CN112313378A - Method for treating dissolving pulp by using soluble polysaccharide monooxygenase - Google Patents

Abstract

The present invention relates to the treatment of dissolving pulp with a soluble polysaccharide monooxygenase. The treatment with the solubilising polysaccharide monooxygenase leads to a reduction in viscosity and/or an improvement in viscosity control and/or 5 an increase in reactivity of the solubilised pulp during production thereof.

Description

Method for treating dissolving pulp by using soluble polysaccharide monooxygenase

Reference to sequence listing

The present application contains a sequence listing in computer readable form. This computer readable form is incorporated herein by reference.

Technical Field

The present invention relates to the treatment of dissolving pulp with one or more enzymes. The enzymatic treatment results in a reduction of viscosity and/or an improvement of viscosity control during the production of the dissolving pulp and/or an increase of reactivity of the final dissolving pulp.

Background

Commercial dissolving pulp or dissolving grade pulp is chemically bleached pulp having a sufficiently high cellulose content to be suitable for the production of regenerated cellulose and cellulose derivatives. Commercial dissolving pulp has special characteristics such as a high level of brightness and a uniform molecular weight distribution. Commercial dissolving pulp is manufactured for uses requiring high chemical cellulose purity and particularly low hemicellulose content, as chemically similar hemicellulose may interfere with subsequent processes. Dissolving pulp is so named because it is not made into paper, but is dissolved in a solvent or derivatized into a homogeneous solution, which makes it fully chemically accessible and removes any residual fibrous structure. Once dissolved, it can be spun into textile fibers, such as viscose or Lyocell (Lyocell), or chemically reacted to produce derivatized cellulose, such as cellulose triacetate, plastic-like materials forming fibers or films, or cellulose ethers such as methyl cellulose, used as thickeners.

Conventional viscose production using dissolving pulp as raw material requires improvement in its environmental impact and its production cost. There is a need in the art to provide a method for treating dissolving pulp with reduced cost and less environmental impact.

The present invention provides a solution based on a soluble polysaccharide monooxygenase that reduces viscosity and/or improves viscosity control in the production of dissolving pulp (e.g., kraft dissolving pulp and sulfite dissolving pulp). With conventional methods used (these methods are directed to many side reactions such as oxygen, hydrogen peroxide, ozone, hypochlorous acidSodium and acid hydrolysis are not selective) the enzyme solution described in the present invention allows more selective depolymerization of cellulose and thus better control of pulp viscosity. Furthermore, the reactivity of the dissolving pulp according to the invention is improved, whereby the amount of chemicals used in the viscose production process is reduced and/or the processability in the viscose spinning filterability during viscose manufacture is improved. Saving chemicals (such as carbon disulfide (CS)) used in the production of regenerated cellulose in viscose manufacturing processes₂) Amount) will reduce cost and environmental impact. Similarly, it is expected that the increased reactivity of the dissolving pulp is beneficial to the production process of cellulose derivatives, such as the subsequent esterification and etherification processes.

Disclosure of Invention

The present invention provides a method for treating a lysis slurry comprising the step of subjecting the lysis slurry to a solubilising polysaccharide monooxygenase.

Compared to a dissolving pulp obtained by the same process, wherein the dissolving polysaccharide monooxygenase (LPMO) treatment is omitted, the process of the present invention results in a dissolving pulp having a reduced viscosity and/or improved viscosity control during the production of the dissolving pulp, and/or an increased reactivity for viscose manufacture, and/or an increased content of oxidizing groups. The dissolving pulp is kraft dissolving pulp and/or sulfite dissolving pulp.

The invention further provides dissolving pulp prepared by the method of the invention.

The invention further provides textile fibers or derived cellulose made from the dissolving pulp of the invention.

The invention further provides the use of a solubilising polysaccharide monooxygenase for the treatment of solubilised pulp.

Definition of

Dissolving pulp:dissolving pulps are high grade cellulose pulps with low hemicellulose, lignin and resin contents. The pulp has special characteristics such as high level of brightness and uniform molecular weight distribution. It is used to make products including rayon and acetate textile fibers, cellophane, film and various chemical additives. To a large extent, the use of dissolving wood pulp depends on its purity (cellulose)Content) depending mainly on the production process. In order to obtain high quality products, these so-called "specialty" pulps must meet certain requirements, such as high cellulose content, low hemicellulose content, uniform molecular weight distribution, and high cellulose reactivity. Most commercial dissolving pulps meet these requirements to some extent. Nevertheless, due to the compact and complex structure that cellulose presents, it is not easy to obtain high cellulose accessibility and solvent and reagent reactivity. About 77% of the total dissolving pulp is used for the production of cellulose fibres (rayon and acetate).

Two basic processes are used to produce dissolving pulp: (a) a sulfite process; and b) sulfate process (kraft paper).

For the manufacture of dissolving grade pulp, the removal of hemicellulose from wood fibers is crucial, since it affects the filterability of viscose, the xanthation of cellulose and the strength of the final product during viscose production. During wood cooking and subsequent bleaching, hemicellulose is removed. In sulfite pulping, the acidic conditions used are responsible for removing most of the hemicellulose, whereas in the sulfate/kraft process a prehydrolysis step is usually required to remove the hemicellulose. Another method of removing hemicellulose is by treating the pulp with an enzyme that reacts only with the hemicellulose portion of the pulp.

Kraft dissolving pulp: "kraft dissolving pulp" is synonymous with "sulfate dissolving pulp". One preferred example is prehydrolyzed kraft dissolving pulp. Kraft dissolving pulp is produced by digesting wood chips with a solution of sodium hydroxide and sodium sulfide at a temperature above about 120 ℃. Some kraft pulping has also been carried out in which sodium sulfide is enhanced by oxygen or anthraquinone. Kraft pulping is particularly useful for the pulping of softwood, which contains a higher percentage of lignin than hardwood, as compared to soda pulping. The term "kraft dissolving pulp" is synonymous with "kraft dissolving cellulose" and "kraft dissolving grade pulp" and refers to pulp having a high cellulose content. The cellulose content of the kraft dissolving pulp is preferably at least 90% (weight/weight), such as at least 91%, at least 92%, at least 93%, at least 94%, to95% less, at least 96%, at least 97%, at least 98% or at least 99% (w/w). Kraft dissolving pulp is manufactured for uses requiring high chemical purity, and particularly low hemicellulose content. The hemicellulose content of the dissolving pulp is preferably less than 10% (w/w), 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). Kraft dissolving pulp can be used, for example, for the production of regenerated cellulose or for the production of cellulose derivatives. "kraft dissolving grade pulp" may also be defined as pulp that has been sufficiently purified for use in the production of viscose, rayon, cellulose ethers, or cellulose esters with organic or inorganic acids.

Sulfite dissolving pulp: the sulfite process extracts lignin from wood chips in large pressure vessels (called digesters) by using various salts of sulfurous acid to produce wood pulp as nearly pure cellulose fibers. The salt used in the pulping process is Sulfite (SO) depending on the pH₃ ^2-) Or bisulfite (HSO)₃ ^-). The counter ion may be sodium (Na)⁺) Calcium (Ca)²⁺) Potassium (K)⁺) Magnesium (Mg)²⁺) Or ammonium (NH)⁴⁺)。

Sulfite pulping is carried out at a pH between 1.5 and 5 depending on the counter ion of the sulfite (bisulfite) and the ratio of alkali to sulfurous acid. The pulp is contacted with pulping chemicals for 4 to 14 hours and at a temperature in the range of 130 ℃ to 160 ℃ (266 ° F to 320 ° F), also depending on the chemicals used.

Most intermediates involved in delignification in sulfite pulping are resonance-stabilized carbenium ions formed by protonation of carbon-carbon double bonds or acidic cleavage of ether linkages linking many lignin components. The latter reaction is responsible for most of the lignin degradation in the sulfite process. It is expected that the sulfite process will not degrade lignin to the same extent as the kraft process and lignosulfonates from the sulfite process are useful by-products.

Spent cooking liquor from sulfite pulping is commonly referred to as brown liquor, but the terms red liquor, thick liquor and sulfite liquor (as compared to black liquor in the kraft process) are also used. A countercurrent washer is used to remove spent cooking chemicals and degraded lignin and hemicellulose.

Bleaching "Is the removal of color from the pulp, primarily the removal of traces of lignin that remain bound to the fiber after the primary pulping operation. Bleaching typically involves the use of oxidizing agents such as chlorine (C stage), chlorine dioxide (D stage), oxygen (O stage), hydrogen peroxide (P stage), ozone (Z stage), and peracetic acid (CH stage)₃CO₃H; the Paa stage); or treatment with a reducing agent such as sodium dithionite (Y stage). In the presence of chlorine (Cl)₂(ii) a C-stage) processes, such as elemental chlorine-free (ECF) bleaching, in which mainly chlorine dioxide (ClO) is used₂(ii) a A D stage) and is typically followed by an alkaline extraction stage. Totally Chlorine Free (TCF) bleaching is another process in which oxygen based chemicals are mainly used. The pulp bleaching process therefore typically comprises a series of bleaching steps and washes between the bleaching steps to remove degradation products resulting from the bleaching reaction.

Cold Caustic Extraction (CCE):cold caustic extraction, also known as Cold Caustic Extraction (CCE), is a method for removing short-chain non-cellulosic carbohydrates (cellulose purification) based on physical effects such as swelling and dissolution. Typically, the CCE phase is carried out at temperatures below 45 ℃ and very high NaOH doses are used, which can reach values up to 100g/L in the liquid phase. Depending on the pulp consistency in use, this will determine the amount of NaOH per dry weight of pulp. Typical conditions for the CCE stage may be 5% -10% w/w NaOH in the liquid phase for at least 10 min.

Hot Caustic Extraction (HCE): the term "hot caustic extraction" (HCE) is synonymous with "hot caustic extraction". HCE is a process used to remove short-chain hemicellulose and amorphous cellulose from pulp. The Hot Caustic Extraction (HCE) stage is a purification process based on chemical reactions, in particular alkaline stripping of hemicelluloses, which is carried out at higher temperatures and lower NaOH concentrations than CCE.

ISO brightness:ISO brightness is ISO 2470-1 (for measuring pulp and paper)ISO brightness method for sheets and cardboard) which is the intrinsic emissivity [ reflection coefficient ] measured with a reflectometer having the characteristics described in ISO 2469]A factor.

Viscosity of pulp: measured by dissolving the pulp in a suitable cellulose solvent such as Copper Ethylenediamine (CED) and measuring the solution viscosity. This measurement gives an indication of the average degree of polymerisation of the cellulose. This property may 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 T230.

Unbleached or partially bleached or alkaline extracted kraft dissolving pulp: is produced by kraft-based cooking processes such as prehydrolyzed kraft (PHK) cooking, but is not fully bleached and purified until it becomes a commercial kraft dissolving pulp, and thus it is not a finished product. Typically, it has an ISO brightness of less than 90% (e.g. less than 85%, such as less than 80%, such as less than 75%, such as less than 70%, such as less than 65%, such as less than 60%, such as less than 55%, such as less than 50%, such as less than 45%, such as less than 40%, such as less than 35%, and such as less than 30%).

Unbleached or partially bleached or alkaline extracted sulfite dissolving pulp:produced by a sulfite-based cooking process, but is not completely bleached and purified until it becomes a commercial sulfite dissolving pulp, and thus it is not a finished product. Typically, it has an ISO brightness of less than 90% (e.g. less than 85%, such as less than 80%, such as less than 75%, such as less than 70%, such as less than 65%, such as less than 60%, such as less than 55%, such as less than 50%, such as less than 45%, such as less than 40%, such as less than 35%, and such as less than 30%).

Bleached kraft dissolving pulp and bleached sulfite dissolving pulp:produced by a cooking process based on kraft dissolving pulp or sulfite, but not completely bleached and purified until it becomes a commercial dissolving pulp. Typically, it has more than 90% (e.g. more than 91%, e.g. more than 92%, e.g. more than 93%, e.g. more than 94%, e.g. more than 95% >, for exampleSuch as higher than 96%, e.g. higher than 97%, e.g. higher than 98%, e.g. higher than 99%, e.g. higher than 100%).

Sequence identity: the degree of relatedness between two amino acid sequences or between two nucleotide sequences is described by the parameter "sequence identity".

For The purposes of The present invention, The sequence identity between two amino acid sequences is determined using The Needman-Wunsch algorithm (Needman-Wunsch algorithm) (Needleman and Wunsch,1970, J.Mol.biol. [ J.McBiobiol ]48: 443-. The parameters used are the gap opening penalty of 10, the gap extension penalty of 0.5, and the EBLOSUM62 (EMBOSS version of BLOSUM 62) substitution matrix. The output of the "longest identity" of the nidel label (obtained using the non-reduced (-nobrief) option) was used as a percentage of identity and was calculated as follows:

(same residue x 100)/(alignment Length-total number of vacancies in alignment)

Detailed Description

The present invention relates to a method for treating dissolving pulp, comprising the step of subjecting the dissolving pulp to a dissolving polysaccharide monooxygenase (LPMO).

In a preferred embodiment, the method of the invention further comprises the step of subjecting the dissolving pulp to cellulase enzymes.

In a preferred method of the invention, the steps of subjecting the solubilised pulp to the solubilised polysaccharide monooxygenase and subjecting the solubilised pulp to the cellulase are performed simultaneously or sequentially in any order. In a further preferred embodiment of the invention, the solubilised polysaccharide monooxygenase is added to the solubilised pulp together with the cellulase. In a further preferred embodiment of the invention, the solubilised polysaccharide monooxygenase is added to the solubilised pulp before the cellulase enzyme is added. In a further preferred embodiment of the invention, the solubilised polysaccharide monooxygenase is added to the solubilised pulp after the addition of the cellulase enzyme.

In a preferred embodiment, the present method of the invention comprises the step of bleaching the dissolving pulp. In a preferred method of the invention, the step of subjecting the solubilised pulp to the solubilised polysaccharide monooxygenase and the step of bleaching the solubilised pulp are performed simultaneously or sequentially in any order.

In a preferred method of the invention, the step of bleaching the dissolving pulp is performed using a chemical selected from the group consisting of: ClO₂、O₂、O₃、H₂O₂、CH₃CO₃H and NaOCl.

In a preferred embodiment, the process of the invention further comprises a step of alkaline extraction. In the method of the invention, the alkaline extraction is the E, HCE or CCE stage. Specific alkaline purification treatments such as HCE or CCE treatments can produce higher cellulose levels in sulfite and kraft processes. In the case of sulfite pulp, HCE is typically used to further purify the sulfite digested pulp.

In a preferred embodiment, the method of the invention further comprises the step of subjecting the solubilised slurry to solubilising polysaccharide monooxygenase and its electron donor, preferably ascorbic acid, gallic acid, pyrogallol or cysteine. The electron donor may be present in the dissolving pulp to be treated. In one embodiment, no or a small amount of electron donor is added to the dissolving paste. In another embodiment, an effective amount of an electron donor is added to the dissolving paste.

In a preferred method of the invention, the dissolving pulp is unbleached, partially bleached, bleached or alkaline extracted dissolving pulp. In a preferred method of the invention, the dissolving pulp is kraft pulp or sulfite pulp.

In one embodiment, the solubilized polysaccharide monooxygenase added in the method of the invention has at least 60% [ e.g. at least 65%, e.g. at least 70%, e.g. at least 75%, e.g. at least 80%, e.g. at least 85%, e.g. at least 90%, e.g. at least 95%, e.g. at least 99% ] sequence identity with the mature polypeptide of SEQ ID NO:1, the mature polypeptide of SEQ ID NO:2, the mature polypeptide of SEQ ID NO: 3. In one embodiment, the cellulase added in the method of the invention has at least 60% (e.g., at least 65%, e.g., at least 70%, e.g., at least 75%, e.g., at least 80%, e.g., at least 85%, e.g., at least 90%, e.g., at least 95%, e.g., at least 99%) sequence identity to the mature polypeptide of SEQ ID NO. 4, SEQ ID NO. 5, SEQ ID NO. 6, or SEQ ID NO. 7.

In a preferred embodiment, the soluble polysaccharide monooxygenase added in the process of the invention comprises or consists of: 1 or a mature polypeptide thereof, or 2 or a mature polypeptide thereof, or 3 or a mature polypeptide thereof; or the cellulase comprises or consists of: SEQ ID NO. 4, SEQ ID NO. 5 or a mature polypeptide thereof, SEQ ID NO. 6 or a mature polypeptide thereof, or SEQ ID NO. 7 or a mature polypeptide thereof.

In another preferred embodiment, the solubilized polysaccharide monooxygenase added in the method of the invention comprises amino acids 19 to 226 of SEQ ID NO. 1, or a homologous sequence thereof, or an allelic variant thereof, or a functional fragment thereof. In another preferred embodiment, the solubilized polysaccharide monooxygenase added in the method of the invention comprises amino acid amino acids 20 to 254 of SEQ ID NO. 2, or a homologous sequence thereof, or an allelic variant thereof, or a functional fragment thereof. In another preferred embodiment, the solubilized polysaccharide monooxygenase added in the method of the invention comprises amino acids 22 to 249 of SEQ ID NO. 3, or a homologous sequence thereof, or an allelic variant thereof, or a functional fragment thereof. In another preferred embodiment, the cellulase added in the method of the invention comprises SEQ ID No. 4 or its full length, or its homologous sequence, its allelic variant, or a functional fragment thereof. In another preferred embodiment, the cellulase added in the method of the invention comprises amino acids 22-305 of SEQ ID NO. 5, or a homologous sequence thereof, or an allelic variant thereof, or a functional fragment thereof. In another preferred embodiment, the cellulase added in the method of the invention comprises amino acids 22 to 293 of SEQ ID NO. 6, or a homologous sequence thereof, or an allelic variant thereof, or a functional fragment thereof. In another preferred embodiment, the cellulase added in the method of the invention comprises amino acids 19 to 409 of SEQ ID NO. 7, or a homologous sequence thereof, or an allelic variant thereof, or a functional fragment thereof.

The concentration of the soluble polysaccharide monooxygenase added in the process of the invention is preferably from 0.05mg/kg of oven dried pulp to 100000mg/kg of oven dried pulp, for example a concentration selected from the group consisting of: from 0.05mg/kg of oven-dried pulp to 250mg/kg of oven-dried pulp, from 0.25mg/kg of oven-dried pulp to 1000mg/kg of oven-dried pulp, from 0.25mg/kg of oven-dried pulp to 2000mg/kg of oven-dried pulp, from 1.0mg/kg of oven-dried pulp to 5000mg/kg of oven-dried pulp, from 5.0mg/kg of oven-dried pulp to 10000mg/kg of oven-dried pulp, from 10.0mg/kg of oven-dried pulp to 15000mg/kg of oven-dried pulp, from 15.0mg/kg of oven-dried pulp to 20000mg/kg of oven-dried pulp, from 20.0mg/kg of oven-dried pulp to 30000mg/kg of oven-dried pulp, from 30.0mg/kg of oven-dried pulp to 40000mg/kg of oven-dried pulp, from 40.0mg/kg of oven-dried pulp to 60000mg/kg of oven-dried pulp, from 60.0mg/kg of oven-dried pulp to 80000mg/kg of oven-dried pulp, and from 10000mg/kg of oven-dried pulp, or any combination of these intervals.

