JP2006207086A - Fiber-treating agent using thermophilic endoglucanase and method for treating fiber - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a fiber-treating agent with which cellulosic fibers can efficiently be bio-washed, and to provide a method for treating the fibers. <P>SOLUTION: This fiber-treating agent contains a thermophilic endoglucanase as an active ingredient. The method for treating the fibers comprises making both a thermophilic endoglucanase and a thermophilic amylase to act on cellulosic fibers to simultaneously subject the cellulosic fibers to both a desizing treatment and a bio-washing treatment. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a fiber treatment agent and a fiber treatment method capable of efficiently performing biowash (enzyme washing) of cellulosic fibers.

  Conventionally, fibers and sewing products have been processed using an enzyme at the time of finishing, and cellulosic fibers (for example, jeans) have been biowashed (enzyme washed) using cellulase (see Patent Document 1). ). This biowash is generally performed using acid cellulase and neutral cellulase, but since the optimum temperature range of the enzyme used is a so-called medium temperature type enzyme agent of 40 ° C to 60 ° C, the processing temperature is also high. It is carried out at a medium temperature of 40 ° C to 60 ° C. However, the treatment efficiency of a conventional biowash using cellulase is not always satisfactory.

Some types of cellulosic fibers have many warp pastes attached to them. Since the presence of such a paste reduces the action of cellulase in biowash processing, it is necessary to perform a desizing process on cellulosic fibers to which the paste is attached. In this desizing process, an enzyme process using a thermophilic amylase acting at 70 to 100 ° C. is usually performed. As described above, amylase used in the desizing process is significantly different from cellulase used in conventional biowash in terms of enzyme reaction conditions such as operating temperature. It is impossible to simultaneously perform the punching process and the biowash process in the same system. Therefore, conventionally, in the treatment of the cellulosic fiber to which the paste is adhered, a two-step process of performing a biowash process after the paste removal process is performed, which is extremely inefficient.
Registered Utility Model No. 3043579

  The object of the present invention is to solve the above-mentioned problems of the prior art. More specifically, an object of the present invention is to provide a fiber treatment agent and a fiber treatment method capable of efficiently performing biowash on cellulosic fibers. Furthermore, an object of this invention is to provide the fiber processing method which can process a cellulosic fiber more efficiently by implementing a desizing process and biowash simultaneously.

  The present inventors have intensively studied to solve the above-mentioned problems. As a result, thermophilic endoglucanase, particularly thermophilic endoglucanase derived from Pyrococcus horikoshii, has been used in conventional biowash. It has been found that biowashing can be performed on cellulosic fibers more efficiently than the above. Further, it was confirmed that the thermophilic endoglucanase can act under the same conditions as the amylase used in the desizing process. Furthermore, by allowing the thermophilic endoglucanase and the thermophilic amylase to act on the cellulosic fibers simultaneously in the same system, it becomes possible to carry out desizing treatment and biowash at the same time, resulting in extremely efficient cellulose. It has been found that the processing of the fiber can be realized. The present invention has been completed by making further improvements based on such findings.

That is, this invention provides the fiber processing agent and fiber processing method of the aspect hung up below.
Item 1. A fiber treatment agent comprising a thermophilic endoglucanase.
Item 2. A method for treating cellulosic fibers, which comprises causing thermophilic endoglucanase to act on cellulosic fibers.
Item 3. Item 3. The treatment method according to Item 2, wherein the thermophilic endoglucanase and the thermophilic amylase are allowed to act on the cellulosic fiber simultaneously.

  Hereinafter, the present invention will be described in detail.

  The fiber treatment agent of the present invention is a treatment agent used for biowash (enzyme washing) which is a finishing process of cellulosic fibers.

  Cellulosic fibers to be treated in the present invention include fabrics used in jeans such as denim, cellulose fibers such as cotton and hemp; regenerated cellulose fibers such as viscose rayon, polynosic rayon and cupra; Tencel (trade name) And refined cellulose fibers such as Modal (trade name); semi-synthetic cellulose fibers such as acetate rayon; mixed-spun fibers containing these fibers; or semi-sewn products and sewn products thereof. The form of the fiber may be any form such as yarn, cotton, woven fabric, knitted fabric, and non-woven fabric.

