CN104499337B

CN104499337B - One kind bleaching complex enzyme and preparation method thereof

Info

Publication number: CN104499337B
Application number: CN201410715635.XA
Authority: CN
Inventors: 邵素英
Original assignee: Ningxia Tian Green Health Intellectual Property Operation Co Ltd
Current assignee: Guangzhou Longli biotechnology limited company
Priority date: 2014-11-30
Filing date: 2014-11-30
Publication date: 2018-02-16
Anticipated expiration: 2034-11-30
Also published as: CN104499337A

Abstract

The invention discloses one kind bleaching complex enzyme and preparation method thereof, using high temperature resistant alkalescent xylanase as primary raw material, science compounds bacillus alcalophilus culture, laccase and its medium, dextranase, EDTA, protective agent, activator, seminase, lipase, tannase, pectase, whiteness stabilizer, nonionic surfactant, antioxidant etc., while paper cellulose is fully retained, dissolved lignin and degraded to greatest extent, it is complete to be prepared for a kind of enzyme system, bleaching effect is good, it is easy to maintain, the good bleaching complex enzyme enzyme of storage stability, to leaf wood, needlebush and the yellow slurry of reed are remarkably improved paper output and quality, whiteness is respectively increased 10.94%, 11.97% and 11.10%；YI yellow index reduces by 39.84%, 37.31% and 24.32% respectively；Conventional chemical articles for use amount 50 60% can be saved；Being finally reached reduces the purpose of cost and environmental protection.

Description

One kind bleaching complex enzyme and preparation method thereof

Technical field

The present invention relates to paper grade (stock) complex enzyme, and in particular to one kind bleaching complex enzyme and preparation method thereof.

Background technology

In the industrial production, pulping process is under conditions of alkalescence, delignification is removed with the method for thermophilic digestion, to make Dissolving pulping fibre element purity reaches 98%, and this just needs substantial amounts of naoh treatment paper pulp, and it is dirty to cause serious environment Dye, transfer to attempt biological treatment slurrying for this many scholar, and the zytase for being used for pulping bleaching should be heat-resisting and alkaline-resisting. Feng Jianliang etc. (Kimura et al.2000) compares examination with xylanase pretr eatment wheat straw and conventional chemistry slurrying Test, xylanase pretr eatment improves the delignification degree of raw material, can also reduce the Kappa number of slurrying.Zytase is aiding in There is wide and in-depth study in terms of bleaching, and achieve very big achievement.Research finds, either softwood pulp, leaf wood Slurry or bamboo pulp, straw pulp, zytase can degrade remaining lignin in paper pulp, improve whiteness (AL BALAA et al,2006；Meek and Lipman,1922).Zytase for association with pulp bleaching must have high temperature resistant feature, Khasin Alkaline bacterial strain Bacillus sterarothermophilus T26 zytases are derived from etc. having obtained one, the zytase There is best bleaching effect (Khasin et al, 1993) to paper pulp in pH 9.0 and 65 DEG C, many zytases do not possess The characteristics of such, limit commercial applications.Caused waste paper quantity is surprising every year in the whole world, is recycled after waste paper recovery Work just becomes particularly important, can make also to protect environment while resource reuses.1991, by South Korea scholar Report and ink can be removed from news secondary stock using zytase first.Hereafter, people, which begin one's study, utilizes xylan Enzyme carries out deinking, so that the environmental pollution (Cao Junwei etc., 2004) that Chemical Deinking is brought is mitigated or eliminated.

At present, it is each to have become the world for the paper pulp green cleaning and bleaching using element-free chlorine and Totally-chlorine-free bleaching technology as representative The inexorable trend of state's paper-making industry association with pulp bleaching development, zytase enzyme process help drift new technology in more than 30 families in Europe and North America Large-scale paper plant is applied, and turns into biotechnology and applies most successful one in paper industry.Wherein Canada has about 10% Sulfate process pulp mill employs this new technology.The more enzyme preparations such as Novozymes Company of Denmark and this chemical company of U.S.'s mountain pass Manufacturer, zytase and cellulase new product dedicated for slurrying processing are proposed one after another, but up to the present, industrially Zytase applied to association with pulp bleaching is neutral meta-acid mostly, and optimal reactive temperature is mostly at 50 DEG C or so, it is well known that Pulp cooking and bleaching are substantially what is carried out under conditions of high temperature and highly basic so that existing low temperature acid zytase production Application of the product in this field is extremely limited.

But because xylanase treatment technique is to remove the modes such as the redeposited hemicellulose in paper pulp surface by degrading To help chemistry drift agent bleaching, zytase can only play a part of helping drift, it is impossible to really substitute chemistry drift agent.Therefore, it is raw Thing pre-bleaching can not substitute chemical bleaching completely, and can reduce pollution but can not finally eliminate pollution, be inherently eliminated The pollution of chlorine bleach liquor, bio-bleaching need to be finally realized, i.e., removes the lignin remained in paper pulp using biological means completely, because Presence for lignin is the basic reason of yellowing reversion of pulp, the color development that chemical bleaching directly can act on lignin and destroy in paper pulp Group.Ligninase can be done directly on lignin, and lignin is degraded, predominantly the lignin peroxide of whiterot fungi secretion Enzyme, manganese peroxidase, laccase and cellobiose dehydrogenase etc., the focus studied in recent years are concentrated mainly on the biology drift of laccase Bai Shang, laccase are a kind of blue multicopper oxidase, also referred to as polyphenol oxidase, lignin structure unit of the main oxidation containing phenol, but naturally Wood lignin is only less than 20% structural units containing phenol, and another generally fungal laccase molecular weight larger (about 70000Da) is difficult Lignin molecule is acted in wood to penetrate, wants to give full play to its enzymatic activity, it is necessary to add laccase mediators, usually violuric acid (VA) and 1- hydroxy benzo triazoles (HBT), selected depending on different bleaching process, and the yield of laccase is smaller, stability compared with Difference.

