BR112020022593A2 - method for treating dissolving pulp, dissolving pulp, textile fiber or derivatized cellulose, and, using a lytic polysaccharide monoxygenase. - Google Patents

Abstract

METHOD FOR TREATING PULP OF DISSOLUTION, DISSOLUTION PULP, TEXTILE FIBER OR DERIVATIZED CELLULOSIS, AND, USE OF A POLYESACARIDE LITHIC MONOXYGENASE. The present invention relates to the treatment of dissolving pulp with a polysaccharide lytic monoxygenase. Monoxygenase treatment polysaccharide lytic results in reduced viscosity and / or control improved viscosity in the dissolution pulp production process and / or greater reactivity of the dissolving pulp.

Description

1/36 METHOD FOR TREATING DISSOLUTION PULP, DISSOLUTION PULP, TEXTILE FIBER OR DERIVATIZED CELLULOSIS, AND,

USE OF A POLYESACARIDE LITHIC MONOXYGENASE Reference to sequence LISTING

[001] This application contains a Sequence Listing in a computer-readable format. The computer-readable format is incorporated into this document by reference.

FIELD OF THE INVENTION

[002] The present invention relates to the treatment of dissolving pulp with one or more enzymes. The enzymatic treatment results in reduced viscosity and / or improved viscosity control in the process of dissolving pulp production and / or greater reactivity of the final dissolving pulp.

BACKGROUND OF THE INVENTION

[003] Commercial dissolution pulp or commercial dissolution grade pulp is a chemical bleached pulp with a cellulose content high enough to be suitable for the production or regeneration of cellulose and cellulose derivatives. The commercially dissolving pulp has specific properties, such as a high level of gloss and uniform molecular weight distribution. Commercial dissolving pulp is manufactured for uses that require a high purity of chemical cellulose and, specifically, a low hemicellulose content, since chemically similar hemicellulose can interfere with subsequent processes. The dissolving pulp is so named because it is not transformed into paper, but rather dissolved or in a solvent or by derivatization in a homogeneous solution, which makes it completely chemically accessible and removes any remaining fibrous structure. Once dissolved, it can be spun into textile fibers, such as viscose or Lyocell, or chemically reacted to produce derivatized celluloses, such as triacetate.

2/36 cellulose, a plastic-like material formed from fibers or films, or cellulose ethers, such as methylcellulose, used as a thickener.

[004] The manufacture of conventional viscose using dissolving pulps as a raw material requires an improvement in terms of its environmental impact, as well as its production cost. There is a need in the art to provide a method for treating dissolving pulp with reduced costs and less environmental impact.

[005] The present invention provides a solution based on polysaccharide lytic monoxygenase that reduces viscosity and / or improves viscosity control in the production of dissolving pulp, for example, kraft dissolving pulp and sulfite. The enzyme solution described in this invention allows for more selective depolymerization of cellulose and thus better control of pulp viscosity compared to conventional methods in use which are non-selective with many side reactions, such as oxygen hydrolysis, peroxide hydrogen, ozone, sodium hypochlorite and acid. In addition, the reactivity of the dissolving pulp in the present invention is improved, thereby reducing the amount of chemicals used in the viscose production process and / or improving the processability in terms of viscose grease filterability in the manufacturing process. viscose. Savings in the amount of chemicals used in the production of regenerated cellulose as carbon disulfide (CS2) in the viscose manufacturing process will reduce costs and the environmental impact. Likewise, it is expected that the increase in the reactivity of the dissolving pulp will benefit the production process of cellulose derivatives, as well as the subsequent esterification and etherification processes.

SUMMARY OF THE INVENTION

[006] The invention provides a method for treating the dissolving pulp, comprising a step of subjecting the dissolving pulp to a lytic polysaccharide monoxygenase.

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[007] The method of the present invention generates dissolving pulp with reduced viscosity and / or improved viscosity control in the dissolution pulp production process and / or greater reactivity for making viscose and / or higher content of oxidized groups, in comparison with the dissolving pulp obtained by the same process in which treatment with lytic polysaccharide monoxygenase (LPMO) is omitted. Said dissolving pulp is kraft dissolving pulp and / or sulfite dissolving pulp.

[008] The present invention further provides a dissolving pulp made by the method of the present invention.

[009] The present invention further provides a textile fiber or a derivatized cellulose made from the dissolving pulp of the present invention.

[0010] The present invention further provides the use of a lytic polysaccharide monoxygenase for the treatment of dissolving pulp.

DEFINITIONS

[0011] Dissolving pulp: Dissolving pulp is a high-grade cellulose pulp, with a low hemicellulose, lignin and resin content. This pulp has special properties, such as a high level of gloss and uniform molecular weight distribution. It is used to manufacture products that include acetate and rayon textile fibers, cellophane, photographic film and various chemical additives. To a large extent, the use of dissolving wood pulp depends on its purity (cellulose content), which depends mainly on the production process. In order to obtain high quality products, these so-called “special” pulps must meet certain requirements, such as high cellulose content, low hemicellulose content, a uniform molecular weight distribution and high cellulose reactivity. Most commercial dissolving slurries meet these demands to a certain extent. However, achieving high cellulose accessibility as well as solvent and reagent reactivity is not an easy task due to the structure

4/36 complex and compact presented by cellulose. About 77% of all dissolving pulp is used in the manufacture of cellulosic fibers (rayon and acetate).

[0012] Two basic processes are used to produce dissolution pulp: (a) the sulfite process; and b) the sulfate (kraft) process.

[0013] To manufacture dissolution grade pulps, the removal of hemicelluloses from wood fiber is crucial, due to the fact that hemicelluloses can affect the viscose filterability, cellulose xanthanation and the strength of the final product during the production of viscose. Hemicelluloses are removed during wood cooking and subsequent bleaching. In sulfite pulping, the acidic conditions used are responsible for removing most of the hemicellulose, while in the sulfate / kraft process, a prehydrolysis step is usually necessary to remove hemicelluloses. Another method for removing hemicelluloses is by treating pulps with enzymes that react only with the hemicellulose portion of the pulp.

[0014] Kraft dissolving pulp: “Kraft dissolving pulp” is synonymous with “sulfate dissolving pulp”. A preferred example is a pre-hydrolysis kraft dissolving pulp. Dissolving kraft pulp is produced by digesting wood chips at temperatures above about 120 ° C with a solution of sodium hydroxide and sodium sulfide. Some kraft pulp productions are also carried out in which the amount of sodium sulfide is increased by oxygen or anthraquinone. In comparison to the production of soda pulp, the production of kraft pulp is particularly useful for the production of pulp from softwoods, which contain a higher percentage of lignin than hardwoods. The term "kraft dissolving pulp" is synonymous with "kraft dissolving pulp" and "kraft dissolving grade pulp" and refers to pulp that has a high cellulose content. The cellulose content of the kraft dissolving pulp is

5/36 preferably at least 90% (weight / weight), such as at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97% at least 98% or at least 99% (w / w). The dissolving kraft pulp is manufactured for uses that require a high chemical price, and particularly a low hemicellulose content. The hemicellulose content of the dissolving pulp is preferably less than 10% (weight / weight) such as less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4 %, less than 3%, less than 2% or less than 1% (w / w). The kraft dissolving pulp can, for example, be used for the generation of regenerated cellulose or for the generation of cellulose derivatives. “Kraft dissolution grade pulp” can also be defined as pulp that has been purified sufficiently for use in the production of viscose rayon, cellulose ethers or cellulose esters with organic or inorganic acids.

[0015] Sulfite dissolving pulp: The sulphite process produces wood pulp which are almost pure cellulose fibers through the use of various sulfurous acid salts to extract lignin from wood chips in large pressure vessels called digesters. The salts used in the pulping process are sulfites (SO32−) or bisulfites (HSO3−), depending on the pH. The counterion can be sodium (Na +), calcium (Ca2 +), potassium (K +), magnesium (Mg2 +) or ammonium (NH4 +).

[0016] Sulphite pulping is carried out between pH 1.5 and 5, depending on the counterion for sulfite (bisulfite) and the ratio between base and sulfurous acid. The pulp is in contact with the pulping chemicals for 4 to 14 hours and at temperatures in the range of 130 to 160 ° C (266 to 320 ° F), again depending on the chemicals used.

[0017] Most of the intermediates involved in delignification in sulfite pulping are resonance-stabilized carbocations formed by carbon-carbon double bond protonation or acid cleavage of

6/36 ether bonds that connect many of the constituents of lignin. It is the last reaction that is responsible for most of the degradation of lignin in the sulfite process. The sulfite process is not expected to degrade lignin to the same extent as the kraft process and the lignosulfonates of the sulfite process are useful by-products.

[0018] Sulphite pulping spent cooking liquor is usually called brown liquor, but the terms red liquor, thick liquor and sulfite liquor are also used (compared to black liquor in the kraft process). Pulp washers, which use countercurrent flow, remove spent cooking chemicals and degraded lignin and hemicellulose.

