CN105441412A - Paper pulp bleaching compound enzyme and preparation method thereof - Google Patents
Paper pulp bleaching compound enzyme and preparation method thereof Download PDFInfo
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- CN105441412A CN105441412A CN201410515623.2A CN201410515623A CN105441412A CN 105441412 A CN105441412 A CN 105441412A CN 201410515623 A CN201410515623 A CN 201410515623A CN 105441412 A CN105441412 A CN 105441412A
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- prozyme
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- pulp bleaching
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- 238000004076 pulp bleaching Methods 0.000 title claims abstract description 51
- 238000002360 preparation method Methods 0.000 title claims abstract description 24
- 102000004190 Enzymes Human genes 0.000 title abstract description 102
- 108090000790 Enzymes Proteins 0.000 title abstract description 102
- 229920001131 Pulp (paper) Polymers 0.000 title abstract description 34
- 150000001875 compounds Chemical class 0.000 title abstract description 7
- 101710121765 Endo-1,4-beta-xylanase Proteins 0.000 claims abstract description 42
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Landscapes
- Paper (AREA)
Abstract
The invention discloses a paper pulp bleaching compound enzyme and a preparation method thereof. Through scientific compounding of high temperature-resistant alkaline xylanase as a main raw material and bacillus thermophilus culture, laccase and its medium, glucanase, EDTA, a protective agent, an activator, seminase, lipase, tannase, pectase, a whiteness stabilizing agent, a nonionic surfactant and an anti-oxidant, the paper pulp bleaching compound enzyme is prepared. The paper pulp bleaching compound enzyme completely retains paper pulp cellulose, maximally dissolves and degrades lignin, has the advantages of complete enzyme system, good bleaching effects, storage easiness and good storage stability, can substantially improve a yield and quality of paper pulp of broad leaf wood, needlebush and reed yellow waste slurry, respectively improves whiteness by 8.18%, 8.13% and 8.23%, respectively reduces a yellow index by 38.35%, 35.82% and 31.76%, saves a conventional chemical article use mount by 50-60%, reduces a cost and protects the environment.
Description
Technical field
The present invention relates to paper grade (stock) prozyme, be specifically related to a kind of association with pulp bleaching prozyme and preparation method thereof.
Background technology
In the industrial production, pulping process is under the condition of alkalescence, delignification is removed by the method for high temperature steaming, 98% is reached for making dissolving pulping fibre element purity, this just needs a large amount of sodium-hydroxide treatment paper pulp, cause serious environmental pollution, many scholars slurrying then trial is carried out a biological disposal upon for this reason, and should be heat-resisting and alkaline-resisting for the zytase of pulping bleaching.Feng Jianliang etc. (Kimuraetal.2000) xylanase pretr eatment wheat straw and conventional chemistry slurrying compare test, and xylanase pretr eatment improves the delignification degree of raw material, can also reduce the Kappa number of slurrying.Zytase has more extensive and deep research in auxiliary bleaching, and achieves very large achievement.Research finds, no matter is softwood pulp, hardwood pulp or bamboo pulp, straw pulp, and zytase all can be degraded xylogen remaining in paper pulp, improves whiteness (ALBALAAetal, 2006; MeekandLipman, 1922).Zytase for association with pulp bleaching must have high temperature resistant feature, Khasin etc. obtain one and derive from alkaline bacterial strain BacillussterarothermophilusT26 zytase, this zytase has best bleaching effect (Khasinetal when pH9.0 and 65 DEG C to paper pulp, 1993), a lot of zytase does not possess such feature, limits commercial applications.The waste paper quantity that the whole world produces every year is surprising, and after waste paper reclaims, recycling work just becomes particularly important, can make also to protect environment while resource recycling.1991, ink can remove by application zytase from news paper waste by Korea S scholar reported first.After this, people begin one's study and utilize zytase to carry out deinking, to alleviate or to eliminate the environmental pollution (Cao Junwei etc., 2004) that Chemical Deinking brings.
At present, the green cleaning and bleaching of the paper pulp being representative with element-free chlorine and Totally-chlorine-free bleaching technology has become the inexorable trend of countries in the world paper-making industry association with pulp bleaching development, zytase enzyme process helps drift novel process to be applied in more than 30 large-scale paper plants of family of Europe and North America, becomes biotechnology in the most successful example of paper industry application.Wherein Canada have an appointment 10% sulphate process pulp mill have employed this novel process.Novozymes Company of Denmark and U.S.'s mountain pass this chemical company Deng Duojia zymin manufacturer, be proposed the zytase and cellulase product innovation that are specifically designed to slurrying process one after another, but up to the present, the industrial zytase being applied to association with pulp bleaching is neutral meta-acid mostly, optimal reactive temperature is mostly at about 50 DEG C, as everyone knows, pulp cooking and bleaching are substantially all carry out under the condition of high temperature and highly basic, make existing low temperature acid zytase product be subject to great restriction in the application in this field.
But because xylanase treatment technique helps chemistry drift agent bleaching by modes such as the surperficial hemicelluloses deposited again of degraded removing paper pulp, zytase can only play the effect helping drift, really can not substitute chemistry and float agent.Therefore, biological pre-bleaching can not substitute chemical bleaching completely, can decreasing pollution but can not finally decontamination, fundamentally to eliminate the pollution of chlorine bleach liquor, finally need realize bio-bleaching, namely adopt xylogen residual in biological means removing paper pulp completely, because the existence of xylogen is the basic reason of yellowing reversion of pulp, chemical bleaching can direct effect xylogen and the chromophoric group that destroys in paper pulp.Ligninase can directly act on xylogen, xylogen is degraded, be mainly the lignin peroxidase of whiterot fungi secretion, manganese peroxidase, laccase and cellobiose dehydrogenase etc., the focus studied in recent years mainly concentrates on the bio-bleaching of laccase, laccase is a kind of blue multicopper oxidase, also polyphenoloxidase is claimed, main oxidation is containing the lignin structure unit of phenol, but natural timber xylogen be only less than 20% containing phenol structure unit, another fungal laccase molecular weight usually comparatively large (being about 70000Da) is difficult to penetrate in wood and acts on lignin molecule, want to give full play to its enzymic activity, laccase mediators must be added, be generally violuric acid (VA) and 1-hydroxy benzo triazole (HBT), select depending on different bleaching process, and the output of laccase is less, less stable.
Chinese patent CN102978986A discloses a kind of method that biological enzyme prepares paper pulp, the mass percentage content that described biological enzyme prepares biological enzyme component in liquid is: cellulase 20%, hemicellulase 10%, lignin degradation enzyme 10%, amylase 2 0%, lipase 5%, polygalacturonase and laccase 20%; Cellulase wherein can be degraded the effective constituent Mierocrystalline cellulose in paper pulp, wayward, have a strong impact on into the physical and mechanical properties of paper, meanwhile, above-mentioned biological enzyme is applicable to pulping process, but not bleaching process, further, single use laccase, its enzyme activity is difficult to play, can not act on xylogen very well, yellowing reversion of pulp possibility increases.Chinese patent CN103555701A discloses a kind of production method of association with pulp bleaching complex enzyme liquid, it is characterized in that by each component enzyme through weighing, composite tank mixed preparing, anticorrosion, inspection, packaging after deposit in 5 DEG C of storehouses, described each component enzyme is alkaline pectase, zytase, mannase and lignoenzyme; Above-mentioned complex enzyme liquid enzyme class and characteristic is single, lignoenzyme vigor is low and the concrete kind of not openly lignoenzyme, industrialization is difficult to realize, and still fundamentally thoroughly can not eliminate xylogen, bleaching effect is still undesirable, and not easily preservation, poor storage stability, Commercialization application is limited.