The A of Chinese patent CN 102978986 disclose a kind of method that biological enzyme prepares paper pulp, prepared by the biology enzyme The mass percentage content of biology enzyme component is in liquid：Cellulase 20%, hemicellulase 10%, lignin degradation enzyme 10%, Amylase 2 0%, lipase 5%, pectase and laccase 20%；The active ingredient that cellulase therein can degrade in paper pulp is fine Dimension element, is difficult to control, and has a strong impact on into the physical and mechanical properties of paper, meanwhile, above-mentioned biology enzyme is applied to pulping process, rather than drift White technique, also, single use laccase, its enzyme activity are difficult to play, it is impossible to act on lignin, yellowing reversion of pulp possibility very well Increase.The A of Chinese patent CN 103555701 disclose a kind of production method of association with pulp bleaching complex enzyme liquid, it is characterized in that will Each component enzyme is deposited in after weighing, compounding tank mixed preparing, anti-corrosion, inspection, packaging in 5 DEG C of storehouses, and each component enzyme is Alkaline pectase, zytase, mannase and lignoenzyme；Above-mentioned complex enzyme liquid enzyme class and characteristic are single, lignin Enzyme activity is low and is difficult to realize, still fundamentally can not thoroughly eliminate without the specific species of open lignoenzyme, industrialization Lignin, bleaching effect is still undesirable, and not easy to maintain, poor storage stability, Commercialization application are limited.

To sum up, preparing the association with pulp bleaching enzyme that a kind of enzyme system is complete, bleaching effect is good, easy to maintain, storage stability is good has extensively Wealthy market prospects and huge market potential value.

The content of the invention

Technical problem solved by the invention is the defects of overcoming existing association with pulp bleaching enzyme, with high temperature resistant alkalescent xylanase For primary raw material, science compounding bacillus alcalophilus culture, laccase and its medium, dextranase, EDTA, protective agent, activation Agent, seminase, lipase, tannase, pectase, whiteness stabilizer, nonionic surfactant, antioxidant etc., prepare The association with pulp bleaching enzyme that a kind of enzyme system is complete, bleaching effect is good, easy to maintain, storage stability is good, is being fully retained paper cellulose Meanwhile dissolved lignin and degraded to greatest extent, fundamentally reduce content of lignin in paper pulp, enhancing delignification effect Fruit, while appropriateness compounding whiteness stabilizer, can make up the deficiency of bio-bleaching, strive reaching permanent bleaching, prevent yellowing reversion of pulp, The yield and quality of paper are improved into, being finally reached reduces cost and guarantor in the application process of paper industry to promote bio-bleaching The purpose in retaining ring border.

In order to achieve the above object, the present invention uses following technical scheme：

One kind bleaching complex enzyme, is prepared by the raw material of following parts by weight：

Alkalescent xylanase 10-20 parts, bacillus alcalophilus culture 8-10 parts, laccase 8-10 parts, dextranase 5-8 Part, 1- hydroxy benzo triazole 3-8 parts, EDTA 3-8 parts, protective agent 4-7 parts, activator 4-7 parts, seminase 4-6 parts, fat Fat enzyme 3-5 parts, tannase 3-5 parts, pectase 3-5 parts, whiteness stabilizer 2-5 parts, nonionic surfactant 2-4 parts, antioxygen Agent 1-3 parts.

Alkalescent xylanase and the bacillus alcalophilus culture is by the thermophilic bud of production high temperature resistant alkalescent xylanase Spore bacillus (Bacillus thermophilus) 701 (deposit number is CCTCC M 2013537) is starting strain, and is passed through Optimal Medium form and zymotechnique and prepare；

The alkalescent xylanase high temperature resistant, alkali resistance is strong, action condition is wide in range, without cellulase activity, more The bio-bleaching and chemistry for being adapted to paper pulp help drift；

The zymologic property of the high temperature resistant alkalescent xylanase is as follows：

(1) temperature：The enzyme Acclimation temperature wider range, Applicable temperature scope are 40-75 DEG C, 65 DEG C of optimal reactive temperature, In 75 DEG C of enzyme activity complete stabilities；

(2)pH：It is 5.0-11.0 that the enzyme, which is applicable pH value in reaction scope, the enzyme activity complete stability when pH value is 11.0, most suitable PH value in reaction is 10.0；

(3) enzymatic activity：Zymotic fluid alkalescent xylanase specific activity of enzyme is higher, is 350-420IU/ml；

(4) heat endurance：The enzyme remains to keep more than 80% enzyme activity after preserving 1h under the conditions of 70 DEG C, is protected under the conditions of 75 DEG C More than 70% enzyme activity is kept after depositing 1h；

(5) storage stability：Enzyme activity loss is less than 3.0% after the enzyme preserves 12 months at room temperature.

