CN1155306A - Method for desizing cellulose-containing fabric - Google Patents

Abstract

Cellulose-containing fabrics or textile may be desized without essentially damaging the fabric or textile by subjecting the fabric or textile to a treatment with a certain type of cellulolytic enzyme, e.g. a cellulolytic enzyme having an activity on carboxymethylcellulose (CMC) and a catalytic activity on cellotriose at pH 8.5 corresponding to kcat of at least 0.01 s<-1> and preferably being of microbial origin, more preferably being obtainable by or derived from a strain of Humicola, Trichoderma, Myceliophthora, Penicillium, Irpex, Aspergillus, Scytalidium or Fusarium.

Description

Process for desizing cellulose-containing fabrics

The present invention relates to a process for desizing cellulose-containing fabrics or cloths, and to an enzyme composition suitable for use in the process.

Background

During the weaving of the fabric, the threads may besubjected to great mechanical tensions. To prevent breakage, the wire is usually reinforced by applying (pasting) a gel-like substance (paste). The most common coating agents are starches in native or modified form, which may also be enriched with other polymers such as polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), polyacrylic acid (PAA) or derivatives of cellulose (e.g. carboxymethylcellulose (CMC), hydroxyethylcellulose, hydroxypropylcellulose or methylcellulose). Small amounts of, for example, fats and oils may also be added to the slurry for the purpose of lubricating the surfaces.

As a result of the sizing, the threads cannot absorb a sufficient amount of water or finishing agent or composition (bleaching, dyeing, anti-wrinkling, etc.). Only after removal of the size from the cloth (so-called desizing) is a uniform and durable finish obtained.

In the case of slurries containing significant amounts of carboxymethylcellulose (CMC) or other cellulose derivatives, the desizing process may be carried out using a cellulolytic enzyme, alone or in combination with other substances, optionally in combination with other enzymes, for example starch degrading enzymes such as amylases. However, in the case of a size-coated cloth or fabric containing cellulose or cellulose derivatives, such treatment with cellulolytic enzymes can decompose not only the size but also the fabric or cloth in whole or in part, thus producing a desized cloth or fabric with a certain loss of strength and/or weight compared to the strength or weight of the fabric or cloth not coated with size.

It is an object of the present invention to provide a method of desizing cellulose-containing fabrics or cloths without substantially damaging the fabric, i.e. without causing loss of strength or loss of weight or both of the fabric. Summary of the invention

Surprisingly, it has now been found that by treating a fabric or cloth with a certain type of cellulolytic enzyme, it is possible to desize a fabric or cloth containing cellulose without substantially damaging the fabric or cloth.

Accordingly, the present invention provides a process for desizing cellulose-containing cloth or fabric by treating the cloth or fabric with a cellulolytic enzyme having a certain activity towards carboxymethylcellulose (CMC) and a certain catalytic activity towards cellotriose at ph8.5, corresponding to at least 0.01Kcat per second.

In another aspect, there is provided a composition suitable for use in the method of the invention, the composition comprising a cellulolytic enzyme having an activity towards carboxymethylcellulose (CMC) and a catalytic activity towards cellotriose at pH8.5, corresponding to at least 0.01Kcat per second. Of course, the composition does not contain any cellulase components that can cause fabric damage.

By using the process of the invention, it is possible to obtain desized cellulose-containing cloths or fabrics which have substantially no loss of strength or weight compared with unpulped cloths or fabrics. Detailed Description

The term "desizing" is understood to mean the removal of size from a fabric in the conventional manner.

In the present specification and claims, the terms "cellulolytic enzyme", "cellulase" and "cellulase component" refer to an enzyme capable of hydrolyzing cellulose, cellulolytic enzyme, cellulase or cellulase components may be used as a component in a cellulase system produced by a given microorganism, such cellulase system containing primarily a number of different cellulase components, including those commonly identified, e.g., cellulolytic enzyme, exo-cellulolytic enzyme, endo- β -1, 4-glucanase, β -glucosidase.

The native or unmodified cellulase or cellulase components may be derived from known cellulolytic enzyme producing microorganisms, for example, strains of the following microorganisms: humicola, Thermomves, Bacillus, Trichoderma, crescent spore genus, Mvceliophthora, Phanerochaete, Rapex, Scvtalidium, Schizophyllum, Penicillium, Aspergillus and Geotrichum. The derived components may be homologous or non-homologous. The components are preferably homologous. But non-homologous components which are immunoreactive with antibodies directed against highly purified cellulase components and which are derivable from a particular microorganism are also preferred.

Useful cellulases suitable for use in the process of the invention are preferably derived from a microorganism. In a preferred embodiment of the invention, the cellulolytic enzyme is obtained or derived from a strain of Humicola, Trichoderma, Mvceliophthora, Penicillium, Irpex, Aspergillus, Scvtalidium or Scutellaria, more preferably from a strain of Humicola insolens, Fusarium oxysporum or Trichoderma reesei.