[0019] The "bleaching" is the removal of color from the pulp, mainly the removal of traces of lignin that remain attached to the fiber after the primary pulp production operation. Bleaching generally involves treatment with oxidizing agents, such as chlorine (C stage), chlorine dioxide (D stage), oxygen (O stage), hydrogen peroxide (P stage), ozone (Z stage) and paracetic acid (CH3CO3H; Paa stage) or a reducing agent, such as sodium dithionite (Y stage). There are chlorine free processes (Cl2; C stage), such as elemental chlorine free bleaching (ECF) in which chlorine dioxide (ClO2; D stage) is used primarily and is typically followed by an alkaline extraction stage. Completely chlorine free bleaching (TCF) is another process in which mainly oxygen-based chemicals are used. The pulp bleaching process typically comprises a sequence of bleaching steps with washing between them to remove degradation products from bleaching reactions. '

[0020] Cold Caustic Extraction (CCE): A cold alkaline extraction, also called Cold Caustic Extraction (CCE), is a method used

7/36 to remove short-chain non-cellulosic carbohydrates (cellulose purification) which is based on physical effects such as swelling and solubilization. Generally, a stage of CCE occurs at temperatures below 45oC and with the use of a very high dose of NaOH which, in the liquid phase, can reach values of up to 100 g / l. Depending on the consistency of the pulp in use, this will determine the amount of NaOH per dry pulp weight. Typical conditions for a CCE stage can be 5 to 10% w / w NaOH in the liquid phase for at least 10 minutes.

[0021] Hot Caustic Extraction (HCE): the term “hot caustic extraction” (HCE) is synonymous with “hot alkaline extraction”. HCE is a method for removing short-chain hemicellulose and amorphous cellulose into pulps. A hot caustic extraction (HCE) stage is a purification process that is based on chemical reactions, in particular, alkaline desquamation of hemicelluloses, which is performed at higher temperatures and a lower NaOH concentration compared to CCE.

[0022] ISO brightness: ISO brightness is defined in ISO 2470-1 (method of measuring the ISO brightness of pulp, paper and boards), and is the intrinsic radiance factor [reflectance] measured with a reflectometer that has the characteristics described in ISO 2469.

[0023] Pulp viscosity: is measured by dissolving the pulp in a suitable cellulose solvent as in cupri-ethylenediamine (CED) and measuring the solution viscosity. This measurement provides an indication of the average degree of cellulose polymerization. This property can be referred to as intrinsic viscosity in ml / g and measured according to ISO 5351 or as TAPPI viscosity in cP and measured according to TAPPI T 230. '

[0024] Kraft dissolving pulp unbleached or partially bleached or extracted alkaline: it is produced by a kraft-based cooking process such as pre-hydrolysis kraft cooking (PHK), but not

8/36 completely bleached and purified until it becomes a commercial kraft dissolving pulp and, therefore, it is not a finished product. Typically, it has an ISO brightness below 90% (such as below 85%, such as below 80%, such as below 75%, such as below 70%, such as below 65%, such as below 60%, such as below 55%, such as below 50%, such as below 45%, such as below 40%, such as below 35% and such as below 30%). '

[0025] Sulphite dissolving pulp unbleached or partially bleached or extracted alkaline: it is produced by a sulphite-based cooking process, but not completely bleached and purified until it becomes a commercial sulphite dissolving pulp and, thus, it is not a finished product. Typically, it has an ISO brightness below 90% (such as below 85%, such as below 80%, such as below 75%, such as below 70%, such as below 65%, such as below 60%, such as below 55%, such as below 50%, such as below 45%, such as below 40%, such as below 35% and such as below 30%). '

[0026] Bleached kraft dissolving paste and bleached sulfite dissolving paste: it is produced by a kraft dissolving paste or a sulphite-based cooking process, but fully bleached and purified until it becomes a commercial dissolving pulp. Typically, it has an ISO brightness above 90% (above 91%, above 92%, above 93%, above 94%, above 95%, above 96%, above 97%, as above 98%, such as above 99%, such as 100%).

[0027] Sequence identity: The relationship between two amino acid sequences or between two nucleotide sequences is described by the parameter "sequence identity".

[0028] For purposes of the present invention, the sequence identity between two amino acid sequences is determined using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol.

9/36 48: 443-453) as implemented in the Needle program of the EMBOSS package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, Trends Genet. 16: 276-277), preferably version 5.0 .0 or later. The parameters used are a gap opening penalty of 10, a gap extension penalty of 0.5 and the replacement matrix EBLOSUM62 (EMBOSS version of BLOSUM62). The Needle result marked “longest identity” (obtained using the –nobrief option) is used as the identity percentage and is calculated as follows: (Identical Residues x 100) / (Alignment Length - Total Number of Gaps in Alignment )

DETAILED DESCRIPTION OF THE INVENTION

[0029] The invention relates to a method for treating the dissolving pulp, comprising a step of subjecting the dissolving pulp to a lytic polysaccharide monoxygenase (LPMO).

[0030] In a preferred embodiment, the method of the present invention further comprises a step of subjecting the dissolving pulp to a cellulase.

[0031] In a preferred method of the present invention, the step of subjecting the dissolving pulp to a lytic polysaccharide monoxygenase and the step of subjecting the dissolving pulp to a cellulase are carried out simultaneously or sequentially in any order. In a further preferred embodiment of the present invention, a lytic polysaccharide monoxygenase is added to the dissolving pulp together with the cellulase. In a further preferred embodiment of the present invention, a lytic polysaccharide monoxygenase is added to the dissolving pulp prior to the addition of a cellulase. In a further preferred embodiment of the present invention, a lytic polysaccharide monoxygenase is added to the dissolving pulp after the addition of a cellulase.

[0032] In a preferred embodiment, the present method of the present

The invention comprises a step of bleaching the dissolving pulp. In a preferred method of the present invention, the step of subjecting the dissolving pulp to a lytic polysaccharide monoxygenase and the step of bleaching the dissolving pulp are carried out simultaneously or sequentially in any order.

[0033] In a preferred method of the present invention, the bleaching step of the dissolving pulp is carried out using a chemical selected from the group consisting of ClO2, O2, O3, H2O2, CH3CO3H and NaOCl.

[0034] In a preferred embodiment, the method of the present invention further comprises the step of Alkaline Extraction. In the method of the present invention, the Alkaline Extraction is an E, HCE or CCE stage. Special alkaline purification treatments such as HCE or CCE treatments can yield higher levels of cellulose in sulfite and kraft processes. In the case of sulfite pulps, HCE is typically employed as further purifying the pulp after cooking sulfite.

[0035] In a preferred embodiment, the method of the present invention further comprises a step of subjecting the dissolving pulp to a lytic polysaccharide monoxygenase and an electron donor thereof, preferably ascorbic acid, gallic acid, pyrogallol or cysteine. The electron donor may exist in the dissolving pulp to be treated. In one embodiment, none or a small amount of electron donor is added to the dissolving pulp. In another embodiment, an effective amount of electron donor is added to the dissolving pulp.

[0036] In a preferred method of the present invention, the dissolution paste is an unbleached, partially bleached, bleached or alkaline extracted dissolution paste. In a preferred method of the present invention, the dissolving pulp is kraft pulp or sulfite pulp.

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[0037] In one embodiment, the polysaccharide lytic monoxygenase added in the method of the present invention has a sequence identity of at least 60% identity [such as at least 65%, such as at least 70%, such as at least 75 %, such as at least 80%, such as at least 85%, such as at least 90%, such as at least 95%, such as at least 99%] with the mature polypeptide of SEQ ID NO: 1, the polypeptide mature from SEQ ID NO: 2, the mature polypeptide from SEQ ID NO: 3. In one embodiment, the cellulase added in the method of the present invention has a sequence identity of at least 60% identity (such as at least 65%, such as at least 70%, such as at least 75%, such as at least 80%, such as at least 85%, such as at least 90%, such as at least 95%, such as at least 99%) with SEQ ID NO: 4, the mature polypeptide of SEQ ID NO: 5, the mature polypeptide of SEQ ID NO: 6, or mature polypeptide of SEQ ID NO: 7.

[0038] In a preferred embodiment, the polysaccharide lytic monoxygenase added in the method of the present invention comprises or consists of SEQ ID NO: 1 or its mature polypeptide, or SEQ ID NO: 2 or its mature polypeptide, or SEQ ID NO: 3 or its mature polypeptide; or cellulase comprises or consists of SEQ ID NO: 4, SEQ ID NO: 5 or its mature polypeptide, SEQ ID NO: 6 or its mature polypeptide, or SEQ ID NO: 7 or its mature polypeptide.