The concentration of cellulase added in the present invention is from 0.05mg/kg of oven dried pulp to 100mg/kg of oven dried pulp, for example a concentration selected from the group consisting of: from 0.05mg/kg of oven-dried pulp to 80.0mg/kg of oven-dried pulp, from 0.25mg/kg of oven-dried pulp to 60.0mg/kg of oven-dried pulp, from 0.25mg/kg of oven-dried pulp to 40.0mg/kg of oven-dried pulp, from 0.25mg/kg of oven-dried pulp to 20.0mg/kg of oven-dried pulp, from 0.25mg/kg of oven-dried pulp to 10.0mg/kg of oven-dried pulp, from 0.25mg/kg of oven-dried pulp to 5.0mg/kg of oven-dried pulp, from 0.50mg/kg of oven-dried pulp to 85.0mg/kg of oven-dried pulp, from 0.50mg/kg of oven-dried pulp to 65.0mg/kg of oven-dried pulp, from 0.50mg/kg of oven-dried pulp to 45.0mg/kg of oven-dried pulp, from 0.50mg/kg of oven-dried pulp to 25.0mg/kg of oven-dried pulp, from 0.50mg/kg of oven-dried pulp to 15.0mg/kg of oven-dried pulp, from 0.50mg/kg of oven-dried pulp to 5mg/kg of oven-dried pulp, or any combination of these intervals.

The method according to the invention results in improved viscosity control, thereby allowing a reduction of production of dissolving pulp outside the final viscosity specification/target, typically reducing production of off-grade dissolving pulp with respect to viscosity by more than 50% (e.g. more than 60% or more than 70%). In one embodiment, the method according to the invention results in an increase in the reactivity of kraft and/or sulfite dissolving pulp, in particular an increase of at least 10% (e.g. at least 20% or at least 30%) in the reactivity of kraft dissolving pulp.

In preferred embodiments, the method results in a reduction in viscosity and/or an improvement in viscosity control during dissolving pulp production; and/or the method results in an increase in reactivity of the dissolving pulp, preferably Fock reactivity related to viscose manufacturing process, allowing CS savings₂And thus reduce cost and environmental impact; and/or the process results in an increase in the content of oxidizing groups of the dissolving pulp. This increase in oxidizing groups can increase the reactivity of the dissolving pulp, not only in terms of fiber swelling and chemical accessibility, but also in view of the fact that more anchor points (carbonyl and/or carboxyl groups) in the cellulose are available for subsequent derivatization processes in the production of cellulose derivatives.

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

Dissolving pulp produced by the above method is also part of the present invention. Textile fibres or derived cellulose made from the dissolving pulp described above are also part of the invention.

The invention also relates to the use of a solubilized polysaccharide monooxygenase for treating a lysis slurry.

Soluble polysaccharide monooxygenase (LPMO)

The term "soluble polysaccharide monooxygenase" means an enzyme that oxidizes sp (3) carbon in polysaccharides such as chitin, cellulose and starch in the presence of an external electron donor and, according to current speculation, utilizes copper at the active site to activate molecular oxygen. As defined in the database of carbohydrate-active enzymes (http:// www.cazy.org /), those enzymes currently belong to the auxiliary activity families AA9, AA10, AA11, AA13, AA14 and AA 15.

In a first aspect, the LPMO comprises the following motif:

[ ILMV ] -P-x (4,5) -G-x-Y- [ ILMV ] -x-R-x- [ EQ ] -x (4) - [ HNQ ] and [ FW ] - [ TF ] -K- [ AIV ],

wherein x is any amino acid, x (4,5) is any four or five consecutive amino acids, and x (4) is any four consecutive amino acids.

The LPMO comprising the motif shown above may further comprise:

H-x(1,2)-G-P-x(3)-[YW]-[AILMV]、

[ EQ ] -x-Y-x (2) -C-x- [ EHQN ] - [ FILV ] -x- [ ILV ], or

H-x (1,2) -G-P-x (3) - [ YW ] - [ AILMV ] and [ EQ ] -x-Y-x (2) -C-x- [ EHQN ] - [ FILV ] -x- [ ILV ],

wherein x is any amino acid, x (1,2) is any one or two consecutive amino acids, x (3) is any three consecutive amino acids, and x (2) is any two consecutive amino acids.

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

In a second aspect, the LPMO comprises the following motif:

[ILMV]-P-x(4,5)-G-x-Y-[ILMV]-x-R-x-[EQ]-x(3)-A-[HNQ]，

wherein x is any amino acid, x (4,5) is any 4 or 5 consecutive amino acids, and x (3) is any 3 consecutive amino acids. In the above motifs, the accepted IUPAC single letter amino acid abbreviation is used.

In one aspect, the LPMO comprises an amino acid sequence that is identical to SEQ ID NO 1 (Thielavia terrestris), SEQ ID NO 2 (Lentinus silmilis), 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 (Thermomyces thermophila 18 (Thermomyces terrestris), SEQ ID NO 19 (myceliophthora thermophila), SEQ ID NO 20 (myceliophthora thermophila), 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 27 (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), and SEQ ID NO 33 (Thielavia terrestris), The mature polypeptide of SEQ ID NO:34 (Thielavia terrestris), SEQ ID NO:35 (Thielavia terrestris), SEQ ID NO:36 (Thermoascus crustus), SEQ ID NO:37 (Thermoascus), or SEQ ID NO:38 (Thermoascus) has at least 50%, such as 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% sequence identity.

In another aspect, the LPMO is an artificial variant comprising 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 19, SEQ ID NO 32, SEQ ID NO 33, SEQ ID NO 25, SEQ ID NO 17, SEQ ID NO 19, 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, 35, 36, 37, or 38; or a homologous sequence thereof, an allelic variant thereof, or a functional fragment thereof.

Preferably, the amino acid change has a minor property, i.e., a conservative amino acid substitution or insertion that does not significantly affect the folding and/or activity of the protein; typically a small deletion of one to about 30 amino acids; small amino-terminal or carboxy-terminal extensions, such as an amino-terminal methionine residue; a small linker peptide of up to about 20-25 residues; or a small extension that facilitates purification by altering the net charge or another function (such as a polyhistidine segment, an epitope, or a binding domain).

Examples of conservative substitutions are within the following groups: 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 which do not generally alter specific activity are known in The art and are described, for example, by H.Neurath and R.L.Hill,1979, in The Proteins, Academic Press, N.Y.. The most commonly occurring exchanges are Ala/Ser, Val/Ile, Asp/Glu, Thr/Ser, Ala/Gly, 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.

Alternatively, the amino acid changes have a property that: altering the physicochemical properties of the polypeptide. For example, amino acid changes can improve the thermostability of the polypeptide, change substrate specificity, change the pH optimum, and the like.

The essential amino acids in a parent polypeptide can be identified according to procedures known in the art, such as site-directed mutagenesis or alanine-scanning mutagenesis (Cunningham and Wells,1989, Science [ Science ]244: 1081-1085). In the latter technique, a single alanine mutation is introduced in each residue in 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. [ J.Biol ]271: 4699-4708. The active site of an enzyme or other biological interaction can also be determined by physical analysis of the structure, as determined by the following technique: nuclear magnetic resonance, crystallography, electron diffraction, or photoaffinity labeling, along with mutating putative contact site (contact site) amino acids. See, e.g., de Vos et al, 1992, Science [ Science ]255: 306-); smith et al, 1992, J.mol.biol. [ J.Mol.224: 899-); wlodaver et al, 1992, FEBS Lett. [ Provisions of the European Association of biochemistry ]309: 59-64. The identity of the essential amino acids can also be inferred from an identity analysis of the polypeptide with which the parent polypeptide is associated.

Single or multiple amino acid substitutions, deletions and/or insertions can be made and tested using known mutagenesis, recombination and/or shuffling methods, followed by relevant screening procedures such as those described by Reidhaar-Olson and Sauer,1988, Science [ Science ]241: 53-57; bowie and Sauer,1989, Proc. Natl. Acad. Sci. USA [ Proc. Natl. Acad. Sci. ]86: 2152-2156; WO 95/17413; or those disclosed in WO 95/22625. Other methods that can be used include error-prone PCR, phage display (e.g., Lowman et al, 1991, Biochemistry [ Biochemistry ]30: 10832-.

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

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, SEQ ID NO 10, 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 32, Or the total number of amino acid substitutions, deletions and/or insertions of the mature LPMO of SEQ ID NO. 38 is NO more than 10, such as 1,2, 3,4,5, 6,7,8, 9 or 10.

In one aspect, the LPMO is used in the presence of a soluble activated divalent metal cation (e.g., copper sulfate) as described in WO 2008/151043.

In one aspect, the LPMO is used in the presence of its electron donor. The electron donor may be present in the dissolving pulp to be treated. In one embodiment, no or a small amount of electron donor may be added to the dissolving paste. In another embodiment, an effective amount of an electron donor may be added to the dissolving paste.

In the present invention, the electron donor may be a dioxy compound, a bicyclic compound, a heterocyclic compound, a nitrogen-containing compound, or a sulfur-containing compound.

The dioxy compounds may include any suitable compound containing two or more oxygen atoms. In some aspects, the dioxy compounds contain substituted aryl moieties as described herein. The dioxy compounds may contain one or more (several) hydroxyl groups and/or hydroxyl derivatives and include substituted aryl moieties lacking hydroxyl groups and hydroxyl derivatives. Non-limiting examples of dioxy compounds include catechol 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; sinapic acid; 3, 5-dihydroxybenzoic acid; 4-chloro-1, 2-benzenediol; 4-nitro-1, 2-benzenediol; tannic acid; 4, gallic acid ethyl ester; methyl glycolate; dihydroxy fumaric acid; 2-butyne-1, 4-diol; (croconic acid; 1, 3-propanediol; tartaric acid; 2, 4-pentanediol; 3-ethoxy-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 salts or solvates thereof.

The bicyclic compound can include any suitable substituted fused ring system as described herein. The compounds may contain one or more (several) additional rings and are not limited to a specific number of rings unless otherwise specified. In one 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 anthocyanin ion (flavylion), such as an optionally substituted anthocyanidin or an optionally substituted anthocyanin, or derivatives thereof. Non-limiting examples of bicyclic compounds include epicatechin; (ii) quercetin; myricetin; taxifolin; kaempferol; morin; robinia pseudoacacia extract; naringenin; isorhamnetin; apigenin; cyanidin; cyanidin glycoside; black bean polyphenol; anthocyanins rhamnoside; or a salt or solvate thereof.

The heterocyclic compound may be any suitable compound as described herein, such as an optionally substituted aromatic or non-aromatic ring comprising a heteroatom. In one aspect, the heterocycle is a compound comprising an optionally substituted heterocycloalkyl moiety or an optionally substituted heteroaryl moiety. In another aspect, the optionally substituted heterocycloalkyl moiety or optionally substituted heteroaryl moiety is an optionally substituted 5-membered heterocycloalkyl or optionally substituted 5-membered heteroaryl moiety. In another aspect, the optionally substituted heterocycloalkyl or optionally substituted heteroaryl moiety is an optionally substituted moiety selected from the group consisting of: pyrazolyl, furyl, imidazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrrolyl, pyridyl, pyrimidinyl, pyridazinyl, thiazolyl, triazolyl, thienyl, dihydrothienopyrazolyl, thianaphthenyl, carbazolyl, benzimidazolyl, benzothienyl, benzofuranyl, indolyl, quinolyl, benzotriazolyl, benzothiazolyl, benzoxazolyl, benzimidazolyl, isoquinolyl, isoindolyl, acridinyl, benzisoxazolyl, dimethylhydantoin, pyrazinyl, tetrahydrofuryl, pyrrolinyl, pyrrolidinyl, morpholinyl, indolyl, diazepine, azepinyl, thiepinyl, piperidyl and oxazepinyl. In another aspect, the optionally substituted heterocycloalkyl moiety or optionally substituted heteroaryl moiety is an optionally substituted furyl. Non-limiting examples of heterocyclic compounds include (1, 2-dihydroxyethyl) -3, 4-dihydroxyfuran-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; ribono gamma-lactone; hexuronic acid (aldohexuronaldheuronic acid) γ -lactone; glucono delta-lactone; 4-hydroxycoumarin; dihydrobenzofuran; 5- (hydroxymethyl) furfural; bi-furfural; 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.

The nitrogen-containing compound may be any suitable compound having one or more nitrogen atoms. In one aspect, the nitrogen-containing compound comprises an amine, imine, hydroxylamine, or nitroxide moiety. Non-limiting examples of nitrogen-containing compounds include acetoxime; purple uric acid; pyridine-2-aldoxime; 2-aminophenol; 1, 2-phenylenediamine; 2,2,6, 6-tetramethyl-1-piperidinyloxy; 5,6,7, 8-tetrahydrobiopterin; 6, 7-dimethyl-5, 6,7, 8-tetrahydropterin; and maleic acid amide; or a salt or solvate thereof.

The quinone compound can be any suitable compound comprising a quinone moiety 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(ii) a2, 3,5, 6-tetramethyl-1, 4-benzoquinone or duroquinone; 1, 4-dihydroxyanthraquinone; 3-hydroxy-1-methyl-5, 6-indolinedione or adrenaline red; 4-tert-butyl-5-methoxy-1, 2-benzoquinone; pyrroloquinoline quinone; or a salt or solvate thereof.

The sulfur-containing compound may be any suitable compound comprising one or more sulfur atoms. In one aspect, the sulfur comprises a moiety selected from the group consisting of: sulfinyl, thioether, sulfinyl, sulfonyl, sulfamide, sulfonamide, sulfonic acid, and sulfonate ester. Non-limiting examples of sulfur-containing compounds include ethanethiol; 2-propanethiol; 2-propene-1-thiol; 2-mercaptoethanesulfonic acid; thiophenol; benzene-1, 2-dithiol; (ii) cysteine; methionine; glutathione; cystine; or a salt or solvate thereof.

Cellulase enzymes

Cellulases or cellulolytic enzymes are enzymes involved in the hydrolysis of cellulose. In the hydrolysis of native cellulose, three main types of cellulases are known to be involved, namely cellobiohydrolases (1,4- β -D-glucan cellobiohydrolase, EC 3.2.1.91, e.g. cellobiohydrolase I and cellobiohydrolase II), endo- β -1, 4-glucanases (endo-1,4- β -D-glucan 4-glucanohydrolase, EC3.2.1.4) and β -glucosidase (EC 3.2.1.21).

To be effective, the digestion of cellulose and hemicellulose may require several types of enzymes to act together. At least three classes of enzymes are necessary for converting cellulose to fermentable sugars: endoglucanases that cleave cellulose chains randomly (EC 3.2.1.4); cellobiohydrolases (EC 3.2.1.91) which cleave cellobiose units from the ends of cellulose chains and beta-glucosidases (EC 3.2.1.21) which convert cellobiose and soluble cellodextrins to glucose. Among the three enzymes involved in the biodegradation of cellulose, cellobiohydrolases are key enzymes in the degradation of natural crystalline cellulose. The term "cellobiohydrolase I" is defined herein as a cellulose 1, 4-beta-cellobiosidase (also referred to as exoglucanase, exo-cellobiohydrolase or 1, 4-beta-cellobiohydrolase) activity, as defined in enzyme class EC 3.2.1.91, which catalyzes the hydrolysis of 1, 4-beta-D-glucoside bonds in cellulose and cellotetraose by releasing cellobiose from the non-reducing end of the chain. The term "cellobiohydrolase II activity" is defined identically, except that cellobiohydrolase II attacks from the reducing end of the chain.

Endoglucanases (EC No. 3.2.1.4) catalyse the endo-hydrolysis of β -1,4 linkages in cellulose, cellulose derivatives (such as carboxymethyl cellulose and hydroxyethyl cellulose), 1,4- β -D-glycosidic linkages in lichen starch, mixed β -1,3 glucans such as cereal β -D-glucans or xyloglucans and other plant materials comprising a cellulose fraction. The name examined is endo-1, 4-beta-D-glucan 4-glucan hydrolase (endo-1, 4-beta-D-glucan 4-glucan hydrosase), but the abbreviated term endoglucanase is used in this specification. In a preferred embodiment of the invention, the cellulase used in the invention is an endoglucanase.

Cellulases may comprise carbohydrate-binding modules (CBMs) that enhance the binding of the enzyme to cellulose-containing fibers and increase the efficacy of the enzyme-catalytically active moiety. CBM is defined as a contiguous amino acid sequence in a carbohydrate-active enzyme that has a discreet (discreet) fold of carbohydrate-binding activity. For further information on CBM, see CAZy Internet Server (supra) or Tomme et al (1995) in enzymic Degradation of Insoluble Polysaccharides (Saddler, J.N. and Penner, eds.), cell-binding domains: classification and properties [ Cellulose binding domains: classification and Properties page 142-163, American Chemical Society, Washington.

In a preferred embodiment, the cellulase may be a formulation as defined in WO 2008/151079 (which is hereby incorporated by reference). The cellulase preparation may further comprise a beta-glucosidase, such as a fusion protein as disclosed in US 60/832,511. In an embodiment, the cellulase preparation further comprises CBH II, preferably thielaviopsis terrestris cellobiohydrolase II CEL 6A. In an embodiment, the cellulase preparation further comprises a cellulase preparation, preferably a cellulase preparation derived from trichoderma reesei. Cellulases can be synthesized by a large number of microorganisms including fungi, actinomycetes, slime molds and eubacteria, and also by plants. In particular, endoglucanases of various specificities have been identified.

In a preferred embodiment, the cellulase activity may be derived from a fungal source, such as a Trichoderma (Trichoderma) strain, preferably a Trichoderma reesei strain; humicola (Humicola) strains, such as Humicola insolens (Humicola insolens) strains; or a Chrysosporium (Chrysosporium) strain, preferably a Chrysosporium lucknowense strain, or a Thielavia strain, preferably Thielavia terrestris.

In one aspect, the cellulase comprises an amino acid sequence having at least 50%, e.g., 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% sequence identity to the mature polypeptide of SEQ ID NO. 4,5, 6, or 7. In another aspect, the cellulase is an artificial variant comprising a substitution, deletion, and/or insertion of one or more (or several) 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 a homologous sequence thereof, an allelic variant thereof, or a functional fragment thereof. 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 does not exceed 10, such as 1,2, 3,4,5, 6,7,8, 9 or 10.

Fungi and bacteria produce a series of cellulolytic enzymes (cellulases) that can be divided into different glycosyl hydrolase families based on sequence similarity (hydrophobic cluster analysis) [ Henrissat B and Bairoch a; j. [ J. Biochem.1993293781-788 ]. Cellulases belonging to families 5,6,7,8, 9, 10, 12, 26, 44, 45, 48, 60 and 61 of glycosyl hydrolases are known at present.