In the present specification, the term "derivable" or "derived from" refers not only to cellulases produced by strains of the organism in question, but also to cellulases encoded by DNA sequences isolated from these strains and produced in a host organism transformed with said DNA sequences. Furthermore, the term also refers to cellulases encoded by DNA sequences of synthetic and/or cDNA origin and having the same properties as the cellulase in question.

The cellulolytic enzyme used in the method of the invention is preferably a recombinant cellulase, i.e. a cellulase which is substantially free of other proteins or cellulase proteins. For the skilled person, a recombinant cellulase component may be cloned and expressed according to conventional standard techniques.

It is advantageously possible touse parent cellulases of fungal origin, for example cellulases derived from the fungi Humicola or from strains of the genus Sinomonas. For example, maternal cellulases can be derived from the fungus h.insolens or fusarium oxysporum, many of which have been characterized and have been described for their full amino acid sequence.

In a preferred embodiment the parent cellulase is selected from the group consisting of h.insolens, fusarium oxysporum and Trichoderma reesei cellulases, or is a functional analogue of any one of said parent cellulases i) having an amino acid sequence which is at least 60% homologous to the amino acid sequence of the parent cellulase, ii) reacting with an antibody directed against the parent cellulase, and/or iii) being encoded by a DNA sequence which is hybridizable to a probe identical to the DNA sequence encoding the parent cellulase.

The nature i) of the analogue refers to the degree of identity between the analogue and the parent cellulase, indicating the derivation of the first sequence from the second. In particular, a polypeptide is considered homologous to a parent cellulase if a comparison of the corresponding amino acid sequences shows an identity of greater than about 60%, such as greater than 70%, 80%, 85%, 90% or even 95%. Sequence comparison can be performed by known algorithms, such as one described by Lipman and Pearson (1985).

The homologous cellulase may be a genetically engineered cellulase, e.g. a cellulase prepared for improving one or several properties, such as thermostability, acid/base stability, temperature or pH optimum, etc.

Other properties of the parent cellulase analogues ii) and iii) can be determined as follows: property ii), i.e. immunological cross-reactivity, can be determined using an antibody directed against or reactive with at least one epitope of the maternal cellulase. Antibodies, which maybe monoclonal or polyclonal, may be prepared by methods well known in the art, such as those described by Hudson et al, 1989. Immunological cross-reactivity can be determined using assays well known in the art, examples of which are Western blotting or radial immunodiffusion assays, e.g., as described by Hudson et al, 1989.

In accordance with property iii) defined above, the oligonucleotide probes used in the characterization of the analogs may suitably be prepared on the basis of the nucleotide or amino acid sequence of all or part of the parent cellulase. Hybridization can be carried out under any suitable conditions that allow for hybridization of DNA sequences, for example, under conditions such as those involving presubscription in 5 XSSC and prehybridization in a solution of 20% formamide, 5XDenhardt's solution, 50mM sodium phosphate, pH6.8 and 50. mu.g of denatured sonicated calf thymus DNA at-40 ℃ for 1 hour followed by hybridization in the same solution supplemented with 100. mu.M ATP at-40 ℃ for 18 hours, or other methods as described by Sambrook et al, 1989.

Examples of cellulolytic enzymes suitable for use according to the invention are:

an endoglucanase component which is immunoreactive with an antibody directed against a highly purified-50 KD endoglucanase derived from Humicola insolens.dsm1800, or a homologue or derivative of the-50 KD endoglucanase which exhibits cellulase activity. A preferred endoglucanase component has an amino acid sequence disclosed in PCT patent application WO 91/17244, FIGS. 14A-E, shown in the accompanying SEQ ID NO: 2, or a variant of said endoglucanase having an amino acid sequence which is at least 60%, preferably at least 70%, better still at least 75%, more preferably at least 80%, better still at least 85%, especially at least 90% homologous to said sequence. Hereinafter the endoglucanase component is referred to as EGI; and

an endoglucanase component which is immunoreactive with an antibody directed against a highly purified-50 KD (apparent molecular weight, amino acid composition corresponding to 45KD with 2n glycosylation sites) endoglucanase derived from fusarium oxysporum, DSM2672, or a homologue or derivative of the-50 KD endoglucanase which exhibits cellulase activity. A preferred endoglucanase component has an amino acid sequence as disclosed in PCT patent application No. WO 91/17244, FIG. 13. This sequence is set forth in the appended SEQ ID NO: 3, or a variant of said endoglucanase having an amino acid sequence which is at least 60%, preferably at least 70%, more preferably at least 75%, more preferably at least 80%, more preferably at least 85%, especially at least 90% homologous to said sequence. Hereinafter this endoglucanase component is referred to as EG I-F. The EG I-F cellulase component may comprise a sequence corresponding to SEQ ID NO: 3 and prepared using a conventional Taka promoter and AMG terminator. EG I-F can be purified by cation chromatography to homogeneity and have a pI>9. pI was 9 calculated from the composition of amino acids using pHKa value in "New Adv. protein Chem." 17, pages 69-165 (1962) (C.Tanford), molar extinction coefficient was 58180 calculated, and

