WO2016015570A1 - Composition d'oxydase de fibre utilisée pour modifier et améliorer la blancheur du papier, procédé de fabrication de papier, et applications de la composition - Google Patents

Composition d'oxydase de fibre utilisée pour modifier et améliorer la blancheur du papier, procédé de fabrication de papier, et applications de la composition Download PDF

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WO2016015570A1
WO2016015570A1 PCT/CN2015/084455 CN2015084455W WO2016015570A1 WO 2016015570 A1 WO2016015570 A1 WO 2016015570A1 CN 2015084455 W CN2015084455 W CN 2015084455W WO 2016015570 A1 WO2016015570 A1 WO 2016015570A1
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fiber
oxidase
paper
pulp
composition
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PCT/CN2015/084455
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Chinese (zh)
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王祥槐
谢焱
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瑞辰星生物技术（广州）有限公司
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Priority claimed from CN201410366879.1A external-priority patent/CN104342424B/zh
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/18—Carboxylic ester hydrolases (3.1.1)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/88—Lyases (4.)
    • D—TEXTILES; PAPER
    • D21—PAPER-MAKING; PRODUCTION OF CELLULOSE
    • D21H—PULP COMPOSITIONS; PREPARATION THEREOF NOT COVERED BY SUBCLASSES D21C OR D21D; IMPREGNATING OR COATING OF PAPER; TREATMENT OF FINISHED PAPER NOT COVERED BY CLASS B31 OR SUBCLASS D21G; PAPER NOT OTHERWISE PROVIDED FOR
    • D21H17/00—Non-fibrous material added to the pulp, characterised by its constitution; Paper-impregnating material characterised by its constitution

Definitions

  • the present invention relates to the field of biological enzymes and the use of biological enzymes to improve the physical and chemical properties of fibers, and in particular to a composition and papermaking method and application for modifying and improving the surface properties of fibers with fiber oxidase.
  • China's paper industry maintained a high growth rate for 10 consecutive years from 2000 to 2010. By 2011, China's paper and paperboard production and consumption exceeded 100 million tons, ranking first in the world.
  • the raw materials of paper or paperboard are mainly fiber. Because pulping and papermaking require a lot of raw materials and have certain influence on the environment, the paper industry in the world is compressing pulp production to ensure forest resources and domestic raw material forest. The construction of the base is sluggish and the supply of materials is limited. The development of non-wood pulp is affected by the lagging development of new technologies for clean production. The constraints on resources, energy and environment faced by the development of China's paper industry are increasingly prominent.
  • U.S. Patent 5,725,732 teaches the use of cellulase and hemicellulase to address the hair loss of fibers.
  • U.S. Patent No. 6,066,233 the use of cellulase and pectinase in combination with water filtration through pulp.
  • U.S. Patent No. 5,582,681 teaches the use of a mixed enzyme preparation such as cellulase, hemicellulase and lipolytic enzyme to improve the flexibility of toilet paper.
  • treatment of the pulp with cellulase or hemicellulase can also improve the compressibility of the fiber, reduce the microporosity of the paper, increase the density, and improve the transparency.
  • fiber-modified enzymes currently used in the paper industry are all utilizing cellulolytic enzymes, including endocellulases and exocellulases, on the fibers (-1,4 glycosidic bonds (b-1,4). -linked D-glucose units) undergo hydrolysis reaction and depolymerization into polysaccharides of smaller molecular weight.
  • cellulose hydrolase has a certain effect on reducing the energy consumption of refining and improving the dehydration of the net.
  • the degradation rate of the fiber can be as high as 1%, which not only causes fiber loss, reduces the comprehensive utilization rate of the fiber, but also greatly increases the COD content in the white water, which is not conducive to energy conservation and emission reduction.
  • Cellulolytic enzymes are "very picky" on the substrate and only have a significant effect on purer cellulose, even on a certain cellulose. For example, most of the existing cellulases have a significant effect on bleaching chemical softwood pulp and less on bleaching chemical hardwood pulp. If the fiber content is lignin, such as unbleached needle pulp, surface lignin The effect of cellulolytic enzymes is small. For mechanical pulps containing high lignin, cellulose hydrolase loses its effect.