[0039] In another preferred embodiment, the polysaccharide lytic monoxygenase added in the method of the present invention comprises 19 to 226 amino acids of SEQ ID NO: 1, or a homologous sequence, or an allelic variant, or a functional fragment thereof. In another preferred embodiment, the polysaccharide lytic monoxygenase added in the method of the present invention comprises 20 to 254 amino acids of SEQ ID NO: 2, or a homologous sequence, or an allelic variant, or a functional fragment thereof. In another preferred embodiment, the

12/36 lytic polysaccharide monoxygenase added in the method of the present invention comprises 22 to 249 amino acids of SEQ ID NO: 3, or a homologous sequence, or an allelic variant, or a functional fragment thereof. In another preferred embodiment, the cellulase added in the method of the present invention comprises SEQ ID NO: 4 or its total length, or a homologous sequence, an allelic variant or functional fragment thereof. In another preferred embodiment, the cellulase added in the method of the present invention comprises 22-305 amino acids of SEQ ID NO: 5 or a homologous sequence, or an allelic variant, or a functional fragment thereof. In another preferred embodiment, the cellulase added in the method of the present invention comprises 22 to 293 amino acids of SEQ ID NO: 6 or a homologous sequence, or an allelic variant, or a functional fragment thereof. In another preferred embodiment, the cellulase added in the method of the present invention comprises 19 to 409 amino acids of SEQ ID NO: 7 or a homologous sequence, or an allelic variant, or a functional fragment thereof.

[0040] The concentration of lytic polysaccharide monoxygenase added in the method of the present invention is preferably 0.05 mg / kg of oven dried pulp to 100000 mg / kg of oven dried pulp, such as a concentration selected from the group that consists of 0.05 mg / kg of kiln-dried pulp to 250 mg / kg of kiln-dried pulp, 0.25 mg / kg of kiln-dried pulp to 1000 mg / kg of kiln-dried pulp, from 0, 25 mg / kg of kiln-dried pulp to 2000 mg / kg of kiln-dried pulp, 1.0 mg / kg of kiln-dried pulp to 5000 mg / kg of kiln-dried pulp, 5.0 mg / kg oven dried pulp to 10,000 mg / kg oven dried pulp, 10.0 mg / kg oven dried pulp to 15000 mg / kg oven dried pulp, 15.0 mg / kg dried pulp in oven at 20000 mg / kg of oven dried pulp, from 20.0 mg / kg of oven dried pulp at 30000 mg / kg of oven dried pulp, from 30.0 mg / kg of oven dried pulp at 40000 mg / kg of oven-dried pulp, 40.0

13/36 mg / kg of oven-dried pulp to 60000 mg / kg of oven-dried pulp, 60.0 mg / kg of oven-dried pulp to 80000 mg / kg of oven-dried pulp and 80.0 mg / kg of oven dried pulp at 100000 mg / kg of oven dried pulp, or any combination of these intervals.

[0041] The cellulase concentration added in the present invention is 0.05 mg / kg of oven dried pulp to 100 mg / kg of oven dried pulp, such as a concentration selected from the group consisting of 0.05 mg / kg of kiln-dried pulp at 80.0 mg / kg of kiln-dried pulp, from 0.25 mg / kg of kiln-dried pulp to 60.0 mg / kg of kiln-dried pulp, from 0.25 mg / kg of oven-dried pulp at 40.0 mg / kg of oven-dried pulp, from 0.25 mg / kg of oven-dried pulp to 20.0 mg / kg of oven-dried pulp, from 0.25 mg / kg of oven-dried pulp to 10.0 mg / kg of oven-dried pulp, from 0.25 mg / kg of oven-dried pulp to 5.0 mg / kg of oven-dried pulp, from 0.50 mg / kg of oven-dried pulp at 85.0 mg / kg of oven-dried pulp, from 0.50 mg / kg of oven-dried pulp to 65.0 mg / kg of oven-dried pulp, from 0.50 mg / kg of oven-dried pulp at 45.0 mg / kg of oven-dried pulp, from 0.50 mg / kg of oven-dried pulp to 25.0 mg / kg of oven-dried pulp, from 0.50 mg / kg of oven-dried pulp at 15.0 mg / kg of powder oven dried pulp and 0.50 mg / kg oven dried pulp to 5.0 mg / kg oven dried pulp, or any combination of these ranges.

[0042] The method according to the invention results in an improved viscosity control, thus allowing the reduction in the production of dissolving pulp outside the final / target viscosity specification, typically more than 50% (such as more than 60% or more than 70%) reduction in out-of-grade dissolving pulp production in relation to viscosity. In one embodiment, the method according to the invention results in increased reactivity of the kraft and / or sulfite dissolving pulp, particularly the kraft dissolving pulp which has an increased reactivity of at least 10% (such as at least 20% or at least 30%).

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[0043] In a preferred embodiment, the method results in reduced viscosity and / or improved viscosity control in the dissolving pulp production process; and / or the method results in greater reactivity of the dissolving pulp, preferably greater Fock reactivity related to the viscose manufacturing process and, therefore, allowing savings in CS2 and, therefore, reducing costs and the environmental impact; and / or the method results in a higher content of oxidized groups in the dissolving pulp. This increase in oxidized groups may increase the reactivity of the dissolving pulp not only in terms of fiber swelling and chemical accessibility, but also considering that more anchoring points (carbonyl and / or carboxyl groups) in the cellulose will be available for subsequent derivatization processes in the production of cellulose derivatives.

[0044] In a preferred embodiment, the method of the present invention further comprises subjecting the dissolving pulp to a xylanase and / or a mannanase and / or a lipase and / or laccase and / or peroxidase.

[0045] A dissolving pulp produced using the method described above is also part of the invention. A textile fiber or derivatized cellulose produced from the dissolving pulp described above is also part of the invention.

[0046] The invention also relates to the use of a lytic polysaccharide monoxygenase for the treatment of dissolving pulp. Polysaccharide lytic monoxygenase (LPMO)

[0047] The term "polysaccharide lytic monoxygenase" means an enzyme that oxidizes sp (3) carbons to polysaccharides such as chitin, cellulose and starch in the presence of an outside electron donor and, as currently hypothesized, uses copper at the active site for activate molecular oxygen. At present, those enzymes that belong to the Auxiliary Activity families AA9, AA10, AA11, AA13, AA14 and AA15, as

15/36 defined in the database of active carbohydrate enzymes (http://www.cazy.org/).

[0048] In a first aspect, the LPMO comprises the following reasons: [ILMV] -Px (4,5) -GxY- [ILMV] -xRx- [EQ] -x (4) - [HNQ] and [FW] - [TF] -K- [AIV], where x is any amino acid, x (4,5) is any four or five contiguous amino acids, ex (4) is any four contiguous amino acids.

[0049] The LPMO, including the reasons mentioned above, can also comprise: Hx (1,2) -GPx (3) - [YW] - [AILMV], [EQ] -xYx (2) -Cx- [EHQN] - [FILV] -x- [ILV], or Hx (1,2) -GPx (3) - [YW] - [AILMV] and [EQ] -xYx (2) -Cx- [EHQN] - [FILV] - x- [ILV], where x is any amino acid, x (1,2) is any one or two contiguous amino acids, x (3) is any three contiguous amino acids and ex (2) is any two contiguous amino acids.

[0050] In a preferred aspect, the LPMO further comprises H- x (1,2) -G-P-x (3) - [YW] - [AILMV]. In a preferred aspect, the LPMO further comprises [EQ] -x-Y-x (2) -C-x- [EHQN] - [FILV] -x- [ILV]. In another preferred aspect, the LPMO further comprises Hx (1,2) -GPx (3) - [YW] - [AILMV] and [EQ] -xYx (2) -Cx- [EHQN] - [FILV] -x - [ILV].

[0051] In a second aspect, the LPMO comprises the following reasons: [ILMV] -Px (4,5) -GxY- [ILMV] -xRx- [EQ] -x (3) -A- [HNQ], in that x is any amino acid, x (4,5) is any four or five contiguous amino acids, ex (3) is any three contiguous amino acids. In the above reason, the abbreviation for amino acids IUPAC is used with

16/36 a single letter.