Additional enzymes

Any enzyme having xylanase, mannanase, lipase, laccase, and/or peroxidase activity may be used as the additional enzyme in the uses and methods of the invention. The additional enzyme and the soluble polysaccharide monooxygenase are added simultaneously or sequentially in any order. Some non-limiting examples of such additional enzymes are listed below. Enzymes written in uppercase letters are commercial enzymes available from Novozymes corporation (Novozymes A/S), Crogshoejvej 36 (Krogshoejej 36), DK-2880 Baggesveld (DK-2880Bagsvaerd), Denmark (Denmark). The activity of any of those additional enzymes can be assayed using any method known in the art for the enzyme in question, including the methods mentioned in the cited references.

An example of a xylanase is PULPZYME HC hemicellulase.

Examples of mannanases are

Et al, J.Biotechnol. [ J.Biotech journal of Biotechnology ]]29(1993), 229-242.

An example of a lipase is RESINASE A2X lipase. An example of a xylanase is PULPZYME HC hemicellulase.

Examples of peroxidases and laccases are disclosed in EP 730641; WO 01/98469; EP 719337; EP 765394; EP 767836; EP 763115; and EP 788547. Such compounds are considered to be included herein whenever reference is made to a peroxidase or laccase that requires or benefits from the presence of a receptor (e.g., oxygen or hydrogen peroxide), an enhancer, a mediator, and/or an activator. 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 by defining a laccase or peroxidase system as a combination of the enzyme in question with its receptor, and optionally also with an enhancer and/or mediator for the enzyme in question.

Temperature for the process of the invention

The temperature used in the process of the invention is typically from 20 ℃ to 100 ℃, such as a temperature interval selected from the group consisting of: from 20 ℃ to 30 ℃, from 30 ℃ to 40 ℃, from 40 ℃ to 50 ℃, from 50 ℃ to 60 ℃, from 60 ℃ to 70 ℃, from 70 ℃ to 80 ℃, from 80 ℃ to 90 ℃, from 90 ℃ to 100 ℃, or any combination of these intervals.

Incubation time for the method of the invention

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

Pulp used and produced in the process according to the invention:

dissolving pulp for use in the present invention may be wood pulp, for example derived from softwood trees (such as spruce, pine, fir, larch and hemlock) and/or hardwood (such as eucalyptus, poplar and birch) or other plant sources such as bamboo.

In a preferred embodiment, the dissolving pulp is selected from the group consisting of: dissolving hardwood pulp and dissolving softwood pulp, or mixtures thereof.

In one embodiment, the present invention relates to dissolving pulp made by the method according to the present invention. In one embodiment, the invention relates to kraft or sulfite dissolving pulp produced by the method according to the invention.

The invention further relates to the use of the dissolving pulp according to the invention for producing textile fibres. The dissolving pulp produced can be used for the manufacture of regenerated cellulose, such as viscose, rayon, lyocell and modal fibres.

The process of the invention is carried out in the presence of one or more surfactants

The process of the present invention may be carried out in the presence of one or more surfactants, such as one or more anionic surfactants, and/or one or more nonionic surfactants, and/or one or more cationic surfactants.

In one embodiment, the surfactant may include a poly (alkylene glycol) based surfactant, an ethoxylated dialkyl phenol, an ethoxylated alcohol, and/or a silicone based surfactant.

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

Specific presently preferred anionic surfactant materials useful in the practice of the present invention comprise, for example, those available under the trade name ALPHA-STEP^TM-ML40 commercially available methyl ester of alpha-sodium sulfonate laurate (which may include some ethyl ester of alpha-sulfolaurate); for example, it can be used under the trade name STEPANATE^TM-X sodium xylene sulfonate commercially available; for example, it can be used under the trade name STEPANOL^TM-WAT commercially available triethanolammonium lauryl sulfate; for example, available under the trade name STEPAN^TM-disodium lauryl sulfosuccinate available from Mild SL 3; further blends of various anionic surfactants, such as the aforementioned ALPHA-STEP, may also be used^TMAnd STEPANATE^TM50% -50% or 25% -75% of the material, or the above ALPHA-STEP^TMAnd STEPANOL^TM20% -80% blend of materials (all of the aforementioned commercially available materials can be obtained from Stepan Company Spandex]Obtained by norsfield, il.).

A specific presently preferred nonionic surfactant material useful in the practice of the present invention comprises cocoa diethanolamide, such as may be available under the NINOL tradename^TM-11CM commercially available; alkyl polyoxyalkylene glycol ethers, e.g. from Stepan Company]May be given the trade name TOXIMUL^TM8320 commercially available higher molecular weight butyl ethylene oxide-propylene oxide block copolymer. Additional alkyl polyoxyalkylene glycol ethers may be selected, for example, as disclosed in U.S. Pat. No. 3,078,315. Blends of various nonionic surfactants may also be used, such as the NINOL described previously^TMAnd TOXIMUL^TM50-50% or 25-75% of the materialA compound (I) is provided.

Specific currently preferred anionic/nonionic surfactant blends for use in the practice of the present invention include various mixtures of the foregoing materials, such as the aforementioned ALPHA-STEP^TMAnd NINOL^TM50% -50% blend of materials, or stephanates as described previously^TMAnd TOXIMUL^TM25% -75% blend of materials.

Preferably, the various anionic, nonionic and anionic/nonionic surfactant blends used in the practice of the present invention have a solids or active content of up to about 100% by weight, and preferably have an active content in the range of from about 10% to about 80%. Of course, other blends or other solids (actives) levels may also be used, and these anionic surfactants, nonionic surfactants, and mixtures thereof may also be used with known pulping chemicals such as, for example, anthraquinone and its derivatives, and/or other typical papermaking chemicals such as caustic, defoamers, and the like.

The invention described and claimed herein is not to be limited in scope by the specific embodiments herein disclosed, since these embodiments are intended as illustrations of several aspects of the invention. Any equivalent embodiments are intended to be within the scope of the present invention. Indeed, various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. In case of conflict, the present disclosure, including definitions, will control.

Various references are cited herein, the disclosure of which is incorporated by reference herein in its entirety.

Examples of the invention

Materials and methods

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

Pulp viscosity was measured by mViPr according to WO 2011/107472 a 9.

The amount of aldehyde groups (CHO content) based on the reaction of 2,3, 5-triphenyltetrazolium chloride (TTC) with aldehyde groups, leading to the formation of formazan (red colorant) was measured spectrophotometrically according to the procedure described by Obolnskaya et al, "Determination of aldehyde groups in oxidized pulps ]", Laboratory management in Wood and Cellulose Chemistry, Ecologia [ ecology ], Mosco, 211-212, 1991.

Fock reactivity is a known measure of how much pulp is present with CS as a small scale simulation of the viscose manufacturing process₂The magnitude of the reaction, and it was performed at 9% NaOH.

Example 1: LPMO treatment of unbleached hardwood kraft dissolving pulp

Unbleached hardwood kraft dissolving pulp having a kappa number of 6.8(TAPPI T236 procedure), an ISO brightness of 51%, and an intrinsic viscosity of 1025mL/g, produced by a prehydrolysis kraft pulping process and further treated with cold caustic extraction stage from the dissolving pulp production process, was used. The pulp was treated with several LPMO in a small scale assay for 20 hours at 0.4% consistency, 45 ℃ and pH 5.0 (acetate buffer, 50mM) using 24mg of oven dried fiber. The enzymatic treatment (denoted X stage) was carried out at a dose of 5mg EP (enzyme protein)/g odp (dried pulp).

The pulp suspension with a consistency of 0.4% was disintegrated with a magnetic bar in a glass test tube placed in a heated block at 45 ℃. Ascorbic acid, gallic acid or pyrogallol as electron donor was added to a final concentration of 1mM in suspension followed by the addition of LPMO enzyme to a final volume of 6 mL. After an incubation time of 20 hours after enzyme addition, the tubes were cooled in ice, and then 6mL of Copper Ethylenediamine (CED) was added to dissolve the fibers. The pulp was dissolved in a rotary stirrer at room temperature of 25 ℃ for 25 min. Control experiments were performed in the same manner, but without the addition of enzyme.

After the dissolution time, the pulp viscosity was measured by mViPr. The mviprr pipette consists of a modified Gilson Concept C300 pipette equipped with a pressure sensor and a Diamond D300 Gilson tip. The samples were kept at a constant temperature within ± 0.1 ℃. A volume of 200 μ L of the lysis slurry was aspirated and dispensed into and out of the pipette, respectively, while recording the pressure in the pipette headspace. The applied pipette speed was 4. There was a 2s delay after aspiration and a 5s delay after dispensing. Each sample measurement consisted of 15 aspirate-dispense cycles and the pressure results were the average of 15 aspirate or dispense pressures, respectively.

Table 1 presents the suction pressure results from the mviprr measurements. It can be seen that LPMO can reduce the average size of cellulose, expressed as a decrease in solution viscosity as measured by a decrease in suction pressure. The performance of each LPMO also depends on the particular electron donor used. Furthermore, the electron donor (particularly ascorbic acid) itself also degrades cellulose compared to the original pulp. Tt LPMO is very effective in reducing the viscosity of the pulp with all electron donors.

TABLE 1 suction pressure for different unbleached pulps dissolved in CED

Example 2: LPMO treatment of bleached hardwood kraft dissolving pulp

An acetate grade bleached hardwood kraft never-dried pulp produced by a prehydrolysis kraft pulping process having an ISO brightness of 93.7% and an intrinsic viscosity of 684mL/g was used. The pulp was treated with several LPMOs with three different electron donors using the same conditions and procedure as in example 1.

Table 2 presents the suction pressure results from mviprr measurements for bleached dissolving pulp. LPMO can reduce the average size of cellulose molecules, expressed as a decrease in solution (dissolving pulp) viscosity as measured by a decrease in suction pressure. When LPMO is used with gallic acid and pyrogallol, a higher reduction in bleached pulp viscosity is achieved compared to the control (no enzyme) compared to ascorbic acid, which itself also significantly reduces viscosity. Tt LPMO is LPMO more powerful in viscosity reduction among all electron donors used.

TABLE 2 suction pressure of different bleached pulps dissolved in CED

Example 3: effect of LPMO treatment with/without endoglucanase on bleached dissolving pulp viscosity, reactivity and CHO content at moderate pulp consistencies

Bleached hardwood kraft dissolving pulp produced by the viscose grade prehydrolysis kraft pulping process was used with an intrinsic viscosity of 512 mL/g. The pulp was treated with several LPMO in a Distek vessel (Distek model Symphony 7100) using gallic acid (1mM) as an electron donor at 1.5% consistency, with heating and continuous overhead stirring.

Once the pulp is disintegrated and at the temperature set point of 45 ℃, gallic acid is added followed by the addition of enzymes (LPMO and/or endoglucanase). The pulp was enzymatically treated at pH 5.0 (acetate buffer, 50mM) using 2mg EP/g odp of LPMO and 1.2mg EP/kg odp of endoglucanase for 25.5 hours. After the enzyme treatment, the pulp was filtered and washed with 1L of tap water in three successive steps. A portion of the pulp sample was dried before measuring CHO content and Fock reactivity, and a portion of the sample was kept wet in a refrigerator to test intrinsic viscosity.

It can be seen in table 3 that a smaller reduction in viscosity reduction can be obtained with the LPMO tested at a lower dose of 2mg EP/g odp compared to example 1 and example 2 with a higher dose of LPMO. However, when combined with endoglucanases, the viscosity reduction is enhanced and has a surprising synergistic effect. The same synergy between LPMO and endoglucanase was seen in Fock reactivity and the amount of CHO groups.

TABLE 3 intrinsic viscosity, Fock reactivity and CHO content of bleached dissolving pulp

Example 4: effect of combined LPMO and endoglucanase treatment on unbleached hardwood kraft dissolving pulp

The pulp was dissolved using unbleached hardwood kraft paper as in example 1 and treated with 24mg of oven dried fiber at 0.4% consistency, 50 ℃, pH 6.0 (phosphate buffered saline, 50mM) for 20 hours using several LPMOs with gallic acid (1mM) as electron donor in combination with several endoglucanases in a small scale assay. LPMO treatment was carried out at a dose of 2.5mg EP (enzyme protein)/g odp (high dose) or 0.5mg EP/g odp (low dose). The endoglucanase treatment was performed at a dose of 0.5mg EP/kg odp. The determinations and viscosity measurements were performed using the same conditions and procedures as described in example 1.

Table 4 presents the suction pressure results from the mviprr measurements. It can be seen that the higher the dose of LPMO, the higher the reduction in pulp viscosity. The high dose of LPMO unexpectedly gave higher viscosity reduction compared to the endoglucanase treatment. The treatment with the combination of LPMO and endoglucanase showed better viscosity reducing performance than the treatment with LPMO or endoglucanase alone. With such high viscosity pulps, a promoting effect on the viscosity reduction by the combined use of LPMO and endoglucanase was observed, which allows to significantly reduce the amount of LPMO required. For example, at low TtLPMO doses (0.5mg EP/g odp), the co-addition of endoglucanases 2 and 3 had an even higher viscosity reduction than the addition of TtLPMO alone at high doses (2.5mg EP/g odp).

TABLE 4 suction pressure for different unbleached pulps dissolved in CED

Sequence listing

<110> Novozymes corporation (Novozymes A/S)

<120> method for treating dissolving pulp

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<150> EP18175341.9

<151> 2018-05-31

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<170> PatentIn 3.5 edition

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<212> PRT

<213> Thielavia terrestris

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Met Leu Ala Asn Gly Ala Ile Val Phe Leu Ala Ala Ala Leu Gly Val

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Ser Gly His Tyr Thr Trp Pro Arg Val Asn Asp Gly Ala Asp Trp Gln