Preferably, the preparation method of the high temperature resistant alkalescent xylanase is as follows：By intact bacillus alcalophilus CCTCC M 2013537 slant strains are activated and expand culture (including one, two, three seed culture and one-level step by step Seed tank culture) seed liquor is obtained, fermentation tank, 37-45 DEG C of cultivation temperature, mixing speed 200- are accessed with 6% inoculum concentration 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, incubated 10-12h；Continue that now, first class seed pot is fermented to 8-10 DEG C with 1-2 DEG C/h rate of temperature fall slow cooling Liquid adds access fermentation tank, incubated 7-12h with 4% inoculum concentration；15- is finally to slowly warm up to 1-2 DEG C/h heating rates 20 DEG C, incubated 10-12h；Continue to be to slowly warm up to 37-45 DEG C with 1-2 DEG C/h heating rates, incubated 20-30h；Hair Zymotic fluid is filtered, concentration, allotment, refined filtration, dry high temperature resistant alkalescent xylanase, it is total that the allocation process adds concentration enzyme liquid Weight 0.5-5% Chinese herbal medicine powders；

The slant medium forms：Beef extract 3-10g, sodium chloride 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 the Chinese herbal medicine powder is as follows：In terms of parts by weight, Radix Astragali 20-30 parts, Radix Codonopsis 10-18 are weighed Part, radix bupleuri 10-15 parts, radix scutellariae 10-15 parts, cordate houttuynia 8-12 parts, Radix Angelicae Sinensis 8-10 parts；Said herbal medicine is crushed to particle diameter respectively For less than 2 millimeters, then uniformly mixed in container and add the water of 3-6 times of weight, control temperature 70 C~90 DEG C holding 2~ 4h, 45-60 DEG C is then cooled to, adds mixed enzyme and digested, be 5.5-6.8 with newborn acid for adjusting pH value, digest 2-4h, finally Add the mixture of 0.5-3 times of w ethanol of mixed material and propyl alcohol, control temperature to 60 DEG C~78 DEG C 3~4h of holding, filtering； Freeze-drying obtains Chinese herbal medicine powder after filter vacuum concentration；

The mixed enzyme addition is the 5-10% of mixed material gross weight；

The parts by weight of the mixed enzyme form：Endo-beta-glucanase 10-20 parts, outer 1,4 beta-glucanase 10-20 parts, β- Glucuroide 10-15 parts, zytase 15-20 parts, pentosanase 15-20 parts, Pullulanase 20-30 parts, beta amylase 10- 15 parts, neutral proteinase 10-15 parts, acid protease 10-15 parts, superoxide dismutase 5-10 parts, glucose oxidase 5- 10 parts, acid phosphatase 5-10 parts；

The mass ratio of the ethanol and propyl alcohol is 1：1-1.5；

The one-level, two level, three-level seed culture medium weight composition are：

Dusty yeast 0.3-0.5%, glucose 1-1.5%, peptone 0.3-0.5%, beef extract 0.5-0.8%, phosphoric acid hydrogen Dipotassium 0.8-1.5%, Chinese herbal medicine powder 1.5-2%, trehalose 1-3%, calcium sulfate 0.1%, magnesium chloride 0.2%, sodium citrate 0.1-0.3%, insufficient section pure water are supplied, pH value 5-11,121-123 DEG C of sterilizing 30-40min；

The seed tank culture base weight forms：

Maltodextrin 5-15%, dusty yeast 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%, sodium 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；

The seeding tank zymotic fluid cell concentration is 7.0x 10⁸-8.0x 10⁸Individual/ml；

The fermentation medium forms：Maltodextrin 50-150g, corn flour 50-60g, beancake powder 15-25g, Chinese herbal medicine Pulvis 30-50g, trehalose 30-40g, birch xylan 5-15g, dusty yeast 4-8g, corn steep liquor 1-5g, ammonium sulfate 1-3g, phosphoric acid Hydrogen dipotassium 1-2g, potassium dihydrogen phosphate 1-2g, sodium citrate 1-5g, defoamer 0.1-1g, pure water l000mL, pH value 5-11,121 DEG C sterilizing 20min；

The concocting method of the fermentation medium is：Raw material is accurately weighed in proportion, by the pure water in raw material, corn In powder, beancake powder input material-compound tank, pH value 5-11 is adjusted, adds medium temperature amylase (3u/g corn flour) and alpha-amylase (30u/g corn flour), while then warming while stirring is to slowly warm up to 90 DEG C of insulation 15- to 70 DEG C of insulation 15-30min 30min is liquefied, and is eventually adding other raw materials, is stirred, and adjusts initial pH5-11, and 121-123 DEG C of sterilizing 30-40min is standby With.

