule or inhibiting the enzyme reaction.

The immobilized enzyme obtained by the above method can be in a water-soluble and water-insoluble state depending on the hydrogen ion concentration of the solution, but the hydrogen ion concentration range depends on the type of amino acid (co) polymer used. Choose timely. For example,
When polyglutamic acid or polyaspartic acid is used as the amino acid (co) polymer, the immobilized enzyme becomes water-insoluble when the pH of the solution is 3 or less, and the immobilized enzyme becomes water-soluble when the pH is 4 or more. Therefore, the enzymatic reaction can be efficiently proceeded in a homogeneous system in a neutral or alkaline solution, and the immobilized enzyme can be easily separated from the reaction system by acidifying the reaction system after the reaction for a predetermined time. Not only can the post-treatment be simplified, but the separated immobilized enzyme can be reused as the immobilized enzyme by returning it to the neutral or alkaline reaction system again. Also,
When polylysine is used as the amino acid (co) polymer, it becomes alkaline and water-insoluble, and becomes neutral or acidic and water-soluble contrary to the case of polyglutamic acid.

The pH of the immobilized enzyme solution can be adjusted by any existing method, but in order to make it acidic, inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid and boric acid, acetic acid,
Organic acids such as citric acid, oxalic acid, and lactic acid can be used. For alkalinity, amines such as sodium hydroxide, potassium hydroxide, ammonia, and ethanolamine can be used. When adjusting the pH, use of a high concentration of acid or alkali may locally deactivate the enzyme, so it is preferable to use the acid or alkali at a concentration of about 0.1 to 2N.

The fiber to be treated in the method of the present invention is not particularly limited as long as it is a fiber which undergoes enzymatic decomposition, and specifically, natural fibers such as cotton, wool and silk,
Examples include mixed fibers of these natural fibers and various synthetic fibers such as polyester, nylon, acrylic, vinyl chloride, rayon, and recycled fibers of these natural fibers. These fibers can be degreased with alcohol or the like before contacting with the immobilized enzyme. In the present invention, the fiber and the immobilized enzyme are brought into contact with each other in water, but it is desired to select an amino acid (co) polymer that is soluble in water at a pH at which the reaction activity of the enzyme used is highest. The conditions such as the ratio of the immobilized enzyme to the fiber, the treatment time of the fiber, the treatment temperature, etc. are appropriately adjusted according to the type of enzyme used. The fiber treated with the immobilized enzyme is washed with water or alcohol. In the present invention, after the treatment of the fiber with the immobilized enzyme is completed, the immobilized enzyme is separated from the fiber in a state of being dissolved in water, and the amino acid (co) polymer using the pH of the recovered reaction solution is treated with water. If the enzyme is adjusted to be insoluble, the immobilized enzyme becomes insoluble in water, so that the immobilized enzyme can be easily recovered by filtration or the like. The treatment method of the present invention can be applied not only to adjusting the thickness of the fiber to improve the texture and feel of the fiber, but also to washing and dyeing the fiber using an enzyme.

[0011]

The present invention will be described in detail below with reference to examples. Example 1 (1) Synthesis of Immobilized Enzyme γ-Methyl-L-glutamic acid-N-carboxylic acid anhydride was polymerized in dichloromethane at room temperature for 3 days using triethylamine as a polymerization initiator. The obtained poly-γ-methyl-L-glutamic acid (PMLG) was hydrolyzed with an aqueous ethanolic sodium hydroxide solution to give poly-L-glutamic acid (PL
G) was obtained. 0.2 g of the obtained PLG was dissolved in 10 ml of a phosphate buffer (pH 7.4), and cellulase (Tricoderma r) was dissolved in the phosphate buffer at a concentration of 5% by weight.
eesei, Sigma) was mixed with 0.5 ml. 50 ml of a 20 wt% glutaraldehyde aqueous solution was gradually added dropwise to the mixed solution, and the reaction was continued at 4 ° C. for 2 hours to synthesize PLG-immobilized cellulase. (2) Fiber treatment Cotton thread for sewing (narrow mouth for household use, 30/3, Y-TK257
(0, manufactured by Kawaguchi Co., Ltd.) was cut into a length of 30 cm, 10 pieces were bundled, both ends thereof were tied, both ends were immersed in methanol overnight, and then dried in a vacuum dryer for 7 hours to obtain a sample for enzyme treatment. This cotton yarn bundle was placed in a 15 ml test tube, and 20 units of the above PLG-immobilized cellulase prepared at pH 5 with an acetate buffer was added with the enzyme (1 unit was carboxymethylcellulose as a substrate, pH 5 and temperature 37 ° C., Enzyme amount for producing 1 μmol glucose), 55
Shake at 160 ° C./min for 8 hours. After processing,
Take the cotton thread out of the reaction solution, and twice with 0.1 M / liter phosphate buffer (pH 8) and three times with pure water.
After washing twice with methanol, it was dried under reduced pressure for 7 hours. The weight and tensile strength of the fibers before and after treatment with the immobilized enzyme were then measured. The weight reduction rate and the tensile strength reduction rate were measured by the following methods. The results are shown in Table 1.
Shown in Weight reduction rate Weight of cotton yarn after treatment with immobilized enzyme ÷ Weight of cotton yarn before treatment with immobilized enzyme × 100 (%) The weight of each cotton yarn is measured after drying under reduced pressure for 7 hours. Tensile strength decrease rate Tensile strength of cotton yarn after treatment with immobilized enzyme / Tensile strength of cotton yarn before treatment with immobilized enzyme × 100 (%) In addition, the tensile strength is measured by a tensile tester (NRM-2010J-
CW, manufactured by Fudo Kogyo Co., Ltd., was used at a temperature of 20 ° C., a humidity of 65%, a gripping interval of 10 cm, and a pulling speed of 5 cm / min. (3) Recovery of immobilized enzyme Add 1N hydrochloric acid to the reaction solution after taking out the fiber to adjust the pH.
Was reduced to 3 and the PLG-immobilized cellulase was precipitated and collected. Next, water was added to the recovered PLG-immobilized cellulase to adjust the pH to 5 with a 1N aqueous sodium hydroxide solution, and the solution was dissolved.
When the amount of glucose produced was measured at H5 and temperature of 37 ° C., 98% of the activity was retained as compared with PLG-immobilized cellulase which was not used for fiber treatment.