20 25 30

Gln Val Arg Lys Ala Asp Asn Trp Gln Asp Asn Gly Tyr Val Gly Asp

35 40 45

Val Thr Ser Pro Gln Ile Arg Cys Phe Gln Ala Thr Pro Ser Pro Ala

50 55 60

Pro Ser Val Leu Asn Thr Thr Ala Gly Ser Thr Val Thr Tyr Trp Ala

65 70 75 80

Asn Pro Asp Val Tyr His Pro Gly Pro Val Gln Phe Tyr Met Ala Arg

85 90 95

Val Pro Asp Gly Glu Asp Ile Asn Ser Trp Asn Gly Asp Gly Ala Val

100 105 110

Trp Phe Lys Val Tyr Glu Asp His Pro Thr Phe Gly Ala Gln Leu Thr

115 120 125

Trp Pro Ser Thr Gly Lys Ser Ser Phe Ala Val Pro Ile Pro Pro Cys

130 135 140

Ile Lys Ser Gly Tyr Tyr Leu Leu Arg Ala Glu Gln Ile Gly Leu His

145 150 155 160

Val Ala Gln Ser Val Gly Gly Ala Gln Phe Tyr Ile Ser Cys Ala Gln

165 170 175

Leu Ser Val Thr Gly Gly Gly Ser Thr Glu Pro Pro Asn Lys Val Ala

180 185 190

Phe Pro Gly Ala Tyr Ser Ala Thr Asp Pro Gly Ile Leu Ile Asn Ile

195 200 205

Tyr Tyr Pro Val Pro Thr Ser Tyr Gln Asn Pro Gly Pro Ala Val Phe

210 215 220

Ser Cys

225

<210> 2

<211> 254

<212> PRT

<213> shiitake mushroom

<400> 2

Met Lys Tyr Ser Ile Leu Gly Leu Thr Ala Leu Ser Phe Val Ala Ser

1 5 10 15

Ala Ala Ala His Thr Leu Val Trp Gly Val Trp Val Asn Gly Val Asp

20 25 30

Gln Gly Asp Gly Arg Asn Ile Tyr Ile Arg Ser Pro Pro Asn Asn Asn

35 40 45

Pro Val Lys Asn Leu Thr Ser Pro Asp Met Thr Cys Asn Val Asp Asn

50 55 60

Arg Val Val Pro Lys Ser Val Pro Val Asn Ala Gly Asp Thr Leu Thr

65 70 75 80

Phe Glu Trp Tyr His Asn Thr Arg Asp Asp Asp Ile Ile Ala Ser Ser

85 90 95

His His Gly Pro Ile Ala Val Tyr Ile Ala Pro Ala Ala Ser Asn Gly

100 105 110

Gln Gly Asn Val Trp Val Lys Leu Phe Glu Asp Ala Tyr Asn Val Thr

115 120 125

Asn Ser Thr Trp Ala Val Asp Arg Leu Ile Thr Ala His Gly Gln His

130 135 140

Ser Val Val Val Pro His Val Ala Pro Gly Asp Tyr Leu Phe Arg Ala

145 150 155 160

Glu Ile Ile Ala Leu His Glu Ala Asp Ser Leu Tyr Ser Gln Asn Pro

165 170 175

Ile Arg Gly Ala Gln Phe Tyr Ile Ser Cys Ala Gln Ile Thr Ile Asn

180 185 190

Ser Ser Asp Asp Ser Thr Pro Leu Pro Ala Gly Val Pro Phe Pro Gly

195 200 205

Ala Tyr Thr Asp Ser Thr Pro Gly Ile Gln Phe Asn Ile Tyr Thr Thr

210 215 220

Pro Ala Thr Ser Tyr Val Ala Pro Pro Pro Ser Val Trp Ser Gly Ala

225 230 235 240

Leu Gly Gly Ser Ile Ala Gln Val Gly Asp Ala Ser Leu Glu

245 250

<210> 3

<211> 249

<212> PRT

<213> Thermoascus aurantiacus

<400> 3

Met Ser Phe Ser Lys Ile Ile Ala Thr Ala Gly Val Leu Ala Ser Ala

1 5 10 15

Ser Leu Val Ala Gly His Gly Phe Val Gln Asn Ile Val Ile Asp Gly

20 25 30

Lys Asn Tyr Gly Gly Tyr Leu Val Asn Gln Tyr Pro Tyr Met Ser Asn

35 40 45

Pro Pro Glu Val Ile Ala Trp Ser Thr Thr Ala Thr Asp Leu Gly Phe

50 55 60

Val Asp Gly Thr Gly Tyr Gln Thr Pro Asp Ile Ile Cys His Arg Gly

65 70 75 80

Ala Lys Pro Gly Ala Leu Thr Ala Pro Val Ser Pro Gly Gly Thr Val

85 90 95

Glu Leu Gln Trp Thr Pro Trp Pro Asp Ser His His Gly Pro Val Ile

100 105 110

Asn Tyr Leu Ala Pro Cys Asn Gly Asp Cys Ser Thr Val Asp Lys Thr

115 120 125

Gln Leu Glu Phe Phe Lys Ile Ala Glu Ser Gly Leu Ile Asn Asp Asp

130 135 140

Asn Pro Pro Gly Ile Trp Ala Ser Asp Asn Leu Ile Ala Ala Asn Asn

145 150 155 160

Ser Trp Thr Val Thr Ile Pro Thr Thr Ile Ala Pro Gly Asn Tyr Val

165 170 175

Leu Arg His Glu Ile Ile Ala Leu His Ser Ala Gln Asn Gln Asp Gly

180 185 190

Ala Gln Asn Tyr Pro Gln Cys Ile Asn Leu Gln Val Thr Gly Gly Gly

195 200 205

Ser Asp Asn Pro Ala Gly Thr Leu Gly Thr Ala Leu Tyr His Asp Thr

210 215 220

Asp Pro Gly Ile Leu Ile Asn Ile Tyr Gln Lys Leu Ser Ser Tyr Ile

225 230 235 240

Ile Pro Gly Pro Pro Leu Tyr Thr Gly

245

<210> 4

<211> 278

<212> PRT

<213> Artificial

<220>

<223> Q120H variant of mature endoglucanase shown as SEQ ID NO:9 of WO 96/29397

<400> 4

Ala Ser Gly Ser Gly Gln Ser Thr Arg Tyr Trp Asp Cys Cys Lys Pro

1 5 10 15

Ser Cys Ala Trp Pro Gly Lys Ala Ala Val Ser Gln Pro Val Tyr Ala

20 25 30

Cys Asp Ala Asn Phe Gln Arg Leu Ser Asp Phe Asn Val Gln Ser Gly

35 40 45

Cys Asn Gly Gly Ser Ala Tyr Ser Cys Ala Asp Gln Thr Pro Trp Ala

50 55 60

Val Asn Asp Asn Leu Ala Tyr Gly Phe Ala Ala Thr Ser Ile Ala Gly

65 70 75 80

Gly Ser Glu Ser Ser Trp Cys Cys Ala Cys Tyr Ala Leu Thr Phe Thr

85 90 95

Ser Gly Pro Val Ala Gly Lys Thr Met Val Val Gln Ser Thr Ser Thr

100 105 110

Gly Gly Asp Leu Gly Ser Asn His Phe Asp Ile Ala Met Pro Gly Gly

115 120 125

Gly Val Gly Ile Phe Asn Gly Cys Ser Ser Gln Phe Gly Gly Leu Pro

130 135 140

Gly Ala Gln Tyr Gly Gly Ile Ser Ser Arg Asp Gln Cys Asp Ser Phe

145 150 155 160

Pro Ala Pro Leu Lys Pro Gly Cys Gln Trp Arg Phe Asp Trp Phe Gln

165 170 175

Asn Ala Asp Asn Pro Thr Phe Thr Phe Gln Gln Val Gln Cys Pro Ala

180 185 190

Glu Ile Val Ala Arg Ser Gly Cys Lys Arg Asn Asp Asp Ser Ser Phe

195 200 205

Pro Val Phe Thr Pro Pro Ser Gly Gly Asn Gly Gly Thr Gly Thr Pro

210 215 220

Thr Ser Thr Ala Pro Gly Ser Gly Gln Thr Ser Pro Gly Gly Gly Ser

225 230 235 240

Gly Cys Thr Ser Gln Lys Trp Ala Gln Cys Gly Gly Ile Gly Phe Ser

245 250 255

Gly Cys Thr Thr Cys Val Ser Gly Thr Thr Cys Gln Lys Leu Asn Asp

260 265 270

Tyr Tyr Ser Gln Cys Leu

275

<210> 5

<211> 305

<212> PRT

<213> Humicola insolens

<400> 5

Met Arg Ser Ser Pro Leu Leu Pro Ser Ala Val Val Ala Ala Leu Pro

1 5 10 15

Val Leu Ala Leu Ala Ala Asp Gly Arg Ser Thr Arg Tyr Trp Asp Cys

20 25 30

Cys Lys Pro Ser Cys Gly Trp Ala Lys Lys Ala Pro Val Asn Gln Pro

35 40 45

Val Phe Ser Cys Asn Ala Asn Phe Gln Arg Ile Thr Asp Phe Asp Ala

50 55 60

Lys Ser Gly Cys Glu Pro Gly Gly Val Ala Tyr Ser Cys Ala Asp Gln

65 70 75 80

Thr Pro Trp Ala Val Asn Asp Asp Phe Ala Leu Gly Phe Ala Ala Thr

85 90 95

Ser Ile Ala Gly Ser Asn Glu Ala Gly Trp Cys Cys Ala Cys Tyr Glu

100 105 110

Leu Thr Phe Thr Ser Gly Pro Val Ala Gly Lys Lys Met Val Val Gln

115 120 125

Ser Thr Ser Thr Gly Gly Asp Leu Gly Ser Asn His Phe Asp Leu Asn

130 135 140

Ile Pro Gly Gly Gly Val Gly Ile Phe Asp Gly Cys Thr Pro Gln Phe

145 150 155 160

Gly Gly Leu Pro Gly Gln Arg Tyr Gly Gly Ile Ser Ser Arg Asn Glu

165 170 175

Cys Asp Arg Phe Pro Asp Ala Leu Lys Pro Gly Cys Tyr Trp Arg Phe

180 185 190

Asp Trp Phe Lys Asn Ala Asp Asn Pro Ser Phe Ser Phe Arg Gln Val

195 200 205

Gln Cys Pro Ala Glu Leu Val Ala Arg Thr Gly Cys Arg Arg Asn Asp

210 215 220

Asp Gly Asn Phe Pro Ala Val Gln Ile Pro Ser Ser Ser Thr Ser Ser

225 230 235 240

Pro Val Asn Gln Pro Thr Ser Thr Ser Thr Thr Ser Thr Ser Thr Thr

245 250 255

Ser Ser Pro Pro Val Gln Pro Thr Thr Pro Ser Gly Cys Thr Ala Glu

260 265 270

Arg Trp Ala Gln Cys Gly Gly Asn Gly Trp Ser Gly Cys Thr Thr Cys

275 280 285

Val Ala Gly Ser Thr Cys Thr Lys Ile Asn Asp Trp Tyr His Gln Cys

290 295 300

Leu

305

<210> 6

<211> 293

<212> PRT

<213> Neurospora tetraspora

<400> 6

Met Arg Ser Ser Thr Val Leu Gln Thr Gly Leu Val Ala Ala Leu Pro

1 5 10 15

Phe Ala Val Gln Ala Ala Ser Gly Ser Gly Gln Ser Thr Arg Tyr Trp

20 25 30

Asp Cys Cys Lys Pro Ser Cys Ser Trp Ser Gly Lys Ala Ser Val Asn

35 40 45

Arg Pro Val Leu Ala Cys Asp Ala Asn Asn Asn Pro Leu Ser Asp Ala

50 55 60

Ser Val Lys Ser Gly Cys Asp Gly Gly Ser Ala Tyr Thr Cys Ala Asn

65 70 75 80

Asn Ser Pro Trp Ala Val Asn Asp Gln Leu Ser Tyr Gly Phe Ala Ala

85 90 95

Thr Lys Leu Ser Gly Gly Thr Glu Ser Ser Trp Cys Cys Ala Cys Tyr

100 105 110

Ala Leu Thr Phe Thr Ser Gly Pro Val Ala Gly Lys Thr Met Val Val

115 120 125

Gln Ser Thr Ser Thr Gly Gly Asp Leu Gly Ser Asn His Phe Asp Ile

130 135 140

Asn Met Pro Gly Gly Gly Val Gly Leu Phe Asp Gly Cys Thr Arg Gln

145 150 155 160

Phe Gly Gly Leu Pro Gly Ala Gln Tyr Gly Gly Ile Ser Ser Arg Ser

165 170 175

Gln Cys Asp Ser Phe Pro Ala Ala Leu Lys Pro Gly Cys Gln Trp Arg

180 185 190

Phe Asp Trp Phe Gln Asn Ala Asp Asn Pro Asn Phe Thr Phe Lys Gln

195 200 205

Val Gln Cys Pro Ser Glu Leu Thr Ser Arg Thr Gly Cys Lys Arg Asn

210 215 220

Asp Asp Ser Gln Phe Pro Val Phe Thr Pro Pro Ser Gly Gly Gly Thr

225 230 235 240

Asn Pro Ser Thr Pro Thr Thr Pro Pro Ser Ser Gly Gly Gly Ser Gly

245 250 255

Cys Thr Ala Asp Lys Tyr Ala Gln Cys Gly Gly Ser Gly Trp Ser Gly

260 265 270

Cys Thr Asn Cys Pro Ser Gly Ser Thr Cys Lys Thr Ile Asn Asp Tyr

275 280 285

Tyr His Gln Cys Ala

290

<210> 7

<211> 409

<212> PRT

<213> Talaromyces thermophilus

<400> 7

Met Lys Phe Ser Asn Val Ile Leu Ala Ala Ser Ala Ser Ser Leu Val

1 5 10 15

Leu Ala Ala Pro Lys Ser Lys Thr Lys Arg Thr Ser Ala Phe Gln Trp

20 25 30

Phe Gly Ala Asn Glu Ser Gly Ala Glu Phe Gly Asn Gln Asn Ile Pro

35 40 45

Gly Thr Leu Gly Thr Asp Tyr Thr Trp Pro Asp Thr Ser Thr Ile Gln

50 55 60

Thr Leu Arg Asn Ala Gly Met Asn Ile Phe Arg Val Pro Phe Leu Met

65 70 75 80

Glu Arg Leu Val Pro Asn Gln Met Thr Gly Ser Pro Asp Pro Thr Tyr

85 90 95

Leu Ala Asp Leu Lys Ser Thr Val Asn Phe Ile Thr Gly Thr Gly Ala

100 105 110

Tyr Ala Val Val Asp Pro His Asn Tyr Gly Arg Tyr Tyr Asn Asn Ile

115 120 125

Ile Thr Ser Thr Ser Asp Phe Ala Ala Phe Trp Thr Thr Val Ala Ser

130 135 140

Gln Phe Ala Ser Asn Pro Arg Val Ile Phe Asp Thr Asn Asn Glu Tyr

145 150 155 160

Asn Asn Met Asp Gln Thr Leu Val Leu Asn Leu Asn Gln Ala Ala Ile

165 170 175

Asn Ala Ile Arg Ala Ala Gly Ala Thr Ser Gln Tyr Ile Phe Ala Glu

180 185 190

Gly Asn Ser Trp Thr Gly Ala Trp Thr Trp Thr Ser Val Asn Asp Asn

195 200 205

Met Lys Gln Leu Thr Asp Pro Ser Asn Lys Leu Val Tyr Glu Met His

210 215 220

Gln Tyr Leu Asp Ser Asp Gly Ser Gly Thr Ser Asp Gln Cys Val Asn

225 230 235 240

Ser Thr Ile Gly Tyr Asp Arg Ile Val Ser Ala Thr Gln Trp Leu Gln

245 250 255

Ala Asn Gly Lys Val Ala Phe Leu Gly Glu Phe Ala Gly Gly Ser Asn

260 265 270

Ser Val Cys Glu Ala Ala Val Thr Gly Met Leu Asp Tyr Met Glu Gln

275 280 285

Asn Ser Asp Val Trp Leu Gly Ala Glu Trp Trp Ala Ala Gly Pro Trp

290 295 300

Trp Gly Asn Tyr Ile Tyr Ser Met Glu Pro Pro Ser Gly Ile Ala Tyr

305 310 315 320

Gln Asn Tyr Leu Ser Ile Leu Glu Pro Tyr Phe Pro Gly Gly Ser Tyr

325 330 335

Ser Gly Gly Thr Gly Ser Gly Ser Gly Ser Thr Thr Thr Thr Ala Thr

340 345 350

Thr Thr Thr Thr Lys Val Pro Pro Thr Ser Thr Thr Ser Ser Ala Ser

355 360 365

Ser Thr Gly Thr Gly Val Ala Gln His Trp Gly Gln Cys Gly Gly Gln

370 375 380

Gly Trp Thr Gly Pro Thr Thr Cys Val Ser Pro Tyr Thr Cys Gln Glu

385 390 395 400

Leu Asn Pro Tyr Tyr Tyr Gln Cys Leu

405

<210> 8

<211> 326

<212> PRT

<213> Thielavia terrestris

<400> 8

Met Lys Ser Phe Thr Ile Ala Ala Leu Ala Ala Leu Trp Ala Gln Glu

1 5 10 15

Ala Ala Ala His Ala Thr Phe Gln Asp Leu Trp Ile Asp Gly Val Asp

20 25 30

Tyr Gly Ser Gln Cys Val Arg Leu Pro Ala Ser Asn Ser Pro Val Thr

35 40 45

Asn Val Ala Ser Asp Asp Ile Arg Cys Asn Val Gly Thr Ser Arg Pro

50 55 60

Thr Val Lys Cys Pro Val Lys Ala Gly Ser Thr Val Thr Ile Glu Met

65 70 75 80

His Gln Gln Pro Gly Asp Arg Ser Cys Ala Asn Glu Ala Ile Gly Gly

85 90 95

Asp His Tyr Gly Pro Val Met Val Tyr Met Ser Lys Val Asp Asp Ala

100 105 110

Val Thr Ala Asp Gly Ser Ser Gly Trp Phe Lys Val Phe Gln Asp Ser

115 120 125

Trp Ala Lys Asn Pro Ser Gly Ser Thr Gly Asp Asp Asp Tyr Trp Gly

130 135 140

Thr Lys Asp Leu Asn Ser Cys Cys Gly Lys Met Asn Val Lys Ile Pro

145 150 155 160

Glu Asp Ile Glu Pro Gly Asp Tyr Leu Leu Arg Ala Glu Val Ile Ala

165 170 175

Leu His Val Ala Ala Ser Ser Gly Gly Ala Gln Phe Tyr Met Ser Cys

180 185 190

Tyr Gln Leu Thr Val Thr Gly Ser Gly Ser Ala Thr Pro Ser Thr Val

195 200 205

Asn Phe Pro Gly Ala Tyr Ser Ala Ser Asp Pro Gly Ile Leu Ile Asn

210 215 220

Ile His Ala Pro Met Ser Thr Tyr Val Val Pro Gly Pro Thr Val Tyr

225 230 235 240

Ala Gly Gly Ser Thr Lys Ser Ala Gly Ser Ser Cys Ser Gly Cys Glu

245 250 255

Ala Thr Cys Thr Val Gly Ser Gly Pro Ser Ala Thr Leu Thr Gln Pro

260 265 270

Thr Ser Thr Ala Thr Ala Thr Ser Ala Pro Gly Gly Gly Gly Ser Gly

275 280 285

Cys Thr Ala Ala Lys Tyr Gln Gln Cys Gly Gly Thr Gly Tyr Thr Gly

290 295 300

Cys Thr Thr Cys Ala Ser Gly Ser Thr Cys Ser Ala Val Ser Pro Pro

305 310 315 320

Tyr Tyr Ser Gln Cys Leu

325

<210> 9

<211> 239

<212> PRT

<213> Thielavia terrestris

<400> 9

Met Arg Phe Asp Ala Leu Ser Ala Leu Ala Leu Ala Pro Leu Val Ala

1 5 10 15

Gly His Gly Ala Val Thr Ser Tyr Ile Ile Gly Gly Lys Thr Tyr Pro

20 25 30

Gly Tyr Glu Gly Phe Ser Pro Ala Ser Ser Pro Pro Thr Ile Gln Tyr

35 40 45

Gln Trp Pro Asp Tyr Asn Pro Thr Leu Ser Val Thr Asp Pro Lys Met

50 55 60

Arg Cys Asn Gly Gly Thr Ser Ala Glu Leu Ser Ala Pro Val Gln Ala

65 70 75 80

Gly Glu Asn Val Thr Ala Val Trp Lys Gln Trp Thr His Gln Gln Gly

85 90 95

Pro Val Met Val Trp Met Phe Lys Cys Pro Gly Asp Phe Ser Ser Ser