The bacillus alcalophilus culture contains stronger bioactivity, passes through a side of fermenting in bio-bleaching process Face can produce high temperature resistant alkalescent xylanase, persistently provide for system, be on the other hand can be made by the dynamic response of fermentation it is thermophilic Hot bacillus is uniform, is sufficiently dispersed to paper cellulose, hemicellulose and lignin interstitial, abundant softening cellulose and drop Hemicellulose (xylan, glucan, mannose etc.) is solved, makes other compositions (such as laccase, medium enzyme preparation, the chela of complex enzyme Mixture, protective agent, activator, whiteness stabilizer etc.) uniformly, fully penetrate into paper pulp plant cell interstitial, give full play to group The effect of part, to improve paper pulp recovery rate and quality, shorten bleaching time, and go the removal of impurity and enhancing bio-bleaching effect；

Preferably, the preparation method of the bacillus alcalophilus culture is the above-mentioned zymotic fluid for preparing alkalescent xylanase Through being concentrated under reduced pressure, being freeze-dried, obtained from low-temperature grinding；

The bacillus alcalophilus culture viable bacteria content is 7x10¹⁰-9x10¹⁰CFU/g。

The 1- hydroxy benzo triazoles can activate the vigor of macromolecular laccase, form the compound amboceptor of strong oxidation, The enzymolysis effect of laccase is improved, degradation of the laccase to lignin is given full play to, is inherently eliminated lignin, prevents paper pulp Brightness reversion.

The EDTA can most of transition metal ions in chelated pulp (copper ion, ferrous ion, ferric ion, Divalent manganesetion, bivalent nickel ion etc.) and micro heavy, acted on excluding its suppression and elimination to biology enzyme enzyme activity, Make enzyme component fully, effectively play a role；

The whiteness stabilizer has the double action of bleaching and stable whiteness, can both effectively improve pulp brightness, again may be used Prevent yellowing reversion of pulp, and will not oxygen and transition metal ions in by paper pulp (copper ion, ferrous ion, ferric ion, Divalent manganesetion, bivalent nickel ion etc.) influence, the use of whiteness stabilizer can effectively make up residual lignin after bio-bleaching Caused yellowing reversion of pulp；

The whiteness stabilizer is mercaptan, thioether, trihydroxy methyl phosphoric acid (THP), tetra methylol phosphate (THPC), three hydroxyls One or more in double phosphorus derivants (BBHPE) of methyl acid phosphate are uniformly mixed with arbitrary proportion；

Preferably, the whiteness stabilizer is double phosphorus derivants (BBHPE) of trihydroxy methyl phosphoric acid；

The nonionic surfactant can promote complex enzyme component to permeate during enzyme bleaching, fully free wooden Element, hemicellulose, pectin, tannin, fat, etc. substrate, promote and extend each component enzyme-to-substrate action time, make in paper pulp Harmful components fully degraded, improve the smoothness of paper pulp fiber and bleaching the uniformity；

Preferably, the nonionic surfactant is environment-friendly type nonionic surfactant, readily biodegradable, without residual Stay, it is without any side effects to human body, biology and environment；

It is highly preferred that the quality component of the nonionic surfactant is：Alkyl polyglycoside (APG) 30-40 parts, alkyl Portugal Grape sugar acid amides (AGA) 15-20 parts, N- dodecyl ethylenediamine triacetic acid sodium (ED3A) 10-15 parts, isomery alcohol ether carboxylate AEC- 110710-15 parts, Lauric Acid Monoethanolamide Ether Carboxylate (LAEC) 8-10 parts, peregal c-1253-5 parts, JFC 3-5 parts；Above-mentioned raw materials It is solid, powdery commodity purchased in market.

The activator is uniformly mixed by the inorganic salts of following quality component：Sodium sulphate 15-18 parts, zinc chloride 6- 8 parts, magnesium chloride 5-8 parts, calcium chloride 3-5 parts.

The protective agent is made up of the raw material of following parts by weight:Trehalose 30-50 parts, GL-B 12-18 parts, NaCl10-15 parts, (NH₄)₂SO₄5-9 parts, cysteine 3-5 parts.

The antioxidant is any or several in grape pip procyanidin, Rosmarinus officinalis extract and apricot leaf extract Kind combination；

Preferably, the antioxidant be grape pip procyanidin, Rosmarinus officinalis extract and apricot leaf extract in mass ratio 3-5:1-3:1-2 is uniformly mixed；

The grape pip procyanidin, Rosmarinus officinalis extract and apricot leaf extract are commodity purchased in market

The preparation method of above-mentioned bleaching complex enzyme, comprises the following steps：

The protective agent, activator are distinguished into ultramicro grinding first, uniformly mixing, immediately sequentially adds nonionic table Face activating agent, bacillus alcalophilus culture uniformly mix 20-30min, then sequentially add alkalescent xylanase, laccase, Portugal Dextranase, seminase, lipase, tannase, pectase uniformly mix, finally sequentially add 1- hydroxy benzo triazoles, EDTA, whiteness stabilizer, antioxidant, packed after well mixed and produce bleaching complex enzyme.

The present invention is another object is that application of the bleaching complex enzyme in papermaking bio-bleaching technique.

Application method：

1. complex enzyme is added by the yellow slurry of drift before Huang starches chlorination or after chlorination or after alkalization；

2. complex enzyme action condition：Adapt to pH value 5-11, optimum pH 7-9,40-70 DEG C of adaptive temperature, optimal reaction temperature 45-55 DEG C of degree；

3. application method：1) by complex enzyme and 50 DEG C of water in mass ratio 1:10 uniformly mixing, adjustment pH value is 7-9, activation 20-30min, added by measuring pump by the yellow slurry of drift, it is well mixed with Huang slurry and carry out enzyme digestion reaction；

2) addition of complex enzyme accounts for the 0.05-0.3 ﹪ of the yellow slurry quality of over dry；

3) concentration of yellow slurry is 5-15 ﹪；

4) enzymolysis time 35-60min, after the completion of enzymolysis, routinely processing step is carried out its chemical bleaching, each step chemical Product dosage is the 40-50 ﹪ of conventional amount used.