any cellulase disclosed in published European patent application No. EP-A2-271,004, i.e., an alkalophilic cellulase having a non-degradation index (NDI) of not less than 500 and having an optimum pH of not less than 7 or a cellulase having a relative activity of 50% or more of that under optimum conditions at a pH of not less than 8 when carboxymethylcellulose (CMC) is used as a substrate; the cellulase is preferably selected from the group consisting of alkaline cellulase K (produced by Bacillus sp. KSM-635, FERM BP 1485); alkaline cellulase K-534 (produced by Bacillus sp.KSM-534, FERMBP 1508); alkaline cellulase K-539 (produced by Bacillus sp. KSM-539, FERM BP 1509); alkaline cellulase K-577 (produced by Bacillus sp.KSM-577, FERM BP 1510); alkaline cellulase K-521 (produced by Bacillus sp. FERM BP 1507); alkaline cellulase K-580 (produced by Bacillus sp. KSM-580, FERM BP 1511); alkaline cellulase K-588 (produced by Bacillus sp. KSM-588, FERM BP 1513); alkaline cellulase K-597 (produced by Bacillus sp. KSM-597, FERM BP 1514); alkaline cellulase K-522 (produced by Bacillus sp. KSM-522, FERM BP 1512); carboxymethyl cellulase I, carboxymethyl cellulase II (both produced by bacillus sp. ksm-635, FERM BP 1485); alkaline cellulase E-II and alkaline cellulase E-III (both produced by Bacillus sp. KSM-522, FERM BP 1512).

The terms "cellulose-containing fabric/cloth" and "cellulosic fabric/cloth" refer to any type of fabric, especially textile, prepared from cellulose-containing raw materials and containing cellulose or cellulose derivatives, such as textiles made from wood pulp, and cotton. The main component of the cellulose or cellulose derivatives present on the fabric is usually a size, wherein the yarn, usually a wound yarn, has been coated before weaving. In the present invention, the term "fabric" also includes garments and other types of processed fabrics. Examples of cellulosic fabrics are cotton, viscose (cellulose rayon); a lyocell; all blended fabrics of viscose, cotton or lyocell with other fabrics such as polyester; viscose, cotton or mixtures of lyocell with other fibres (such as polyester), lyocell/cotton blends, viscose/wool blends, lyocell/wool blends, cotton/wool blends; linen, ramie and other fabrics based on cellulose fabrics, all blends of fabrics comprising cellulose with other fabrics such as wool, polyamide, acrylic and polyester fabrics, for example viscose/cotton/polyester blends, wool/cotton/polyester blends, linen/cotton blends and the like. Process conditions

It goes without saying that the process of the invention can be carried out according to any suitable desizing method well known in the art, such as, for example, the methods described by Olson, E.S, (1983), and Peter, m., and Rouette H, K (1988). Thus, the process conditions employed in the practice of the present invention may be selected to suit a particular apparatus or a particular form of process required for use. Preferred examples of the types of processes suitable for use in connection with the present invention include the types Jigger/Winch, Pad-Roll and Red-Stem. These types are discussed in detail below.

The process of the invention may be carried out as a batch process, a semi-continuous process or a continuous process, which may comprise the following steps, as examples: (a) immersing the fabric in a desizing bath containing (minimally) a cellulase having an activity towards carboxymethylcellulose (CMC) and a catalytic activity towards cellotriose at ph8.5 corresponding to Kcat of at least 0.01 per second, then squeezing out excess liquid to maintain the amount of liquid necessary to carry out the reaction (typically 60% to 120% by weight of the dry fabric), (b) steaming the impregnated fabric to bring the fabric to the desired reaction temperature, typically between 20 ° and 120 °, and (c) holding the garment in a J-Box, U-Box, carpet machine, or the like by winding or pleating for a sufficient period of time (typically between minutes and hours) to effect desizing.

The cellulase enzymes suitable for use may conveniently be mixed with other components conventionally used in desizing processes prior to treatment. Examples of these components are othersPreferred are those commercially available amylases which are usually used for desizing, such as microbial amylases, in particular amylases producible with Bacillus (e.g.Bacillus licheniformis, Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus stearothermophilus) or Aspergillus; or by mutants of these genera, for example amylases produced as described in WO 91/00353 or WO 91/16423.Examples of commercial amylases for desizing are Aquazym, Aquazymeultra, Dezyme, Thermozyme from Novo Nordisk A/S^TMOther preferred amylases are oxidation-stable α -amylase mutants disclosed in international patent application PCT/DK 94/00371, which is incorporated herein by reference.

Thus, it may be advantageous to add an α -amylase enzyme with improved acid stability to the composition of the invention, so as to allow the composition to be used in a combined desizing and bleaching process (carried out in a single step), for example, a process using sodium chlorite in combination with a strong base, a surfactant, an activator, an enzyme that promotes amylolysis and optionally a cellulolytic enzyme, or a process using sodium tetraborate hexahydrate as a buffer in a bath containing hydrogen peroxide, a sequestering agent, a surfactant, an enzyme that promotes amylolysis and optionally a cellulolytic enzyme.

Other components required to carry out the process may be added separately. Examples of such components include stabilizers and wetting agents. The stabilizing agent may be an agent that stabilizes the cellulolytic enzyme.