The fiber treatment agent of the present invention contains a thermophilic endoglucanase as an active ingredient. The thermophilic endoglucanase has an action of catalyzing the hydrolytic cleavage of β-1,4-glucoside bond between D-glucose of cellulose, and is stable at a high temperature around 90 to 100 ° C. It is an enzyme whose temperature range is optimal. The thermophilic endoglucanase is preferably exemplified by those having the following characteristics;
(a) The estimated molecular weight by SDS-PAGE is about 43 kDa.
(b) The optimum pH is 5.4 to 6.0. The optimal pH of enzyme activity was measured by adjusting 0.5% substrate (carboxymethylcellulose) solution up to pH 4-9 with 100 mM sodium acetate buffer, 100 mM phosphate buffer and 100 mM borate buffer, and It is determined by measuring the initial rate of hydrolysis activity. The activity is detected by quantifying the reducing end generated after hydrolysis by the Somogy Nelson method.
(c) The optimum temperature is 95 ° C or higher and 100 ° C or lower. Using 0.5% carboxymethylcellulose as a substrate, adding a certain amount of enzyme in 100 mM sodium acetate buffer (pH 5.6), and reacting at each temperature for 15 minutes, the activity is the same as in the above (b), somogy It is detected by quantification by the Nelson method.
(d) Regarding temperature stability, it is stable at about 97 ° C. for 3 hours. Specifically, the enzyme solution (0.1 mg / mL) was heat-treated in 100 mM sodium acetate buffer (pH 5.6) at 97 ° C. for 3 hours, and then 0.5% carboxymethylcellulose was used as a substrate, and 100 mM sodium acetate. The residual enzyme activity measured at 85 ° C. in a buffer solution (pH 5.6) is 80% or more.

  The thermophilic endoglucanase has a natural amino acid sequence described in SEQ ID NO: 1 in the sequence listing, for example. Alternatively, the amino acid sequence of SEQ ID NO: 1 in the sequence listing includes an amino acid sequence in which 1 to 100 amino acid residues are deleted, substituted, or added, and is substantially different from a natural thermophilic endoglucanase. Polypeptides having equivalent activity are also included in the thermophilic endoglucanase. Further, in the amino acid sequence shown in SEQ ID NO: 1 in the sequence listing, 28 residues from the N-terminal side are considered to be a signal sequence, and (i) has the amino acid sequence of positions 29 to 458 of SEQ ID NO: 1. Or (ii) a natural thermophilic endoglucanase comprising an amino acid sequence in which 1 to 100 amino acid residues are deleted, substituted, or added in the amino acid sequence of positions 29 to 458 of SEQ ID NO: 1 A polypeptide having a polypeptide having substantially the same activity as the above is also included in the thermophilic endoglucanase.

  The thermophilic endoglucanase is a prokaryote, preferably an archaebacterium Pyrococcus genus, more preferably a hyperthermophilic bacterium, a thermophilic bacterium Pyrococcus holicosi with an optimum growth temperature of 98 ° C. Pyrococcus horikoshii, registration number JCM9974, JCM microorganism strain catalog 7th edition, published in January 1999). Further, the thermophilic endoglucanase can be obtained by obtaining a gene encoding the thermophilic endoglucanase from these microorganisms, preparing a recombinant microorganism into which the gene has been introduced, and culturing the recombinant microorganism.

  The fiber treatment agent of the present invention may contain other cellulases, amylases, proteases, etc. in addition to the above thermophilic endoglucanases, as long as they do not interfere with the action of the endoglucanases.

  The fiber treatment agent of the present invention is used as an enzyme agent in biowash (enzyme washing), which is a finishing process for cellulosic fibers. That is, biowash (enzyme washing), which is a finishing process of cellulosic fibers, can be performed by allowing the thermophilic endoglucanase to act on cellulosic fibers using the fiber treatment agent.

  As a method for causing the thermophilic endoglucanase to act on the cellulosic fiber, for example, the cellulosic fiber can be immersed in a treatment solution to which the thermophilic endoglucanase is added. As specific conditions, a thermophilic endoglucanase of about 10 to 5000 units, preferably 100 to 500 units, more preferably about 250 to 500 units is added to a 1 L aqueous solution adjusted to pH 4.5 to 7.5, preferably 5.5 to 6.5. A treatment liquid is prepared by immersing about 30-50 g of cellulose fiber in this treatment liquid, and it is about 70-100 ° C., preferably about 90-100 ° C., for 10-60 minutes, preferably 30-60 minutes. The degree processing is exemplified.