Described yellow slurry manufactured yellow slurry after stock, slurrying, washing, screening for paper making raw material；

Described yellow slurry is wood pulp or one kind in non-wood pulp or combination；

Described non-wood pulp is prepared by raw material wheat straw or straw or reed or bagasse；

When the complex enzyme adds before chlorination, the addition of complex enzyme accounts for the 0.15-0.3 ﹪ of the yellow slurry quality of over dry；It is multiple When synthase adds after chlorination, the addition of complex enzyme accounts for the 0.1-0.15 ﹪ of the yellow slurry quality of over dry；Complex enzyme adds after alkalization Fashionable, the addition of complex enzyme accounts for the 0.05-0.1 ﹪ of the yellow slurry quality of over dry.

Beneficial effect：

Present invention bleaching complex enzyme is using high temperature resistant alkalescent xylanase as primary raw material, science compounding bacillus alcalophilus training Support thing, laccase and its medium, dextranase, EDTA, protective agent, activator, seminase, lipase, tannase, pectase, Whiteness stabilizer, nonionic surfactant, antioxidant etc., prepare a kind of enzyme system is complete, bleaching effect is good, it is easy to maintain, storage The good association with pulp bleaching enzyme of stability, while paper cellulose is fully retained, dissolved lignin and dropped to greatest extent Solution, content of lignin in paper pulp is fundamentally reduced, strengthen delignification effect, while appropriateness compounding whiteness stabilizer, can make up The deficiency of bio-bleaching, to reach permanent bleaching, yellowing reversion of pulp is prevented, improve into the yield and quality of paper, floated with Market Pulp White specific enzyme is compared, and present invention bleaching complex enzyme is remarkably improved paper output and matter to the yellow slurry of leaf wood, needlebush and reed Amount：Yield is respectively increased 8.94%, 8.80% and 10.05%；8.58%, 9.49% and is respectively increased in alpha-cellulose content 16.41%；Whiteness is respectively increased 10.94%, 11.97% and 11.10%；YI yellow index reduces by 39.84%, 37.31% and respectively 24.32%；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%；So as to promote bio-bleaching in the application process of paper industry, be finally reached reduce cost and The purpose of environmental protection, its advantage are the comprehensive embodiments of complex enzyme each component synergy, are in particular in：

1. the alkalescent xylanase high temperature resistant of the present invention, alkali resistance is strong, action condition is wide in range, without cellulase activity Property, it is more suitable for the bio-bleaching of paper pulp and chemistry helps drift.

2. the bacillus alcalophilus culture of the present invention contains stronger bioactivity, pass through hair in bio-bleaching process On the one hand ferment can produce high temperature resistant alkalescent xylanase, persistently provided for system, be on the other hand the dynamic response by fermentation Can make bacillus alcalophilus uniformly, be sufficiently dispersed to paper cellulose, hemicellulose and lignin interstitial, abundant softening fiber Element and degradation of hemicellulose (xylan, glucan, mannose etc.), make other compositions (such as laccase, medium enzyme system of complex enzyme Agent, chelating agent, protective agent, activator, whiteness stabilizer etc.) uniformly, fully penetrate into paper pulp plant cell interstitial, fully hair The effect of component is waved, to improve paper pulp recovery rate and quality, shortens bleaching time, and goes the removal of impurity and enhancing bio-bleaching effect Fruit.

3. the present invention compounds laccase and its medium 1- hydroxy benzo triazoles science, compound Jie of strong oxidation is formed Body, the enzymolysis effect of laccase is improved, fully, effectively realize degradation of the laccase to lignin, be inherently eliminated wooden Element, prevent yellowing reversion of pulp.

4. the present invention can most of transition metal ions (copper ion, ferrous ion, trivalent in chelated pulp using EDTA Iron ion, divalent manganesetion, bivalent nickel ion etc.) and micro heavy, to exclude its suppression to biology enzyme enzyme activity and disappear Except effect, make enzyme component fully, effectively play a role.

5. whiteness stabilizer of the present invention has the double action of bleaching and stable whiteness, pulp brightness can be both effectively improved, It can prevent yellowing reversion of pulp again, and will not oxygen and transition metal ions (copper ion, ferrous ion, ferric iron in by paper pulp Ion, divalent manganesetion, bivalent nickel ion etc.) influence, the use of whiteness stabilizer remains after can effectively making up bio-bleaching Yellowing reversion of pulp caused by lignin.

6. the main science of nonionic surfactant of the present invention has compounded the non-ionic surface active of environment-friendly type superior performance Agent, readily biodegradable, noresidue are without any side effects to human body, biology and environment；It is able to can promote during enzyme bleaching Complex enzyme component permeates, fully free lignin, hemicellulose, pectin, tannin, fat, etc. substrate, promote and extend each component The action time of enzyme-to-substrate, make the harmful components fully degraded in paper pulp, carry the smoothness of paper pulp fiber and the bleaching uniformity.