To sum up, prepare that a kind of enzyme system is complete, bleaching effect good, easy preservation, association with pulp bleaching enzyme that stability in storage is good have wide market outlook and huge market potential value.
Summary of the invention
Technical problem solved by the invention is the defect overcoming existing association with pulp bleaching enzyme, with high temperature resistant alkalescent xylanase for main raw material, the composite bacillus acidocldarius culture of science, laccase and medium thereof, dextranase, EDTA, protective material, activator, seminase, lipase, tannase, polygalacturonase, whiteness stablizer, nonionogenic tenside, antioxidant etc., prepare a kind of enzyme system complete, bleaching effect is good, easy preservation, the association with pulp bleaching enzyme that stability in storage is good, while retaining paper cellulose completely, dissolved lignin being degraded to greatest extent, fundamentally reduce content of lignin in paper pulp, strengthen delignification effect, appropriate composite whiteness stablizer simultaneously, the deficiency of bio-bleaching can be made up, strive reaching permanent bleaching, prevent yellowing reversion of pulp, improve into the seed output and quality of paper, to advance bio-bleaching in the application process of paper industry, finally reach the object reduced costs with protection of the environment.
In order to achieve the above object, the present invention is by the following technical solutions:
A kind of association with pulp bleaching prozyme, prepared by the raw material of following parts by weight:
Alkalescent xylanase 20-40 part, bacillus acidocldarius culture 10-30 part, laccase 10-20 part; dextranase 5-8 part, 1-hydroxy benzo triazole 3-8 part, EDTA3-8 part; protective material 4-7 part, activator 4-7 part, seminase 4-6 part; lipase 3-5 part; tannase 3-5 part, polygalacturonase 3-5 part, whiteness stablizer 2-5 part; nonionogenic tenside 2-4 part, antioxidant 1-3 part.
Described alkalescent xylanase and bacillus acidocldarius culture are starting strain by producing the bacillus acidocldarius (Bacillusthermophilus) 701 (deposit number is CCTCCM2013537) of high temperature resistant alkalescent xylanase, and prepared by Optimal Medium composition and zymotechnique;
Described alkalescent xylanase is high temperature resistant, alkali resistance is strong, action condition is wide in range, do not have cellulase activity, and the bio-bleaching and the chemistry that are more applicable to paper pulp help drift;
The zymologic property of described high temperature resistant alkalescent xylanase is as follows:
(1) temperature: this enzyme thermal adaptation a wider range, Applicable temperature scope is 40-75 DEG C, optimal reactive temperature 65 DEG C, at 75 DEG C of enzymes complete stability alive;
(2) pH: it is 5.0-11.0 that this enzyme is suitable for pH value in reaction scope, the enzyme complete stability alive when pH value is 11.0, optimal reaction pH value is 10.0;
(3) enzymic activity: fermented liquid alkalescent xylanase specific activity of enzyme is higher is 350-420IU/ml;
(4) thermostability: this enzyme still can keep more than 80% enzyme to live preserve 1h under 70 DEG C of conditions after, keeps more than 70% enzyme to live after preserving 1h under 75 DEG C of conditions;
(5) storage stability: this enzyme is at room temperature preserved the loss alive of enzyme after 12 months and is less than 3.0%.
Preferably, the preparation method of described high temperature resistant alkalescent xylanase is as follows: by activated for the slant strains of intact bacillus acidocldarius CCTCCM2013537 and step by step enlarged culturing (comprise one, two, three seed culture and first class seed pot cultivate) obtain seed liquor, with 6% inoculum size access fermentor tank, culture temperature 37-45 DEG C, stirring velocity 200-700rpm, ventilation (V/V) 1:1-3, incubation time 25-30h; Then with 1-2 DEG C/h rate of temperature fall slow cooling to 15-20 DEG C, constant temperature culture 10-12h; Continue with 1-2 DEG C/h rate of temperature fall slow cooling to 8-10 DEG C, now, first class seed pot fermented liquid is added access fermentor tank with 4% inoculum size, constant temperature culture 7-12h; Finally slowly be warming up to 15-20 DEG C, constant temperature culture 10-12h with 1-2 DEG C/h temperature rise rate; Continue slowly to be warming up to 37-45 DEG C, constant temperature culture 20-30h with 1-2 DEG C/h temperature rise rate; Fermentation liquor is filtered, concentrated, allotment, essence filter, dry high temperature resistant alkalescent xylanase, described allocation process adds concentrated enzyme liquid gross weight 0.5-5% Chinese herbal medicine powder;
Described slant medium consists of: extractum carnis 3-10g, sodium-chlor 5-12g, peptone 10-20g, glucose 2-5g, agar 15-20g, Chinese herbal medicine powder 5-10g, distilled water l000mL, pH value 5-11,121 DEG C of sterilizing 20min;
The preparation method of described Chinese herbal medicine powder is as follows: with weight parts, takes Radix Astragali 20-30 part, Radix Codonopsis 10-18 part, radix bupleuri 10-15 part, root of large-flowered skullcap 10-15 part, Herba Houttuyniae 8-12 part, Radix Angelicae Sinensis 8-10 part; Respectively said herbal medicine being crushed to particle diameter is less than 2 millimeters, then Homogeneous phase mixing add the water of 3-6 times of weight in container, control temperature 70 DEG C ~ 90 DEG C keeps 2 ~ 4h, then being cooled to 45-60 DEG C, adding mixed enzyme and carry out enzymolysis, is 5.5-6.8 by lactic acid adjust ph, enzymolysis 2-4h, finally add the mixture of mixture 0.5-3 times of w ethanol and propyl alcohol, control temperature to 60 DEG C ~ 78 DEG C keeps 3 ~ 4h, filters; Filter vacuum concentrates postlyophilization and obtains Chinese herbal medicine powder;
Described mixed enzyme addition is the 5-10% of mixture gross weight;
The parts by weight of described mixed enzyme consist of: endo-beta-glucanase 10-20 part, outer beta-glucanase 10-20 part, beta-glucosidase 10-15 part, zytase 15-20 part, pentosanase 15-20 part, Pullulanase 20-30 part, beta-amylase 10-15 part, neutral protease 10-15 part, aspartic protease 10-15 part, superoxide-dismutase 5-10 part, glucose oxidase 5-10 part, acid phosphatase 5-10 part;
The mass ratio of described ethanol and propyl alcohol is 1:1-1.5;
Described one-level, secondary, three grades of seed culture medium weight consist of:
Yeast powder 0.3-0.5%, glucose 1-1.5%, peptone 0.3-0.5%, extractum carnis 0.5-0.8%, dipotassium hydrogen phosphate 0.8-1.5%, Chinese herbal medicine powder 1.5-2%, trehalose 1-3%, calcium sulfate 0.1%, magnesium chloride 0.2%, Trisodium Citrate 0.1-0.3%, insufficient section pure water is supplied, pH value 5-11,121-123 DEG C of sterilizing 30-40min;
Described seed tank culture basic weight amount consists of:
Maltodextrin 5-15%, yeast powder 0.4-0.8%, Chinese herbal medicine powder 1.5-2%, trehalose 1-3%, peptone 0.1-0.5%, corn steep liquor 0.1-0.5%, dipotassium hydrogen phosphate 0.8-1.5%, magnesium sulfate 0.05-0.1%, Trisodium Citrate 0.1-0.5%, birch xylan 0.1-0.8%, insufficient section pure water is supplied, pH value 5-11,121-123 DEG C of sterilizing 30-40min;
Described seeding tank fermented liquid cell concentration is 7.0x10
8-8.0x10
8individual/ml;
Described fermention medium consists of: maltodextrin 50-150g, Semen Maydis powder 50-60g, soybean cake powder 15-25g, Chinese herbal medicine powder 30-50g, trehalose 30-40g, birch xylan 5-15g, yeast powder 4-8g, corn steep liquor 1-5g, ammonium sulfate 1-3g, dipotassium hydrogen phosphate 1-2g, potassium primary phosphate 1-2g, Trisodium Citrate 1-5g, defoamer 0.1-1g, pure water l000mL, pH value 5-11,121 DEG C of sterilizing 20min;
The concocting method of described fermention medium is: accurately take raw material in proportion, pure water in raw material, Semen Maydis powder, soybean cake powder are dropped in material-compound tank, adjust ph 5-11, add middle temperature amylase (3u/g Semen Maydis powder) and alpha-amylase (30u/g Semen Maydis powder), DEG C insulation 15-30min of warming while stirring to 70 simultaneously, then be slowly warming up to 90 DEG C of insulation 15-30min to liquefy, finally add other raw material, stir, adjust initial pH5-11,121-123 DEG C of sterilizing 30-40min for subsequent use.