Example 2 (1) Synthesis of Immobilized Enzyme γ-Benzyl-L-glutamic acid-N-carboxylic acid anhydride was used as a polymerization initiator in a dichloromethane at room temperature 3 using polybutadiene having a molecular weight of 5,000, which was amine-modified at one end. Polymerized day. The obtained poly-γ-benzyl-L-glutamic acid-polybutadiene copolymer was hydrolyzed with bromic acid / acetic acid to obtain an amino acid copolymer. Then, in the same manner as in Example 1, the amino acid copolymer obtained above and cellulase were immobilized. The obtained immobilized enzyme is designated as immobilized enzyme A. (2) Fiber treatment Example 1 except that 20 units of immobilized enzyme A was used in place of PLG-immobilized cellulase in Example 1 (2).
The cotton yarn bundle was treated in the same manner as (2). Then, the weight reduction rate and the tensile strength reduction rate of the treated cotton yarn were determined as in Example 1.
It measured similarly to. The results are shown in Table 1. (3) Recovery of immobilized enzyme The immobilized enzyme was recovered in the same manner as in Example 1 (3), and the decomposition activity of the recovered immobilized enzyme was measured in the same manner as in Example 1 (3). The results are shown in Table 1.

Example 3 (1) Synthesis of immobilized enzyme ε-Benzyl-L-lysine-N-carboxylic acid anhydride was polymerized in dichloromethane with triethylamine as a polymerization initiator at room temperature for 3 days. The resulting poly-ε-benzyl-L-lysine (PBLL) was hydrolyzed with bromic acid / acetic acid to give poly-L-
Lysine (PLL) was obtained. The obtained PLL and cellulase were immobilized in the same manner as in Example 1 (1) to synthesize PLL-immobilized cellulase. (2) Fiber treatment A cotton yarn bundle was treated in the same manner as in Example 1 (2) except that 20 units of PLL-immobilized cellulase was used in place of PLG-immobilized cellulase in Example 1 (2). Then, the weight reduction rate and the tensile strength reduction rate of the treated cotton yarn were measured in the same manner as in Example 1. The results are shown in Table 1. (3) Recovery of Immobilized Enzyme To the reaction solution after taking out the fiber, 1N sodium hydroxide aqueous solution was added to adjust the pH to 10, and PLL immobilized cellulase was precipitated and recovered. The degradation activity of the recovered immobilized enzyme was measured in the same manner as in Example 1 (3). The results are shown in Table 1.
Shown in

Comparative Example 1 Cotton yarn was treated in the same manner as in Example 1 (2) except that 20 units of cellulase not immobilized on a carrier was used in place of the immobilized enzyme in Example 1 (2). The weight reduction rate and the tensile strength reduction rate of the cotton yarn after the treatment are shown in Example 1.
It measured similarly to. The results are shown in Table 1.

[0015]

[Table 1]

[0016]

EFFECTS OF THE INVENTION According to the present invention, the weight reduction of the fiber after contact with the enzyme is moderate, and the tensile strength is not significantly reduced even after the treatment. Further, according to the present invention, the weight of the fiber can be reduced without significantly lowering the fiber strength, and the texture and feel of the fiber can be significantly improved. Furthermore, according to the present invention, the immobilized enzyme can be easily recovered by a simple operation of changing the pH of the reaction solution, so that the immobilized enzyme can be reused, and an expensive enzyme can be efficiently used, and a uniform enzyme can be obtained. Since the reaction can be performed in the system, the reaction efficiency can be improved as compared with the conventional immobilized enzyme.

Claims (1)

[Claims] A method for treating a fiber, which comprises bringing an immobilized enzyme in which an amino acid (co) polymer and an enzyme are bound into contact with the fiber.