  When processing cellulosic fibers to which glue such as starch or polyvinyl alcohol is adhered (for example, when 3 wt% or more of glue is adhered to the total weight of cellulosic fibers), usually the fibers The paste removal process is performed on In this case, prior to the treatment with the thermophilic endoglucanase, a desizing treatment may be performed by a known method, but together with the thermophilic endoglucanase, a thermophilic amylase for desizing is simultaneously applied to the cellulosic fiber. By making it act, it is desirable to carry out the desizing process and biowash simultaneously in the same process. Thermophilic amylase, which is usually used for desizing of cellulosic fibers, can act under the same conditions as the above thermophilic endoglucanase, so that both enzymes can act simultaneously to reduce desizing and biowash. Since it can be carried out simultaneously in the same process, more efficient fiber processing becomes possible.

  When the desizing process and biowash are performed simultaneously in the same process, the thermophilic amylase for desizing is not particularly limited as long as it is normally used for desizing. Specifically, those having amylase activity at 70 to 100 ° C., preferably 80 to 100 ° C. and pH 5.0 to 8.0, preferably 6.5 to 7.5 are mentioned. Examples of such thermophilic amylase include those derived from microorganisms such as Bacillus subtilis and Bacillus licheniformis. Such thermophilic amylases are generally commercially available, and commercially available products may be used. Specific examples of such commercially available products include kristase (manufactured by Daiwa Kasei Co., Ltd.), terma meal (manufactured by Novozyme), SVA-300 (manufactured by Toto Kasei Co., Ltd.), and the like.

  Conditions for simultaneously performing the desizing treatment and biowashing in the same process vary depending on the type and amount of the paste adhering to the cellulosic fiber. For example, in the treatment liquid 1L to which a thermophilic endoglucanase is added. Further, the above thermophilic amylase may be added in an amount of about 300 to 150,000 units, preferably about 3000 to 15000 units, more preferably about 6000 to 15000 units, and treated under the same conditions as described above.

  In this specification, endoglucanase and cellulase activity of 1 unit was carried out by adding 1 ml of enzyme solution to 9 ml of substrate 1.11 wt% CMC (carboxymethylcellulose Na) solution (pH 5.0) and reacting at the optimum temperature. At this time, the amount of enzyme required to produce a reducing sugar amount equivalent to 1 micromole of glucose per minute is defined as 1 unit.

  In addition, 1 unit of amylase activity follows the modified method of Mr. Walgemuth. That is, in the present invention, the activity unit of amylase is 1.786 times the activity value according to the bacterial amylase measurement method (JIS K 7001 JIS liquefaction power method, a method for measuring the decrease in starch viscosity).

  The fiber treatment agent of the present invention uses a thermophilic endoglucanase, and performs biowashing on cellulosic fibers more efficiently than a conventional fiber treatment agent using cellulase that acts in the middle temperature range. be able to. Further, since the fiber treatment agent of the present invention can be used without causing dye contamination, crying, fiber damage, etc., its practical value is high.

  Furthermore, the thermophilic endoglucanase used in the fiber treatment agent of the present invention can act simultaneously under the same conditions as the thermophilic amylase for desizing. Therefore, by making the thermophilic amylase and the thermophilic endoglucanase act on the cellulosic fiber to which the paste is attached, the desizing treatment and the biowash can be simultaneously performed, and the cellulosic fiber is more efficiently performed. Can be realized.

EXAMPLES Hereinafter, although an Example is given and this invention is demonstrated, this invention is not limited to these Examples.
Example 1
Thermophilic endoglucanase (specific activity 50 units / g) consisting of the amino acid sequence shown in SEQ ID NO: 1 in the sequence listing derived from Pyrococcus horikoshi (registration number JCM9974) or cellulase derived from Trichoderma (optimum temperature 50 ° C) (specific activity) 50 unit / g) was added to 1N acetate buffer (pH 6) to a concentration of 2 g / L to prepare a treatment solution. In this treatment liquid (70) L, 6 jeans (approximately 2300 g) were immersed and treated for 60 minutes to perform biowash. The treatment temperature was 95 ° C. for thermophilic endoglucanase and 50 ° C. for cellulase derived from Trichoderma. After the treatment, the jeans were taken out from the treatment liquid, the weight of the treated jeans was measured, and the ratio of the weight (weight loss rate:%) reduced compared with that before the treatment was determined. For comparison, the same test as described above was performed at a treatment temperature of 95 ° C. using a 1N acetate buffer solution (pH 6) to which no enzyme was added as a treatment solution.

  The obtained results are shown in Table 1. From this result, the thermophilic endoglucanase derived from Pyrococcus horikoshi has a high weight loss rate compared to the cellulase acting in the middle temperature range conventionally used for biowash, and can perform an efficient biowash. It was confirmed. In addition, the jeans after treatment with the thermophilic endoglucanase derived from Pyrococcus horikoshi showed no defects such as fiber damage.