Described bacillus acidocldarius culture contains stronger biological activity, high temperature resistant alkalescent xylanase can be produced on the one hand by fermentation in bio-bleaching process, for system continues to provide, bacillus acidocldarius can be made even by the dynamic response of fermentation on the other hand, be distributed to paper cellulose fully, hemicellulose and xylogen interstitial, abundant softening Mierocrystalline cellulose and degradation of hemicellulose (xylan, dextran, seminose etc.), make other composition of prozyme (as laccase, the zymins such as medium, sequestrant, protective material, activator, whiteness stablizer etc.) evenly, penetrate into paper pulp vegetable cell interstitial fully, give full play to the effect of component, to improve paper pulp recovery rate and quality, shorten bleaching time, and remove impurity and strengthen bio-bleaching effect,
Preferably, the preparation method of described bacillus acidocldarius culture is that above-mentioned fermentation liquor concentrating under reduced pressure, lyophilize, the pulverize at low temperature preparing alkalescent xylanase obtains;
Described bacillus acidocldarius culture viable bacteria content is 7x10
10-9x10
10cFU/g.
Described 1-hydroxy benzo triazole can activate the vigor of macromole laccase, forms the compound amboceptor of strong oxidation, improves the enzymolysis effect of laccase, gives full play to the Degradation of laccase to xylogen, fundamentally eliminate xylogen, prevent yellowing reversion of pulp.
Described EDTA can most of transition metal ion (cupric ion, ferrous ion, ferric ion, divalent manganesetion, bivalent nickel ion etc.) and micro heavy in chelated pulp, to get rid of it to the suppression of biological enzyme enzyme activity and eliminating effect, enzyme component fully, is effectively played a role;
Described whiteness stablizer has the dual function of bleaching and stable whiteness, both effectively pulp brightness can be improved, yellowing reversion of pulp can be prevented again, and can not by the impact of oxygen in paper pulp and transition metal ion (cupric ion, ferrous ion, ferric ion, divalent manganesetion, bivalent nickel ion etc.), the yellowing reversion of pulp that after the use of whiteness stablizer can effectively make up bio-bleaching, residual lignin causes;
Described whiteness stablizer is that one or more in two phosphorus derivants (BBHPE) of mercaptan, thioether, trishydroxymethyl phosphoric acid (THP), tetra methylol phosphoric acid salt (THPC), trishydroxymethyl phosphoric acid are with arbitrary proportion Homogeneous phase mixing;
Preferably, described whiteness stablizer is two phosphorus derivants (BBHPE) of trishydroxymethyl phosphoric acid;
Described nonionogenic tenside can promote that in enzyme bleaching process prozyme component permeates, fully free xylogen, hemicellulose, pectin, tannin, fat, etc. substrate, promote and extend the action time of each component enzyme-to-substrate, objectionable constituent in paper pulp are fully degraded, improves the smoothness of paper pulp fiber and bleaching uniformity coefficient;
Preferably, described nonionogenic tenside is environment-friendly type nonionogenic tenside, easy biological degradation, and noresidue is without any side effects to human body, biology and environment;
More preferably, the quality component of described nonionogenic tenside is: alkyl polyglycoside (APG) 20-40 part, methyl glucamine (AGA) 20-30 part, N-dodecyl ethylenediamine triacetic acid sodium (ED3A) 10-15 part, isomery alcohol ether carboxylate AEC-110710-15 part, Lauric Acid Monoethanolamide Ether Carboxylate (LAEC) 8-10 part, peregal c-1253-5 part, JFC3-5 part; Above-mentioned raw materials is commercial solid, powdery commodity.
Described activator is formed by the inorganic salt Homogeneous phase mixing of following quality component: sodium sulfate 10-15 part, zinc chloride 6-8 part, magnesium chloride 5-8 part, calcium chloride 3-5 part.
Described protective material is made up of the raw material of following parts by weight: trehalose 20-40 part, ganoderan 12-18 part, NaCl10-15 part, (NH
4)
2sO
45-9 part, halfcystine 3-5 part.
Described antioxidant is any or several combination in grape pip procyanidin, rosemary extract and apricot leaf extract;
Preferably, described antioxidant is grape pip procyanidin, rosemary extract and apricot leaf extract 2-4:1-3:1-2 Homogeneous phase mixing in mass ratio;
Described grape pip procyanidin, rosemary extract and apricot leaf extract are commercial commodity
The preparation method of above-mentioned association with pulp bleaching prozyme, comprises the steps:
First by described protective material, activator micronizing respectively; Homogeneous phase mixing; immediately add nonionogenic tenside, bacillus acidocldarius culture Homogeneous phase mixing 20-30min successively; then alkalescent xylanase, laccase, dextranase, seminase, lipase, tannase, polygalacturonase Homogeneous phase mixing is added successively; finally add 1-hydroxy benzo triazole, EDTA, whiteness stablizer, antioxidant successively, pack after mixing and obtain association with pulp bleaching prozyme.
Another object of the present invention is the application of described association with pulp bleaching prozyme in papermaking bio-bleaching technique.
Using method:
1. prozyme adds by the yellow slurry of drift before the chlorination of Huang slurry or after chlorination or after alkalization;
2. prozyme action condition: adapt to pH value 5-11, optimum pH 7-9, adaptive temperature 40-70 DEG C, optimal reactive temperature 45-55 DEG C;
3. using method: 1) by prozyme and 50 DEG C of water 1:10 Homogeneous phase mixing in mass ratio, adjusted to ph is 7-9, activation 20-30min, is added by the yellow slurry of drift by volume pump, make its with yellow starch to mix carry out enzyme digestion reaction;
2) addition of prozyme accounts for the 0.05-0.3 ﹪ of over dry Huang slurry quality;
3) concentration of yellow slurry is 5-15 ﹪;
4) enzymolysis time 35-60min, after enzymolysis completes, its chemical bleaching routinely processing step carries out, and each step chemical product consumption is the 40-50 ﹪ of conventional amount used.