[0052] In one aspect, the LPMO comprises an amino acid sequence that has a sequence identity with the mature polypeptide of SEQ ID NO: 1 (Thielavia terrestris), SEQ ID NO: 2 (Lentinus similis), SEQ ID NO: 3 (Thermoascus aurantiacus), SEQ ID NO: 8 (Thielavia terrestris), SEQ ID NO: 9 (Thielavia terrestris), SEQ ID NO: 10 (Thielavia terrestris), SEQ ID NO: 11 (Thielavia terrestris), SEQ ID NO: 12 (Thielavia terrestris), SEQ ID NO: 13 (Thielavia terrestris), SEQ ID NO: 14 (Trichoderma reesei), SEQ ID NO: 15 (Myceliophthora thermophila), SEQ ID NO: 16 (Myceliophthora thermophila), SEQ ID NO: 17 (Myceliophthora thermophila), SEQ ID NO: 18 (Myceliophthora thermophila), SEQ ID NO: 19 (Myceliophthora thermophila), SEQ ID NO: 20 (Thermoascus aurantiacus), SEQ ID NO: 21 (Aspergillus fumigatus), SEQ ID NO: 22 (Penicillium pinophilum), SEQ ID NO: 23 (Thermoascus sp.), SEQ ID NO: 24 (Penicillium sp.), SEQ ID NO: 25 (Thielavia terrestris), SEQ ID NO: 26 (Thielavia terrestris), SEQ ID NO : two 7 (Thielavia terrestris), SEQ ID NO: 28 (Thielavia terrestris), SEQ ID NO: 29 (Thielavia terrestris), SEQ ID NO: 30 (Thielavia terrestris), SEQ ID NO: 31 (Thielavia terrestris), SEQ ID NO: 32 (Thielavia terrestris), SEQ ID NO: 33 (Thielavia terrestris), SEQ ID NO: 34 (Thielavia terrestris), SEQ ID NO: 35 (Thielavia terrestris), SEQ ID NO: 36 (Thermoascus crustaceus), SEQ ID NO: 37 (Thermoascus crustaceus), or SEQ ID NO: 38 (Talaromyces stipitatus) of at least 50%, for example, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, or at least 95%, at least 96%, at least 97% , at least 98%, at least 99%, or at least 100%.

[0053] In another aspect, the LPMO is an artificial variant comprising a substitution, deletion and / or insertion of one or more (or

17/36 various) amino acids of the mature polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, or SEQ ID NO: 38; or its homologous sequence, its allelic variant or its functional fragment.

[0054] Preferably, the amino acid changes are of a secondary nature, that is, substitutions of conservative amino acids or inserts that do not significantly affect the folding and / or activity of the protein; small deletions, usually from one to about 30 amino acids; small extensions of the amino or carboxyl terminus, such as a methionine residue at the amino terminus; a small peptide linker of about 20-25 residues; or a small extension that facilitates purification by altering the total charge or other function, such as a polyhistidine tract, an antigenic epitope or a binding domain.

[0055] Examples of conservative substitutions are within the groups of basic amino acids (arginine, lysine and histidine), acidic amino acids (glutamic acid and aspartic acid), polar amino acids (glutamine and asparagine), hydrophobic amino acids (leucine, isoleucine and valine), aromatic amino acids (phenylalanine, tryptophan and tyrosine) and small amino acids (glycine, alanine, serine, threonine and methionine). Amino acid substitutions that do not generally alter specific activity are known in the art and are described, for example, by H. Neurath and R.L. Hill, 1979, Em, The Proteins, Academic Press, New York. The most common swaps are Ala / Ser, Val / Ile, Asp / Glu, Thr / Ser, Ala / Gly,

18/36 Ala / Thr, Ser / Asn, Ala / Val, Ser / Gly, Tyr / Phe, Ala / Pro, Lys / Arg, Asp / Asn, Leu / Ile, Leu / Val, Ala / Glu and Asp / Gly .

[0056] Alternatively, the amino acid changes are of such a nature that the physical and chemical properties of the polypeptides are altered. For example, changes in amino acids can improve the thermal stability of the polypeptide, change substrate specificity, change the optimum pH, and the like.

[0057] The essential amino acids in a parental polypeptide can be identified according to procedures known in the art, such as site-directed mutagenesis or Alanine screening mutagenesis (Cunningham and Wells, 1989, Science 244: 1081-1085). In the latter technique, simple alanine mutations are introduced into all residues of the molecule, and the resulting mutant molecules are tested for cellulolytic enhancing activity to identify amino acid residues that are critical to the activity of the molecule. See also, Hilton et al., 1996, J. Biol. Chem. 271: 4,699 to 4,708. The active site of the enzyme or other biological interaction can also be determined by physical analysis of the structure, as determined by such techniques as nuclear magnetic resonance, crystallography, electron diffraction, or photo-affinity tagging, in conjunction with contact site amino acid mutation putative. See, for example, de Vos et al., 1992, Science 255: 306-312; Smith et al., 1992, J. Mol. Biol. 224: 899-904; Wlodaver et al., 1992, FEBS Lett. 309: 59-64. The identities of essential amino acids can also be inferred from the analysis of identities with polypeptides that are related to the parental polypeptide.

[0058] Single or multiple amino acid substitutions, deletions and / or insertions can be made and tested using known methods of mutagenesis, recombination and / or shuffling, followed by a relevant screening procedure, such as those disclosed by Reidhaar-

19/36 Olson and Sauer, 1988, Science 241: 53-57; Bowie and Sauer, 1989, Proc. Natl. Acad. Sci. USA 86: 2152-2156; WO 95/17413; or WO 95/22625. Other methods that can be used include error-prone PCR, phage display (for example, Lowman et al., 1991, Biochemistry 30: 10,832 to 10,837; U.S. Patent No. 5,223,409; WO 92/06204), and site mutagenesis -directed (Derbyshire et al., 1986, Gene 46: 145; Ner et al., 1988, DNA 7: 127).

[0059] Mutagenesis / shuffling methods can be combined with high productivity, automated screening methods to detect cloned polypeptide activity, submitted to mutagenesis expressed by host cells (Ness et al., 1999, Nature Biotechnology 17: 893-896) . Mutagenized DNA molecules encoding active polypeptides can be recovered from host cells and quickly sequenced using standard methods in the art. These methods allow for the rapid determination of the importance of individual amino acid residues in a polypeptide.

[0060] The total number of amino acid substitutions, deletions and / or insertions of the mature LPMO of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, or SEQ ID NO: 38 is not more than 10, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0061] In one aspect, LPMO is used in the presence of a soluble bivalent metallic activation cation as described in WO 2008/151043, for example, copper sulfate.

[0062] In one aspect, the LPMO is used in the presence of your

20/36 electron donor. The electron donor may exist in the dissolving pulp to be treated. In one embodiment, none or a small amount of electron donors can be added to the dissolving pulp. In another embodiment, an effective amount of electron donor can be added to the dissolving pulp.

[0063] In the present invention, the electron donor can be a dioxy compound, a bicyclic compound, a heterocyclic compound, a nitrogen-containing compound, or a sulfur-containing compound.

[0064] The dioxy compound can include any suitable compound containing two or more oxygen atoms. In some respects, the dioxy compounds contain a substituted aryl fraction as described here. Dioxy compounds can comprise one or more (several) hydroxyl groups and / or hydroxyl derivatives, but also include substituted aryl fractions having no hydroxyl and hydroxyl derivatives. Non-limiting examples of dioxy compounds include pyrocatechol or catechol, caffeic acid; 3,4-dihydroxybenzoic acid, 4-tert-butyl-5-methoxy-1,2-benzenediol; ascorbic acid, pyrogallol; gallic acid; methyl-3,4,5-trihydroxybenzoate; 2,3,4-trihydroxybenzophenone; 2,6-dimethoxyphenol; synapinic acid; 3,5-dihydroxybenzoic acid; 4-chloro-1,2-benzenediol; 4-nitro-1,2-benzenediol; tannic acid; ethyl gallate; methyl glycolate; dihydroxyfumaric acid; 2-butino-1,4-diol; crochonic acid; 1,3-propanediol; Tartaric acid; 2,4-pentanediol, 3-ethioxy-1,2-propanediol; 2,4,4'-trihydroxybenzophenone; cis-2-butene-1,4-diol; 3,4-dihydroxy-3-cyclobutene-1,2-dione; dihydroxyacetone; acrolein acetal; methyl-4-hydroxybenzoate; 4-hydroxybenzoic acid; and methyl-3,5-dimethoxy-4-hydroxybenzoate; or a salt or solvate thereof.

[0065] The bicyclic compound can include any suitable substituted fused ring system as described herein. The compounds may comprise one or more (several) additional rings, and are not limited to a specific number of rings unless otherwise noted. On a

21/36 aspect, the bicyclic compound is a flavonoid. In another aspect, the bicyclic compound is an optionally substituted isoflavonoid. In another aspect, the bicyclic compound is an optionally substituted flavilium ion, such as an optionally substituted anthocyanidin or optionally substituted anthocyanin, or a derivative thereof. Non-limiting examples of bicyclic compounds include epicatechin; quercetin; myricetin; taxifoline; campferol; morina; acacetin; naringenin; isoramnetine; apigenin; cyanidin; cyanine; curomanine; keracyanine, or a salt or solvate thereof.