Wetting agents serve to improve the wettability of the fabric so as to obtain a fast and smooth desizing process. The wetting agent is preferably oxidation-stable.

In a preferred embodiment of the invention, the cellulase enzyme is used in an amount of more than 1 g/l, preferably in an amount of 1-20 g/l, such as 1-10 g/l, 1-5 g/l or 1-3 g/l, corresponding to a cellulase activity in the range of 10 to 5000ECU per liter of desized liquor, preferably 50 to 500ECU per liter of desized liquor. The amount of cellulose used will of course depend on the composition of the cellulase product in question.

Regardless of the particular type of process used for the desizing of the present invention, theprocess of the present invention is generally carried out at a temperature in the range of 30 to 100 ℃, such as 35 to 60 ℃, and at a pH in the range of 3 to 11, preferably 7 to 9. However, the actual process conditions may vary widely within these ranges, as will be apparent from the following disclosure.

Preferred examples of process conditions for use in the present invention include: batch processes, for example of the type Jigger/Winch, using 1 to 5 g/l of a suitable cellulase and 0.25 to 5 g/l of a wetting agent, for example Arbyl R, a product commercially available from Grunau, Germany, are carried out at a pH in the range from 7 to 9 (which can be achieved by addition of NaOH) and a temperature in the range from 40 to 55 ℃ for generally 1 to 2 hours.

A semi-continuous process, for example of the Pad-Roll type, using 1 to 5 g/l of a suitable cellulase and 0.25 to 5 g/l of a wetting agent, for example Arbyl R, is carried out at a pH in the range from 7 to 9 (which can be achieved by addition of NaOH) and at a temperature in the range from 30 to 50 ℃ for generally 12 to 24 hours.

Of course, the process can be carried out in any apparatus which is sufficiently tolerant of the conditions of the process.

Furthermore, it is clear that such cellulases should be effective at a pH greater than 3 (e.g., greater than about pH7), except that Kcat limits the cellulases used in the methods of the invention. The cellulase is preferably more active in the pH range 7-9.

The process of the invention may be used before or after another desizing treatment step which may be supplemented with cellulase treatment, preferably an enzymatic desizing treatment with amylase. Compositions of the invention

Although suitable cellulases may be added directly, they are preferably combined to form a suitable desizing composition, which preferably further comprises other desizing enzymes, such as the amylases described above.

The desizing composition of the invention can be in the form of a granulate, preferably a dust-free granulate, a liquid, in particular a stabilized liquid, a slurry or a mixture of the individual enzymes in a protected form. Dust-free granules can be prepared as disclosed in U.S. Pat. No. 4,106,991 and U.S. Pat. No. 4,661,452 (both Novo Nordisk A/S) and can optionally be coated by methods known in the art.

For example, liquid enzyme preparations may be stabilized according to established methods by adding a polyol (e.g. propylene glycol), a sugar or sugar alcohol or acetic acid. Other enzyme stabilizers are well known in the art and protected enzymes may be prepared according to the method disclosed in European patent EP 238216.

In principle, the composition of the invention comprises a cellulolytic enzyme having an activity towards carboxymethylcellulose (CMC) and a catalytic activity towards cellotriose at pH8.5 corresponding to a Kcat of at least 0.01 per second, and the amylase may also comprise any other agent used in the combined process of the invention. However, it is preferred that the composition is free of bleach and other strong oxidants.

The composition of the invention may comprise one or several further ingredients selected from the group consisting of wetting agents, dispersing agents, sequestering agents and emulsifiers. Examples of suitable wetting agents are disclosed above. The emulsifier functions to emulsify hydrophobic impurities present on the fabric. The function of the dispersant is to prevent the extracted impurities from redepositing onto the fabric. The sequestering agent functions to remove ions such as Ca, Mg and Fe, which may have a negative effect on the process and preferred examples include caustic soda (sodium hydroxide) and soda ash (sodium carbonate). Determination of Cellulose Activity by cellulase

Under the term Kcat (S)^-1) The cellulase activity expressed for cellotriose can be determined by a coupling assay:

cellotriose → glucose + cellobiose (catalyst: cellulase)

(catalyst: glucose oxidase)

(catalyst: peroxidase) followed by 418nm (ABTS)^OxMaximum absorptionAt 418nm) was analyzed spectrophotometrically. The method comprises the following steps:

an God-Perid test kit (commercially available from Boehringer Mannhein, art 124036) was used. The buffer-enzyme solution in the test kit was dissolved in 500 ml of milli Q water and the pH of the solution was adjusted to 8.5 (NaOH).

80 mg of ABTS^R(commercially available from Boehringer Mannheim, art 756407) was dissolved in 10 ml of God-Perid, corresponding to ABTS^RThe total concentration was 10 mg/ml.

A substrate stock solution of 5 mmol (2.52 mg/mL) cellotriose (art 24741 commercially available from Merck) was prepared. Dilute aqueous solutions corresponding to 1000 micromolar, 500 micromolar, 376 micromolar, 250 micromolar, 100 micromolar and 60 micromolar were prepared.