Described Huang slurry for paper making raw material through getting the raw materials ready, slurrying, washing, the Huang slurry made after screening;
Described Huang is starched as the one in wood pulp or non-wood pulp or combination;
Described non-wood pulp is prepared by raw material wheat straw or straw or reed or bagasse;
Described prozyme adds fashionable before chlorination, and the addition of prozyme accounts for the 0.15-0.3 ﹪ of over dry Huang slurry quality; Prozyme adds fashionable after chlorination, and the addition of prozyme accounts for the 0.1-0.15 ﹪ of over dry Huang slurry quality; Prozyme adds fashionable after alkalization, and the addition of prozyme accounts for the 0.05-0.1 ﹪ of over dry Huang slurry quality.
Beneficial effect:
Association with pulp bleaching prozyme of the present invention with high temperature resistant alkalescent xylanase for main raw material, the composite bacillus acidocldarius culture of science, laccase and medium thereof, dextranase, EDTA, protective material, activator, seminase, lipase, tannase, polygalacturonase, whiteness stablizer, nonionogenic tenside, antioxidant etc., it is complete to prepare a kind of enzyme system, bleaching effect is good, easy preservation, the association with pulp bleaching enzyme that stability in storage is good, while retaining paper cellulose completely, dissolved lignin being degraded to greatest extent, fundamentally reduces content of lignin in paper pulp, strengthen delignification effect, appropriate composite whiteness stablizer, can make up the deficiency of bio-bleaching, to reach permanent bleaching simultaneously, prevent yellowing reversion of pulp, improve into the seed output and quality of paper, compared with bleaching specific enzyme with Market Pulp, association with pulp bleaching prozyme of the present invention is to leaf wood, needlebush and reed Huang slurry can significantly improve paper output and quality: yield improves 8.94% respectively, 8.80% and 10.05%, alpha-cellulose content improves 8.58%, 9.49% and 16.41% respectively, whiteness improves 10.2%, 10.72% and 10.59% respectively, YI yellow index reduces by 38.35%, 35.82% and 31.76% respectively, viscosity reduces by 1.20%, 1.18% and 1.43% respectively, ash content reduces by 25%, 26% and 28% respectively, can save conventional chemical articles for use amount 50-60%, thus advance bio-bleaching in the application process of paper industry, finally reach the object reduced costs with protection of the environment, its beneficial effect is the synergistic comprehensive embodiment of prozyme each component, is in particular in:
1. alkalescent xylanase of the present invention is high temperature resistant, alkali resistance is strong, action condition is wide in range, do not have cellulase activity, and the bio-bleaching and the chemistry that are more applicable to paper pulp help drift.
2. bacillus acidocldarius culture of the present invention contains stronger biological activity, high temperature resistant alkalescent xylanase can be produced on the one hand by fermentation in bio-bleaching process, for system continues to provide, bacillus acidocldarius can be made even by the dynamic response of fermentation on the other hand, be distributed to paper cellulose fully, hemicellulose and xylogen interstitial, abundant softening Mierocrystalline cellulose and degradation of hemicellulose (xylan, dextran, seminose etc.), make other composition of prozyme (as laccase, the zymins such as medium, sequestrant, protective material, activator, whiteness stablizer etc.) evenly, penetrate into paper pulp vegetable cell interstitial fully, give full play to the effect of component, to improve paper pulp recovery rate and quality, shorten bleaching time, and remove impurity and strengthen bio-bleaching effect.
3. the present invention by laccase and medium 1-hydroxy benzo triazole science composite, form the compound amboceptor of strong oxidation, improve the enzymolysis effect of laccase, fully, effectively realize the Degradation of laccase to xylogen, fundamentally eliminate xylogen, prevent yellowing reversion of pulp.
4. the present invention uses EDTA can most of transition metal ion (cupric ion, ferrous ion, ferric ion, divalent manganesetion, bivalent nickel ion etc.) and micro heavy in chelated pulp, to get rid of it to the suppression of biological enzyme enzyme activity and eliminating effect, enzyme component fully, is effectively played a role.
5. whiteness stablizer of the present invention has the dual function of bleaching and stable whiteness, both effectively pulp brightness can be improved, yellowing reversion of pulp can be prevented again, and can not by the impact of oxygen in paper pulp and transition metal ion (cupric ion, ferrous ion, ferric ion, divalent manganesetion, bivalent nickel ion etc.), the yellowing reversion of pulp that after the use of whiteness stablizer can effectively make up bio-bleaching, residual lignin causes.
6. the main science of nonionogenic tenside of the present invention the is composite nonionogenic tenside of environment-friendly type superior performance, easy biological degradation, noresidue, without any side effects to human body, biology and environment; Can promote that in enzyme bleaching process prozyme component permeates, fully free xylogen, hemicellulose, pectin, tannin, fat, etc. substrate, promote and extend the action time of each component enzyme-to-substrate, objectionable constituent in paper pulp are fully degraded, puies forward the smoothness of paper pulp fiber and bleaching uniformity coefficient.
7. the science of association with pulp bleaching prozyme antioxidant of the present invention is composite, prozyme enzyme molecular structure effectively can be prevented to be oxidized and to cause loss of enzyme activity, improve enzyme activity stability.12 months are stored under 0 DEG C and 40 DEG C of conditions, in prozyme, the loss alive of single enzyme enzyme is respectively 0.41% and 0.56%, 18.0% and 65% is reduced respectively than comparative example, effectively to prevent from links such as packaging, storage, transport, uses, because environment change, working method are improper and cause the inactivation of enzyme, especially can preventing the enzyme deactivation that high temperature causes.
8. in association with pulp bleaching prozyme of the present invention, protectant science is composite, effectively slow down the moisture regain of compound enzymic preparation; Can strengthen prozyme simultaneously resistance toly to freeze, resistance toheat, keep identical enzyme activity, its heat resisting temperature can improve 20-30 DEG C, resistance to freezing temp can reduce 10-15 degree Celsius, effectively prevent the loss of prozyme enzyme activity in transport, preservation and use procedure, extend the quality guaranteed period of prozyme, reach same enzyme activity, can 2-3 be extended than the like product quality guaranteed period.
9. association with pulp bleaching prozyme of the present invention adds inorganic salt as activator, creates the top condition of enzyme catalysis, has given full play to the vigor of plant enzyme and microbial enzyme, improve prozyme action potency.
Embodiment
The present invention is described below by specific embodiment.Unless stated otherwise, technique means used in the present invention is method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, but not limits the scope of the invention, and the spirit and scope of the invention only limited by claims.To those skilled in the art, under the prerequisite not deviating from essence of the present invention and scope, the various change carry out the material component in these embodiments and consumption or change also belong to protection scope of the present invention.