  • U.S. Patent Nos. 6,294,366 and 6,635,146 disclose the use of truncated cellulase (CBD-truncated cellulose) to treat pulp, truncated enzymes are lacking in order to avoid the drawbacks of conventional cellulase to excessively shear the fibers to reduce fiber strength.
  • the cellulose binding domain (CBD) which is treated with such cellulase, can avoid such fiber strength loss.
  • US 5,916,799 discloses cellulase compositions containing cellobiohydrolases and endoglucanases, which have been subjected to limited proteolysis, thereby The cellulose binding domains were separated and the resulting enzyme composition was found to reduce back-staining.
  • WO 96/2 3928 discloses the treatment of cellulose-containing fabrics using truncated cellulases, which have been found to reduce redeposition of dyes and increase wear.
  • the Chinese patent "A composition for changing and improving the surface properties of fibers and a papermaking method” proposes a combination of cellulase and fibronectin (CBP) and cellulase treatment of fiber pulp through The fibronectin is selectively adsorbed on the surface of the amorphous region of the fiber, protecting the region from the action of cellulase, thereby avoiding excessive shearing of the fibrous amorphous fiber by the cellulase.
  • CBP fibronectin
  • One of the objects of the present invention is to provide a fiber oxidase composition for modifying and improving the properties of a pulping and papermaking fiber material, which can significantly improve the bonding force between fibers and improve the paper by treating the pulp raw material with the composition. Quality and pulp dewatering efficiency.
  • a fiber oxidase composition for modifying and improving fiber properties comprising:
  • a biological enzyme component having an oxidizing function on the fiber the biological enzyme component being a fiber oxidase which changes the surface properties of the fiber by a redox reaction
  • auxiliary component having an increased catalytic activity for said cell oxidase, said auxiliary component being a protein having a promoting effect on cell oxidase;
  • the weight ratio of the biological enzyme component to the auxiliary component is 1:0.4-50.
  • the cell oxidase is a glycanoxygenase having a copper-dependent small molecular metalloenzyme having a molecular weight of 20-50 KDa; or/and
  • the protein having a promoting effect on the fiber oxidase is a catalytically active biological enzyme or a catalytically inactive cellulose binding protein.
  • the catalytically active biological enzyme is an oxidoreductase, a non-cellulolytic enzyme hydratase or a cellulolytic enzyme;
  • the oxidoreductase is selected from the group consisting of cellobiose dehydrogenase, laccase One or more of glucose oxidase, hydrogen peroxide dismutase, alcohol dehydrogenase or lignin peroxidase
  • the non-cellulolytic enzyme hydratase is selected from the group consisting of amylase, xylanase and fruit One or more of a gelase or an esterase.
  • the oxidoreductase is cellobiose dehydrogenase and laccase, and the cellobiose dehydrogenase and laccase are in a weight ratio of 1:1-5.
  • the catalytically inactive protein is a fibronectin and/or a fibroin.
  • the weight ratio of the bio-enzyme component to the auxiliary component is 1:0.4-30.
  • the weight ratio of the bio-enzyme component to the auxiliary component is 1:0.4-10.
  • the weight ratio of the biological enzyme component to the auxiliary component is 1:0.4-5.
  • the cell oxidase composition further comprises a scavenger of a cell oxidase reaction product; the fiber oxidase and scavenger are present in a weight ratio of 1:1-5.
  • the scavenger is selected from one or more of the group consisting of ascorbic acid, gallic acid, lignin, manganese (II) salt, copper (II) salt, or iron (II) salt.
  • the reaction substrate of the cell oxidase is an oxygen, air or oxygen releasing compound; the oxygen releasing compound is a peroxide or ozone.
  • the peroxide is at least one of hydrogen peroxide, sodium peroxide, calcium peroxide, or potassium peroxide.
  • Another object of the present invention is to provide a new papermaking process which utilizes fiber raw materials more efficiently than the prior art, improves the performance and quality of paper products, improves production efficiency and reduces energy consumption.
  • a papermaking method mainly comprises the following steps:
  • the amount of cell oxidase is from 0.01 to 10 kilograms per ton of dry slurry, and the amount of the auxiliary component is from 0.05 to 10 kilograms per ton of dry slurry.
  • the amount of cell oxidase is from 0.05 to 10 kilograms per ton of dry slurry, and the amount of the auxiliary component is from 0.1 to 5 kilograms per ton of dry slurry.