The heterocyclic compound can be any suitable compound, such as an optionally substituted aromatic or non-aromatic ring comprising a heteroatom, as described herein. In one aspect, the heterocyclic compound is a compound comprising an optionally substituted heterocycloalkyl moiety or an optionally substituted heteroaryl moiety. In another aspect, the optionally substituted heterocycloalkyl fraction or the optionally substituted heteroaryl fraction is an optionally substituted 5-membered heterocycloalkyl or 5-membered heteroaryl fraction. In another aspect, the optionally substituted heterocycloalkyl or optionally substituted heteroaryl fraction is an optionally substituted fraction selected from pyrazolyl, furanyl, imidazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrrolyl, pyridyl, pyrimidyl, pyridazinyl, thiazolyl, thiazolyl, thiazolyl, thiazolyl, thiazolyl, thiazolyl, thiazolyl, thiazolyl, thiazolyl, thiazolyl, thiazolyl, pyrazolyl, tianaftenyl, carbazolyl, benzimidazolyl, benzothienyl, benzofuranyl, indolyl, quinolinyl, benzotriazolyl, benzothiazolyl, benzo-oxazolyl, benzimidazolyl, isoquinolinyl, isoindolyl, acridinyl, pyridine, pyridyl, pyridine, pyro azepinila, tiepinila, piperidinila and oxepinila. In another aspect, the optionally substituted heterocycloalkyl fraction or the optionally substituted heteroaryl fraction is an optionally substituted furanyl. Non-limiting examples of

22/36 heterocyclic compounds include (1,2-dihydroxyethyl) -3,4-dihydroxyifuran-2 (5H) -one; 4-hydroxy-5-methyl-3-furanone; 5-hydroxy-2 (5H) -furanone; [1,2-dihydroxyethyl] -furan-2,3,4 (5H) -trione; α-hydroxy-γ-butyrolactone; ribonic γ-lactone; γ-lactone of aldohexuronicaldohexuronic acid; gluconic acid δ-lactone, 4-hydroxycoumarin; hydrobenzofuran; 5- (hydroxymethyl) furfural; furoin; 2 (5H) -furanone; 5,6-dihydro-2H-pyran-2-one; and 5,6-dihydro-4-hydroxy-6-methyl-2H-pyran-2-one; or a salt or solvate thereof.

[0067] The nitrogen-containing compound can be any suitable compound with one or more nitrogen atoms. In one aspect, the nitrogen-containing compound comprises an amine, imine, hydroxylamine, or nitroxide fraction. Non-limiting examples of nitrogen-containing compounds include acetone oxime; violuric acid; pyridine-2-aldoxime; 2-aminophenol; 1,2-benzenediamine; 2,2,6,6-tetramethyl-1-piperidinyloxy; 5,6,7,8-tetrahydrobiopterin; 6,7-dimethyl-5,6,7,8-tetrahydropterin; and maleamic acid; or a salt or solvate thereof.

[0068] The quinone compound can be any suitable compound comprising a quinone fraction as described herein. Non-limiting examples of quinone compounds include 1,4-benzoquinone; 1,4- naphthoquinone; 2-hydroxy-1,4-naphthoquinone; 2,3-dimethoxy-5-methyl-1,4-benzoquinone or coenzyme Q0; 2,3,5,6-tetramethyl-1,4-benzoquinone or duroquinone; 1,4-dihydroxyanthraquinone; 3-hydroxy-1-methyl-5,6-indolinodione or adrenochrome; 4-tert-butyl-5-methoxy-1,2-benzoquinone; quinone pyrroloquinoline; or a salt or solvate thereof.

The sulfur-containing compound can be any suitable compound comprising one or more sulfur atoms. In one aspect, the sulfur-containing compound comprises a selected fraction of thionyl, thioether, sulfinyl, sulfonyl, sulfamide, sulfonamide, sulfonic acid, and sulfonic ester. Non-limiting examples of sulfur-containing compounds include ethanethiol; 2-propanethiol; 2-propene-1-thiol; 2-mercaptoethanesulfonic acid;

23/36 benzenethiol; benzene-1,2-dithiol; cysteine; methionine; glutathione; cystine; or a salt or solvate thereof. Cellulases

[0070] Cellulases or cellulolytic enzymes are enzymes involved in cellulose hydrolysis. In the hydrolysis of native cellulose, it is known that there are three main types of cellulase enzymes involved, namely, cellobiohydrolase (1,4-β-D-glucan cellobiohydrolase, EC 3.2.1.91, for example, cellobio- hydrolase I and cellobiohydrolase II), endo-β-1,4-glucanase (endo-1,4-β-D-glucan 4-glucan-hydrolase, EC 3.2.1.4) and β-glucosidase (EC 3.2.1.21 ).

[0071] In order to be efficient, the digestion of cellulose and hemicellulose requires several types of enzymes that work in cooperation. At least three categories of enzymes are needed to convert cellulose into fermentable sugars: endo-glucanases (EC 3.2.1.4) that cut cellulose chains at random; cellobiohydrolases (EC 3.2.1.91) that cleave cellobiosil units of the cellulose chain and beta-glucosidases (EC 3.2.1.21) that convert cellobiose and soluble cellodextrins into glucose. Among these three categories of enzymes involved in cellulose biodegradation, cellobiohydrolases are the key enzymes for the degradation of native crystalline cellulose. The term “cellobiohydrolase I” is defined in this document as a 1,4-beta-cellobiosidase cellulose (also called exo-glucanase, exo-cellobiohydrolase or 1,4-beta-cellobiohydrolase activity), as defined in the enzyme class EC 3.2.1.91, which catalyzes the hydrolysis of 1,4-beta-D-glucoside bonds in cellulose and cellotetraose, through the release of cellobiosis from the non-reducing ends of the chains. The definition of the term "cellobiohydrolase II activity" is identical, except that cellobiohydrolase II attacks from the reducing ends of the chains.

[0072] Endoglucanases (EC No. 3.2.1.4) catalyze the hydrolysis of 1,4-beta -D-glycosidic bonds in cellulose, cellulose derivatives (such as carboxymethylcellulose and hydroxyethylcellulose), lichenine, beta-1,4 bonds in

24/36 beta-1,3 glucans mixed as cereal beta-D-glucans or xyloglucans and other plant material containing cellulosic parts. The authorized name is endo-1,4-beta-D-glucan 4-glucan hydrolase, but the abbreviated term endoglucanase is used in this specification. In a preferred embodiment of the present invention, the cellulase used in the present invention is an endoglucanase.

[0073] Cellulases may comprise a carbohydrate binding module (CBM) that enhances the enzyme's binding to a cellulose-containing fiber and increases the effectiveness of the active catalytic part of the enzyme. A CBM is defined as a sequence of contiguous amino acids in an active carbohydrate enzyme with a discrete fold having carbohydrate binding activity. For additional information on CBMs, see the CAZy Internet server (Supra) or Tomme et al., (1995) in Enzymatic Degradation of Insoluble Polysaccharides (Saddler, JN & Penner, M., eds.), Cellulose- binding domains: classification and properties. pages 142 to 163, American Chemical Society, Washington, USA.

[0074] In a preferred embodiment, cellulases can be a preparation as defined in WO 2008/151079, which is incorporated herein by reference. The cellulase preparation can further comprise a beta-glucosidase, like the fusion protein disclosed in US 60 / 832,511. In one embodiment, the cellulase preparation also comprises a CBH II, preferably Thielavia terrestris cellobiohydrolase II CEL6A. In one embodiment, the cellulase preparation also comprises a cellulase enzyme preparation, preferably that derived from Trichoderma reesei. Cellulases are synthesized by a large number of microorganisms that include fungi, actinomycetes, myxobacteria and true bacteria, but also by plants. In particular, endoglucanases of a wide variety of specificities have been identified

25/36

[0075] The cellulase activity can, in a preferred modality, be derived from a fungal source, such as a strain of the genus Trichoderma, preferably a strain of Trichoderma reesei; a strain of the Humicola genus, like a strain of Humicola insolens; or a strain of Chrysosporium, preferably a strain of Chrysosporium lucknowense or a strain of the genus Thielavia, preferably Thielavia terrestris.

[0076] In one aspect, cellulase comprises an amino acid sequence that has a sequence identity with SEQ ID NO: 4, the mature polypeptide of SEQ ID NO: 5, the mature polypeptide of SEQ ID NO: 6, or the mature polypeptide of SEQ ID NO: 7 of at least 50%, for example, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85 %, at least 90%, at least 91%, at least 92%, at least 93%, or at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 100%. In another aspect, cellulase is an artificial variant comprising a substitution, deletion, and / or insertion of one or more (or more) amino acids of SEQ ID NO: 4, the mature polypeptide of SEQ ID NO: 5, the mature polypeptide of SEQ ID NO: 6, or the mature polypeptide of SEQ ID NO: 7; or its homologous sequence, or its allelic variant, or its functional fragment. The total number of amino acid substitutions, deletions and / or insertions of SEQ ID NO: 4, the mature polypeptide of SEQ ID NO: 5, the mature polypeptide of SEQ ID NO: 6, or the mature polypeptide of SEQ ID NO: 7 it is not more than 10, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0077] Fungi and bacteria produce a spectrum of cellulolytic enzymes (cellulases) that, based on sequence similarities (analysis of hydrophobic clusters), can be classified into different families of glycosyl hydrolases [Henrissat B & Bairoch A; Biochem. J. 1993 293 781-788]. Currently, cellulases belonging to families 5, 6, 7, 8, 9 are known,

26/36 10, 12, 26, 44, 45, 48, 60 and 61 of glycosyl hydrolases. Additional enzymes

Any enzyme with xylanase, mannanase, lipase, laccase and / or peroxidase activity can be used as additional enzymes in the use and process of the invention. Additional enzymes and a polysaccharide lytic monoxygenase are added simultaneously or sequentially in any order. Some non-limiting examples of such additional enzymes are listed below. Capitalized enzymes are commercial enzymes available from Novozymes A / S, Krogshoejvej 36, DK-2880 Bagsvaerd, Denmark. The activity of any of these additional enzymes can be analyzed using any method known in the art for the enzyme in question, including the methods mentioned in the cited references.