Prepared by embodiment 1 raw material
1. the preparation of alkalescent xylanase: comprise the steps
(1) actication of culture
The slant strains of intact bacillus acidocldarius CCTCCM2013537 is inoculated in slant medium, cultivates 36h for 45 DEG C and carry out actication of culture, so activation 3 times;
Described slant medium consists of: extractum carnis 10g, sodium-chlor 12g, peptone 20g, glucose 5g, agar 20g, Chinese herbal medicine powder 10g, distilled water l000mL, pH value 10,121 DEG C of sterilizing 20min;
The preparation method of described Chinese herbal medicine powder is as follows:
With weight parts, take the Radix Astragali 25 parts, Radix Codonopsis 15 parts, radix bupleuri 12 parts, the root of large-flowered skullcap 12 parts, Herba Houttuyniae 10 parts, Radix Angelicae Sinensis 9 parts; Respectively said herbal medicine being crushed to particle diameter is less than 2 millimeters, then Homogeneous phase mixing add the water of 5 times of weight in container, control temperature 80 DEG C keeps 3h, and be then cooled to 55 DEG C, the mixed enzyme adding mixture gross weight 8% carries out enzymolysis, be 6 by lactic acid adjust ph, enzymolysis 3h, finally add the mixture of mixture 2 times of w ethanol and propyl alcohol, the mass ratio of ethanol and propyl alcohol is 1:1.5, control temperature to 70 DEG C keeps 4h, filters; Filter vacuum concentrates postlyophilization and obtains Chinese herbal medicine powder;
The parts by weight of described mixed enzyme consist of: endo-beta-glucanase 15 parts, outer beta-glucanase 15 parts, beta-glucosidase 12 parts, zytase 18 parts, pentosanase 18 parts, Pullulanase 25 parts, beta-amylase 12 parts, neutral protease 12 parts, aspartic protease 12 parts, superoxide-dismutase 8 parts, glucose oxidase 8 parts, acid phosphatase 8 parts;
(2) liquid seeds enlarged culturing
1. first order seed is cultivated: slant strains 2 articulating after step (1) being activated enters in 500 ml shake flasks, substratum loading amount 100 milliliters, rotary shaker 180 revs/min, culture temperature 43 DEG C, incubation time 15h;
2. secondary seed cultivate: by first order seed according to 10% inoculum size access in 500 milliliters of secondary seed shaking flasks, culture condition is identical with first order seed;
3. three grades of seed culture: secondary seed is accessed in 5000 milliliters of three grades of seed flask with 10% inoculum size, substratum loading amount 1000 milliliters, rotary shaker 100 revs/min, culture temperature 43 DEG C, incubation time 15h;
4. first class seed pot is cultivated: the first class seed pot being 150L with 10% inoculum size access cubic capacity by three grades of seeds, fermention medium loading amount 100L, culture temperature 45 DEG C, stirring velocity 400rpm, ventilation (V/V) 1:2, tank pressure 0.05Mpa, incubation time 20h;
Described one-level, secondary, three grades of seed culture medium weight consist of:
Yeast powder 0.5%, glucose 1.5%, peptone 0.5%, extractum carnis 0.8%, dipotassium hydrogen phosphate 1.5%, Chinese herbal medicine powder 2%, trehalose 3%, calcium sulfate 0.1%, magnesium chloride 0.2%, Trisodium Citrate 0.3%, insufficient section pure water is supplied, pH value 10,123 DEG C of sterilizing 40min;
Described seed tank culture basic weight amount consists of:
Maltodextrin 15%, yeast powder 0.8%, Chinese herbal medicine powder 2%, trehalose 3%, peptone 0.5%, corn steep liquor 0.5%, dipotassium hydrogen phosphate 1.5%, magnesium sulfate 0.1%, Trisodium Citrate 0.5%, birch xylan 0.8%, insufficient section pure water is supplied, pH value 10,123 DEG C of sterilizing 40min;
Described seeding tank fermented liquid cell concentration is 8.0x10
8individual/ml;
(3) ferment tank
By first class seed pot fermented liquid in step (2) with 6% inoculum size access fermentor tank, culture temperature 45 DEG C, stirring velocity 700rpm, ventilation (V/V) 1:3, incubation time 30h; Then with 2 DEG C/h rate of temperature fall slow cooling to 20 DEG C, constant temperature culture 12h; Continue with 2 DEG C/h rate of temperature fall slow cooling to 10 DEG C, now, first class seed pot fermented liquid in step (2) is added access fermentor tank with 4% inoculum size, constant temperature culture 12h; Finally slowly be warming up to 20 DEG C, constant temperature culture 12h with 2 DEG C/h temperature rise rate; Continue slowly to be warming up to 45 DEG C, constant temperature culture 30h with 2 DEG C/h temperature rise rate;
Dissolved oxygen controls: by adjustment mixing speed and ventilation, controls dissolved oxygen 30%;
PH controls: by mending ammoniacal liquor or dilute phosphoric acid, controls pH value in fermenting process and remains on 10;
Control of additive raw material: when the reducing sugar content in fermented liquid is down to 3mg/ml-8mg/ml, start to add supplemented medium, feed supplement amount is to maintain fermented liquid reducing sugar content for 2mg/ml-5mg/ml;
Put tank standard: the old and feeble self-dissolving of 80% thalline, enzyme activity increasess slowly;
Described fermention medium consists of: maltodextrin 150g, Semen Maydis powder 60g, soybean cake powder 25g, Chinese herbal medicine powder 50g, trehalose 40g, yeast powder 8g, corn steep liquor 5g, ammonium sulfate 3g, dipotassium hydrogen phosphate 2g, potassium primary phosphate 2g, Trisodium Citrate 5g, birch xylan 15, defoamer 1g, pure water l000mL, pH value 10,121 DEG C of sterilizing 20min;
Described supplemented medium weight consists of: maltodextrin 20-30%, Semen Maydis powder 10-20%, bean powder 15-25%, Chinese herbal medicine powder 5-10%, and insufficient section pure water is supplied, pH value 5-11,121-123 DEG C of sterilizing 30-40min;
The concocting method of described fermention medium is:
Accurately take raw material in proportion, pure water in raw material, Semen Maydis powder, soybean cake powder are dropped in material-compound tank, regulate pH value 10, add middle temperature amylase (3u/g Semen Maydis powder) and alpha-amylase (30u/g Semen Maydis powder), DEG C insulation 30min of warming while stirring to 70 simultaneously, be then slowly warming up to 90 DEG C of insulation 30min and liquefy, finally add other raw material, stir, adjust initial pH value 10,123 DEG C of sterilizing 40min for subsequent use.
(4) fermentation liquor filter, concentrated, allotment, essence filter, dry high temperature resistant alkalescent xylanase;
Described allocation process does not add any sanitas, adds concentrated enzyme liquid gross weight 5% Chinese herbal medicine powder.
2. the preparation of bacillus acidocldarius culture: the preparation method of described bacillus acidocldarius culture be prepare the fermentation liquor concentrating under reduced pressure of alkalescent xylanase in above-mentioned 1, lyophilize, pulverize at low temperature obtain, its viable bacteria content is 9x1010CFU/g.
The mensuration (3,5-dinitrosalicylic Acid Colorimetry, is called for short DNS method) of alkaline xylanase activity power that embodiment 2 is high temperature resistant
1. reagent and solution
1.1 water used are except special requirement, and be the tertiary effluent meeting GB/6682-1992 and specify, pharmaceutical chemicals, except special requirement, is analytical pure.