A reaction mixture was prepared by mixing 1 part of the substrate solution with 1 part of God-Red.

A cellulase solution to be assayed is prepared at a concentration of 1.0 to 3.0 micromolar.

50 microliters of the enzyme solution and 450 microliters of the reaction mixture were mixed.

The measurement was carried out at a constant temperature of 40℃ in a 1 cm cuvette at 418nm using an HP 8452ADiode Array spectrophotometer. The reaction was followed by measuring the oxidation reaction of ABTS every 20 seconds for a total of 600 seconds. And (3) calculating:

the term Kact (S)^-1) The cellulase activity against cellotriose can be expressed from the Lineweaver-Burk curve (1/v vs. 1/[ S])]Curve (d) is calculated to yield: the slope and intersection points were determined by linear regression analysis.

The following constants were used in the calculations: cellulase: e is 66,310M^-1Cm^-1ABTS^Ox: e 0.0323 micromole^-1Cm^-1

The following results were obtained from the analysis:

enzyme	Kcat(S-1)
		EG I	1.5
EG I-F	5.5

The cellulolytic activity of endoglucanases is determined in an analytical standard and may be expressed in ECU units.

The cellulolytic enzyme hydrolyzes the carboxymethyl cellulose, thereby reducing the viscosity ofthe incubation mixture. The result of the viscosity reduction can be determined with a vibration viscometer (for example MIVI 3000 commercially available from Sofraser, france).

The determination of the cellulolytic activity measured by the ECU can be determined according to analytical method AF 301.1, which is available from the Applicant as required.

The ECU determines the amount of catalytic activity a sample has quantified by measuring the ability of the sample to reduce the viscosity of a carboxymethyl cellulose (CMC) solution. The assay was performed at 40 ℃ at pH7.5 using a relative enzyme standard for viscosity reduction of carboxymethyl cellulose substrates.

The invention is further illustrated by the following non-limiting examples. EXAMPLE 1 incubation of Cotton knitwear with Humicola insolens EG I and EGV in a launcher-O-meter

To demonstrate the significantly low activity of endoglucanase I (EG I) obtained from Humicola insolens on cotton fabrics, a comparative study was conducted on endoglucanase V (EGV, 43KDa, disclosed in WO 91/17243) obtained from Humicola insolens. Both cellulases have appreciable activity expressed as ECU and are thus capable of degrading carboxymethylcellulose and a number of other hydrocolloid cellulose derivatives which may be present in the slurry composition.

Desized 100% cotton knits were treated with two different enzymes in a Lannder-O-Meter under the following conditions:

enzyme	Dosage form	Buffering agent
			H.insolens EG I	5.0 ECU/ml	2g/l KH2PO4/NaOH pH 7.3
H.insolens EG V	1.0 ECU/ml	2g/l KH2PO4/NaOH pH 7.0

Fabric: bleached interlockingly knitted 100% cotton (205 g/m)²) 7 g/beaker (2 cloth samples, each approximately 13 cm. times.13 cm). Liquid volume: 140 ml (LQR 1: 20) incubation: 60 minutes at 55 DEG C

To each Launder-O-meter beaker 20 steel balls (14 mm diameter and 11 g weight) were added to increase the mechanical action on the fabric in the cellulase treatment.

The following results were obtained:

enzyme	Dosage form	Weight loss (%)
			H.insolens EG I	5.0 ECU/ml	0.1
H.insolens EG V	1.0 ECU/ml	3.9

Weight loss is defined as follows: percent weight loss (%) [1- (weight after treatment)/(weight before treatment)], 100

The results show that: the EG I cellulase had substantially no weight loss to cotton compared to the weight loss caused by EG V cellulase. Example 2

This example illustrates that the use of a cellulolytic enzyme having the characteristics set out in the claims provides a moderate excess of carboxymethyl cellulose-size removal and results in a significant improvement in the hand/stiffness of the fabric and, in addition, the enzyme has a beneficial effect on fabrics sized with a composition consisting of carboxymethyl cellulose and a starch/starch derivative mixture.

Sizing compositions consisting of pure carboxy cellulose (CMC) or carboxymethyl cellulose (CMC) -rich sizing compositions are generally very water soluble, i.e., the majority of the carboxymethyl cellulose slurry dissolves rapidly by simply contacting the sized fabric with an aqueous solution. However, a small portion of the carboxymethyl cellulose (CMC) size will adhere tightly to the fabric and give the fabric a very stiff hand, which makes the fabric unsuitable for further processing.

The fabric used in these experiments was a 100% cotton interlock sweater, which had been sized prior to desizing with a mixed size consisting of a mixture of pure carboxymethylcellulose CMC (Blanose 7LFD commercially available from Aqualon GmbH, germany) or 1: 1(W/W) carboxymethylcellulose CMC (Blanose 7LFD commercially available from Aqualon GmbH, germany) and carboxymethylated starch (CMC Solvitose C5 commercially available from lamberti s.p.a., italy).