100 105 110

His Gly Asp Gly Lys Gly Trp Phe Lys Ile Asp Gln Leu Gly Leu Trp

115 120 125

Gly Asn Asn Leu Asn Ser Asn Asn Trp Gly Thr Ala Ile Val Tyr Lys

130 135 140

Thr Leu Gln Trp Ser Asn Pro Ile Pro Lys Asn Leu Ala Pro Gly Asn

145 150 155 160

Tyr Leu Ile Arg His Glu Leu Leu Ala Leu His Gln Ala Asn Thr Pro

165 170 175

Gln Phe Tyr Ala Glu Cys Ala Gln Leu Val Val Ser Gly Ser Gly Ser

180 185 190

Ala Leu Pro Pro Ser Asp Tyr Leu Tyr Ser Ile Pro Val Tyr Ala Pro

195 200 205

Gln Asn Asp Pro Gly Ile Thr Val Asp Ile Tyr Asn Gly Gly Leu Thr

210 215 220

Ser Tyr Thr Pro Pro Gly Gly Pro Val Trp Ser Gly Phe Glu Phe

225 230 235

<210> 10

<211> 258

<212> PRT

<213> Thielavia terrestris

<400> 10

Met Leu Leu Thr Ser Val Leu Gly Ser Ala Ala Leu Leu Ala Ser Gly

1 5 10 15

Ala Ala Ala His Gly Ala Val Thr Ser Tyr Ile Ile Ala Gly Lys Asn

20 25 30

Tyr Pro Gly Tyr Gln Gly Phe Ser Pro Ala Asn Ser Pro Asn Val Ile

35 40 45

Gln Trp Gln Trp His Asp Tyr Asn Pro Val Leu Ser Cys Ser Asp Ser

50 55 60

Lys Leu Arg Cys Asn Gly Gly Thr Ser Ala Thr Leu Asn Ala Thr Ala

65 70 75 80

Ala Pro Gly Asp Thr Ile Thr Ala Ile Trp Ala Gln Trp Thr His Ser

85 90 95

Gln Gly Pro Ile Leu Val Trp Met Tyr Lys Cys Pro Gly Ser Phe Ser

100 105 110

Ser Cys Asp Gly Ser Gly Ala Gly Trp Phe Lys Ile Asp Glu Ala Gly

115 120 125

Phe His Gly Asp Gly Val Lys Val Phe Leu Asp Thr Glu Asn Pro Ser

130 135 140

Gly Trp Asp Ile Ala Lys Leu Val Gly Gly Asn Lys Gln Trp Ser Ser

145 150 155 160

Lys Val Pro Glu Gly Leu Ala Pro Gly Asn Tyr Leu Val Arg His Glu

165 170 175

Leu Ile Ala Leu His Gln Ala Asn Asn Pro Gln Phe Tyr Pro Glu Cys

180 185 190

Ala Gln Val Val Ile Thr Gly Ser Gly Thr Ala Gln Pro Asp Ala Ser

195 200 205

Tyr Lys Ala Ala Ile Pro Gly Tyr Cys Asn Gln Asn Asp Pro Asn Ile

210 215 220

Lys Val Pro Ile Asn Asp His Ser Ile Pro Gln Thr Tyr Lys Ile Pro

225 230 235 240

Gly Pro Pro Val Phe Lys Gly Thr Ala Ser Lys Lys Ala Arg Asp Phe

245 250 255

Thr Ala

<210> 11

<211> 226

<212> PRT

<213> Thielavia terrestris

<400> 11

Met Leu Ala Asn Gly Ala Ile Val Phe Leu Ala Ala Ala Leu Gly Val

1 5 10 15

Ser Gly His Tyr Thr Trp Pro Arg Val Asn Asp Gly Ala Asp Trp Gln

20 25 30

Gln Val Arg Lys Ala Asp Asn Trp Gln Asp Asn Gly Tyr Val Gly Asp

35 40 45

Val Thr Ser Pro Gln Ile Arg Cys Phe Gln Ala Thr Pro Ser Pro Ala

50 55 60

Pro Ser Val Leu Asn Thr Thr Ala Gly Ser Thr Val Thr Tyr Trp Ala

65 70 75 80

Asn Pro Asp Val Tyr His Pro Gly Pro Val Gln Phe Tyr Met Ala Arg

85 90 95

Val Pro Asp Gly Glu Asp Ile Asn Ser Trp Asn Gly Asp Gly Ala Val

100 105 110

Trp Phe Lys Val Tyr Glu Asp His Pro Thr Phe Gly Ala Gln Leu Thr

115 120 125

Trp Pro Ser Thr Gly Lys Ser Ser Phe Ala Val Pro Ile Pro Pro Cys

130 135 140

Ile Lys Ser Gly Tyr Tyr Leu Leu Arg Ala Glu Gln Ile Gly Leu His

145 150 155 160

Val Ala Gln Ser Val Gly Gly Ala Gln Phe Tyr Ile Ser Cys Ala Gln

165 170 175

Leu Ser Val Thr Gly Gly Gly Ser Thr Glu Pro Pro Asn Lys Val Ala

180 185 190

Phe Pro Gly Ala Tyr Ser Ala Thr Asp Pro Gly Ile Leu Ile Asn Ile

195 200 205

Tyr Tyr Pro Val Pro Thr Ser Tyr Gln Asn Pro Gly Pro Ala Val Phe

210 215 220

Ser Cys

225

<210> 12

<211> 304

<212> PRT

<213> Thielavia terrestris

<400> 12

Met Lys Gly Leu Phe Ser Ala Ala Ala Leu Ser Leu Ala Val Gly Gln

1 5 10 15

Ala Ser Ala His Tyr Ile Phe Gln Gln Leu Ser Ile Asn Gly Asn Gln

20 25 30

Phe Pro Val Tyr Gln Tyr Ile Arg Lys Asn Thr Asn Tyr Asn Ser Pro

35 40 45

Val Thr Asp Leu Thr Ser Asp Asp Leu Arg Cys Asn Val Gly Ala Gln

50 55 60

Gly Ala Gly Thr Asp Thr Val Thr Val Lys Ala Gly Asp Gln Phe Thr

65 70 75 80

Phe Thr Leu Asp Thr Pro Val Tyr His Gln Gly Pro Ile Ser Ile Tyr

85 90 95

Met Ser Lys Ala Pro Gly Ala Ala Ser Asp Tyr Asp Gly Ser Gly Gly

100 105 110

Trp Phe Lys Ile Lys Asp Trp Gly Pro Thr Phe Asn Ala Asp Gly Thr

115 120 125

Ala Thr Trp Asp Met Ala Gly Ser Tyr Thr Tyr Asn Ile Pro Thr Cys

130 135 140

Ile Pro Asp Gly Asp Tyr Leu Leu Arg Ile Gln Ser Leu Ala Ile His

145 150 155 160

Asn Pro Trp Pro Ala Gly Ile Pro Gln Phe Tyr Ile Ser Cys Ala Gln

165 170 175

Ile Thr Val Thr Gly Gly Gly Asn Gly Asn Pro Gly Pro Thr Ala Leu

180 185 190

Ile Pro Gly Ala Phe Lys Asp Thr Asp Pro Gly Tyr Thr Val Asn Ile

195 200 205

Tyr Thr Asn Phe His Asn Tyr Thr Val Pro Gly Pro Glu Val Phe Ser

210 215 220

Cys Asn Gly Gly Gly Ser Asn Pro Pro Pro Pro Val Ser Ser Ser Thr

225 230 235 240

Pro Ala Thr Thr Thr Leu Val Thr Ser Thr Arg Thr Thr Ser Ser Thr

245 250 255

Ser Ser Ala Ser Thr Pro Ala Ser Thr Gly Gly Cys Thr Val Ala Lys

260 265 270

Trp Gly Gln Cys Gly Gly Asn Gly Tyr Thr Gly Cys Thr Thr Cys Ala

275 280 285

Ala Gly Ser Thr Cys Ser Lys Gln Asn Asp Tyr Tyr Ser Gln Cys Leu

290 295 300

<210> 13

<211> 317

<212> PRT

<213> Thielavia terrestris

<400> 13

Met Lys Gly Leu Ser Leu Leu Ala Ala Ala Ser Ala Ala Thr Ala His

1 5 10 15

Thr Ile Phe Val Gln Leu Glu Ser Gly Gly Thr Thr Tyr Pro Val Ser

20 25 30

Tyr Gly Ile Arg Asp Pro Ser Tyr Asp Gly Pro Ile Thr Asp Val Thr

35 40 45

Ser Asp Ser Leu Ala Cys Asn Gly Pro Pro Asn Pro Thr Thr Pro Ser

50 55 60

Pro Tyr Ile Ile Asn Val Thr Ala Gly Thr Thr Val Ala Ala Ile Trp

65 70 75 80

Arg His Thr Leu Thr Ser Gly Pro Asp Asp Val Met Asp Ala Ser His

85 90 95

Lys Gly Pro Thr Leu Ala Tyr Leu Lys Lys Val Asp Asp Ala Leu Thr

100 105 110

Asp Thr Gly Ile Gly Gly Gly Trp Phe Lys Ile Gln Glu Ala Gly Tyr

115 120 125

Asp Asn Gly Asn Trp Ala Thr Ser Thr Val Ile Thr Asn Gly Gly Phe

130 135 140

Gln Tyr Ile Asp Ile Pro Ala Cys Ile Pro Asn Gly Gln Tyr Leu Leu

145 150 155 160

Arg Ala Glu Met Ile Ala Leu His Ala Ala Ser Thr Gln Gly Gly Ala

165 170 175

Gln Leu Tyr Met Glu Cys Ala Gln Ile Asn Val Val Gly Gly Ser Gly

180 185 190

Ser Ala Ser Pro Gln Thr Tyr Ser Ile Pro Gly Ile Tyr Gln Ala Thr

195 200 205

Asp Pro Gly Leu Leu Ile Asn Ile Tyr Ser Met Thr Pro Ser Ser Gln

210 215 220

Tyr Thr Ile Pro Gly Pro Pro Leu Phe Thr Cys Ser Gly Ser Gly Asn

225 230 235 240

Asn Gly Gly Gly Ser Asn Pro Ser Gly Gly Gln Thr Thr Thr Ala Lys

245 250 255

Pro Thr Thr Thr Thr Ala Ala Thr Thr Thr Ser Ser Ala Ala Pro Thr

260 265 270

Ser Ser Gln Gly Gly Ser Ser Gly Cys Thr Val Pro Gln Trp Gln Gln

275 280 285

Cys Gly Gly Ile Ser Phe Thr Gly Cys Thr Thr Cys Ala Ala Gly Tyr

290 295 300

Thr Cys Lys Tyr Leu Asn Asp Tyr Tyr Ser Gln Cys Gln

305 310 315

<210> 14

<211> 249

<212> PRT

<213> Trichoderma reesei

<400> 14

Met Lys Ser Cys Ala Ile Leu Ala Ala Leu Gly Cys Leu Ala Gly Ser

1 5 10 15

Val Leu Gly His Gly Gln Val Gln Asn Phe Thr Ile Asn Gly Gln Tyr

20 25 30

Asn Gln Gly Phe Ile Leu Asp Tyr Tyr Tyr Gln Lys Gln Asn Thr Gly

35 40 45

His Phe Pro Asn Val Ala Gly Trp Tyr Ala Glu Asp Leu Asp Leu Gly

50 55 60

Phe Ile Ser Pro Asp Gln Tyr Thr Thr Pro Asp Ile Val Cys His Lys

65 70 75 80

Asn Ala Ala Pro Gly Ala Ile Ser Ala Thr Ala Ala Ala Gly Ser Asn

85 90 95

Ile Val Phe Gln Trp Gly Pro Gly Val Trp Pro His Pro Tyr Gly Pro

100 105 110

Ile Val Thr Tyr Val Val Glu Cys Ser Gly Ser Cys Thr Thr Val Asn

115 120 125

Lys Asn Asn Leu Arg Trp Val Lys Ile Gln Glu Ala Gly Ile Asn Tyr

130 135 140

Asn Thr Gln Val Trp Ala Gln Gln Asp Leu Ile Asn Gln Gly Asn Lys

145 150 155 160

Trp Thr Val Lys Ile Pro Ser Ser Leu Arg Pro Gly Asn Tyr Val Phe

165 170 175

Arg His Glu Leu Leu Ala Ala His Gly Ala Ser Ser Ala Asn Gly Met

180 185 190

Gln Asn Tyr Pro Gln Cys Val Asn Ile Ala Val Thr Gly Ser Gly Thr

195 200 205

Lys Ala Leu Pro Ala Gly Thr Pro Ala Thr Gln Leu Tyr Lys Pro Thr

210 215 220

Asp Pro Gly Ile Leu Phe Asn Pro Tyr Thr Thr Ile Thr Ser Tyr Thr

225 230 235 240

Ile Pro Gly Pro Ala Leu Trp Gln Gly

245

<210> 15

<211> 232

<212> PRT

<213> myceliophthora thermophila

<400> 15

Met Lys Phe Thr Ser Ser Leu Ala Val Leu Ala Ala Ala Gly Ala Gln

1 5 10 15

Ala His Tyr Thr Phe Pro Arg Ala Gly Thr Gly Gly Ser Leu Ser Gly

20 25 30

Glu Trp Glu Val Val Arg Met Thr Glu Asn His Tyr Ser His Gly Pro

35 40 45

Val Thr Asp Val Thr Ser Pro Glu Met Thr Cys Tyr Gln Ser Gly Val

50 55 60

Gln Gly Ala Pro Gln Thr Val Gln Val Lys Ala Gly Ser Gln Phe Thr

65 70 75 80

Phe Ser Val Asp Pro Ser Ile Gly His Pro Gly Pro Leu Gln Phe Tyr

85 90 95

Met Ala Lys Val Pro Ser Gly Gln Thr Ala Ala Thr Phe Asp Gly Thr

100 105 110

Gly Ala Val Trp Phe Lys Ile Tyr Gln Asp Gly Pro Asn Gly Leu Gly

115 120 125

Thr Asp Ser Ile Thr Trp Pro Ser Ala Gly Lys Thr Glu Val Ser Val

130 135 140

Thr Ile Pro Ser Cys Ile Asp Asp Gly Glu Tyr Leu Leu Arg Val Glu

145 150 155 160

His Ile Ala Leu His Ser Ala Ser Ser Val Gly Gly Ala Gln Phe Tyr

165 170 175

Ile Ala Cys Ala Gln Leu Ser Val Thr Gly Gly Ser Gly Thr Leu Asn

180 185 190

Thr Gly Ser Leu Val Ser Leu Pro Gly Ala Tyr Lys Ala Thr Asp Pro

195 200 205

Gly Ile Leu Phe Gln Leu Tyr Trp Pro Ile Pro Thr Glu Tyr Ile Asn

210 215 220

Pro Gly Pro Ala Pro Val Ser Cys

225 230

<210> 16

<211> 235

<212> PRT

<213> myceliophthora thermophila

<400> 16

Met Lys Ala Leu Ser Leu Leu Ala Ala Ala Ser Ala Val Ser Ala His

1 5 10 15

Thr Ile Phe Val Gln Leu Glu Ala Asp Gly Thr Arg Tyr Pro Val Ser

20 25 30

Tyr Gly Ile Arg Asp Pro Ser Tyr Asp Gly Pro Ile Thr Asp Val Thr

35 40 45

Ser Asn Asp Val Ala Cys Asn Gly Gly Pro Asn Pro Thr Thr Pro Ser

50 55 60

Ser Asp Val Ile Thr Val Thr Ala Gly Thr Thr Val Lys Ala Ile Trp

65 70 75 80

Arg His Thr Leu Gln Ser Gly Pro Asp Asp Val Met Asp Ala Ser His

85 90 95

Lys Gly Pro Thr Leu Ala Tyr Leu Lys Lys Val Gly Asp Ala Thr Lys

100 105 110

Asp Ser Gly Val Gly Gly Gly Trp Phe Lys Ile Gln Glu Asp Gly Tyr

115 120 125

Asn Asn Gly Gln Trp Gly Thr Ser Thr Val Ile Ser Asn Gly Gly Glu

130 135 140

His Tyr Ile Asp Ile Pro Ala Cys Ile Pro Glu Gly Gln Tyr Leu Leu

145 150 155 160

Arg Ala Glu Met Ile Ala Leu His Ala Ala Gly Ser Pro Gly Gly Ala

165 170 175

Gln Leu Tyr Met Glu Cys Ala Gln Ile Asn Ile Val Gly Gly Ser Gly

180 185 190

Ser Val Pro Ser Ser Thr Val Ser Phe Pro Gly Ala Tyr Ser Pro Asn

195 200 205

Asp Pro Gly Leu Leu Ile Asn Ile Tyr Ser Met Ser Pro Ser Ser Ser

210 215 220

Tyr Thr Ile Pro Gly Pro Pro Val Phe Lys Cys

225 230 235

<210> 17

<211> 323

<212> PRT

<213> myceliophthora thermophila

<400> 17

Met Lys Ser Phe Ala Leu Thr Thr Leu Ala Ala Leu Ala Gly Asn Ala

1 5 10 15

Ala Ala His Ala Thr Phe Gln Ala Leu Trp Val Asp Gly Val Asp Tyr

20 25 30

Gly Ala Gln Cys Ala Arg Leu Pro Ala Ser Asn Ser Pro Val Thr Asp

35 40 45

Val Thr Ser Asn Ala Ile Arg Cys Asn Ala Asn Pro Ser Pro Ala Arg

50 55 60

Gly Lys Cys Pro Val Lys Ala Gly Ser Thr Val Thr Val Glu Met His

65 70 75 80

Gln Gln Pro Gly Asp Arg Ser Cys Ser Ser Glu Ala Ile Gly Gly Ala

85 90 95

His Tyr Gly Pro Val Met Val Tyr Met Ser Lys Val Ser Asp Ala Ala

100 105 110

Ser Ala Asp Gly Ser Ser Gly Trp Phe Lys Val Phe Glu Asp Gly Trp

115 120 125

Ala Lys Asn Pro Ser Gly Gly Ser Gly Asp Asp Asp Tyr Trp Gly Thr

130 135 140

Lys Asp Leu Asn Ser Cys Cys Gly Lys Met Asn Val Lys Ile Pro Ala

145 150 155 160

Asp Leu Pro Ser Gly Asp Tyr Leu Leu Arg Ala Glu Ala Leu Ala Leu

165 170 175

His Thr Ala Gly Ser Ala Gly Gly Ala Gln Phe Tyr Met Thr Cys Tyr

180 185 190

Gln Leu Thr Val Thr Gly Ser Gly Ser Ala Ser Pro Pro Thr Val Ser

195 200 205

Phe Pro Gly Ala Tyr Lys Ala Thr Asp Pro Gly Ile Leu Val Asn Ile

210 215 220

His Ala Pro Leu Ser Gly Tyr Thr Val Pro Gly Pro Ala Val Tyr Ser

225 230 235 240

Gly Gly Ser Thr Lys Lys Ala Gly Ser Ala Cys Thr Gly Cys Glu Ser

245 250 255

Thr Cys Ala Val Gly Ser Gly Pro Thr Ala Thr Val Ser Gln Ser Pro

260 265 270

Gly Ser Thr Ala Thr Ser Ala Pro Gly Gly Gly Gly Gly Cys Thr Val

275 280 285

Gln Lys Tyr Gln Gln Cys Gly Gly Glu Gly Tyr Thr Gly Cys Thr Asn

290 295 300

Cys Ala Ser Gly Ser Thr Cys Ser Ala Val Ser Pro Pro Tyr Tyr Ser

305 310 315 320

Gln Cys Val

<210> 18

<211> 310

<212> PRT

<213> myceliophthora thermophila

<400> 18

Met Lys Pro Phe Ser Leu Val Ala Leu Ala Thr Ala Val Ser Gly His

1 5 10 15

Ala Ile Phe Gln Arg Val Ser Val Asn Gly Gln Asp Gln Gly Gln Leu

20 25 30

Lys Gly Val Arg Ala Pro Ser Ser Asn Ser Pro Ile Gln Asn Val Asn

35 40 45

Asp Ala Asn Met Ala Cys Asn Ala Asn Ile Val Tyr His Asp Ser Thr

50 55 60

Ile Ile Lys Val Pro Ala Gly Ala Arg Val Gly Ala Trp Trp Gln His

65 70 75 80

Val Ile Gly Gly Pro Gln Gly Ala Asn Asp Pro Asp Asn Pro Ile Ala

85 90 95

Ala Ser His Lys Gly Pro Ile Gln Val Tyr Leu Ala Lys Val Asp Asn

100 105 110

Ala Ala Thr Ala Ser Pro Ser Gly Leu Arg Trp Phe Lys Val Ala Glu

115 120 125

Arg Gly Leu Asn Asn Gly Val Trp Ala Val Asp Glu Leu Ile Ala Asn