1.2DNS reagent
Take 3,5-dinitrosalicylic acid 10g adds in 500ml water, adds sodium hydroxide 16g several times, stirring and dissolving (temperature < 45 DEG C), add Seignette salt 300g several times again, be stirred to entirely molten, after cooling, be settled to 1000ml with water.Under room temperature, brown bottle stores dark place placement, uses (using if any after sedimentation and filtration) after one week.
1.30.1mol/L, pH5.0 acetic acid-sodium-acetate buffer
Draw Glacial acetic acid 1.8ml, add suitable quantity of water, add sodium-acetate 5.78g, dissolve, be settled to 1000ml, use after regulating PH to 5.00 ± 0.01.2 months are deposited effectively under room temperature.
1.40.1% be 1mg/ml xylose standard solution
Anhydrous wood sugar dries to constant weight in 80 DEG C, takes 0.1000g in beaker, adds suitable quantity of water and dissolves, to proceed in volumetric flask and to be settled to 100ml.
1.51.0% xylan solution
Take xylan 1.00g in beaker, pasty state is soaked into 2ml0.5mol/L sodium hydroxide, add 40ml damping fluid, heating for dissolving in 60 ~ 70 DEG C of water-baths, pH5.0 is adjusted with 0.5mol/L acetic acid (about 2ml) after cooling, proceed in volumetric flask, be settled to 100ml with damping fluid (4.3).If in refrigerator storage period grown and occurred precipitation, before using should in hot water bath thermosol.
The drafting of 1.6 xylose standard curve
Draw 1mg/ml xylose solution 0,0.2,0.4,0.6,0.8,1.0ml in test tube, moisturizing, to 2ml, adds DNS reagent 3ml, boils 10min, be settled to 15ml after cooling after mixing in boiling water, surveys absorbancy (A) under spectrophotometer 540nm wavelength.Take absorbancy as ordinate zou, wood sugar amount is X-coordinate, drawing standard curve, and the average least square fitting a linear equation y=ax ﹢ b of three revision tests, obtains the relation of absorbancy and wood sugar amount.
2. the process of enzyme sample to be measured
Direct absorption enzyme liquid does suitable dilution, makes mensuration absorbance value in 0.25-0.4 scope.
3. enzyme activity determination
Get 15ml tool plug scale test tube to operate by reaction sequence below, in reaction process, from adding substrate (shaking up before absorption) (1.5), want definitely consistent, 50 DEG C of hydrolysis 10min. to often propping up in test tube the timed interval adding reagent
Reactions steps and reagent, solution usage are in table 1:
Table 1 reactions steps and reagent, solution usage
| Sample | Blank |
| 1. add substrate (1.5) 1.8ml | 1. add substrate (1.5) 1.8ml |
| 2.50 DEG C of preheating 3min | 2.50 DEG C of preheating 3min |
| 3. enzyme-added liquid 0.2ml | 3.50 DEG C of hydrolysis 10min |
| 4. mix | 4. add DNS reagent 3ml |
| 5.50 DEG C of hydrolysis 10min | 5. mix |
| 6. add DNS reagent 3ml | 6. enzyme-added liquid 0.2ml |
| 7. mix | 7. mix |
| 8. 10min in boiling water bath | 8. 10min in boiling water bath |
| 9. cooling is settled to 15ml | 9. cooling is settled to 15ml |
| 10. measure under blank zeroing 540nm | 10. measure under blank zeroing 540nm |
4. enzyme activity unit calculates
In enzyme activity=(S × D × 1000)/(0.2 × 10) × 1.7 μ g/ml (g) .min formula:
S-record photoabsorption wood sugar amount corresponding on typical curve, mg number;
D-enzyme liquid or enzyme powder extension rate;
Reduction factor between 1000-mg and μ g;
0.2-measure enzyme liquid measure of getting, ml number;
10-reaction times, min number;
1.7-method reduction factor.
5. enzyme activity unit definition and conversion
5.1 present method enzymes are lived and are defined: under this condition determination, and per minute hydrolyzed xylan produces the required enzyme amount of 1 μ g reducing sugar (in wood sugar) and is defined as an enzyme activity unit (u).
The definition of international Mei Huo unit: under condition determination, per minute hydrolyzed xylan produces the required enzyme amount of 1 μm of ol reducing sugar (in wood sugar) and is defined as an enzyme activity unit (IU).
The conversion relation of 5.2u and international unit IU:
1u=1 ÷ 150IU (μm ol sugar/ml (g) .min)
150: the molecular weight of wood sugar
The thermal stability analysis of embodiment 3 alkalescent xylanase
The thermostability of enzyme is analyzed, at crude enzyme liquid prepared by embodiment 1 alkalescent xylanase is placed in 40 DEG C, 45 DEG C, 50 DEG C, 55 DEG C, 60 DEG C, 65 DEG C, 70 DEG C respectively, lives every 10 minutes sampling and measuring enzymes.At crude enzyme liquid 40 DEG C, 45 DEG C, 50 DEG C, 55 DEG C, 60 minutes enzymes are lived and are not declined.At 60 DEG C and 65 DEG C, what within 30 minutes, drop to constitutive enzyme work drops to 85% in 95%, 60 minutes.At 70 DEG C, what within 30 minutes, drop to constitutive enzyme work drops to 80% in 85%, 60 minutes.At 75 DEG C, what within 30 minutes, drop to constitutive enzyme work drops to 70% in 80%, 60 minutes, and compared with prior art reach same enzyme and live, tolerable temperature is higher.
Embodiment 4 alkalescent xylanase pH stability analysis
The pH stability of enzyme is analyzed, crude enzyme liquid prepared by embodiment 1 alkalescent xylanase is placed in respectively pH value 4.0,5.0,6.0,7.0,8.0,9.0,10.0,11.0 times, live every 10 minutes sampling and measuring enzymes.Crude enzyme liquid pH value 6.0,7.0,8.0,9.0 times, 60 minutes enzymes are lived and are not declined.PH value 5.0 and 10.0 times, what within 30 minutes, drop to that constitutive enzyme lives drops to 85% in 95%, 60 minutes.PH value 4.0 and 11.0 times, what within 30 minutes, drop to that constitutive enzyme lives drops to 70% in 80%, 60 minutes.It is obviously more wide in range than prior art bacterial strain that similarity condition is issued to same enzyme resistance to pH value alive.