7. the present invention bleaching complex enzyme antioxidant science compounding, can effectively prevent complex enzyme enzyme molecular structure aoxidize and Loss of enzyme activity is caused, improves enzyme activity stability.Stored 12 months under the conditions of 0 DEG C and 40 DEG C, single enzyme enzyme activity in complex enzyme Loss is respectively 0.41% and 0.56%, and 18.0% and 65% are reduced respectively than comparative example, is effectively prevented in packaging, storage, fortune It is defeated, use etc. link because environment changes, operating method is improper and caused by enzyme inactivation, enzyme caused by especially can preventing high temperature loses It is living.

8. protectant science compounding, effectively slow down the moisture regain of complex enzyme formulation in present invention bleaching complex enzyme；Simultaneously Resistance to jelly, the heat resistance of complex enzyme can be strengthened, keep identical enzyme activity, its heat resisting temperature can improve 20-30 DEG C, freeze-resistant temperature Degree can reduce 10-15 degrees Celsius, effectively prevent the loss of complex enzyme enzyme activity during transport, preservation and use, extend The shelf-life of complex enzyme, reach same enzyme activity, can extend 2-3 than the like product shelf-life.

9. present invention bleaching complex enzyme adds inorganic salts as activator, the optimum condition of enzyme catalysis is created, is filled The vigor of phytoenzyme and microbial enzyme has been waved in distribution, improves complex enzyme action potency.

Embodiment

The present invention is described below by specific embodiment.Unless stated otherwise, technological means used in the present invention It is method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, it is not intended to limit the present invention Scope, the spirit and scope of the invention are limited only by the claims that follow.To those skilled in the art, without departing substantially from this The various changes carried out on the premise of invention spirit and scope to the material component in these embodiments and dosage or change Belong to protection scope of the present invention.

It is prepared by the raw material of embodiment 1

1. the preparation of alkalescent xylanase：Comprise the following steps

(1) actication of culture

Intact bacillus alcalophilus CCTCC M 2013537 slant strains are inoculated in slant medium, 45 DEG C culture 36h carry out actication of culture, so activation 3 times；

The slant medium forms：Beef extract 10g, sodium chloride 12g, peptone 20g, glucose 5g, agar 20g, Chinese herbal medicine powder 10g, distilled water l000mL, 10,121 DEG C of sterilizing 20min of pH value；

The preparation method of the Chinese herbal medicine powder is as follows：

In terms of parts by weight, 25 parts of the Radix Astragali, 15 parts of Radix Codonopsis, 12 parts of radix bupleuri, 12 parts of radix scutellariae, 10 parts of cordate houttuynia, Radix Angelicae Sinensis 9 are weighed Part；Said herbal medicine is crushed to particle diameter as less than 2 millimeters respectively, is then uniformly mixed in container and adds 5 times of weight Water, 80 DEG C of holding 3h of control temperature, is then cooled to 55 DEG C, the mixed enzyme for adding mixed material gross weight 8% is digested, and is used Newborn acid for adjusting pH value is 6, digests 3h, finally adds the mixture of 2 times of w ethanols of mixed material and propyl alcohol, ethanol and propyl alcohol Mass ratio is 1：1.5, control temperature to 70 DEG C of holding 4h, filtering；Freeze-drying obtains Chinese herbal medicine powder after filter vacuum concentration Agent；

The parts by weight of the mixed enzyme form：15 parts of endo-beta-glucanase, outer 15 parts of 1,4 beta-glucanase, β-glucose 12 parts of glycosides enzyme, 18 parts of zytase, 18 parts of pentosanase, 25 parts of Pullulanase, 12 parts of beta amylase, 12 parts of neutral proteinase, 12 parts of acid protease, 8 parts of superoxide dismutase, 8 parts of glucose oxidase, 8 parts of acid phosphatase；

(2) liquid seeds expand culture

1. first order seed culture：The ring of slant strains 2 after step (1) is activated is accessed in 500 milliliters of shaking flasks, culture medium dress 100 milliliters of amount, 180 revs/min of rotary shaker, 43 DEG C of cultivation temperature, incubation time 15h；

2. secondary seed culture：First order seed is accessed in 500 milliliters of secondary seed shaking flasks according to 10% inoculum concentration, training The condition of supporting is identical with first order seed；

3. three-level seed culture：Secondary seed is accessed in 5000 milliliters of three-level seed flasks with 10% inoculum concentration, culture 1000 milliliters of base loading amount, 100 revs/min of rotary shaker, 43 DEG C of cultivation temperature, incubation time 15h；

4. first class seed pot culture：Three-level seed is accessed into first class seed pot of the total measurement (volume) as 150L using 10% inoculum concentration, Fermentation medium loading amount 100L, 45 DEG C, mixing speed 400rpm of cultivation temperature, ventilation (V/V) 1:2, tank pressure 0.05Mpa, training Support time 20h；

Dusty yeast 0.5%, glucose 1.5%, peptone 0.5%, beef extract 0.8%, dipotassium hydrogen phosphate 1.5%, medium-height grass Medicine powder 2%, trehalose 3%, calcium sulfate 0.1%, magnesium chloride 0.2%, sodium citrate 0.3%, insufficient section pure water are supplied, 10,123 DEG C of sterilizing 40min of pH value；