  • the amount of cell oxidase is from 0.05 to 2 kilograms per ton of dry slurry, and the amount of the auxiliary component is from 0.1 to 5 kilograms per ton of dry slurry.
  • the reaction is carried out for a period of from 5 to 600 minutes, at a pH of from 3 to 10, and at a temperature of from 20 to 80 °C.
  • the reaction is carried out for a period of from 20 to 300 minutes, a pH of from 5 to 10, and a temperature of from 30 to 65 °C.
  • the reaction is carried out for a period of from 20 to 200 minutes, a pH of from 5 to 7, and a temperature of from 40 to 65 °C.
  • Another object of the present invention is to provide a fiber oxidase composition or fiber oxidase for use as a reinforcing agent or retention and drainage aid in a papermaking process for modifying and improving fiber properties.
  • the problem of pulp pulping in the existing papermaking technology is that the fiber has low activity and low reactivity between fibers.
  • the usual method is to increase the degree of beating, but increasing the refining leads to fiber. It is cut and produces a large amount of fine fibers, which not only reduces the dewatering performance of pulping, but also increases the drying energy consumption of papermaking, and may cause the loss of fine fibers and the utilization of raw materials.
  • the existing "beating enzyme” is a hydrolysis reaction of cellulose by using cellulolytic enzyme.
  • the present invention has the following advantages and benefits:
  • the fiber oxidase and its composition can change and improve the surface properties of the fiber, and determine the optimum components of the fiber oxidase composition and the ratio thereof, the fiber oxidase.
  • the composition or cell oxidase has significant reactivity with different fiber materials, including wood fibers, non-wood fibers and recycled fibers, and the pulp material is treated with the fiber oxidase composition or cell oxidase, such as: wood pulp/chemistry Pulp, mainly bleached and unbleached wood pulp / chemical pulp treatment, including secondary recycled fiber; wood pulp / mechanical pulp, and non-wood pulp (straw, straw, reed and tobacco), during processing,
  • the reaction of fiber oxidase on the surface of the crystal fiber activates the surface activity of the fiber, increases the fiber reactivity, and improves the bonding force between the fibers without breaking the fiber to cause the fiber strength to decrease; using the fiber oxidase and
  • the composition of the above-mentioned pulp can not only improve various physical strengths of the produced paper, such as: degree of decomposition, tensile index, tearing degree, interlaminar bonding force, bursting resistance and Whiteness (especially the whiteness of waste paper) can also improve
  • Example 1 is a schematic flow chart of biological enzyme treatment of wood chips in Example 3.
  • Example 2 is a graph showing the relationship between the freeness of the TMP pulp and the refining energy consumption under different processing conditions in Example 3;
  • Example 3 is a graph showing the effect of the composite biological enzyme-treated TMP slurry on the energy consumption of refining in Example 4;
  • Fig. 4 is a graph showing the effect of the fiber oxidase-treated non-wood pulp on the tensile strength of the paper in Example 6.
  • Cellulolytic enzyme refers to all biological enzymes which degrade cellulose by a hydrolysis reaction, and the terms such as cellulolytic enzyme and cellulase, fiber hydrolase and cellulase are often used. Alternate use. According to the different modes of action of cellulase degradation of substrates, they can be divided into three categories:
  • Endocellulase is also known as endoglucanase (Endoglucanase, EG; EC 3.2.1.4);
  • Exocellulase is also known as cellobiohydrolase (CBOH; EC 3.2.1.91);
  • endocellulase also known as endog-glucanase, endo-1,4- ⁇ -D-glucanase, EC 3.2.1.4
  • endocellulase also known as endog-glucanase, endo-1,4- ⁇ -D-glucanase, EC 3.2.1.4
  • endocellulase also known as endog-glucanase, endo-1,4- ⁇ -D-glucanase, EC 3.2.1.4
  • exocellulase also known as exog-glucanase, exo-1,4- ⁇ -D-glucanase, EC 3.2.1.91
  • exocellulase also known as exog-glucanase, exo-1,4- ⁇ -D-glucanase, EC 3.2.1.91
  • CBH cellobiohydrolase
  • Cellobiase also known as ⁇ -glucosidase, ⁇ -1,4-glucosidase, EC 3.2.1.21
  • BG Cellobiase
  • Such enzymes typically hydrolyze cellobiose or soluble cellodextrin to glucose molecules having a molecular weight of about 76 KD.