[0079] An example of a xylanase is the hemicellulase PULPZYME HC.

[0080] Examples of mannanases are Trichoderma reesei endo-beta-mannanases described in Ståhlbrand et al, J. Biotechnol. 29 (1993), 229-242.

[0081] An example of a lipase is the RESINASE A2X lipase. An example of a xylanase is the hemicellulase PULPZYME HC.

[0082] Examples of peroxidases and laccases are disclosed in EP 730641; WO 01/98469; EP 719337; EP 765394; EP 767836; EP 763115; and EP 788547. In the present context, whenever peroxity or laccase an oxidoreductase enzyme is mentioned that requires or benefits from the presence of acceptors (for example, hydrogen peroxide or oxygen), enhancers, mediators and / or activators, such compounds should be considered to be included. Examples of enhancers and mediators are disclosed in EP 705327; WO 98/56899; EP 677102; EP 781328; and EP 707637. If desired, a distinction can be made through the

27/36 definition of a laccase, or a system of peroxidase enzymes, as the combination of the enzyme in question and its acceptor, and, optionally, also an enhancer and / or mediator for the enzyme in question. Temperature for the method of the present invention

[0083] The temperature for the method of the present invention is typically 20 ° C to 100 ° C as a temperature range selected from the group consisting of 20 ° C to 30 ° C, 30 ° C to 40 ° C, 40 ° C to 50 o C, 50 oC to 60 oC, 60 oC to 70 oC, 70 oC to 80 oC, 80 oC to 90 oC, 90 oC to 100 oC, or any combination of these ranges. Incubation time for the method of the present invention

[0084] The incubation time used for the method of the present invention is typically 1 minute to 60 hours, as is a time interval selected from the group consisting of 1 minute to 15 minutes, 15 minutes to 30 minutes, 30 minutes to 45 minutes, 45 minutes to 60 minutes, 1 hour to 3 hours, 3 hours to 6 hours, 6 hours to 10 hours, 10 hours to 12 hours, 12 hours to 15 hours, 15 hours 20 hours, 20 hours to 22 hours, 22 hours to 25 hours, 25 hours to 30 hours, 30 hours to 40 hours, 40 hours to 50 hours, 50 hours to 60 hours, or any combination of these time intervals. Pulp used and produced in the method according to the invention:

[0085] The dissolving pulp used in the present invention can be wood pulp derived, for example, from softwood trees (such as spruce, pine, spruce, larch and pinaceae) and / or hardwoods (such as eucalyptus) , poplar and birch) or other plant sources, such as bamboo.

[0086] In a preferred embodiment, the dissolution paste is selected from the group consisting of hardwood dissolving pulp and softwood dissolving pulp, or a mixture thereof.

28/36

[0087] The invention relates in one embodiment to a dissolving pulp produced by the method according to the invention. The invention relates in one embodiment to a kraft dissolving pulp or sulfite dissolving pulp produced by the method according to the invention.

[0088] The invention also refers to the use of the dissolving pulp according to the invention for the production of textile fibers. The dissolving pulp produced can be used in the manufacture of regenerated cellulose such as viscose rayon, Lyocell and modal fibers. Carrying out the method of the invention in the presence of one or more surfactants

[0089] The method of the present invention can be carried out in the presence of one or more surfactants, such as one or more anionic surfactants and / or one or more non-ionic surfactants and / or one or more cationic surfactants.

[0090] Surfactants may include, in one embodiment, surfactants based on poly (alkylene glycol), ethoxylated dialkylphenols, ethoxylated dialkylphenols, ethoxylated alcohols and / or silicone based surfactants.

[0091] Examples of surfactants based on poly (alkylene glycol) are polyester (ethylene glycol) alkyl, poly (ethylene glycol) alkylether, homo and copolymers of ethylene oxide / propylene oxide, or alkyl esters or ethers of poly (ethylene oxide) -propylene oxide). Other examples include ethoxylated derivatives of primary alcohols, such as dodecanol, secondary alcohols, poly [propylene oxide], derivatives thereof, ethoxylated tridecyl alcohol phosphate ester and the like.

The currently preferred specific anionic surfactant materials useful in the practice of the invention comprise sodium alpha-sulfo methyl laureate, (which may include some alpha-sulfo ethyl laureate) for example, as commercially available under the trademark ALPHA-STEPTM- ML40; sodium xylenesulfonate, for example, as commercially available under the trademark STEPANATETM-X;

29/36 triethanolammonium lauryl sulfate, for example, as commercially available under the trademark STEPANOLTM-WAT; disodium lauryl sulfosuccinate, for example, as commercially available under the trademark STEPANTM-Mild SL3; additional blends of various anionic surfactants can also be used, for example, a blend of 50% - 50% or 25% -75% of the APHA-STEPTM and STEPANATETM materials mentioned above, or a blend of 20% -80% of the materials of ALPHA-STEPTM and STEPANOLTM mentioned above (all commercially available materials mentioned above can be obtained from Stepan Company, Northfield, Ill.).

[0093] The currently preferred specific non-ionic surfactant materials useful in the practice of the invention comprise cocodiethanolamide, as commercially available under the trademark NINOLTM-11CM; alkyl polyoxyalkylene glycol ethers, such as the relatively high molecular weight butylene oxide ethylene propylene oxide block copolymers commercially available under the trademark TOXIMULTM-8320 from the Stepan Company. Additional alkyl polyoxyalkylene glycol ethers can be selected, for example, as disclosed in U.S. Patent No. 3,078,315. Mixtures of the various non-ionic surfactants can also be used, for example, a mixture of 50% -50% or 25% -75% of the NINOLTM and TOXIMULTM materials mentioned above.

[0094] The presently preferred specific anionic / nonionic surfactant blends useful in the practice of the invention include various mixtures of the above materials, for example, blends of 50% -50% of the ALPHA-STEPTM and NINOLTM materials mentioned above or a blend of 25% -75% of the STEPANATETM and TOXIMULTM materials mentioned above.

[0095] Preferably, the various mixtures of anionic surfactants,

30/36 non-ionic, anionic / non-ionic used in the practice of the invention have a solids or active content of up to about 100% by weight and preferably have an active content in the range of about 10% to about 80%. In fact, another blend content or solid (active) content can also be used and these anionic surfactants, non-ionic surfactants and mixtures thereof can also be used with known pulp-making chemicals, such as, for example, anthraquinone and derivatives thereof and / or other typical paper chemicals, such as caustics, antifoam agents and the like.

[0096] The invention described and claimed in this document is not to be limited in scope by the specific modalities disclosed in this document, since these modalities are intended as illustrations of the various aspects of the invention. Any equivalent modalities are intended to be within the scope of this invention. In fact, various modifications of the invention in addition to those shown and described in this document will become apparent to those skilled in the art from the foregoing description. Such modifications are also intended to reside within the scope of the appended claims. In the event of a conflict, this disclosure including definitions will prevail.

[0097] Several references are cited in this document, whose disclosures are incorporated by reference in their entirety.

EXAMPLES Materials and Methods

[0098] The intrinsic viscosity of the pulp was measured according to ISO 5351 (International Organization for Standardization 5351).

[0099] The viscosity of the pulp was measured by mViPr according to WO 2011/107472 A9.

[00100] The amount of aldehyde groups (CHO content) was measured spectrophotometrically according to the procedure described by

31/36 Obolenskaya et al., “Determination of aldehyde groups in oxidized pulps”, Laboratory Manipulations in Wood and Cellulose Chemistry, Ecology, Moscow, 211-212, 1991, which is based on the 2,3,5- chloride reaction triphenyltetrazolium (TTC) with the aldehyde groups leading to the formation of formazan (red dye).