The seed tank culture base weight forms：

Maltodextrin 15%, dusty yeast 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%, sodium citrate 0.5%, birch xylan 0.8%, insufficient section is pure Water is supplied, 10,123 DEG C of sterilizing 40min of pH value；

The seeding tank zymotic fluid cell concentration is 8.0x 10⁸Individual/ml；

(3) ferment tank

First class seed pot zymotic fluid in step (2) is accessed into fermentation tank, 45 DEG C of cultivation temperature, stirring speed with 6% inoculum concentration Spend 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 is trained Support 12h；Continue with 2 DEG C/h rate of temperature fall slow cooling to 10 DEG C, now, by first class seed pot zymotic fluid in step (2) with 4% Inoculum concentration adds access fermentation tank, incubated 12h；Finally 20 DEG C are to slowly warm up to 2 DEG C/h heating rates, it is incubated 12h；Continue to be to slowly warm up to 45 DEG C with 2 DEG C/h heating rates, incubated 30h；

Dissolved oxygen controls：By adjusting speed of agitator and ventilation, dissolved oxygen 30% is controlled；

PH is controlled：By mending ammoniacal liquor or phosphoric acid,diluted, pH value in fermentation process is controlled to be maintained at 10；

Control of additive raw material：When the content of reducing sugar in zymotic fluid is down to 3mg/ml-8mg/ml, start to add feed-batch culture Base, feed supplement amount is to maintain zymotic fluid content of reducing sugar as 2mg/ml-5mg/ml；

Put tank standard：80% thalline aging self-dissolving, enzyme activity increases slowly；

The fermentation medium forms：Maltodextrin 150g, corn flour 60g, beancake powder 25g, Chinese herbal medicine powder 50g, Trehalose 40g, dusty yeast 8g, corn steep liquor 5g, ammonium sulfate 3g, dipotassium hydrogen phosphate 2g, potassium dihydrogen phosphate 2g, sodium citrate 5g, birch Wooden xylan 15, defoamer 1g, pure water l000mL, 10,121 DEG C of sterilizing 20min of pH value；

The supplemented medium weight forms：Maltodextrin 20-30%, corn flour 10-20%, bean powder 15-25%, in Herbal medicine pulvis 5-10%, insufficient section pure water are supplied, pH value 5-11,121-123 DEG C of sterilizing 30-40min；

The concocting method of the fermentation medium is：

Raw material is accurately weighed in proportion, by the pure water in raw material, corn flour, beancake powder input material-compound tank, adjusts PH Value 10, medium temperature amylase (3u/g corn flour) and alpha-amylase (30u/g corn flour) are added, while warming while stirring is to 70 DEG C insulation 30min, be then to slowly warm up to 90 DEG C insulation 30min liquefied, be eventually adding other raw materials, stir, adjust 10,123 DEG C of sterilizing 40min of initial pH value are standby.

(4) zymotic fluid is filtered, concentration, allotment, refined filtration, dry high temperature resistant alkalescent xylanase；

The allocation process does not add any preservative, adds the Chinese herbal medicine powder of concentration enzyme liquid gross weight 5%.

2. the preparation of bacillus alcalophilus culture：During the preparation method of the bacillus alcalophilus culture is above-mentioned 1 Prepare the zymotic fluid of alkalescent xylanase through being concentrated under reduced pressure, being freeze-dried, low-temperature grinding and be made, its viable bacteria content is 9x10¹⁰CFU/g。

Measure (3,5-dinitrosalicylic Acid Colorimetry, the abbreviation DNS of the high temperature resistant alkalescence xylanase activity power of embodiment 2 Method)

1. reagent and solution

Water used in 1.1 is to meet tertiary effluent as defined in GB/6682-1992 in addition to special requirement, and chemicals is except spy Yao Qiu not be outer, it is that analysis is pure.

1.2DNS reagent

3 are weighed, 5-dinitrosalicylic acid 10g is added in 500ml water, adds sodium hydroxide 16g, stirring and dissolving several times (45 DEG C of temperature ＜), then add sodium potassium tartrate tetrahydrate 300g several times, stir to complete molten, 1000ml is settled to water after cooling.Room The lower brown bottle storage dark place of temperature is placed, and is used and (is used after being filtered if any precipitation) after one week.

1.30.1mol/L, pH5.0 acetic acid-sodium-acetate buffer

Glacial acetic acid 1.8ml is drawn, adds suitable quantity of water, adds sodium acetate 5.78g, is dissolved, is settled to 1000ml, adjusts PH to 5.00 Used after ± 0.01.2 months are deposited at room temperature effectively.

1.40.1% it is 1mg/ml xylose standard solution

Anhydrous xylose dries to constant weight in 80 DEG C, weighs 0.1000g in beaker, adds suitable quantity of water to dissolve, is transferred in volumetric flask And it is settled to 100ml.

1.51.0% xylan solution

Xylan 1.00g is weighed in beaker, is soaked with 2ml 0.5mol/L sodium hydroxides into pasty state, adds 40ml to buffer Liquid, dissolved by heating in 60~70 DEG C of water-baths, adjust pH5.0 with 0.5mol/L acetic acid (about 2ml) after cooling, be transferred in volumetric flask, Buffer solution (4.3) is added to be settled to 100ml.Precipitated if standing time has been grown in refrigerator, should be hot in hot bath before use It is molten.