  • Cellulase Enzyme Activity The enzymatic activity of cellulase in the present invention refers to the glycan of the cellulase component in their ability to degrade cellulose into glucose, cellobiose and disaccharide. Enzyme activity. Cellulase activity is generally determined by a decrease in the viscosity of the solution of carboxymethylcellulose.
  • CBP Cellulose binding protein
  • cellulose-binding protein in the present invention means a cellulose having a special affinity for strongly adsorbing on the surface of cellulose, but itself is not catalyzed by cellulose. Hydrolyzed reactive protein.
  • CBD Cellulose Binding Domain
  • CBM Cellulose Binding Module
  • cellulose adsorption domain Cellulose affinity domain.
  • Fibroswellin Swollenin or Expansin: "Fibrous expanded protein” in the present invention means a protein/polypeptide compound which can loosen the fibrous structure of a natural substrate such as crystalline cellulose and hemicellulose. Cellulose does not have the catalytic function of degradation, but it can improve the hydrolysis ability of cellulase to microcrystalline cellulose. "Fibrous fiber expansion protein”, also known as fiber swelling factor, is called Swollenin and Expansin in English. It is a kind of plant cell wall expansion protein with non-hydrolysis activity of typical cellulase composition (including cellulose binding domain (CBD)). protein.
  • CBD cellulose binding domain
  • ⁇ -expansin has a molecular weight of about 25kDa, amino acid sequence is highly conserved, and homology is 70% to 90%; ⁇ -expansin has a molecular weight of about 29kDa, but the amino acid sequence big change.
  • Fibrous oxidase in the present invention means polysaccharide monooxygenase (PMO), and the "glycan oxygenase” refers to a family having a molecular weight of 20-50 KDa.
  • glycanoxygenase examples include polyglucose oxygenase or polysaccharide monooxygenase (PMO), lytic polysaccharide monooxygenase (LPMO), and glycosidation Enzyme 61 (glycosyl hydrolase 61, GH61).
  • cell oxidase and cellulolytic enzyme degrade cellulose by hydrolysis of ⁇ -1,4-D-14 glycosidic bonds; while cellulolytic enzymes oxidize cellulose.
  • the reduction reaction, the bond changed by the reaction is not limited to the ⁇ -1,4-D-14 glycosidic bond.
  • PMO was discovered as early as 1974, the hydrolysis reaction of PMO to ⁇ -1,4-D-14 glycosidic bonds was found to be very weak due to the analysis method using CMCase (the method for testing endocellulase). The role of this enzyme species and its reaction mechanism have not been further studied.
  • CAZy Carbohydrate Active Enzymes, CAZy, http://www.cazy.org/
  • CAZy Carbohydrate Active Enzymes, CAZy, http://www.cazy.org/
  • GH61 Glycosyl Hydrolases Group 61
  • PMO is actually a copper-dependent monooxygenases (EC 1.14.17.x), which acts on the fibers through oxidation, rather than ordinary
  • the cellulose hydrolyzing enzyme acts on the fiber by a hydrolysis reaction, and thus the PMO is an oxidase rather than a hydrolase.
  • Classical cellulolytic enzymes have many enzyme families. In contrast, PMO has only two families, namely carbohydrate binding module family 33 (CBM33, carbon-water binding domain family 33) and glycosyl.
  • CBM33 carbohydrate binding module family 33
  • glycosyl glycosyl
  • GH61 glycoside hydrolase family 61
  • CAZy recently reclassified PMOs into AA9 and AA10 biological enzymes.
  • the main source of the former is fungus (fungus), while the latter is mainly derived from bacteria, viruses and certain fungi.
  • PMO is present in many lignin-degrading microorganisms, which can be purified and expressed, or recombinantly expressed by modern protein engineering techniques.
  • U.S. Pat. No. 7,273,738, U.S. Patent Application Serial No U.S. Patent No. 8,298,795 and U.S. Patent Application Serial No. 2012/0083019 the disclosure of which is incorporated herein by reference to the entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire all Lignocellulosic materials increase biorefinery efficiency and increase alcohol production. So far, the application of PMO has been in biorefinery, and there is no report on the application of PMO in pulp and paper.