130 135 140

Asn Gly Trp His Tyr Phe Asp Leu Pro Ser Cys Val Ala Pro Gly Gln

145 150 155 160

Tyr Leu Met Arg Val Glu Leu Leu Ala Leu His Ser Ala Ser Ser Pro

165 170 175

Gly Gly Ala Gln Phe Tyr Met Gly Cys Ala Gln Ile Glu Val Thr Gly

180 185 190

Ser Gly Thr Asn Ser Gly Ser Asp Phe Val Ser Phe Pro Gly Ala Tyr

195 200 205

Ser Ala Asn Asp Pro Gly Ile Leu Leu Ser Ile Tyr Asp Ser Ser Gly

210 215 220

Lys Pro Thr Asn Gly Gly Arg Ser Tyr Pro Ile Pro Gly Pro Arg Pro

225 230 235 240

Ile Ser Cys Ser Gly Ser Gly Asp Gly Gly Asn Asn Gly Gly Gly Gly

245 250 255

Asp Asp Asn Asn Asn Asn Asn Gly Gly Gly Asn Asn Gly Gly Gly Gly

260 265 270

Gly Gly Ser Val Pro Leu Tyr Gly Gln Cys Gly Gly Ile Gly Tyr Thr

275 280 285

Gly Pro Thr Thr Cys Ala Gln Gly Thr Cys Lys Val Ser Asn Glu Tyr

290 295 300

Tyr Ser Gln Cys Leu Pro

305 310

<210> 19

<211> 246

<212> PRT

<213> myceliophthora thermophila

<400> 19

Met Lys Leu Ser Leu Phe Ser Val Leu Ala Thr Ala Leu Thr Val Glu

1 5 10 15

Gly His Ala Ile Phe Gln Lys Val Ser Val Asn Gly Ala Asp Gln Gly

20 25 30

Ser Leu Thr Gly Leu Arg Ala Pro Asn Asn Asn Asn Pro Val Gln Asp

35 40 45

Val Asn Ser Gln Asp Met Ile Cys Gly Gln Ser Gly Ser Thr Ser Asn

50 55 60

Thr Ile Ile Glu Val Lys Ala Gly Asp Arg Ile Gly Ala Trp Tyr Gln

65 70 75 80

His Val Ile Gly Gly Ala Gln Phe Pro Asn Asp Pro Asp Asn Pro Ile

85 90 95

Ala Lys Ser His Lys Gly Pro Val Met Ala Tyr Leu Ala Lys Val Asp

100 105 110

Asn Ala Ala Thr Ala Ser Lys Thr Gly Leu Lys Trp Phe Lys Ile Trp

115 120 125

Glu Asp Thr Phe Asn Pro Ser Thr Lys Thr Trp Gly Val Asp Asn Leu

130 135 140

Ile Asn Asn Asn Gly Trp Val Tyr Phe Asn Leu Pro Gln Cys Ile Ala

145 150 155 160

Asp Gly Asn Tyr Leu Leu Arg Val Glu Val Leu Ala Leu His Ser Ala

165 170 175

Tyr Ser Gln Gly Gln Ala Gln Phe Tyr Gln Ser Cys Ala Gln Ile Asn

180 185 190

Val Ser Gly Gly Gly Ser Phe Thr Pro Pro Ser Thr Val Ser Phe Pro

195 200 205

Gly Ala Tyr Ser Ala Ser Asp Pro Gly Ile Leu Ile Asn Ile Tyr Gly

210 215 220

Ala Thr Gly Gln Pro Asp Asn Asn Gly Gln Pro Tyr Thr Ala Pro Gly

225 230 235 240

Pro Ala Pro Ile Ser Cys

245

<210> 20

<211> 354

<212> PRT

<213> Thermoascus aurantiacus

<400> 20

Met Ser Phe Ser Lys Ile Ala Ala Ile Thr Gly Ala Ile Thr Tyr Ala

1 5 10 15

Ser Leu Ala Ala Ala His Gly Tyr Val Thr Gly Ile Val Ala Asp Gly

20 25 30

Thr Tyr Tyr Gly Gly Tyr Ile Val Thr Gln Tyr Pro Tyr Met Ser Thr

35 40 45

Pro Pro Asp Val Ile Ala Trp Ser Thr Lys Ala Thr Asp Leu Gly Phe

50 55 60

Val Asp Pro Ser Ser Tyr Ala Ser Ser Asp Ile Ile Cys His Lys Gly

65 70 75 80

Ala Glu Pro Gly Ala Leu Ser Ala Lys Val Ala Ala Gly Gly Thr Val

85 90 95

Glu Leu Gln Trp Thr Asp Trp Pro Glu Ser His Lys Gly Pro Val Ile

100 105 110

Asp Tyr Leu Ala Ala Cys Asn Gly Asp Cys Ser Thr Val Asp Lys Thr

115 120 125

Lys Leu Glu Phe Phe Lys Ile Asp Glu Ser Gly Leu Ile Asp Gly Ser

130 135 140

Ser Ala Pro Gly Thr Trp Ala Ser Asp Asn Leu Ile Ala Asn Asn Asn

145 150 155 160

Ser Trp Thr Val Thr Ile Pro Ser Thr Ile Ala Pro Gly Asn Tyr Val

165 170 175

Leu Arg His Glu Ile Ile Ala Leu His Ser Ala Gly Asn Thr Asn Gly

180 185 190

Ala Gln Asn Tyr Pro Gln Cys Ile Asn Leu Glu Val Thr Gly Ser Gly

195 200 205

Thr Asp Thr Pro Ala Gly Thr Leu Gly Thr Glu Leu Tyr Lys Ala Thr

210 215 220

Asp Pro Gly Ile Leu Val Asn Ile Tyr Gln Thr Leu Thr Ser Tyr Asp

225 230 235 240

Ile Pro Gly Pro Ala Leu Tyr Thr Gly Gly Ser Ser Gly Ser Ser Gly

245 250 255

Ser Ser Asn Thr Ala Lys Ala Thr Thr Ser Thr Ala Ser Ser Ser Ile

260 265 270

Val Thr Pro Thr Pro Val Asn Asn Pro Thr Val Thr Gln Thr Ala Val

275 280 285

Val Asp Val Thr Gln Thr Val Ser Gln Asn Ala Ala Val Ala Thr Thr

290 295 300

Thr Pro Ala Ser Thr Ala Val Ala Thr Ala Val Pro Thr Gly Thr Thr

305 310 315 320

Phe Ser Phe Asp Ser Met Thr Ser Asp Glu Phe Val Ser Leu Met Arg

325 330 335

Ala Thr Val Asn Trp Leu Leu Ser Asn Lys Lys His Ala Arg Asp Leu

340 345 350

Ser Tyr

<210> 21

<211> 250

<212> PRT

<213> Aspergillus fumigatus

<400> 21

Met Thr Leu Ser Lys Ile Thr Ser Ile Ala Gly Leu Leu Ala Ser Ala

1 5 10 15

Ser Leu Val Ala Gly His Gly Phe Val Ser Gly Ile Val Ala Asp Gly

20 25 30

Lys Tyr Tyr Gly Gly Tyr Leu Val Asn Gln Tyr Pro Tyr Met Ser Asn

35 40 45

Pro Pro Asp Thr Ile Ala Trp Ser Thr Thr Ala Thr Asp Leu Gly Phe

50 55 60

Val Asp Gly Thr Gly Tyr Gln Ser Pro Asp Ile Ile Cys His Arg Asp

65 70 75 80

Ala Lys Asn Gly Lys Leu Thr Ala Thr Val Ala Ala Gly Ser Gln Ile

85 90 95

Glu Phe Gln Trp Thr Thr Trp Pro Glu Ser His His Gly Pro Leu Ile

100 105 110

Thr Tyr Leu Ala Pro Cys Asn Gly Asp Cys Ala Thr Val Asp Lys Thr

115 120 125

Thr Leu Lys Phe Val Lys Ile Ala Ala Gln Gly Leu Ile Asp Gly Ser

130 135 140

Asn Pro Pro Gly Val Trp Ala Asp Asp Glu Met Ile Ala Asn Asn Asn

145 150 155 160

Thr Ala Thr Val Thr Ile Pro Ala Ser Tyr Ala Pro Gly Asn Tyr Val

165 170 175

Leu Arg His Glu Ile Ile Ala Leu His Ser Ala Gly Asn Leu Asn Gly

180 185 190

Ala Gln Asn Tyr Pro Gln Cys Phe Asn Ile Gln Ile Thr Gly Gly Gly

195 200 205

Ser Ala Gln Gly Ser Gly Thr Ala Gly Thr Ser Leu Tyr Lys Asn Thr

210 215 220

Asp Pro Gly Ile Lys Phe Asp Ile Tyr Ser Asp Leu Ser Gly Gly Tyr

225 230 235 240

Pro Ile Pro Gly Pro Ala Leu Phe Asn Ala

245 250

<210> 22

<211> 322

<212> PRT

<213> Penicillium pinophilum

<400> 22

Met Pro Ser Thr Lys Val Ala Ala Leu Ser Ala Val Leu Ala Leu Ala

1 5 10 15

Ser Thr Val Ala Gly His Gly Phe Val Gln Asn Ile Val Ile Asp Gly

20 25 30

Lys Ser Tyr Ser Gly Tyr Leu Val Asn Gln Phe Pro Tyr Glu Ser Asn

35 40 45

Pro Pro Ala Val Ile Gly Trp Ala Thr Thr Ala Thr Asp Leu Gly Phe

50 55 60

Val Ala Pro Ser Glu Tyr Thr Asn Ala Asp Ile Ile Cys His Lys Asn

65 70 75 80

Ala Thr Pro Gly Ala Leu Ser Ala Pro Val Ala Ala Gly Gly Thr Val

85 90 95

Glu Leu Gln Trp Thr Thr Trp Pro Asp Ser His His Gly Pro Val Ile

100 105 110

Ser Tyr Leu Ala Asn Cys Asn Gly Asn Cys Ser Thr Val Asp Lys Thr

115 120 125

Lys Leu Asp Phe Val Lys Ile Asp Gln Gly Gly Leu Ile Asp Asp Thr

130 135 140

Thr Pro Pro Gly Thr Trp Ala Ser Asp Lys Leu Ile Ala Ala Asn Asn

145 150 155 160

Ser Trp Thr Val Thr Ile Pro Ser Thr Ile Ala Pro Gly Asn Tyr Val

165 170 175

Leu Arg His Glu Ile Ile Ala Leu His Ser Ala Gly Asn Ala Asp Gly

180 185 190

Ala Gln Asn Tyr Pro Gln Cys Ile Asn Leu Glu Ile Thr Gly Ser Gly

195 200 205

Thr Ala Ala Pro Ser Gly Thr Ala Gly Glu Lys Leu Tyr Thr Ser Thr

210 215 220

Asp Pro Gly Ile Leu Val Asn Ile Tyr Gln Ser Leu Ser Thr Tyr Val

225 230 235 240

Ile Pro Gly Pro Thr Leu Trp Ser Gly Ala Ala Asn Gly Ala Val Ala

245 250 255

Thr Gly Ser Ala Thr Ala Val Ala Thr Thr Ala Thr Ala Ser Ala Thr

260 265 270

Ala Thr Pro Thr Thr Leu Val Thr Ser Val Ala Pro Ala Ser Ser Thr

275 280 285

Phe Ala Thr Ala Val Val Thr Thr Val Ala Pro Ala Val Thr Asp Val

290 295 300

Val Thr Val Thr Asp Val Val Thr Val Thr Thr Val Ile Thr Thr Thr

305 310 315 320

Val Leu

<210> 23

<211> 444

<212> PRT

<213> Thermoascus species

<400> 23

Met Leu Ser Phe Ala Ser Ala Lys Ser Ala Val Leu Thr Thr Leu Leu

1 5 10 15

Leu Leu Gly Ser Ala Gln Ala His Thr Leu Met Thr Thr Leu Phe Val

20 25 30

Asp Gly Val Asn Gln Gly Asp Gly Val Cys Ile Arg Met Asn Asn Asn

35 40 45

Gly Ser Thr Ala Asn Thr Tyr Ile Gln Pro Val Thr Ser Lys Asp Ile

50 55 60

Ala Cys Gly Ile Gln Gly Glu Ile Gly Ala Ala Arg Val Cys Pro Ala

65 70 75 80

Lys Ala Ser Ser Thr Leu Thr Phe Gln Phe Arg Glu Gln Pro Ser Asn

85 90 95

Pro Asn Ser Ala Pro Leu Asp Pro Ser His Lys Gly Pro Ala Ala Val

100 105 110

Tyr Leu Lys Lys Val Asp Ser Ala Ile Ala Ser Asn Asn Ala Ala Gly

115 120 125

Asp Gly Trp Phe Lys Ile Trp Glu Ser Val Tyr Asp Glu Ser Thr Gly

130 135 140

Lys Trp Gly Thr Thr Lys Met Ile Glu Asn Asn Gly His Ile Ser Val

145 150 155 160

Lys Val Pro Asp Asp Ile Glu Gly Gly Tyr Tyr Leu Ala Arg Thr Glu

165 170 175

Leu Leu Ala Leu His Ala Ala Asn Glu Gly Asp Pro Gln Phe Tyr Val

180 185 190

Gly Cys Ala Gln Leu Phe Ile Asp Ser Ala Gly Thr Ala Lys Pro Pro

195 200 205

Thr Val Ser Ile Gly Glu Gly Thr Tyr Asp Leu Ser Met Pro Ala Met

210 215 220

Thr Tyr Asn Ile Tyr Gln Thr Pro Leu Ala Leu Pro Tyr Pro Met Tyr

225 230 235 240

Gly Pro Pro Val Tyr Thr Pro Gly Ser Gly Ser Gly Ser Gly Ser Gly

245 250 255

Ser Gly Ser Ala Ser Ala Thr Arg Ser Ser Ala Ile Pro Thr Ala Thr

260 265 270

Ala Val Thr Asp Cys Ser Ser Glu Glu Asp Arg Glu Asp Ser Val Met

275 280 285

Ala Thr Gly Val Pro Val Ala Arg Ser Thr Leu Arg Thr Trp Val Asp

290 295 300

Arg Leu Ser Trp His Gly Lys Ala Arg Glu Asn Val Lys Pro Ala Ala

305 310 315 320

Arg Arg Ser Ala Leu Val Gln Thr Glu Gly Leu Lys Pro Glu Gly Cys

325 330 335

Ile Phe Val Asn Gly Asn Trp Cys Gly Phe Glu Val Pro Asp Tyr Asn

340 345 350

Asp Ala Glu Ser Cys Trp Ala Ala Ser Asp Asn Cys Trp Lys Gln Ser

355 360 365

Asp Ser Cys Trp Asn Gln Thr Gln Pro Thr Gly Tyr Asn Asn Cys Gln

370 375 380

Ile Trp Gln Asp Gln Lys Cys Lys Pro Ile Gln Asp Ser Cys Ser Gln

385 390 395 400

Ser Asn Pro Thr Gly Pro Pro Asn Lys Gly Lys Asp Ile Thr Pro Thr

405 410 415

Trp Pro Pro Leu Glu Gly Ser Met Lys Thr Phe Thr Lys Arg Thr Val

420 425 430

Ser Tyr Arg Asp Trp Ile Met Lys Arg Lys Gly Ala

435 440

<210> 24

<211> 253

<212> PRT

<213> Penicillium species

<400> 24

Met Leu Ser Ser Thr Thr Arg Thr Leu Ala Phe Thr Gly Leu Ala Gly

1 5 10 15

Leu Leu Ser Ala Pro Leu Val Lys Ala His Gly Phe Val Gln Gly Ile

20 25 30

Val Ile Gly Asp Gln Phe Tyr Ser Gly Tyr Ile Val Asn Ser Phe Pro

35 40 45

Tyr Glu Ser Asn Pro Pro Pro Val Ile Gly Trp Ala Thr Thr Ala Thr

50 55 60

Asp Leu Gly Phe Val Asp Gly Thr Gly Tyr Gln Gly Pro Asp Ile Ile

65 70 75 80

Cys His Arg Asn Ala Thr Pro Ala Pro Leu Thr Ala Pro Val Ala Ala

85 90 95

Gly Gly Thr Val Glu Leu Gln Trp Thr Pro Trp Pro Asp Ser His His

100 105 110

Gly Pro Val Ile Thr Tyr Leu Ala Pro Cys Asn Gly Asn Cys Ser Thr

115 120 125

Val Asp Lys Thr Thr Leu Glu Phe Phe Lys Ile Asp Gln Gln Gly Leu

130 135 140

Ile Asp Asp Thr Ser Pro Pro Gly Thr Trp Ala Ser Asp Asn Leu Ile

145 150 155 160

Ala Asn Asn Asn Ser Trp Thr Val Thr Ile Pro Asn Ser Val Ala Pro

165 170 175

Gly Asn Tyr Val Leu Arg His Glu Ile Ile Ala Leu His Ser Ala Asn

180 185 190

Asn Lys Asp Gly Ala Gln Asn Tyr Pro Gln Cys Ile Asn Ile Glu Val

195 200 205

Thr Gly Gly Gly Ser Asp Ala Pro Glu Gly Thr Leu Gly Glu Asp Leu

210 215 220

Tyr His Asp Thr Asp Pro Gly Ile Leu Val Asp Ile Tyr Glu Pro Ile

225 230 235 240

Ala Thr Tyr Thr Ile Pro Gly Pro Pro Glu Pro Thr Phe

245 250

<210> 25

<211> 223

<212> PRT

<213> Thielavia terrestris

<400> 25

Met Lys Leu Ser Ser Gln Leu Ala Ala Leu Thr Leu Ala Ala Ala Ser

1 5 10 15

Val Ser Gly His Tyr Ile Phe Glu Gln Ile Ala His Gly Gly Thr Lys

20 25 30

Phe Pro Pro Tyr Glu Tyr Ile Arg Arg Asn Thr Asn Tyr Asn Ser Pro

35 40 45

Val Thr Ser Leu Ser Ser Asn Asp Leu Arg Cys Asn Val Gly Gly Glu

50 55 60

Thr Ala Gly Asn Thr Thr Val Leu Asp Val Lys Ala Gly Asp Ser Phe

65 70 75 80

Thr Phe Tyr Ser Asp Val Ala Val Tyr His Gln Gly Pro Ile Ser Leu

85 90 95

Tyr Met Ser Lys Ala Pro Gly Ser Val Val Asp Tyr Asp Gly Ser Gly

100 105 110

Asp Trp Phe Lys Ile His Asp Trp Gly Pro Thr Phe Ser Asn Gly Gln

115 120 125

Ala Ser Trp Pro Leu Arg Asp Asn Tyr Gln Tyr Asn Ile Pro Thr Cys

130 135 140

Ile Pro Asn Gly Glu Tyr Leu Leu Arg Ile Gln Ser Leu Ala Ile His

145 150 155 160

Asn Pro Gly Ala Thr Pro Gln Phe Tyr Ile Ser Cys Ala Gln Val Arg

165 170 175

Val Ser Gly Gly Gly Ser Ala Ser Pro Ser Pro Thr Ala Lys Ile Pro

180 185 190

Gly Ala Phe Lys Ala Thr Asp Pro Gly Tyr Thr Ala Asn Ile Tyr Asn

195 200 205

Asn Phe His Ser Tyr Thr Val Pro Gly Pro Ala Val Phe Gln Cys

210 215 220

<210> 26

<211> 246

<212> PRT

<213> Thielavia terrestris

<400> 26

Met Lys Phe Ser Leu Val Ser Leu Leu Ala Tyr Gly Leu Ser Val Glu

1 5 10 15

Ala His Ser Ile Phe Gln Arg Val Ser Val Asn Gly Gln Asp Gln Gly

20 25 30

Leu Leu Thr Gly Leu Arg Ala Pro Ser Asn Asn Asn Pro Val Gln Asp

35 40 45

Val Asn Ser Gln Asn Met Ile Cys Gly Gln Ser Gly Ser Lys Ser Gln

50 55 60

Thr Val Ile Asn Val Lys Ala Gly Asp Arg Ile Gly Ser Leu Trp Gln

65 70 75 80

His Val Ile Gly Gly Ala Gln Phe Ser Gly Asp Pro Asp Asn Pro Ile

85 90 95

Ala His Ser His Lys Gly Pro Val Met Ala Tyr Leu Ala Lys Val Asp

100 105 110

Asn Ala Ala Ser Ala Ser Gln Thr Gly Leu Lys Trp Phe Lys Ile Trp

115 120 125

Gln Asp Gly Phe Asp Thr Ser Ser Lys Thr Trp Gly Val Asp Asn Leu

130 135 140

Ile Lys Asn Asn Gly Trp Val Tyr Phe His Leu Pro Gln Cys Leu Ala

145 150 155 160

Pro Gly Gln Tyr Leu Leu Arg Val Glu Val Leu Ala Leu His Ser Ala