The amount of size on the fabric was about 6.5% for the carboxymethyl cellulose-size (by weight of the fabric) and about 5.6% for the carboxymethyl cellulose/carboxymethylated starch-size (by weight of the fabric).

The fabric was cut into a 12 cm by 14 cm swatch (approximately 3.54 g/swatch weight without sizing).

After the climatic experiment the swatches were weighed and then incubated in a glass beaker containing 200 ml of 2 g/l potassium phosphate buffer (including enzymes as in the following table) ph7.0 for 30 minutes at 50 ℃:

serial number	Slurry material	Cellulase enzymes	Amylase
				1	CMC	Is free of	Is free of

2	CMC	20 ECU/gram fabric	Is free of
				3	CMC/CMS	Is free of	100 KNU/gram fabric
4	CMC/CMS	20 ECU/gram fabric	100 KNU/gram fabric

Cellulase: EG I from h.insolens (as described in example I). Amylase: aquazym 120L (activity: 120 KNU/g), a bacterial amylase commercially available from Novo Nordisk A/S.

After incubation the cloths were dried in an oven at 103 ℃ for 60 minutes, climatized and then weighed to evaluate the removal of the slurry, the average data for which are given in the following table:

serial number	Average gram of slurry/cloth sample	Average grams of removed slurry/cloth sample	Removing percent
				1	0.23	0.21	91％
2	0.23	0.23	100％
				3	0.20	0.15	75％
4	0.20	0.17	85％

As can be seen, the cellulase does facilitate the removal of the pulp in both cases (carboxymethyl cellulose-pulp and carboxymethyl cellulose/carboxymethylated starch-pulp, respectively).

For carboxymethyl cellulose-size, it was found that a large amount of size could also be removed when the swatches were incubated in buffer without cellulase, however it is clear from the fabric feel that the remaining carboxymethyl cellulose (approximately 20 mg/swatch) has a significant effect on the stiffness of the fabric.

To illustrate this effect, a panel of 5 persons experienced in the evaluation of the hand of the fabric, arranged the swatches in each series on a scale from 1 to 4, with notation 1 representing the hardest fabric and notation 4 representing the softest (best desizing).

This effect was so pronounced that all members of the panel gave identical ratings, as shown in the following table:

serial number

Mark (average)

1	2
		2	4
3	1
		4	3

Comparing series 1 with series 2 and series 3 with series 4 revealed that a significant reduction in fabric stiffness and a correspondingly better desizing effect were obtained from the treatment with cellulolytic enzymes.

References cited in the specification

Lipman and Pearson, science 227, 1435 (1985);

hudson, L., and Hay, F., Utility immunology, third edition (1989),

blackwell scientific publishing company;

sambrook et al, molecular cloning: a laboratory manual, 2 nd edition, cold spring harbor, 1989;

olson, e.s. fabric wetting method, volume 1, Noyes publishing company, ParkRidge, new jersey, usa (1983);

paper und h.k.pouette, grundgen der Textilveredlung, Deutsche, fachvlag GmbH, Frankfurt am Main, germany (1988);

sequence listing SEQ ID NO: 1, information:

(i) sequence characteristics:

(A) length: 415 amino acid

(B) Type (2): amino acids

(C) Chain conditions: sheet

(D) Topology: linearity

(ii) Molecular type: protein

(vi) The source is as follows:

(A) an organism: humicola insolens

(B) The strain is as follows: the DSM1800 sequence describes: SEQ ID NO: 1Gln Lys Pro Gly Glu Thr Lys Glu Val His Pro Gln Leu Thr Thr Phe 151015 Arg Cys Thr Lys Arg Gly Gly Cys Lys Pro Ala Thr Asn Phe Ile Val

20 25 30Leu Asp Ser Leu Ser His Pro Ile His Arg Ala Glu Gly Leu Gly Pro

35 40 45Gly Gly Cys Gly Asp Trp Gly Asn Pro Pro Pro Lys Asp Val Cys Pro

50 55 60Asp Val Glu Ser Cys Ala Lys Asn Cys Ile Met Glu Gly Ile Pro Asp65 70 75 80Tyr Ser Gln Tyr Gly Val Thr Thr Asn Gly Thr Ser Leu Arg Leu Gln

85 90 95His Ile Leu Pro Asp Giy Arg Val Pro Ser Pro Arg Val Tyr Leu Leu

100 105 110Asp Lys Thr Lys Arg Arg Tyr Glu Met Leu His Leu Thr Gly Phe Glu

115 120 125Phe Thr Phe Asp Val Asp Ala Thr Lys Leu Pro Cys Gly Met Asn Ser

130 135 140Ala Leu Tyr Leu Ser Glu Met His Pro Thr Gly Ala Lys Ser Lys Tyr145 150 155 160Asn Pro Gly Gly Ala Tyr Tyr Gly Thr Gly Tyr Cys Asp Ala Gln Cys