  • Fibrous oxidase composition in the present invention means that the composition contains the following two components, one component being a polysaccharide monooxygenase, and the other component being capable of Maintaining or increasing an adjuvant to the activity of the oxidative enzyme, wherein the auxiliary may be one or more compounds of the following compounds, (1) a reaction substrate of a cell oxidase, oxygen or oxygen precursors, (2) Reagents (radical scavengers), and (3) proteins that promote fiber oxidase.
  • reaction substrate of cellulase means an oxygen acceptor which is necessary for the reaction of a glycan oxidase in addition to a fiber; the reaction substrate can be provided by: releasing oxygen, air or oxygen
  • the compound, the oxygen releasing compound in the present invention means a peroxide or ozone, wherein the peroxide includes hydrogen peroxide, sodium peroxide, calcium peroxide, potassium peroxide.
  • the "promoting protein for cell oxidase" of the present invention refers to a biological enzyme, a cellulose-binding protein, and a fibroin protein which can increase the activity of a glycan oxidase; among the organisms which promote the glycan oxidase Enzyme refers to other hydrolase enzymes (including amylase, pectinase, lipase, esterase or xylanase) and oxidoreductases (including cellobiose dehydrogenase, laccase, etc.) other than cellulolytic enzymes. Glucose oxidase, hydrogen peroxide dismutase, alcohol dehydrogenase Or lignin peroxidase).
  • reaction product scavenger of cellulase means a compound capable of reducing the concentration of hydrogen peroxide of a reaction product of glycan oxidase, including ascorbate, gallate, lignin, copper.
  • the salt is a divalent copper salt, an iron (II) salt, that is, a divalent iron salt, a manganese (II) salt, that is, a divalent manganese salt, and a zinc (II) salt, that is, a divalent zinc salt.
  • the invention relates to the use of fiber oxidase to treat fiber raw materials for pulping and papermaking, to change the chemical properties of the fiber surface, to improve the reactivity of the fiber, increase the bonding force between the fibers, thereby improving the strength index of the paper and increasing the dewatering and drying efficiency. Reduce energy consumption, save production costs for paper companies, and increase economic profits.
  • the cellulose oxidase involved in the present invention may be separately added to the papermaking process as a product of its individual enzyme components, so that the two components can be separately added to different positions or mixed into one product formula to be added to the agreed position in the papermaking process. To achieve the best results.
  • the object and effect of using the fiber oxidase according to the present invention is to cause a bond having a high surface reactivity by an oxidase to react with a crystal region on the surface of the fiber to increase the binding force between the fibers. Moreover, the reaction of fiber oxidase on the fiber can be controlled on the surface of the fiber without excessive reaction to the fiber, thereby achieving selective fiber surface modification and repair. Therefore, the problem of degrading fibers of the conventional cellulolytic enzyme is avoided as compared with the conventional cellulolytic enzyme.
  • the unit "kg/T" in the present invention refers to the number of kilograms of material added per ton of dry pulp.
  • Fibrin oxidase is a GH61 supplied by the American company Dyadic, which is prepared from the strain Myceliophthora thermophila, and its amino acid sequence is described in detail in U.S. Patent No. 8,298,795 and U.S. Patent Application Serial No. 2012/0083019.
  • Laccase and cellobiose dehydrogenase are commercial products of the DuPont Group Genencor.
  • Pulping 200 g of pulp plate and 1300 g of deionized water were placed in a 10 liter hydropulper, slurried for 10 minutes, and then 2500 g of water was added to obtain a 5% slurry.
  • Control group 1 was a blank sample: no drug was added
  • Control group 1 was: 100g/T, 400g/T laccase treatment
  • Control group 2 was: 100 g/T, 250 g/T cellobiose dehydrogenase treatment;
  • the experimental group is an enzyme-treated sample, wherein
  • Experimental group 1 was: 250 g/T (absolute dry pulp) fiber oxidase (PMO) treatment;
  • Experimental group 2 is treated with a fiber oxidase + laccase composition: 100 g / T, 250 g / T fiber oxidase (PMO), plus laccase treatment together, the amount of laccase is 400 g / T;
  • Experimental group 3 was treated with a fiber oxidase + cellobiose dehydrogenase composition: 250 g/T cell oxidase (PMO), plus 100 g/T cellobiose dehydrogenase;
  • PMO cell oxidase
  • Experimental group 4 was treated with a fiber oxidase + laccase + cellobiose dehydrogenase composition: 100 g / T fiber oxidase + 200 g / T laccase + 100 g / T cellobiose dehydrogenase;
  • reaction conditions of the above respective groups were as follows: a reaction temperature of 50 ° C, a reaction time of 150 minutes, a reaction pH of 5.7-6.0, and constant stirring.