165 170 175

Tyr Gln Gln Gly Gln Ala Gln Phe Tyr Gln Ser Cys Ala Gln Ile Asn

180 185 190

Val Ser Gly Ser Gly Ser Phe Ser Pro Ser Gln Thr Val Ser Ile Pro

195 200 205

Gly Val Tyr Ser Ala Thr Asp Pro Ser Ile Leu Ile Asn Ile Tyr Gly

210 215 220

Ser Thr Gly Gln Pro Asp Asn Gly Gly Lys Ala Tyr Asn Pro Pro Gly

225 230 235 240

Pro Ala Pro Ile Ser Cys

245

<210> 27

<211> 334

<212> PRT

<213> Thielavia terrestris

<400> 27

Met Arg Thr Thr Phe Ala Ala Ala Leu Ala Ala Phe Ala Ala Gln Glu

1 5 10 15

Val Ala Gly His Ala Ile Phe Gln Gln Leu Trp His Gly Ser Ser Cys

20 25 30

Val Arg Met Pro Leu Ser Asn Ser Pro Val Thr Asn Val Gly Ser Arg

35 40 45

Asp Met Ile Cys Asn Ala Gly Thr Arg Pro Val Ser Gly Lys Cys Pro

50 55 60

Val Lys Ala Gly Gly Thr Val Thr Val Glu Met His Gln Gln Pro Gly

65 70 75 80

Asp Arg Ser Cys Asn Asn Glu Ala Ile Gly Gly Ala His Trp Gly Pro

85 90 95

Val Gln Val Tyr Leu Ser Lys Val Glu Asp Ala Ser Thr Ala Asp Gly

100 105 110

Ser Thr Gly Trp Phe Lys Ile Phe Ala Asp Thr Trp Ser Lys Lys Ala

115 120 125

Gly Ser Ser Val Gly Asp Asp Asp Asn Trp Gly Thr Arg Asp Leu Asn

130 135 140

Ala Cys Cys Gly Lys Met Gln Val Lys Ile Pro Ala Asp Ile Pro Ser

145 150 155 160

Gly Asp Tyr Leu Leu Arg Ala Glu Ala Leu Ala Leu His Thr Ala Gly

165 170 175

Gln Val Gly Gly Ala Gln Phe Tyr Met Ser Cys Tyr Gln Ile Thr Val

180 185 190

Ser Gly Gly Gly Ser Ala Ser Pro Ala Thr Val Lys Phe Pro Gly Ala

195 200 205

Tyr Ser Ala Asn Asp Pro Gly Ile His Ile Asn Ile His Ala Ala Val

210 215 220

Ser Asn Tyr Val Ala Pro Gly Pro Ala Val Tyr Ser Gly Gly Thr Thr

225 230 235 240

Lys Val Ala Gly Ser Gly Cys Gln Gly Cys Glu Asn Thr Cys Lys Val

245 250 255

Gly Ser Ser Pro Thr Ala Thr Ala Pro Ser Gly Lys Ser Gly Ala Gly

260 265 270

Ser Asp Gly Gly Ala Gly Thr Asp Gly Gly Ser Ser Ser Ser Ser Pro

275 280 285

Asp Thr Gly Ser Ala Cys Ser Val Gln Ala Tyr Gly Gln Cys Gly Gly

290 295 300

Asn Gly Tyr Ser Gly Cys Thr Gln Cys Ala Pro Gly Tyr Thr Cys Lys

305 310 315 320

Ala Val Ser Pro Pro Tyr Tyr Ser Gln Cys Ala Pro Ser Ser

325 330

<210> 28

<211> 227

<212> PRT

<213> Thielavia terrestris

<400> 28

Met Lys Leu Ser Val Ala Ile Ala Val Leu Ala Ser Ala Leu Ala Glu

1 5 10 15

Ala His Tyr Thr Phe Pro Ser Ile Gly Asn Thr Ala Asp Trp Gln Tyr

20 25 30

Val Arg Ile Thr Thr Asn Tyr Gln Ser Asn Gly Pro Val Thr Asp Val

35 40 45

Thr Ser Asp Gln Ile Arg Cys Tyr Glu Arg Asn Pro Gly Thr Gly Ala

50 55 60

Gln Gly Ile Tyr Asn Val Thr Ala Gly Gln Thr Ile Asn Tyr Asn Ala

65 70 75 80

Lys Ala Ser Ile Ser His Pro Gly Pro Met Ser Phe Tyr Ile Ala Lys

85 90 95

Val Pro Ala Gly Gln Thr Ala Ala Thr Trp Asp Gly Lys Gly Ala Val

100 105 110

Trp Thr Lys Ile Tyr Gln Asp Met Pro Lys Phe Gly Ser Ser Leu Thr

115 120 125

Trp Pro Thr Met Gly Ala Lys Ser Val Pro Val Thr Ile Pro Arg Cys

130 135 140

Leu Gln Asn Gly Asp Tyr Leu Leu Arg Ala Glu His Ile Ala Leu His

145 150 155 160

Ser Ala Ser Ser Val Gly Gly Ala Gln Phe Tyr Leu Ser Cys Ala Gln

165 170 175

Leu Thr Val Ser Gly Gly Ser Gly Thr Trp Asn Pro Lys Asn Arg Val

180 185 190

Ser Phe Pro Gly Ala Tyr Lys Ala Thr Asp Pro Gly Ile Leu Ile Asn

195 200 205

Ile Tyr Tyr Pro Val Pro Thr Ser Tyr Ser Pro Pro Gly Pro Pro Ala

210 215 220

Glu Thr Cys

225

<210> 29

<211> 368

<212> PRT

<213> Thielavia terrestris

<400> 29

Met Pro Ser Phe Ala Ser Lys Thr Leu Leu Ser Thr Leu Ala Gly Ala

1 5 10 15

Ala Ser Val Ala Ala His Gly His Val Ser Asn Ile Val Ile Asn Gly

20 25 30

Val Ser Tyr Gln Gly Tyr Asp Pro Thr Ser Phe Pro Tyr Met Gln Asn

35 40 45

Pro Pro Ile Val Val Gly Trp Thr Ala Ala Asp Thr Asp Asn Gly Phe

50 55 60

Val Ala Pro Asp Ala Phe Ala Ser Gly Asp Ile Ile Cys His Lys Asn

65 70 75 80

Ala Thr Asn Ala Lys Gly His Ala Val Val Ala Ala Gly Asp Lys Ile

85 90 95

Phe Ile Gln Trp Asn Thr Trp Pro Glu Ser His His Gly Pro Val Ile

100 105 110

Asp Tyr Leu Ala Ser Cys Gly Ser Ala Ser Cys Glu Thr Val Asp Lys

115 120 125

Thr Lys Leu Glu Phe Phe Lys Ile Asp Glu Val Gly Leu Val Asp Gly

130 135 140

Ser Ser Ala Pro Gly Val Trp Gly Ser Asp Gln Leu Ile Ala Asn Asn

145 150 155 160

Asn Ser Trp Leu Val Glu Ile Pro Pro Thr Ile Ala Pro Gly Asn Tyr

165 170 175

Val Leu Arg His Glu Ile Ile Ala Leu His Ser Ala Glu Asn Ala Asp

180 185 190

Gly Ala Gln Asn Tyr Pro Gln Cys Phe Asn Leu Gln Ile Thr Gly Thr

195 200 205

Gly Thr Ala Thr Pro Ser Gly Val Pro Gly Thr Ser Leu Tyr Thr Pro

210 215 220

Thr Asp Pro Gly Ile Leu Val Asn Ile Tyr Ser Ala Pro Ile Thr Tyr

225 230 235 240

Thr Val Pro Gly Pro Ala Leu Ile Ser Gly Ala Val Ser Ile Ala Gln

245 250 255

Ser Ser Ser Ala Ile Thr Ala Ser Gly Thr Ala Leu Thr Gly Ser Ala

260 265 270

Thr Ala Pro Ala Ala Ala Ala Ala Thr Thr Thr Ser Thr Thr Asn Ala

275 280 285

Ala Ala Ala Ala Thr Ser Ala Ala Ala Ala Ala Gly Thr Ser Thr Thr

290 295 300

Thr Thr Ser Ala Ala Ala Val Val Gln Thr Ser Ser Ser Ser Ser Ser

305 310 315 320

Ala Pro Ser Ser Ala Ala Ala Ala Ala Thr Thr Thr Ala Ala Ala Ser

325 330 335

Ala Arg Pro Thr Gly Cys Ser Ser Gly Arg Ser Arg Lys Gln Pro Arg

340 345 350

Arg His Ala Arg Asp Met Val Val Ala Arg Gly Ala Glu Glu Ala Asn

355 360 365

<210> 30

<211> 330

<212> PRT

<213> Thielavia terrestris

<400> 30

Met Pro Pro Ala Leu Pro Gln Leu Leu Thr Thr Val Leu Thr Ala Leu

1 5 10 15

Thr Leu Gly Ser Thr Ala Leu Ala His Ser His Leu Ala Tyr Ile Ile

20 25 30

Val Asn Gly Lys Leu Tyr Gln Gly Phe Asp Pro Arg Pro His Gln Ala

35 40 45

Asn Tyr Pro Ser Arg Val Gly Trp Ser Thr Gly Ala Val Asp Asp Gly

50 55 60

Phe Val Thr Pro Ala Asn Tyr Ser Thr Pro Asp Ile Ile Cys His Ile

65 70 75 80

Ala Gly Thr Ser Pro Ala Gly His Ala Pro Val Arg Pro Gly Asp Arg

85 90 95

Ile His Val Gln Trp Asn Gly Trp Pro Val Gly His Ile Gly Pro Val

100 105 110

Leu Ser Tyr Leu Ala Arg Cys Glu Ser Asp Thr Gly Cys Thr Gly Gln

115 120 125

Asn Lys Thr Ala Leu Arg Trp Thr Lys Ile Asp Asp Ser Ser Pro Thr

130 135 140

Met Gln Asn Val Ala Gly Ala Gly Thr Gln Gly Glu Gly Thr Pro Gly

145 150 155 160

Lys Arg Trp Ala Thr Asp Val Leu Ile Ala Ala Asn Asn Ser Trp Gln

165 170 175

Val Ala Val Pro Ala Gly Leu Pro Thr Gly Ala Tyr Val Leu Arg Asn

180 185 190

Glu Ile Ile Ala Leu His Tyr Ala Ala Arg Lys Asn Gly Ala Gln Asn

195 200 205

Tyr Pro Leu Cys Met Asn Leu Trp Val Asp Ala Ser Gly Asp Asn Ser

210 215 220

Ser Val Ala Ala Thr Thr Ala Ala Val Thr Ala Gly Gly Leu Gln Met

225 230 235 240

Asp Ala Tyr Asp Ala Arg Gly Phe Tyr Lys Glu Asn Asp Pro Gly Val

245 250 255

Leu Val Asn Val Thr Ala Ala Leu Ser Ser Tyr Val Val Pro Gly Pro

260 265 270

Thr Val Ala Ala Gly Ala Thr Pro Val Pro Tyr Ala Gln Gln Ser Pro

275 280 285

Ser Val Ser Thr Ala Ala Gly Thr Pro Val Val Val Thr Arg Thr Ser

290 295 300

Glu Thr Ala Pro Tyr Thr Gly Ala Met Thr Pro Thr Val Ala Ala Arg

305 310 315 320

Met Lys Gly Arg Gly Tyr Asp Arg Arg Gly

325 330

<210> 31

<211> 236

<212> PRT

<213> Thielavia terrestris

<400> 31

Met Lys Thr Phe Thr Ala Leu Leu Ala Ala Ala Gly Leu Val Ala Gly

1 5 10 15

His Gly Tyr Val Asp Asn Ala Thr Ile Gly Gly Gln Phe Tyr Gln Asn

20 25 30

Pro Ala Val Leu Thr Phe Phe Gln Pro Asp Arg Val Ser Arg Ser Ile

35 40 45

Pro Gly Asn Gly Pro Val Thr Asp Val Thr Leu Ile Asp Leu Gln Cys

50 55 60

Asn Ala Asn Ser Thr Pro Ala Lys Leu His Ala Thr Ala Ala Ala Gly

65 70 75 80

Ser Asp Val Ile Leu Arg Trp Thr Leu Trp Pro Glu Ser His Val Gly

85 90 95

Pro Val Ile Thr Tyr Met Ala Arg Cys Pro Asp Thr Gly Cys Gln Asp

100 105 110

Trp Met Pro Gly Thr Ser Ala Val Trp Phe Lys Ile Lys Glu Gly Gly

115 120 125

Arg Asp Gly Thr Ser Asn Thr Trp Ala Asp Thr Pro Leu Met Thr Ala

130 135 140

Pro Thr Ser Tyr Thr Tyr Thr Ile Pro Ser Cys Leu Lys Lys Gly Tyr

145 150 155 160

Tyr Leu Val Arg His Glu Ile Ile Ala Leu His Ala Ala Tyr Thr Tyr

165 170 175

Pro Gly Ala Gln Phe Tyr Pro Gly Cys His Gln Leu Asn Val Thr Gly

180 185 190

Gly Gly Ser Thr Val Pro Ser Ser Gly Leu Val Ala Phe Pro Gly Ala

195 200 205

Tyr Lys Gly Ser Asp Pro Gly Ile Thr Tyr Asp Ala Tyr Lys Ala Gln

210 215 220

Thr Tyr Gln Ile Pro Gly Pro Ala Val Phe Thr Cys

225 230 235

<210> 32

<211> 250

<212> PRT

<213> Thielavia terrestris

<400> 32

Met Ala Leu Leu Leu Leu Ala Gly Leu Ala Ile Leu Ala Gly Pro Ala

1 5 10 15

His Ala His Gly Gly Leu Ala Asn Tyr Thr Val Gly Asn Thr Trp Tyr

20 25 30

Arg Gly Tyr Asp Pro Phe Thr Pro Ala Ala Asp Gln Ile Gly Gln Pro

35 40 45

Trp Met Ile Gln Arg Ala Trp Asp Ser Ile Asp Pro Ile Phe Ser Val

50 55 60

Asn Asp Lys Ala Leu Ala Cys Asn Thr Pro Ala Thr Ala Pro Thr Ser

65 70 75 80

Tyr Ile Pro Ile Arg Ala Gly Glu Asn Ile Thr Ala Val Tyr Trp Tyr

85 90 95

Trp Leu His Pro Val Gly Pro Met Thr Ala Trp Leu Ala Arg Cys Asp

100 105 110

Gly Asp Cys Arg Asp Ala Asp Val Asn Glu Ala Arg Trp Phe Lys Ile

115 120 125

Trp Glu Ala Gly Leu Leu Ser Gly Pro Asn Leu Ala Glu Gly Met Trp

130 135 140

Tyr Gln Lys Ala Phe Gln Asn Trp Asp Gly Ser Pro Asp Leu Trp Pro

145 150 155 160

Val Thr Ile Pro Ala Gly Leu Lys Ser Gly Leu Tyr Met Ile Arg His

165 170 175

Glu Ile Leu Ser Ile His Val Glu Asp Lys Pro Gln Phe Tyr Pro Glu

180 185 190

Cys Ala His Leu Asn Val Thr Gly Gly Gly Asp Leu Leu Pro Pro Asp

195 200 205

Glu Phe Leu Val Lys Phe Pro Gly Ala Tyr Lys Glu Asp Asn Pro Ser

210 215 220

Ile Lys Ile Asn Ile Tyr Ser Asp Gln Tyr Ala Asn Thr Thr Asn Tyr

225 230 235 240

Thr Ile Pro Gly Gly Pro Ile Trp Asp Gly

245 250

<210> 33

<211> 478

<212> PRT

<213> Thielavia terrestris

<400> 33

Met Met Pro Ser Leu Val Arg Phe Ser Met Gly Leu Ala Thr Ala Phe

1 5 10 15

Ala Ser Leu Ser Thr Ala His Thr Val Phe Thr Thr Leu Phe Ile Asn

20 25 30

Gly Val Asp Gln Gly Asp Gly Thr Cys Ile Arg Met Ala Lys Lys Gly

35 40 45

Ser Val Cys Thr His Pro Ile Ala Gly Gly Leu Asp Ser Pro Asp Met

50 55 60

Ala Cys Gly Arg Asp Gly Gln Gln Ala Val Ala Phe Thr Cys Pro Ala

65 70 75 80

Pro Ala Gly Ser Lys Leu Ser Phe Glu Phe Arg Met Trp Ala Asp Ala

85 90 95

Ser Gln Pro Gly Ser Ile Asp Pro Ser His Leu Gly Ser Thr Ala Ile

100 105 110

Tyr Leu Lys Gln Val Ser Asn Ile Ser Ser Asp Ser Ala Ala Gly Pro

115 120 125

Gly Trp Phe Lys Ile Tyr Ala Glu Gly Tyr Asp Thr Ala Ala Lys Lys

130 135 140

Trp Ala Thr Glu Lys Leu Ile Asp Asn Gly Gly Leu Leu Ser Ile Glu

145 150 155 160

Leu Pro Pro Thr Leu Pro Ala Gly Tyr Tyr Leu Ala Arg Ser Glu Ile

165 170 175

Val Thr Ile Gln Asn Val Thr Asn Asp His Val Asp Pro Gln Phe Tyr

180 185 190

Val Gly Cys Ala Gln Leu Phe Val Gln Gly Pro Pro Thr Thr Pro Thr

195 200 205

Val Pro Pro Asp Arg Leu Val Ser Ile Pro Gly His Val His Ala Ser

210 215 220

Asp Pro Gly Leu Thr Phe Asn Ile Trp Arg Asp Asp Pro Ser Lys Thr

225 230 235 240

Ala Tyr Thr Val Val Gly Pro Ala Pro Phe Ser Pro Thr Ala Ala Pro

245 250 255

Thr Pro Thr Ser Thr Asn Thr Asn Gly Gln Gln Gln Gln Gln Gln Gln

260 265 270

Gln Ala Ile Lys Gln Thr Asp Gly Val Ile Pro Ala Asp Cys Gln Leu

275 280 285

Lys Asn Ala Asn Trp Cys Gly Ala Glu Val Pro Ala Tyr Ala Asp Glu

290 295 300

Ala Gly Cys Trp Ala Ser Ser Ala Asp Cys Phe Ala Gln Leu Asp Ala

305 310 315 320

Cys Tyr Thr Ser Ala Pro Pro Thr Gly Ser Arg Gly Cys Arg Leu Trp

325 330 335

Glu Asp Trp Cys Thr Gly Ile Gln Gln Gly Cys Arg Ala Gly Arg Trp

340 345 350

Arg Gly Pro Pro Pro Phe His Gly Glu Gly Ala Ala Ala Glu Thr Ala

355 360 365

Ser Ala Gly Arg Gly Gly Ala Arg Ile Ala Ala Val Ala Gly Cys Gly

370 375 380

Gly Gly Thr Gly Asp Met Val Glu Glu Val Phe Leu Phe Tyr Trp Asp

385 390 395 400

Ala Cys Ser Gly Trp Arg Arg Ser Arg Gly Gly Gly Ser Ile Leu Ala

405 410 415

Arg Leu Ile Leu His Val Leu Leu Pro Leu Leu Arg Pro Arg Arg Ala

420 425 430

Pro Arg Val His Leu Leu Leu Phe His Leu Tyr Leu Asn Phe Cys Tyr

435 440 445

Pro Gly Thr Ser Gly Phe Tyr Asn Arg Leu Ser Ile Lys Leu Gly Ile

450 455 460

Trp Pro Ser Lys Met Ser Pro Asp Val Ala His Tyr Val Lys

465 470 475

<210> 34

<211> 230

<212> PRT

<213> Thielavia terrestris

<400> 34

Met Gln Leu Leu Val Gly Leu Leu Leu Ala Ala Val Ala Ala Arg Ala

1 5 10 15

His Tyr Thr Phe Pro Arg Leu Val Val Asn Gly Gln Pro Glu Asp Lys

20 25 30

Asp Trp Ser Val Thr Arg Met Thr Lys Asn Ala Gln Ser Lys Gln Gly

35 40 45

Val Gln Asp Pro Thr Ser Pro Asp Ile Arg Cys Tyr Thr Ser Gln Thr

50 55 60

Ala Pro Asn Val Ala Thr Val Pro Ala Gly Ala Thr Val His Tyr Ile

65 70 75 80

Ser Thr Gln Gln Ile Asn His Pro Gly Pro Thr Gln Tyr Tyr Leu Ala

85 90 95

Lys Val Pro Ala Gly Ser Ser Ala Lys Thr Trp Asp Gly Ser Gly Ala

100 105 110

Val Trp Phe Lys Ile Ser Thr Thr Met Pro Tyr Leu Asp Asn Asn Lys

115 120 125