[00101] Fock reactivity is a measure of how much of a known amount of pulp reacts with CS2 as a small scale simulation of the viscose manufacturing process and was performed with 9% NaOH. Enzyme Description LPMO Thielavia terrestris Tt LPMO shown as the mature polypeptide of SEQ ID (Tt LPMO) NO: 1 here, also shown as the mature polypeptide of SEQ ID NO: 8 of WO 2010 / 080532A LPMO Lentinus similis Ls LPMO shown as the polypeptide mature of SEQ ID (Ls LPMO) NO: 2 here, also shown as the mature polypeptide of SEQ ID NO: 6 of WO2014 / 066141A LPMO Thermoascus aurantiacus Ta LPMO shown as the mature polypeptide of SEQ ID (Ta LPMO) NO: 3 here , also shown as the mature polypeptide of SEQ ID NO: 2 of WO2005 / 074656A Endoglucanase-1 Endoglucanase shown as SEQ ID NO: 4 here, which is a Q120H variant of the mature endoglucanase shown as SEQ ID NO: 9 of WO 96 / 29397A Endoglucanase-2 Endoglucanase shown as SEQ ID NO: 5 here, which is a cellulase shown as the mature polypeptide of SEQ ID NO: 2 in WO1991 / 017243A Endoglucanase-3 Endoglucanase shown as SEQ ID NO: 6 here, which is a GH45 cellulase shown as the mature polypeptide of SEQ ID NO: 2 in WO 2015 / 058700A Endoglucanase-4 Endoglucanase shown as SEQ ID NO: 7 here, which is a GH5 endoglucanase shown as the mature polypeptide of SEQ ID NO: 2 in WO2013 / 019780A Example 1: LPMO treatment of wood kraft dissolving pulp unbleached hardwood

[00102] Unbleached hardwood kraft dissolving paste with a cover number of 6.8 (TAPPI T 236 procedure), ISO brightness of 51% and an intrinsic viscosity of 1025 mL / g produced by a kraft pulping process of pre-hydrolysis and subsequently treated with a cold caustic extraction stage of a dissolving pulp production process. This pulp was treated with several LPMO in a small scale trial using 24 mg of kiln dried fiber at 0.4% consistency,

32/36 45 ° C, pH 5.0 (acetate buffer, 50 mM) for 20 hours. The enzymatic treatment (called stage X) was performed at a dosage of 5 mg EP (enzymatic protein) / g odp (kiln-dried pulp).

[00103] The pulp suspension with a consistency of 0.4% was disintegrated with a magnetic bar in glass test tubes placed in a heating block at 45 ° C. Ascorbic acid, gallic acid or pyrogallol was added as an electron donor to a final concentration of 1 mM in the suspension, followed by the addition of the LPMO enzyme to a final volume of 6 mL. After the incubation time of 20 hours after adding the enzyme, the tubes were cooled on ice and then 6 ml of cupriethylenediamine solution (CED) were added to dissolve the fibers. The pulp was dissolved on a rotary shaker at an ambient temperature of 25 ° C for 25 min. Control experiments were done in the same way, but without the addition of the enzyme.

[00104] After the dissolution time, the viscosity of the pulp was measured by mViPr. The mViPr pipette consists of a modified Gilson Concept C300 pipette equipped with a Gilson Diamond D300 pressure sensor and tips. The samples were kept at a constant temperature within ± 0.1 ° C. A volume of 200 µL of dissolved pulp was aspirated and dispensed inside and outside the pipette, respectively, while recording the pressure in the empty space of the pipette. A pipette speed of

4. Aspirations were followed by a delay of 2s and dispensation was followed by a delay of 5s. Each sample measurement consisted of 15 aspiration dispensing cycles, and the pressure results were an average of 15 aspiration or dispensing pressures, respectively.

[00105] The results of the suction pressure of the mViPr measurements are shown in Table 1. It can be seen that the LPMO can reduce the average cellulose size which is expressed as the reduction of the viscosity of the solution measured by the reduction of the suction pressure. The performance of each

33/36 LPMO also depends on the specific electron donor used. In addition, the electron donor itself also degrades cellulose when compared to the original pulp, especially ascorbic acid. Tt LPMO is very effective in reducing the viscosity of the pulp with all electron donors. Table 1. Aspiration pressure of the different unbleached pulps dissolved in CED Enzyme Aspiration pressure (Pa) Reduction vs. control (%) Original -1900.9 --- Control ascorbic acid -1408.8 0 Ta LPMO + Ascorbic acid -1195.9 15 Ls LPMO + Ascorbic acid -1135.8 19 Tt LPMO + Ascorbic acid -656.6 53 Control Gallic Acid -1753.8 0 Ta LPMO + Gallic Acid -1089.5 38 Ls LPMO + Gallic Acid -1091.6 38 Tt LPMO + Gallic Acid -580.2 67 Control Pyrogallol -1605.9 0 Ta LPMO + Pyrogallol -923.7 42 Ls LPMO + Pyrogallol -232.9 85 Tt LPMO + Pyrogallol -655.9 59 Example 2: LPMO treatment of bleached hardwood kraft dissolving pulp

[00106] It was used kraft pulp of bleached hardwood of dry acetate grade produced by a pre-hydrolysis kraft pulping process having an ISO gloss of 93.7% and an intrinsic viscosity of 684 mL / g. This pulp was treated with several LPMO with three different electron donors using the same conditions and procedures as in Example

1.

[00107] The suction pressure results of the mViPr measurements are shown in Table 2 for bleached dissolving pulps. PMLs can reduce the average size of cellulose molecules as expressed by reducing the solution's viscosity (dissolved pulp), measured by reducing the suction pressure. Greater reduction in viscosity of bleached pulp compared to controls (without enzyme) was achieved when PMLs were used together with gallic acid and pyrogallol in

34/36 compared to ascorbic acid, which also significantly reduces viscosity itself. Tt LPMO is a more robust LPMO in terms of reducing viscosity with all used electron donors. Table 2. Aspiration pressure of the different bleached pulps dissolved in CED Enzyme Aspiration pressure (Pa) Reduction vs. control (%) Original -450.9 --- Control ascorbic acid -345.1 0 Ta LPMO + Ascorbic acid -339.5 2 Ls LPMO + Ascorbic acid -338.2 2 Tt LPMO + Ascorbic acid -244.5 29 Control Gallic Acid -523.1 0 Ta LPMO + Gallic Acid -245.3 53 Ls LPMO + Gallic Acid -235.2 55 Tt LPMO + Gallic Acid -160.3 69 Control Pyrogallol -433.7 0 Ta LPMO + Pyrogallol -260.9 40 Ls LPMO + Pyrogallol -88.8 80 Tt LPMO + Pyrogallol -43.7 90 Example 3: Effect of LPMO treatment with / without endoglucanase on the medium pulp consistency on viscosity, reactivity and CHO content of bleached dissolving pulp

[00108] Bleached hardwood kraft dissolving pulp produced by a viscose grade prehydrolysis kraft pulping process with an intrinsic viscosity of 512 mL / g was used. This pulp was treated with several LPMO using gallic acid (1 mM) as an electron donor at 1.5% consistency in Distek vessels (Distek model Symphony 7100) with heating and continuous aerial agitation.

[00109] As soon as the pulps were disintegrated and at the temperature set point of 45 ºC, gallic acid was added and then the enzyme (LPMO and / or endoglucanase). The enzymatic treatment of the pulp lasted 25.5 hours at pH 5.0 (50 mM acetate buffer) using 2 mg EP / g odp of LPMO and 1.2 mg EP / kg odp of endoglucanase. After the enzymatic treatment, the pulps were filtered and washed in three consecutive steps with 1 L of running water. Part of the pulp sample was dried before measuring the CHO content and Fock reactivity and part of the sample was kept moist in the refrigerator

35/36 to test intrinsic viscosity.

[00110] In Table 3, it can be seen that with a lower dosage of 2 mg EP / g odp a small reduction in viscosity reduction is obtained with the tested LPMO, in comparison with Examples 1 and 2 with a higher dosage of LPMOs. However, when combined with an endoglucanase, the reduction in viscosity is increased with a surprising synergistic effect. The same synergy between LPMO and endoglucanase is seen in terms of Fock reactivity and the number of CHO groups. Table 3. Intrinsic viscosity, Fock reactivity and CHO content of bleached dissolving pulps Enzyme Intrinsic viscosity Fock reactivity CHO content (ml / g) (%) (mmol / kg odp) Original 512 16 17.8 Gallic acid Control 482 16 16.3 Tt LPMO + Gallic acid 479 20 19.8 Ta LPMO + Gallic acid 471 16 22.9 Endoglucanase 480 15 19.7 Tt LPMO + endoglucanase + 429 26 27.7 Gallic acid Example 4: Treatment effect combination of LPMO and endoglucanase in dissolving pulp of unbleached hardwood

[00111] The same unbleached hardwood kraft dissolving pulp from Example 1 was used and treated with several LPMO's using gallic acid (1 mM) as an electron donor in combination with several endoglucanases in a small-scale assay using 24 mg oven-dried fiber with 0.4% consistency, 50 ° C, pH 6.0 (phosphate buffer, 50 mM) for 20 hours. The treatments with LPMO were made in the dosages of 2.5 mg EP (enzymatic protein) / g odp (high dosage) or 0.5 mg EP / g odp (low dosage). The treatments with endoglucanase were carried out at a dosage of 0.5 mg EP / kg odp. The viscosity test and measurements were performed using the same conditions and procedures described in Example 1.

[00112] The results of the suction pressure of the mViPr measurements are shown in Table 4. It can be seen that the higher the dosage of

36/36

LPMO, the greater the pulp viscosity reduction.