  • Handsheet preparation and strength measurement Dilute the prepared slurry, add tap water to 1.0%, mix, measure temperature and slurry concentration, and accurately prepare 10-12 sheets of 6.5 g according to TAPPI method. Hand paper. After drying, the handsheets were placed in a constant temperature and constant humidity control box for 24 hours, and then their thickness (density), tensile strength, tear strength, and bond strength were measured according to the TAPPI measurement method.
  • PMO dosage Laccase dosage Cellobiose dehydrogenase Degree of resolution Tensile index Tear degree Interlayer bonding g/T g/T g/T °SR Nm/g mN*m/g Kpa blank 0 0 42.5 18.9 6.55 210.8 0 0 100 41.0 19.2 6.37 220.1 250 0 0 43.0 21.4 7.70 249.5 0 100 0 42.0 19.7 6.70 223.7 0 0 250 42.5 20.4 6.70 227.5 0 400 0 42.5 21.1 7.00 247.6 100 400 0 43.8 21.9 8.12 280.6 250 400 0 45.0 22.7 8.45 270.7 250 0 100 43.5 24.1 8.64 290.0 100 200 100 46.0 26.6 10.76 358.6
  • Example 2 The effect of fiber oxidase and laccase treatment on the strength of paper by OCC pulp
  • the effect of the fiber oxidase composition on the strength of the paper by the ODC pulp was evaluated by comparative analysis.
  • the recovered OCC waste paper pulp was obtained from the Guangdong Tobacco Paper Multi-plate Concentrate Pulp Pool, and the pulp concentration was 4.8-5%;
  • Fibrin oxidase is supplied by Dyadic, USA, GH61 from the strain Myceliophthora Prepared by thermophila, the amino acid sequence of which is described in detail in U.S. Patent No. 8,298,795 and U.S. Patent Application Serial No. 2012/0083019;
  • Laccase is a commercial product of Genencor, a subsidiary of the DuPont Group.
  • This experiment is divided into experimental group and control group;
  • the control group is a blank sample: no drug is added
  • the experimental group is a sample treated with an enzyme or an enzyme and a scavenger, wherein
  • Experimental group 1 is: fiber oxidase (PMO) 250g / T + iron sulfate dosage is 500g / T;
  • Experimental group 2 is: laccase 250g / T + iron sulfate dosage is 500g / T;
  • Experimental group 3 is: fiber oxidase 250g / T + laccase 250g / T + iron sulfate dosage is 500g / T;
  • the prepared slurry was diluted with tap water to 1.0% strength, mixed, and the temperature and slurry concentration were measured. According to the TAPPI method, 10-12 sheets of 6.5 g weight handsheets were accurately prepared. After drying, the handsheets were placed in a constant temperature and constant humidity control box for 24 hours, and then their thickness (density), tensile strength, tear strength, and bond strength were measured according to the TAPPI measurement method.
  • the paper has a slightly increased bursting resistance, but not as significant as the fiber oxidase; after the addition of ferric sulfate alone, the bursting resistance of the paper made from the fluff pulp or the fluff pulp is almost unchanged; only when the fiber oxidase When combined with laccase and ferric sulfate, the effect is most pronounced. Regardless of long fiber pulp or short fiber pulp, the paper's bursting strength is increased by 25-30%.
  • Example 3 Effect of fiber oxidase and its different compositions on the energy consumption of refining wood chips
  • PMO Cellulose oxidase
  • the cellulolytic enzyme is the commercial product FiberZyme G200 from Dyadic.
  • a paper mill uses southern pine wood to make TMP. When the wood is removed from the bark, it is cut into wood chips by a slicer and enters the wood chip storage bin or stacking yard.
  • Pulping process/process The wood chips prepared above are cleaned and removed by wood chips, then enter the wood chip steam screw press, and then enter the first-stage disc grinding, and then enter the first-stage grinding and decontamination pool, after coarse screening and double After the net press filter presses, it enters the secondary refiner. After the grind is passed through the fine screen, it enters the multi-plate concentrator and finally enters the TMP storage tower.