165 170 175Phe Val Thr Pro Phe Ile Asn Gly Leu Gly Asn Ile Glu Gly Lys Gly

180 185 190Ser Cys Cys Asn Glu Met Asp Ile Trp Glu Ala Asn Ser Arg Ala Ser

195 200 205His Val Ala Pro His Tr Cys Asn Lys Lys Gly Leu Tyr Leu Cys Glu

210 215 220Gly Glu Glu Cys Ala Phe Glu Gly Val Cys Asp Lys Asn Gly Cys Gly225 230 235 240Trp Asn Asn Tyr Arg Val Asn Val Thr Asp Tyr Tyr Gly Arg Gly Glu

245 250 255Glu Phe Lys Val Asn Thr Leu Lys Pro Phe Thr Val Val Thr Gln Phe

260 265 270Leu Ala Asn Arg Arg Gly Lys Leu Glu Lys Ile His Arg Phe Tyr Val

275 280 285Gln Asp Gly Lys Val Ile Glu Ser Phe Tyr Thr Asn Lys Glu Gly Val

290 295 300Pro Tyr Thr Asn Met Ile Asp Asp Glu Phe Cys Glu Ala Thr Gly Ser305 310 315 320Arg Lys Tyr Met Glu Leu Gly Ala Thr Gln Gly Met Gly Glu Ala Leu

325 330 335Thr Arg Gly Met Val Leu Ala Met Ser Ile Trp Trp Asp Gln Gly Gly

340 345 350Asn Met Glu Trp Leu Asp His Gly Glu Ala Gly Pro Cys Ala Lys Gly

355 360365Glu Gly Ala Pro Ser Asn Ile Val Gln Val Glu Pro Phe Pro Glu Val

370 375 380Thr Tyr Thr Asn Leu Arg Trp Gly Glu Ile Gly Ser Thr Tyr Gln Glu385 390 395 400Val Gln Lys Pro Lys Pro Lys Pro Gly His Gly Pro Arg Ser Asp

405410415 SEQ ID NO: 2, information:

(i) sequence characteristics:

(A) length: 409 amino acid

(B) Type (2): amino acids

(C) Chain conditions: sheet

(D) Topology: linearity

(ii) Molecular type: protein

(vi) The source is as follows:

(A) an organism: fusarium oxysporum

(B) The strain is as follows: DSM2672

(xi) Description of the sequence: SEQ ID NO: 2Gln Thr Pro Asp Lys Ala Lys Glu Gln His Pro Lys Leu Glu Thr Tyr 151015 Arg Cys Thr Lys Ala Ser Gly Cys Lys Lys Gln Thr Asn Tyr Ile Val

20 25 30Ala Asp Ala Gly Ile His Gly Ile Arg Arg Ser Ala Gly Cys Gly Asp

35 40 45Trp Gly Gln Lys Pro Asn Ala Thr Ala Cys Pro Asp Glu Ala Ser Cys

50 55 60Ala Lys Asn Cys Ile Leu Ser Gly Met Asp Ser Asn Ala Tyr Lys Asn65 70 75 80Ala Gly Ile Thr Thr Ser Gly Asn Lys Leu Arg Leu Gln Gln Leu Ile

85 90 95Asn Asn Gln Leu Val Ser Pro Arg Val Tyr Leu Leu Glu Glu Asn Lys

100 105 110Lys Lys Tyr Glu Met Leu His Leu Thr Gly Thr Glu Phe Ser Phe Asp

115 120 125Val Glu Met Glu Lys Leu Pro Cys Gly Met Asn Gly Ala Leu Tyr Leu

130 135 140Ser Glu Met Pro Gln Asp Gly Gly Lys Ser Thr Ser Arg Asn Ser Lys145 150 155 160Ala Gly Ala Tyr Tyr Gly Ala Gly Tyr Cys Asp Ala Gln Cys Tyr Val

165 170 175Thr Pro Phe Ile Asn Gly Val Gly Asn Ile Lys Gly Gln Gly Val Cys

180 185 190Cys Asn Glu Leu Asp Ile Trp Glu Ala Asn Ser Arg Ala Thr His Ile

195 200 205Ala Pro His Pro Cys Ser Lys Pro Gly Leu Tyr Gly Cys Thr Gly Asp

210 215 220Glu Cys Gly Ser Ser Gly Ile Cys Asp Lys Ala Gly Cys Gly Trp Asn225 230 235 240His Asn Arg Ile Asn Val Thr Asp Phe Tyr Gly Arg Gly Lys Gln Tyr

245 250 255Lys Val Asp Ser Thr Arg Lys Phe Thr Val Thr Ser Gln Phe Val Ala

260 265 270Asn Lys Gln Gly Asp Leu Ile Glu Leu His Arg His Tyr Ile Gln Asp

275 280 285Asn Lys Val Ile Glu Ser Ala Val Val Asn Ile Ser Gly Pro Pro Lys

290 295 300Ile Asn Phe Ile Asn Asp Lys Tyr Cys Ala Ala Thr Gly Ala Asn Glu305 310 315 320Tyr Met Arg Leu Gly Gly Thr Lys Gln Met Gly Asp Ala Met Ser Arg

325 330 335Gly Met Val Leu AlaMet Ser Val Trp Trp Ser Glu Gly Asp Phe Met

340 345 350Ala Trp Leu Asp Gln Gly Val Ala Gly Pro Cys Asp Ala Thr Glu Gly

355 360 365Asp Pro Lys Asn Ile Val Lys Val Gln Pro Asn Pro Glu Val Thr Phe

370 375 380Ser Asn Ile Arg Ile Gly Glu Ile Gly Ser Thr Ser Ser Val Lys Ala385 390 395 400Pro Ala Tyr Pro Gly Pro His Arg Leu

405

Claims (16)

1. A process for desizing cellulose-containing fabrics by applying a composition having activity towards carboxymethylcellulose CMC and catalytic activity towards cellotriose at pH8.5, corresponding to a Kcat of at least 0.01S^-1The cellulolytic enzyme of (a) is treated.