Gln Leu Val Trp Pro Asn Gln Asn Thr Tyr Thr Thr Val Asn Thr Thr

130 135 140

Ile Pro Ala Asp Thr Pro Ser Gly Glu Tyr Leu Leu Arg Val Glu Gln

145 150 155 160

Ile Ala Leu His Leu Ala Ser Gln Pro Asn Gly Ala Gln Phe Tyr Leu

165 170 175

Ala Cys Ser Gln Ile Gln Ile Thr Gly Gly Gly Asn Gly Thr Pro Gly

180 185 190

Pro Leu Val Ala Leu Pro Gly Ala Tyr Lys Ser Asn Asp Pro Gly Ile

195 200 205

Leu Val Asn Ile Tyr Ser Met Gln Pro Gly Asp Tyr Lys Pro Pro Gly

210 215 220

Pro Pro Val Trp Ser Gly

225 230

<210> 35

<211> 257

<212> PRT

<213> Thielavia terrestris

<400> 35

Met Lys Leu Tyr Leu Ala Ala Phe Leu Gly Ala Val Ala Thr Pro Gly

1 5 10 15

Ala Phe Ala His Gln Ile His Gly Ile Leu Leu Val Asn Gly Thr Glu

20 25 30

Thr Pro Glu Trp Lys Tyr Val Arg Asp Val Ala Trp Glu Gly Ala Tyr

35 40 45

Glu Pro Glu Lys Tyr Pro Asn Thr Glu Phe Phe Lys Thr Pro Pro Gln

50 55 60

Thr Asp Ile Asn Asn Pro Asn Ile Thr Cys Gly Arg Asn Ala Phe Asp

65 70 75 80

Ser Ala Ser Lys Thr Glu Thr Ala Asp Ile Leu Ala Gly Ser Glu Val

85 90 95

Gly Phe Arg Val Ser Trp Asp Gly Asn Gly Lys Tyr Gly Val Phe Trp

100 105 110

His Pro Gly Pro Gly Gln Ile Tyr Leu Ser Arg Ala Pro Asn Asp Asp

115 120 125

Leu Glu Asp Tyr Arg Gly Asp Gly Asp Trp Phe Lys Ile Ala Thr Gly

130 135 140

Ala Ala Val Ser Asn Thr Glu Trp Leu Leu Trp Asn Lys His Asp Phe

145 150 155 160

Asn Phe Thr Ile Pro Lys Thr Thr Pro Pro Gly Lys Tyr Leu Met Arg

165 170 175

Ile Glu Gln Phe Met Pro Ser Thr Val Glu Tyr Ser Gln Trp Tyr Val

180 185 190

Asn Cys Ala His Val Asn Ile Ile Gly Pro Gly Gly Gly Thr Pro Thr

195 200 205

Gly Phe Ala Arg Phe Pro Gly Thr Tyr Thr Val Asp Asp Pro Gly Ile

210 215 220

Lys Val Pro Leu Asn Gln Ile Val Asn Ser Gly Glu Leu Pro Gln Asp

225 230 235 240

Gln Leu Arg Leu Leu Glu Tyr Lys Pro Pro Gly Pro Ala Leu Trp Thr

245 250 255

Gly

<210> 36

<211> 251

<212> PRT

<213> Thermoascus crustaceus

<400> 36

Met Ala Phe Ser Gln Ile Met Ala Ile Thr Gly Val Phe Leu Ala Ser

1 5 10 15

Ala Ser Leu Val Ala Gly His Gly Phe Val Gln Asn Ile Val Ile Asp

20 25 30

Gly Lys Ser Tyr Gly Gly Tyr Ile Val Asn Gln Tyr Pro Tyr Met Ser

35 40 45

Asp Pro Pro Glu Val Val Gly Trp Ser Thr Thr Ala Thr Asp Leu Gly

50 55 60

Phe Val Asp Gly Thr Gly Tyr Gln Gly Pro Asp Ile Ile Cys His Arg

65 70 75 80

Gly Ala Lys Pro Ala Ala Leu Thr Ala Gln Val Ala Ala Gly Gly Thr

85 90 95

Val Lys Leu Glu Trp Thr Pro Trp Pro Asp Ser His His Gly Pro Val

100 105 110

Ile Asn Tyr Leu Ala Pro Cys Asn Gly Asp Cys Ser Thr Val Asp Lys

115 120 125

Thr Gln Leu Lys Phe Phe Lys Ile Ala Gln Ala Gly Leu Ile Asp Asp

130 135 140

Asn Ser Pro Pro Gly Ile Trp Ala Ser Asp Asn Leu Ile Ala Ala Asn

145 150 155 160

Asn Ser Trp Thr Val Thr Ile Pro Thr Thr Thr Ala Pro Gly Asn Tyr

165 170 175

Val Leu Arg His Glu Ile Ile Ala Leu His Ser Ala Gly Asn Lys Asp

180 185 190

Gly Ala Gln Asn Tyr Pro Gln Cys Ile Asn Leu Lys Val Thr Gly Asn

195 200 205

Gly Ser Gly Asn Pro Pro Ala Gly Ala Leu Gly Thr Ala Leu Tyr Lys

210 215 220

Asp Thr Asp Pro Gly Ile Leu Ile Asn Ile Tyr Gln Lys Leu Ser Ser

225 230 235 240

Tyr Val Ile Pro Gly Pro Ala Leu Tyr Thr Gly

245 250

<210> 37

<211> 349

<212> PRT

<213> Thermoascus crustaceus

<400> 37

Met Ser Phe Ser Lys Ile Leu Ala Ile Ala Gly Ala Ile Thr Tyr Ala

1 5 10 15

Ser Ser Ala Ala Ala His Gly Tyr Val Gln Gly Ile Val Val Asp Gly

20 25 30

Ser Tyr Tyr Gly Gly Tyr Met Val Thr Gln Tyr Pro Tyr Thr Ala Gln

35 40 45

Pro Pro Glu Leu Ile Ala Trp Ser Thr Lys Ala Thr Asp Leu Gly Phe

50 55 60

Val Asp Gly Ser Gly Tyr Thr Ser Pro Asp Ile Ile Cys His Lys Gly

65 70 75 80

Ala Glu Pro Gly Ala Gln Ser Ala Lys Val Ala Ala Gly Gly Thr Val

85 90 95

Glu Leu Gln Trp Thr Ala Trp Pro Glu Ser His Lys Gly Pro Val Ile

100 105 110

Asp Tyr Leu Ala Ala Cys Asp Gly Asp Cys Ser Ser Val Asp Lys Thr

115 120 125

Ala Leu Lys Phe Phe Lys Ile Asp Glu Ser Gly Leu Ile Asp Gly Asn

130 135 140

Gly Ala Gly Thr Trp Ala Ser Asp Thr Leu Ile Lys Asn Asn Asn Ser

145 150 155 160

Trp Thr Val Thr Ile Pro Ser Thr Ile Ala Ser Gly Asn Tyr Val Leu

165 170 175

Arg His Glu Ile Ile Ala Leu His Ser Ala Gly Asn Lys Asp Gly Ala

180 185 190

Gln Asn Tyr Pro Gln Cys Ile Asn Leu Glu Val Thr Gly Ser Gly Thr

195 200 205

Glu Asn Pro Ala Gly Thr Leu Gly Thr Ala Leu Tyr Thr Asp Thr Asp

210 215 220

Pro Gly Leu Leu Val Asn Ile Tyr Gln Gly Leu Ser Asn Tyr Ser Ile

225 230 235 240

Pro Gly Pro Ala Leu Tyr Ser Gly Asn Ser Asp Asn Ala Gly Ser Leu

245 250 255

Asn Pro Thr Thr Thr Pro Ser Ile Gln Asn Ala Ala Ala Ala Pro Ser

260 265 270

Thr Ser Thr Ala Ser Val Val Thr Asp Ser Ser Ser Ala Thr Gln Thr

275 280 285

Ala Ser Val Ala Ala Thr Thr Pro Ala Ser Thr Ser Ala Val Thr Ala

290 295 300

Ser Pro Ala Pro Asp Thr Gly Ser Asp Val Thr Lys Tyr Leu Asp Ser

305 310 315 320

Met Ser Ser Asp Glu Val Leu Thr Leu Val Arg Gly Thr Leu Ser Trp

325 330 335

Leu Val Ser Asn Lys Lys His Ala Arg Asp Leu Ser His

340 345

<210> 38

<211> 436

<212> PRT

<213> Thermoascus crustaceus

<400> 38

Met Leu Ser Phe Ile Pro Thr Lys Ser Ala Ala Leu Thr Thr Leu Leu

1 5 10 15

Leu Leu Gly Thr Ala His Ala His Thr Leu Met Thr Thr Met Phe Val

20 25 30

Asp Gly Val Asn Gln Gly Asp Gly Val Cys Ile Arg Met Asn Asn Asp

35 40 45

Gly Gly Thr Ala Asn Thr Tyr Ile Gln Pro Ile Thr Ser Lys Asp Ile

50 55 60

Ala Cys Gly Ile Gln Gly Glu Ile Gly Ala Ser Arg Val Cys Pro Val

65 70 75 80

Lys Ala Ser Ser Thr Leu Thr Phe Gln Phe Arg Glu Gln Pro Asn Asn

85 90 95

Pro Asn Ser Ser Pro Leu Asp Pro Ser His Lys Gly Pro Ala Ala Val

100 105 110

Tyr Leu Lys Lys Val Asp Ser Ala Ile Ala Ser Asn Asn Ala Ala Gly

115 120 125

Asp Ser Trp Phe Lys Ile Trp Glu Ser Val Tyr Asp Glu Ser Thr Gly

130 135 140

Lys Trp Gly Thr Thr Lys Met Ile Glu Asn Asn Gly His Ile Ser Val

145 150 155 160

Lys Val Pro Asp Asp Ile Glu Gly Gly Tyr Tyr Leu Ala Arg Thr Glu

165 170 175

Leu Leu Ala Leu His Ser Ala Asp Gln Gly Asp Pro Gln Phe Tyr Val

180 185 190

Gly Cys Ala Gln Leu Phe Ile Asp Ser Asp Gly Thr Ala Lys Pro Pro

195 200 205

Thr Val Ser Ile Gly Glu Gly Thr Tyr Asp Leu Ser Met Pro Ala Met

210 215 220

Thr Tyr Asn Ile Trp Glu Thr Pro Leu Ala Leu Pro Tyr Pro Met Tyr

225 230 235 240

Gly Pro Pro Val Tyr Thr Pro Gly Ser Gly Ser Gly Ser Val Arg Ala

245 250 255

Thr Ser Ser Ser Ala Val Pro Thr Ala Thr Glu Ser Ser Phe Val Glu

260 265 270

Glu Arg Ala Asn Pro Val Thr Ala Asn Ser Val Tyr Ser Ala Arg Gly

275 280 285

Lys Phe Lys Thr Trp Ile Asp Lys Leu Ser Trp Arg Gly Lys Val Arg

290 295 300

Glu Asn Val Arg Gln Ala Ala Gly Arg Arg Ser Thr Leu Val Gln Thr

305 310 315 320

Val Gly Leu Lys Pro Lys Gly Cys Ile Phe Val Asn Gly Asn Trp Cys

325 330 335

Gly Phe Glu Val Pro Asp Tyr Asn Asp Ala Glu Ser Cys Trp Ala Ala

340 345 350

Ser Asp Asn Cys Trp Lys Gln Ser Asp Ala Cys Trp Asn Lys Thr Gln

355 360 365

Pro Thr Gly Tyr Asn Asn Cys Gln Ile Trp Gln Asp Lys Lys Cys Lys

370 375 380

Val Ile Gln Asp Ser Cys Ser Gly Pro Asn Pro His Gly Pro Pro Asn

385 390 395 400

Lys Gly Lys Asp Leu Thr Pro Glu Trp Pro Pro Leu Lys Gly Ser Met

405 410 415

Asp Thr Phe Ser Lys Arg Thr Ile Gly Tyr Arg Asp Trp Ile Val Arg

420 425 430

Arg Arg Gly Ala

435

Claims (16)

1. A method for treating a dissolving pulp, the method comprising the step of subjecting the dissolving pulp to a soluble polysaccharide monooxygenase.

2. The method according to claim 1, comprising the step of subjecting the dissolving pulp to a cellulase, preferably an endoglucanase.

3. The method according to claim 1 or 2, wherein the steps of subjecting the dissolving pulp to a solubilised polysaccharide monooxygenase and subjecting the dissolving pulp to a cellulase enzyme are performed simultaneously or sequentially in any order.

4. A method according to any one of claims 1 to 3, comprising the step of bleaching the dissolving pulp.

5. The method according to any one of claims 1 to 4, comprising the step of subjecting the solubilised pulp to solubilising polysaccharide monooxygenase and its electron donor, preferably ascorbic acid, gallic acid, pyrogallol or cysteine.

6. The method according to any one of claims 1 to 5, wherein the dissolving pulp is unbleached, partially bleached, bleached or alkaline extracted dissolving pulp; or wherein the dissolving pulp is kraft pulp or sulfite pulp.

7. The method according to any one of claims 1 to 6, wherein the soluble polysaccharide monooxygenase has at least 60% (e.g. at least 65%, e.g. at least 70%, e.g. at least 75%, e.g. at least 80%, e.g. at least 85%, e.g. at least 90%, e.g. at least 95%, e.g. at least 99%) sequence identity with the mature polypeptide of SEQ ID No. 1 or the mature polypeptide of SEQ ID No. 2 or the mature polypeptide of SEQ ID No. 3.

8. The method according to any one of claims 1 to 7, wherein the cellulase has at least 60% (e.g. at least 65%, e.g. at least 70%, e.g. at least 75%, e.g. at least 80%, e.g. at least 85%, e.g. at least 90%, e.g. at least 95%, e.g. at least 99%) sequence identity with the mature polypeptide of SEQ ID No. 4, SEQ ID No. 5, SEQ ID No. 6 or SEQ ID No. 7.

9. The method according to any one of claims 1 to 8, wherein the soluble polysaccharide monooxygenase comprises or consists of: 1 or a mature polypeptide thereof, or 2 or a mature polypeptide thereof, or 3 or a mature polypeptide thereof; or the cellulase comprises or consists of: SEQ ID NO. 4, SEQ ID NO. 5 or a mature polypeptide thereof, SEQ ID NO. 6 or a mature polypeptide thereof, or SEQ ID NO. 7 or a mature polypeptide thereof.

10. The method according to any one of claims 1 to 9, wherein the concentration of the soluble polysaccharide monooxygenase is from 0.05mg/kg of oven dried pulp to 100000mg/kg of oven dried pulp, for example a concentration selected from the group consisting of: from 0.05mg/kg of oven-dried pulp to 250mg/kg of oven-dried pulp, from 0.25mg/kg of oven-dried pulp to 1000mg/kg of oven-dried pulp, from 0.25mg/kg of oven-dried pulp to 2000mg/kg of oven-dried pulp, from 1.0mg/kg of oven-dried pulp to 5000mg/kg of oven-dried pulp, from 5.0mg/kg of oven-dried pulp to 10000mg/kg of oven-dried pulp, from 10.0mg/kg of oven-dried pulp to 15000mg/kg of oven-dried pulp, from 15.0mg/kg of oven-dried pulp to 20000mg/kg of oven-dried pulp, from 20.0mg/kg of oven-dried pulp to 30000mg/kg of oven-dried pulp, from 30.0mg/kg of oven-dried pulp to 40000mg/kg of oven-dried pulp, from 40.0mg/kg of oven-dried pulp to 60000mg/kg of oven-dried pulp, from 60.0mg/kg of oven-dried pulp to 80000mg/kg of oven-dried pulp, and from 10000mg/kg of oven-dried pulp, or any combination of these intervals.

11. The method according to any of claims 1 to 10, wherein the concentration of the cellulase enzyme is from 0.05mg/kg of oven dried pulp to 100mg/kg of oven dried pulp, for example a concentration selected from the group consisting of: from 0.05mg/kg of oven-dried pulp to 80.0mg/kg of oven-dried pulp, from 0.25mg/kg of oven-dried pulp to 60.0mg/kg of oven-dried pulp, from 0.25mg/kg of oven-dried pulp to 40.0mg/kg of oven-dried pulp, from 0.25mg/kg of oven-dried pulp to 20.0mg/kg of oven-dried pulp, from 0.25mg/kg of oven-dried pulp to 10.0mg/kg of oven-dried pulp, from 0.25mg/kg of oven-dried pulp to 5.0mg/kg of oven-dried pulp, from 0.50mg/kg of oven-dried pulp to 85.0mg/kg of oven-dried pulp, from 0.50mg/kg of oven-dried pulp to 65.0mg/kg of oven-dried pulp, from 0.50mg/kg of oven-dried pulp to 45.0mg/kg of oven-dried pulp, from 0.50mg/kg of oven-dried pulp to 25.0mg/kg of oven-dried pulp, from 0.50mg/kg of oven-dried pulp to 15.0mg/kg of oven-dried pulp, from 0.50mg/kg of oven-dried pulp to 5mg/kg of oven-dried pulp, or any combination of these intervals.

12. The method according to any one of claims 1 to 11, wherein the method results in a reduction of viscosity and/or an improvement of viscosity control during dissolving pulp production; and/or wherein the process results in an increase in the reactivity of the dissolving pulp, preferably an increase in Fock reactivity; and/or wherein the process results in an increase in the content of oxidizing groups of the dissolving pulp.

13. The method according to any one of claims 1 to 12, wherein 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 produced by the method according to any one of claims 1 to 13.

15. A textile fiber or derived cellulose made from the dissolving pulp of claim 14.

16. Use of a solubilising polysaccharide monooxygenase for the treatment of solubilised pulp.