  • Fig. 2 The results are shown in Fig. 2. It can be seen from Fig. 2 that the refining performance of the wood chips after the treatment with the fiber hydrolase (enzyme A) has little change; after the treatment with the fiber oxidase (enzyme B), the refining performance is obviously improved, and the refining is improved. The energy consumption is reduced; and the best effect is to use both fiber oxidase and cellulolytic enzyme, and the energy consumption required to achieve the same freeness is reduced by more than 100 KWH/T compared with the blank.
  • Example 4 Effect of fiber oxidase and its different compositions on the energy consumption of refining wood chips
  • PMO Cellulose oxidase
  • the cellulolytic enzyme is the commercial product FiberZyme G200 from Dyadic.
  • the wood chips After the wood chips are cleaned and sanded, they enter the wood chip steam screw press, and then enter the first-stage disc grinding. After grinding, they enter the first-stage grinding and decontamination pool. After the coarse slurry and the double mesh filter press are squeezed, they enter the secondary refiner. After grinding, the slurry enters the multi-plate concentrator after fine screening, and finally enters the TMP storage tower.
  • the biological enzyme is added to the first-stage refining decontamination tank, the pulp concentration is about 3.5%, the temperature is 75-80 ° C, the residence time is 45-60 minutes; the fiber oxidase composition (fibrous oxidase and fiber) The amount of the enzyme 1:3) is 0.30-0.6 kg/T.
  • Fig. 3 The experimental results are shown in Fig. 3. From Fig. 3, the change of the total refining energy consumption when the biological enzyme is added; specifically, the fiber oxidase and the cellulase are used in combination with the total refining before and after the deactivation. Energy consumption has dropped by 200-300KWH/T.
  • the effect of the fiber oxidase composition on the whiteness of the paper was evaluated by comparative analysis.
  • Recycled old newspaper (ONP) and old magazine (OMG) waste paper are mixed at a ratio of 80% to 20%.
  • Fibrin oxidase is a GH61 supplied by the American company Dyadic, which is prepared from the strain Myceliophthora thermophila, and its amino acid sequence is described in detail in U.S. Patent No. 8,298,795 and U.S. Patent Application Serial No. 2012/0083019.
  • Glucose Oxidase (GOX) is a commercial product of Genencor, a subsidiary of the DuPont Group.
  • A biological enzyme treatment conditions: taking 600 grams of the above prepared slurry, placed in a 1000ml mixing stirrer, using a thermostat controller to control the temperature of the slurry at 60 ° C;
  • the control group is: adding 1-10 kg/T of caustic soda, 15-30 kg/T of water glass, 10-20 kg/T of hydrogen peroxide, and 0.5-3 kg/T of deinking agent;
  • the experimental group is treated with a fiber oxidase composition, wherein
  • Experimental group 1 is: adding fiber oxidase (PMO) 150-300g / T, deinking agent 0.5kg / T, adding caustic soda 2kg / T, water glass 15kg / T, hydrogen peroxide 10kg / T;
  • PMO fiber oxidase
  • Experimental group 2 is: adding glucose-oxidase 250-500g/T, deinking agent 1kg/T, adding caustic soda 2kg/T, water glass 15kg/T, hydrogen peroxide 10kg/T;
  • the experimental group 3 is: adding a combination of cell oxidase and glucose oxidase in the slurry, PMO 150-300g/T, GOX 250-500g/T, deinking agent 0.5kg/T, adding caustic soda 1-2kg/T , water glass 15kg / T, hydrogen peroxide 10kg / T;
  • reaction conditions were as follows: a temperature of 60 ° C, a reaction time of 90 minutes, a reaction pH of 8-10, and constant stirring.
  • Handsheet preparation and strength measurement Dilute the prepared deinked pulp, add tap water to 1.0%, mix, measure temperature and slurry concentration, and accurately prepare 10-12 sheets of 6.5 g according to TAPPI method. Heavy hand paper. After drying, the handsheets were placed in a constant temperature and constant humidity control box for 24 hours, and then measured for whiteness by the TAPPI measurement method.