Embodiment 5
A kind of association with pulp bleaching prozyme, prepared by the raw material of following parts by weight:
Alkalescent xylanase 30 parts, bacillus acidocldarius culture 20 parts, laccase 15 parts, dextranase 6 parts, 1-hydroxy benzo triazole 5 parts, EDTA5 part, protective material 6 parts, activator 6 parts, seminase 5 parts, 4 parts, lipase, tannase 4 parts, polygalacturonase 4 parts, two phosphorus derivants (BBHPE) 3 parts of trishydroxymethyl phosphoric acid, nonionogenic tenside 3 parts, antioxidant 2 parts;
Described alkalescent xylanase and bacillus acidocldarius culture are prepared by embodiment 1;
The quality component of described nonionogenic tenside is: alkyl polyglycoside (APG) 30 parts, methyl glucamine (AGA) 25 parts, N-dodecyl ethylenediamine triacetic acid sodium (ED3A) 12 parts, isomery alcohol ether carboxylate AEC-110712 part, Lauric Acid Monoethanolamide Ether Carboxylate (LAEC) 9 parts, peregal c-1254 part, JFC4 part;
Described activator is formed by the inorganic salt Homogeneous phase mixing of following quality component: 12 parts, sodium sulfate, zinc chloride 7 parts, 6 parts, magnesium chloride, 4 parts, calcium chloride;
Described protective material is made up of the raw material of following parts by weight: trehalose 30 parts, ganoderan 15 parts, NaCl12 part, (NH
4)
2sO
47 parts, halfcystine 4 parts;
Described antioxidant is grape pip procyanidin, rosemary extract and apricot leaf extract 3:4:1.5 Homogeneous phase mixing in mass ratio;
The preparation method of above-mentioned association with pulp bleaching prozyme, comprise the steps: first by described protective material, activator is micronizing respectively, Homogeneous phase mixing, immediately add nonionogenic tenside successively, bacillus acidocldarius culture Homogeneous phase mixing 25min, then alkalescent xylanase is added successively, laccase, dextranase, seminase, lipase, tannase, polygalacturonase Homogeneous phase mixing, finally add 1-hydroxy benzo triazole successively, EDTA, two phosphorus derivants (BBHPE) of trishydroxymethyl phosphoric acid, antioxidant, pack after mixing and obtain association with pulp bleaching prozyme.
Embodiment 6
A kind of association with pulp bleaching prozyme, prepared by the raw material of following parts by weight:
Alkalescent xylanase 20 parts, bacillus acidocldarius culture 10 parts, laccase 10 parts, dextranase 5 parts, 1-hydroxy benzo triazole 3 parts, EDTA3 part, protective material 4 parts, activator 4 parts, seminase 4 parts, 3 parts, lipase, tannase 3 parts, polygalacturonase 3 parts, trishydroxymethyl phosphoric acid (THP) 2 parts, nonionogenic tenside 2 parts, antioxidant 1 part;
Described alkalescent xylanase and bacillus acidocldarius culture are prepared by embodiment 1;
The quality component of described nonionogenic tenside is: alkyl polyglycoside (APG) 20 parts, methyl glucamine (AGA) 20 parts, N-dodecyl ethylenediamine triacetic acid sodium (ED3A) 10 parts, isomery alcohol ether carboxylate AEC-110710 part, Lauric Acid Monoethanolamide Ether Carboxylate (LAEC) 8 parts, peregal c-1253 part, JFC3 part;
Described activator is formed by the inorganic salt Homogeneous phase mixing of following quality component: 10 parts, sodium sulfate, zinc chloride 6 parts, 5 parts, magnesium chloride, 3 parts, calcium chloride;
Described protective material is made up of the raw material of following parts by weight: trehalose 20 parts, ganoderan 12 parts, NaCl10 part, (NH
4)
2sO
45 parts, halfcystine 3 parts;
Described antioxidant is grape pip procyanidin, rosemary extract and apricot leaf extract 2:1:1 Homogeneous phase mixing in mass ratio;
The preparation method of above-mentioned association with pulp bleaching prozyme is with embodiment 2.
Embodiment 7
A kind of association with pulp bleaching prozyme, prepared by the raw material of following parts by weight:
Alkalescent xylanase 40 parts, bacillus acidocldarius culture 30 parts, laccase 20 parts, dextranase 8 parts, 1-hydroxy benzo triazole 8 parts, EDTA8 part, protective material 7 parts, activator 7 parts, seminase 6 parts, 5 parts, lipase, tannase 5 parts, polygalacturonase 5 parts, whiteness stablizer 5 parts, nonionogenic tenside 4 parts, antioxidant 3 parts;
Described alkalescent xylanase and bacillus acidocldarius culture are prepared by embodiment 1;
Described whiteness stablizer is two phosphorus derivants (BBHPE) 1:2 Homogeneous phase mixing in mass ratio of trishydroxymethyl phosphoric acid (THP) and trishydroxymethyl phosphoric acid;
The quality component of described nonionogenic tenside is: alkyl polyglycoside (APG) 40 parts, methyl glucamine (AGA) 30 parts, N-dodecyl ethylenediamine triacetic acid sodium (ED3A) 15 parts, isomery alcohol ether carboxylate AEC-110715 part, Lauric Acid Monoethanolamide Ether Carboxylate (LAEC) 10 parts, peregal c-1255 part, JFC5 part;
Described activator is formed by the inorganic salt Homogeneous phase mixing of following quality component: 15 parts, sodium sulfate, zinc chloride 8 parts, 8 parts, magnesium chloride, 5 parts, calcium chloride;
Described protective material is made up of the raw material of following parts by weight: trehalose 40 parts, ganoderan 18 parts, NaCl15 part, (NH4) 2SO49 part, halfcystine 5 parts;
Described antioxidant is grape pip procyanidin, rosemary extract and apricot leaf extract 4:3:2 Homogeneous phase mixing in mass ratio;
The preparation method of above-mentioned association with pulp bleaching prozyme is with embodiment 2.
Embodiment 8
A kind of association with pulp bleaching prozyme, prepared by the raw material of following parts by weight:
Alkalescent xylanase 20 parts, bacillus acidocldarius culture 30 parts, laccase 20 parts, dextranase 5 parts, 1-hydroxy benzo triazole 3 parts, EDTA8 part, protective material 7 parts, activator 7 parts, seminase 4 parts, 5 parts, lipase, tannase 5 parts, polygalacturonase 5 parts, 5 parts, mercaptan, nonionogenic tenside 2 parts, grape pip procyanidin 3 parts;
Described alkalescent xylanase and bacillus acidocldarius culture are prepared by embodiment 1;
The quality component of described nonionogenic tenside is: alkyl polyglycoside (APG) 20 parts, methyl glucamine (AGA) 30 parts, N-dodecyl ethylenediamine triacetic acid sodium (ED3A) 15 parts, isomery alcohol ether carboxylate AEC-110710 part, Lauric Acid Monoethanolamide Ether Carboxylate (LAEC) 8 parts, peregal c-1255 part, JFC5 part;
Described activator is formed by the inorganic salt Homogeneous phase mixing of following quality component: 10 parts, sodium sulfate, zinc chloride 8 parts, 8 parts, magnesium chloride, 3 parts, calcium chloride;
Described protective material is made up of the raw material of following parts by weight: trehalose 40 parts, ganoderan 12 parts, NaCl10 part, (NH
4)
2sO
49 parts, halfcystine 3 parts;
The preparation method of above-mentioned association with pulp bleaching prozyme is with embodiment 2.
Embodiment 9 antioxidant is on the impact of association with pulp bleaching prozyme enzyme activity
Adopt association with pulp bleaching prozyme prepared by the embodiment of the present invention 5, other raw material, material component, preparation method are identical, unique difference is that feed composition is not containing antioxidant, form comparative example, 12 months are stored respectively under 0 DEG C and 40 DEG C of conditions, detection method in " GB/T23535-2009 lipase preparation " is adopted to measure the enzyme activity of lipase, calculate enzyme rate of loss alive, enzyme rate of loss alive refers to that the enzyme activity of actual detection and the difference of product annotation enzyme activity account for the percentage marking enzyme activity, and result is as table 2
Lipase activity power rate of loss in table 2 shelf lives prozyme
Above result shows, 12 months are stored under 0 DEG C and 40 DEG C of conditions, lipase in embodiment 2 reduces by 18% and 65% respectively than the lipase activity loss in comparative example, illustrate that the science of antioxidant is composite, significantly improve the vigor of each component enzymes in prozyme, enzyme is lived in losing and is significantly reduced, and effect is more remarkable especially under the high temperature conditions.