A high dosage of LPMO gives a surprisingly greater viscosity reduction than treatments with endoglucanase.

Treatment with a combination of LPMO and an endoglucanase shows better performance in reducing viscosity than individual treatment with LPMO or endoglucanase.

Using this high viscosity pulp, a reinforcing effect is observed in the reduction of viscosity by the use of LPMO combined with endoglucanase, which allows a significant reduction in the amount of LPMO required.

For example, at a low dosage of Tt LPMO (0.5 mg EP / g odp), the addition of an endoglucanase 2 and 3 gives an even greater viscosity reduction than the high dosage of Tt LPMO alone (2.5 mg EP / g odp). Table 4. Aspiration pressure of the different unbleached pulps dissolved in CED Aspiration pressure Reduction vs. control- Enzyme (Pa) le (%) Original -1594.1 --- Control Gallic Acid -1565.3 0 Tt LPMO (high dose) + Gallic acid -821.0 48 Tt LPMO (low dose) + Gallic acid -1361.0 13 Ls LPMO (high dose) + Gallic acid -665.3 57 Ls LPMO (low dose) + Gallic acid -1141.0 27 Endoglucanase 1 -995.9 36 Tt LPMO (high dose) + Gallic acid + endoglucanase 1 -415.1 73 Tt LPMO (low dose) + Gallic acid + endoglucanase 1 -883.2 44 Ls LPMO (high dose) + Gallic acid + endoglucanase 1 -646.2 59 Ls LPMO (low dose) + Gallic acid + endoglucanase 1 -905.4 42 Endoglucanase 2 -1083.7 31 Tt LPMO (high dose) + Gallic acid + endoglucanase 2 -341.9 78 Tt LPMO (low dose) + Gallic acid + endoglucanase 2 -762.6 51 Ls LPMO (high dose) + Gallic acid + endoglucanase 2 -485.7 69 Ls LPMO (low dose) + Gallic acid + endoglucanase 2 -969.1 38 Endoglucanase 3 -1049.2 33 Tt LPMO (high dose) + Gallic acid + endoglucanase 3 -441.7 72 Tt LPMO (low dose) + Gallic acid + endoglucanase 3 -506.1 68 Ls LPMO (high dose) + Gallic acid + endoglucanase 3 -284.6 82 Ls LPMO (low dose) + Gallic acid + endoglucanase 3 -315.3 80 Endoglucanase 4 -1270.7 19 Tt LPMO (high dose) + Acid gallic + endoglucanase 4 -732.9 53 Tt LPMO (low dose) + Gallic acid + endoglucanase 4 -1169.2 25 Ls LPMO (high dose) + Gallic acid + endoglucanase 4 -904.8 42 Ls LPMO (low dose) + Gallic acid + endoglucanase 4 -1186.5 24

Claims (16)

1/4 CLAIMS 1. Method for treating dissolving pulp, characterized by the fact that it comprises a step of subjecting the dissolving pulp to a lytic polysaccharide monoxygenase. Method according to claim 1, characterized in that it comprises a step of subjecting the dissolving pulp to a cellulase, preferably an endoglucanase. Method according to either of claims 1 or 2, characterized in that the step of subjecting the dissolving pulp to a lytic polysaccharide monoxygenase and the step of subjecting the dissolving pulp to a cellulase are carried out simultaneously or sequentially in any order. Method according to any one of claims 1 to 3, characterized in that it comprises a step of bleaching the dissolving pulp. Method according to any one of claims 1 to 4, characterized in that it comprises a step of subjecting the dissolving pulp to a lytic polysaccharide monoxygenase and an electron donor, preferably ascorbic acid, gallic acid, pyrogallol or cysteine. Method according to any one of claims 1 to 5, characterized in that the dissolving pulp is an unbleached, partially bleached, bleached or alkaline-dissolving pulp; or wherein the dissolving pulp is kraft pulp or sulfite pulp. Method according to any one of claims 1 to 6, characterized in that the polysaccharide lytic monoxygenase has a sequence identity of at least 60% (such as at least 65%, such as at least 70%, such as at least 75%, such as at least 80%, such as at least 85%, such as at least 90%, such as at 2/4 minus 95%, such as at least 99%) with the mature polypeptide of SEQ ID NO: 1 or with the mature polypeptide of SEQ ID NO: 2 or with the mature polypeptide of SEQ ID NO: 3. Method according to any one of claims 1 to 7, characterized in that the cellulase has a sequence identity of at least 60% (such as at least 65%, such as at least 70%, such as at least 75%, such as at least 80%, such as at least 85%, such as at least 90%, such as at least 95%, such as at least 99%) with SEQ ID NO: 4, the mature polypeptide from SEQ ID NO: 5, the mature polypeptide of SEQ ID NO: 6 or the mature polypeptide of SEQ ID NO: 7. Method according to any one of claims 1 to 8, characterized in that the lytic polysaccharide monoxygenase comprises or consists of SEQ ID NO: 1 or its mature polypeptide, or SEQ ID NO: 2 or its polypeptide mature, or SEQ ID NO: 3 or its mature polypeptide; or cellulase comprises or consists of SEQ ID NO: 4, SEQ ID NO: 5 or its mature polypeptide, SEQ ID NO: 6 or its mature polypeptide, or SEQ ID NO: 7 or its mature polypeptide. Method according to any one of claims 1 to 9, characterized in that the concentration of lytic polysaccharide monoxygenase is 0.05 mg / kg of oven dried pulp to 100000 mg / kg of oven dried pulp, such as a concentration selected from the group consisting of 0.05 mg / kg of oven dried pulp to 250 mg / kg of oven dried pulp, 0.25 mg / kg of oven dried pulp to 1000 mg / kg oven dried pulp, 0.25 mg / kg oven dried pulp to 2000 mg / kg oven dried pulp, 1.0 mg / kg oven dried pulp to 5000 mg / kg dried pulp in kiln, from 5.0 mg / kg of kiln-dried pulp to 10,000 mg / kg of kiln-dried pulp, from 10.0 mg / kg of kiln-dried pulp to 15000 mg / kg of pulp 3/4 oven dried, from 15.0 mg / kg oven dried pulp to 20000 mg / kg oven dried pulp, from 20.0 mg / kg oven dried pulp to 30000 mg / kg dry pulp in oven, from 30.0 mg / kg of oven dried pulp to 40000 mg / kg of oven dried pulp, from 40.0 mg / kg of oven dried pulp to 60000 mg / kg of oven dried pulp, from 60.0 mg / kg of oven dried pulp to 80000 mg / kg of oven dried pulp and 80.0 mg / kg of oven dried pulp to 100000 mg / kg of oven dried pulp or any combination of these ranges. Method according to any one of claims 1 to 10, characterized in that the cellulase concentration is from 0.05 mg / kg of oven dried pulp to 100 mg / kg of oven dried pulp, such as a concentration selected from the group consisting of 0.05 mg / kg of kiln-dried pulp to 80.0 mg / kg of kiln-dried pulp, 0.25 mg / kg of kiln-dried pulp to 60.0 mg / kiln-dried pulp, 0.25 mg / kg kiln-dried pulp to 40.0 mg / kg kiln-dried pulp, 0.25 mg / kg kiln-dried pulp to 20.0 mg / kg of oven dried pulp, from 0.25 mg / kg of oven dried pulp to 10.0 mg / kg of oven dried pulp, from 0.25 mg / kg of oven dried pulp to 5.0 mg / kg of oven dried pulp, from 0.50 mg / kg of oven dried pulp to 85.0 mg / kg of oven dried pulp, from 0.50 mg / kg of oven dried pulp to 65.0 mg / kg of oven dried pulp, from 0.50 mg / kg of oven dried pulp to 45.0 mg / kg of oven dried pulp, from 0.50 mg / kg of oven dried pulp to 25.0 mg / kg of dried pulp in fo rno, from 0.50 mg / kg of oven dried pulp to 15.0 mg / kg of oven dried pulp and from 0.50 mg / kg of oven dried pulp to 5.0 mg / kg of dried pulp in oven or any combination of these intervals. Method according to any one of claims 1 to 11, characterized in that the method results in reduced viscosity and / or improved viscosity control in the dissolving pulp production process; and / or where the method results in greater reactivity of the dissolving pulp, preferably greater Fock reactivity; and / or where the 4/4 method results in higher content of oxidized groups in the dissolving pulp. Method according to any one of claims 1 to 12, characterized in that the method further comprises subjecting the dissolving pulp to xylanase and / or mannanase and / or lipase and / or laccase and / or peroxidase. 14. Dissolving pulp, characterized by the fact that it is produced by the method, as defined in any one of claims 1 to 13. 15. Textile fiber or derivatized cellulose, characterized by the fact that it is produced from the dissolving pulp, as defined in claim 14. 16. Use of a polysaccharide lytic monoxygenase, characterized by the fact that it is for the treatment of dissolving pulp.