2. The process according to claim 1, wherein the catalytic activity towards cellotriose corresponds to a Kcat of at least 0.1S at pH8.5^-1Preferably at least 1S^-1。

3. The process according to claim 1 or 2, wherein the cellulolytic enzyme is derived from a strain of the genera Humicola, Trichoderma, Mvceliophthora, Penicillium, Irpex, Aspergillus, Scvlalidium or Sinapis.

4. A process according to claim 3, wherein the enzyme is derived from a strain of Humicola insolens, Fusarium oxysporum or Trichoderma reesei.

5. The method according to any one of claims 1-4, wherein the enzyme is a recombinant cellulase, i.e. a cellulase essentially freeof other proteins.

6. A method according to claim 5, wherein the parent cellulolytic enzyme comprises the amino acid sequence of the Humicola insolens endoglucanase shown in SEQ ID NO.1 or is an analogue of said endoglucanase which i) is at least 60% homologous to the sequence shown in SEQ ID NO.1, ii) reacts with an antibody directed against said endoglucanase, and/or iii) is encoded by a DNA sequence which hybridizes with the same probe as the DNA sequence encoding said endoglucanase.

7. A method according to claim 5, wherein the parent cellulolytic enzyme comprises the amino acid sequence of the Fusarium oxysporum endoglucanase shown in SEQ ID No.2 or is an analogue of said endoglucanase i) being at least 60% homologous to the sequence shown in SEQ ID No.2, ii) reacting with an antibody directed against said endoglucanase, and/or iii) encoded by a DNA sequence hybridizing to a probe identical to the DNA sequence encoding said endoglucanase.

8. The method according to any one of claims 1 to 7, wherein the cellulolytic enzyme is used in an amount corresponding to a cellulase activity of 10 to 5000ECU per liter of desized liquor; preferably, an amount corresponding to 1-10 g of enzyme/l, more preferably 1-5 g/l, especially 1-3 g/l is used.

9. The process according to any one of claims 1 to 8, wherein the desizing is carried out at a temperature in the range of 30 to 100 ℃, preferably 30 to 60 ℃, and a pH in the range of 3 to 11, preferably 7 to 9.

10. A method according to any of claims 1 to 9 wherein the fabric is selected from the group consisting of cotton fibres, viscose (rayon) fibres, Lyocell fibres, cotton, viscose or all blends of Lyocell fibres with other fibres such as polyester fibres, viscose/cotton fibre blends, Lyocell/cotton fibre blends, viscose/wool fibre blends, Lyocell, wool fibres, cotton/wool fibre blends, linen, ramie, other fibres including cellulosic fibres, and all blends including cellulosic fibres with other fibres such as wool, polyester, polyamide and acrylic fibres.

11. A desizing composition for desizing cellulose-containing fabrics comprises a carboxymethyl cellulose CMC having an activity and a Kcat corresponding to a catalytic activity for cellotriose at pH8.5 of at least 0.01S^-1And an enzyme that promotes starch decomposition.

12. The desizing composition according to claim 11, wherein the cellulolytic enzyme is obtained or derived from a strain of humicola, trichoderma, Myceliophthora, penicillium, irpex, aspergillus, Scvtalidium or cresosporium.

13. Desizing composition according to claim 12, wherein the enzyme is derived from a strain of humicola insolens, fusarium or Trichoderme reesei.

14. The desizing composition according to claim 13, wherein the parent enzyme comprises the amino acid sequence of a Humicola insolens endoglucanase as shown in SEQ ID No.1 or is an analogue of said endoglucanase which i) is at least 60% homologous to the sequence shown in SEQ ID No.1, ii) reacts with an antibody directed against said endoglucanase, and/or iii) is encoded by a DNA sequence which hybridizes with a probe identical to the DNA sequence encoding said endoglucanase.

15. A desizing composition according to claim 13, wherein the parent enzyme comprises the amino acid sequence of the fusarium oxysporum endoglucanase as shown in SEQ ID No.2 or is an analogue of said endoglucanase i) being at least 60% homologous to the sequence shown in SEQ ID No.2, ii) reacting with an antibody directed against said endoglucanase, and/or iii) encoded by a DNA sequence hybridizing to a probe identical to the DNA sequence encoding said endoglucanase.

16. The desizing composition of any of claims 11 to 15, further comprising at least one additional component selected from the group consisting of wetting agents, dispersing agents, sequestering agents, and emulsifiers.