Embodiment 10 association with pulp bleaching prozyme of the present invention is to the quality influence of different material Huang slurry
One, test site: beautiful paper industry is avenged in Hunan.
Two, test period: on April 22 ,-2014 years on the 12nd January in 2014, last 100 days.
Three, plan design: 1. test as single-factor designs, test group and control group are set, the raw material of control group and test group, technique, equipment, operator, environment and way to manage are all identical, difference is: add association with pulp bleaching prozyme prepared by the embodiment of the present invention 5 in the Huang slurry of test group before chlorination, control group adds Market Pulp bleaching specific enzyme.2. test group and control group all add association with pulp bleaching prozyme by the 0.15-0.3% that yellow slurry over dry is heavy.3. same raw material Huang slurry test group and control group respectively process 10 batches, calculating mean value.4. postorder chemical bleaching routinely technique carry out.5. pair different material Huang slurry carries out detected result as table 3 through the paper pulp correlated quality index that bleaching is obtained
Table 3 association with pulp bleaching prozyme is on the impact of different material Huang slurry quality
Above result shows, compared with bleaching specific enzyme with Market Pulp, association with pulp bleaching prozyme of the present invention can significantly improve paper output and quality to leaf wood, needlebush and reed Huang slurry: yield improves 8.94%, 8.80% and 10.05% respectively; Alpha-cellulose content improves 8.58%, 9.49% and 16.41% respectively; Whiteness improves 10.2%, 10.72% and 10.59% respectively; YI yellow index reduces by 38.35%, 35.82% and 31.76% respectively; Viscosity reduces by 1.20%, 1.18% and 1.43% respectively; Ash content reduces by 25%, 26% and 28% respectively, while raising alpha-cellulose content and output, especially substantially increases pulp brightness, reduces the YI yellow index of paper pulp, effectively prevent paper from yellowing.
Claims (10)
1. an association with pulp bleaching prozyme, is prepared by the raw material of following parts by weight: alkalescent xylanase 20-40 part, bacillus acidocldarius culture 10-30 part, laccase 10-20 part, dextranase 5-8 part, 1-hydroxy benzo triazole 3-8 part, EDTA3-8 part, protective material 4-7 part, activator 4-7 part, seminase 4-6 part, lipase 3-5 part, tannase 3-5 part, polygalacturonase 3-5 part, whiteness stablizer 2-5 part, nonionogenic tenside 2-4 part, antioxidant 1-3 part;
Described alkalescent xylanase and bacillus acidocldarius culture are starting strain by the bacillus acidocldarius CCTCCM2013537 producing high temperature resistant alkalescent xylanase, and are prepared by Optimal Medium composition and zymotechnique;
Described activator is formed by the inorganic salt Homogeneous phase mixing of following quality component: sodium sulfate 10-15 part, zinc chloride 6-8 part, magnesium chloride 5-8 part, calcium chloride 3-5 part;
Described protective material is made up of the raw material of following parts by weight: trehalose 20-40 part, ganoderan 12-18 part, NaCl10-15 part, (NH
4)
2sO
45-9 part, halfcystine 3-5 part.
2. a kind of association with pulp bleaching prozyme as claimed in claim 1, it is characterized in that, the preparation method of described high temperature resistant alkalescent xylanase is as follows: obtain seed liquor by activated for the slant strains of intact bacillus acidocldarius CCTCCM2013537 with one, two, three seed culture and first class seed pot cultivation, with 6% inoculum size access fermentor tank, culture temperature 37-45 DEG C, stirring velocity 200-700rpm, ventilation (V/V) 1:1-3, incubation time 25-30h; Then with 1-2 DEG C/h rate of temperature fall slow cooling to 15-20 DEG C, constant temperature culture 10-12h; Continue with 1-2 DEG C/h rate of temperature fall slow cooling to 8-10 DEG C, now, first class seed pot fermented liquid is added access fermentor tank with 4% inoculum size, constant temperature culture 7-12h; Finally slowly be warming up to 15-20 DEG C, constant temperature culture 10-12h with 1-2 DEG C/h temperature rise rate; Continue slowly to be warming up to 37-45 DEG C, constant temperature culture 20-30h with 1-2 DEG C/h temperature rise rate; Fermentation liquor is filtered, concentrated, allotment, essence filter, dry high temperature resistant alkalescent xylanase.
3. a kind of association with pulp bleaching prozyme as claimed in claim 1, is characterized in that, described bacillus acidocldarius culture viable bacteria content is 7x10
10-9x10
10cFU/g.
4. a kind of association with pulp bleaching prozyme as claimed in claim 1, it is characterized in that, described whiteness stablizer is that one or more in two phosphorus derivants of mercaptan, thioether, trishydroxymethyl phosphoric acid, tetra methylol phosphoric acid salt, trishydroxymethyl phosphoric acid are with arbitrary proportion Homogeneous phase mixing.
5. a kind of association with pulp bleaching prozyme as claimed in claim 1, is characterized in that, described nonionogenic tenside is environment-friendly type nonionogenic tenside.
6. a kind of association with pulp bleaching prozyme as claimed in claim 1, is characterized in that, described antioxidant is any or several combination in grape pip procyanidin, rosemary extract and apricot leaf extract.
7. the preparation method of a kind of association with pulp bleaching prozyme as claimed in claim 1, it is characterized in that, comprise the steps: first by described protective material, activator is micronizing respectively, Homogeneous phase mixing, immediately add nonionogenic tenside successively, bacillus acidocldarius culture Homogeneous phase mixing 20-30min, then alkalescent xylanase is added successively, laccase, dextranase, seminase, lipase, tannase, polygalacturonase Homogeneous phase mixing, finally add 1-hydroxy benzo triazole successively, EDTA, whiteness stablizer, antioxidant, pack after mixing and obtain association with pulp bleaching prozyme.
8. the preparation method of a kind of association with pulp bleaching prozyme as claimed in claim 7, is characterized in that, described whiteness stablizer is two phosphorus derivants of trishydroxymethyl phosphoric acid.
9. the preparation method of a kind of association with pulp bleaching prozyme as claimed in claim 7, it is characterized in that, the quality component of described nonionogenic tenside is: alkyl polyglycoside 20-40 part, methyl glucamine 20-30 part, N-dodecyl ethylenediamine triacetic acid sodium 10-15 part, isomery alcohol ether carboxylate AEC-110710-15 part, Lauric Acid Monoethanolamide Ether Carboxylate 8-10 part, peregal c-1253-5 part, JFC3-5 part.
10. the preparation method of a kind of association with pulp bleaching prozyme as claimed in claim 7, is characterized in that, described antioxidant is grape pip procyanidin, rosemary extract and apricot leaf extract 2-4:1-3:1-2 Homogeneous phase mixing in mass ratio.
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| CN104499337A (en) * | 2014-11-30 | 2015-04-08 | 邵素英 | Bleached compound enzyme and preparation method thereof |
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