Example 2
Example 1 above, except that a thermophilic endoglucanase consisting of the amino acid sequence shown in SEQ ID NO: 1 in the sequence listing derived from Pyrococcus horikoshi (registration number JCM9974) is used and the treatment time is 40, 30 or 20 minutes. The biowash process was performed in the same manner as described above, and the weight loss rate of the treated jeans was determined.

  The obtained results are shown in Table 2. As a result, it was confirmed that by using a thermophilic endoglucanase derived from Pyrococcus horikoshi, the desired biowash treatment can be performed even if the treatment time is shortened to about 20 minutes.

Example 3
Thermophilic endoglucanase (specific activity 50unit / g) consisting of the amino acid sequence shown in SEQ ID NO: 1 in Pyrococcus horikoshi (registration number JCM9974) is 2g / L in 1N acetate buffer (pH 6). In addition, 2g of thermophilic amylase (trade name "SVA-300", manufactured by Nitto Kasei Kogyo Co., Ltd.) (specific activity 3000unit / g) that is commonly used as an amylase for desizing / L was added to obtain a treatment solution. Six jeans (about 2300g) with warp paste adhering at a rate of 5.8% by weight to this treatment solution 70L were soaked and treated for 60 minutes at 95 ° C, and desizing and biowash were performed simultaneously. (In Table 3, described as “Example 3 (one step)”).

  For comparison, according to the method shown below, the jeans were processed by performing biowashing after the desizing process. That is, a thermophilic amylase (trade name “SVA-300”, manufactured by Nitto Kasei Kogyo Co., Ltd.) (specific activity 3000 unit / g) was added to 70 L of 1N acetate buffer (pH 6) added to 2 g / L. The above 6 jeans (about 2300 g) were immersed and desizing was performed at 90 ° C. for 30 minutes. Next, after taking out the jeans and washing with water, they were immersed in 70 L of 1N acetate buffer (pH 6) to which cellulase derived from Trichoderma (optimum temperature 50 ° C., specific activity 50 unit / g) was added to 2 g / L. Then, it was treated at 50 ° C. for 60 minutes to perform biowash (in Table 3, “conventional method (two steps)”).

  After the above treatment, the jeans were taken out from the treatment liquid, the weight of the treated jeans was measured, and the proportion of weight (weight loss rate:%) reduced compared with that before the treatment was determined. Further, the processed jeans were subjected to a complete desizing process, and the weight loss from before the processing was measured, whereby the amount of the glue remaining in the processed jeans was measured. Here, the complete desizing process means a process of completely removing the paste by treating with an excessive amount of medicine for a long time.

  The obtained results are shown in Table 3. As a result, by allowing thermophilic amylase and thermophilic endoglucanase to act simultaneously in the same system, desizing treatment and biowash can be performed at the same time, and biowashing is performed after desizing treatment. In comparison, it was confirmed that the fiber treatment can be performed efficiently and easily.

Example 4
The following tests were conducted to examine the influence of thermophilic endoglucanase on the activity of thermophilic amylase when thermophilic amylase and thermophilic endoglucanase were allowed to act simultaneously.

  3g of cotton broad # 40 raw fabric (warp paste: 4.1% by weight starch and 5.7% by weight polyvinyl alcohol (PVA)) is immersed in 100mL of the treatment solution shown in Table 4 and treated at 95 ° C for 10 or 30 minutes. After the treatment, the raw material was taken out, washed with water and air-dried. Measure iodine color value (index value of residual starch) by dropping N / 50 iodine solution onto cloth and observing blue coloration by iodo-starch reaction. In addition, by dropping a N / 50 borate iodine solution onto the cloth and observing the dark blue-green coloration due to the borate-iodine-PVA reaction, the iodine borate coloration value (index value of remaining PVA) ) Was measured.

  The results obtained are shown in Table 5. From this result, in the treatment time of 10 minutes, the iodine color values were all 0, the thermophilic endoglucanase did not adversely affect the desizing treatment with the thermophilic amylase, and the starch paste was completely removed. It was. In addition, the removal effect of the PVA paste was found to be better when more thermophilic endoglucanase was added.

Claims (3)

A fiber treatment agent comprising a thermophilic endoglucanase. A method for treating cellulosic fibers, which comprises causing thermophilic endoglucanase to act on cellulosic fibers. The processing method according to claim 2, wherein the thermophilic endoglucanase and the thermophilic amylase are allowed to act on the cellulosic fiber simultaneously.