  • Example 6 Effect of cell oxidase on energy consumption and strength of non-wood pulp (tobacco stem) refining
  • PMO Fibrous oxidase
  • Slurry The tobacco/tobacco mixed slurry from the front tank of No. 2 machine of a regenerated tobacco leaf production company, and the simulation experiment was carried out according to the production process and process conditions of the tobacco stem slurry. That is, after one extraction and extrusion of the tobacco stem pulping of the No. 2 machine, the fiber oxidase composition is introduced into the secondary extraction reactor, extracted for 20 minutes, and then squeezed to remove the extract, and then The slurry is refined and then tested for various physical properties of the slurry.
  • the test method is as follows:
  • sample preparation is the same as the test sample of dry strength (ie tensile strength);
  • the degree of decomposability and tensile strength of the slurry after refining increase with the amount of the enzyme product, for example, the amount of the enzyme used when the refining number is 250 rpm.
  • the tensile index of the flakes increased by nearly 100% compared with the blank at 250 rpm, which is similar to the tensile index of 1500 rpm of the blank refining; however, the wet weight of the slurry after bio-enzyme treatment did not change significantly
  • the wet weight drops to 30%, which indicates that the new biological enzyme does not cause any damage to the fiber.
  • test results are shown in Table 5 and Figure 4.
  • Table 5 the wet strength of the paper after treatment with cell oxidase Increase.
  • Fig. 4 the tendency of the tensile index of the paper after the addition of the cell oxidase of the present invention in the process of papermaking of the reconstituted tobacco leaf sheet was observed. It can be seen that the fiber oxidase body significantly increases the strength of the sheet, which is beneficial to the production of paper machines.
  • PMO Fibrous oxidase
  • Slurry taken from the slurry of the straw pulping production plant. The specific steps and conditions are as follows: Weigh 20 pieces of dry slurry, place it in a 1000ml beaker, and then place the beaker in a constant temperature water bath set at 60 °C. In, to make it balanced. Thereafter, a certain amount of the fiber oxidase composition was added according to the desired biological enzyme treatment conditions, reacted for 60 minutes, and continuously stirred to equilibrate the reaction. After that, the degree of resolution of the test slurry was sampled.
  • Table 7 shows the effect of the bioenzyme product on the sheet strength after treating the slurry. Obviously, as the amount of enzyme increases, the strength of the paper increases. When the dosage is 0.5kg/T, the tensile index increases by 16%; when the dosage is 1.0kg/T, the tensile index increases by 36%; and when the dosage increases to 3.0kg/T, the tensile index increases nearly 80%.

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Abstract

L'invention concerne une composition d'oxydase de fibre utilisée pour modifier et améliorer la blancheur d'un papier, un procédé de fabrication de papier, et des applications de la composition. Cette composition d'oxydase de fibre est constituée principalement des constituants suivants : un constituant enzyme, ayant une fonction d'oxydation pour une fibre, le constituant enzyme étant une oxydase de fibre qui modifie les propriétés de surface de la fibre au moyen d'une réaction d'oxydoréduction; et, un constituant auxiliaire possédant des activités catalytiques accrues pour l'oxydase de fibre, ce constituant auxiliaire étant une protéine possédant des effets catalytiques pour l'oxydase de fibre et le rapport en poids constituant enzymatique - constituant auxiliaire étant de 1 : 0,4 à 50.
PCT/CN2015/084455 2014-07-29 2015-07-20 Composition d'oxydase de fibre utilisée pour modifier et améliorer la blancheur du papier, procédé de fabrication de papier, et applications de la composition WO2016015570A1 (fr)

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CN102086611A (zh) * 2010-11-30 2011-06-08 王祥槐 一种用于改变和改善纤维表面性质的组合物和造纸方法
CN103835174A (zh) * 2012-11-27 2014-06-04 广州瑞辰盛达生物技术有限公司 湿强废纸的制浆方法
CN103866604A (zh) * 2012-12-13 2014-06-18 广州瑞辰盛达生物技术有限公司 用于提高生活用纸的松厚度和柔软性的造纸方法
CN104342424A (zh) * 2013-07-29 2015-02-11 瑞辰星生物技术（广州）有限公司 用于改变和改善纤维性质的纤维氧化酶组合物及造纸方法和应用

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CN103835174A (zh) * 2012-11-27 2014-06-04 广州瑞辰盛达生物技术有限公司 湿强废纸的制浆方法
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