The drafting of 1.6 xylose standard curves

1mg/ml xylose solutions 0,0.2,0.4,0.6,0.8,1.0ml are drawn in test tube, moisturizing to 2ml, add DNS reagents 3ml, mix after boiling 10min in boiling water, 15ml is settled to after cooling, absorbance is surveyed under spectrophotometer 540nm wavelength (A).Using absorbance as ordinate, xylose amount is abscissa, draws standard curve, repeats the average least square of experiment three times Method is fitted a linear equation y=ax ﹢ b, obtains the relation of absorbance and xylose amount.

2. the processing of enzyme sample to be measured

Directly draw enzyme liquid and do appropriate dilution, make measure absorbance value in the range of 0.25-0.4.

3. enzyme activity determination

Take 15ml tool plug scale test tubes to be operated by following reaction sequence, during the course of the reaction, (inhaled from substrate is added Shaken up before taking) (1.5) beginning, the time interval that reagent is added into every test tube will definitely unanimously, 50 DEG C of hydrolysis 10min.

Reactions steps and reagent, solution usage are shown in Table 1：

The reactions steps of table 1 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. mixing	4. add DNS reagents 3ml
		5.50 DEG C of hydrolysis 10min	5. mixing
6. add DNS reagents 3ml	6. enzyme-added liquid 0.2ml
		7. mixing	7. mixing
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. determined under blank zeroing 540nm	10. determined under blank zeroing 540nm

4. enzyme activity unit calculates

In enzyme activity=(S × D × 1000)/μ g/ml (g) .min formulas of (0.2 × 10) × 1.7：

S-measure light absorbs corresponding xylose amount, mg numbers on standard curve；

D-enzyme liquid or enzyme powder extension rate；

Conversion coefficient between 1000-mg and μ g；

0.2-measure takes enzyme liquid amount, ml numbers；

10-reaction time, min numbers；

1.7-method conversion coefficient.

5. enzyme activity unit defines and conversion

5.1 this method enzyme activity define:Under this condition determination, hydrolyzed xylan per minute produces 1 μ g reduced sugars (with xylose Meter) needed for enzyme amount be defined as an enzyme activity unit (u).

The definition of international enzyme-activity unit:Under condition determination, hydrolyzed xylan per minute produces 1 μm of ol reduced sugar (with wood Sugar meter) needed for enzyme amount be defined as an enzyme activity unit (IU).

5.2u and international unit IU conversion relation：

1 u=1 ÷ 150IU (μm ol sugar/ml (g) .min)

150:The molecular weight of xylose

The thermal stability analysis of the alkalescent xylanase of embodiment 3

The heat endurance of enzyme is analyzed, crude enzyme liquid prepared by the alkalescent xylanase of embodiment 1 is respectively placed in 40 DEG C, 45 DEG C, 50 DEG C, 55 DEG C, 60 DEG C, 65 DEG C, at 70 DEG C, enzyme activity was measured by sampling every 10 minutes.40 DEG C of crude enzyme liquid, 45 DEG C, 50 DEG C, at 55 DEG C, enzyme activity does not decline within 60 minutes.60 DEG C and at 65 DEG C, drop to original enzyme activity within 30 minutes 95%, drop within 60 minutes 85%.At 70 DEG C, drop to original enzyme activity within 30 minutes 85%, drop to 80% within 60 minutes.At 75 DEG C, original enzyme activity is dropped within 30 minutes 80%, drop to 70% within 60 minutes, reach same enzyme activity compared with prior art, tolerable temperature is higher.

The alkalescent xylanase pH stability analyses of embodiment 4

The pH stability of enzyme is analyzed, crude enzyme liquid prepared by the alkalescent xylanase of embodiment 1 is respectively placed in pH Under value 4.0,5.0,6.0,7.0,8.0,9.0,10.0,11.0, enzyme activity was measured by sampling every 10 minutes.Crude enzyme liquid pH value 6.0, 7.0th, under 8.0,9.0, enzyme activity does not decline within 60 minutes.Under pH value 5.0 and 10.0, drop to original enzyme activity within 30 minutes 95%, 60 Minute drops to 85%.Under pH value 4.0 and 11.0, drop to original enzyme activity within 30 minutes 80%, drop to 70% within 60 minutes.Similarity condition It is substantially more wide in range than prior art bacterial strain to be issued to the resistance to pH value of same enzyme activity.

Embodiment 5

15 parts of alkalescent xylanase, 9 parts of bacillus alcalophilus culture, 9 parts of laccase, 6 parts of dextranase, 1- hydroxy benzos 5 parts of triazole, 5 parts of EDTA, 6 parts of protective agent, 6 parts of activator, 5 parts of seminase, 4 parts of lipase, 4 parts of tannase, pectase 4 parts, 3 parts of double phosphorus derivants (BBHPE) of trihydroxy methyl phosphoric acid, 3 parts of nonionic surfactant, 2 parts of antioxidant；

Alkalescent xylanase and the bacillus alcalophilus culture is prepared by embodiment 1；

The quality component of the nonionic surfactant is：35 parts of alkyl polyglycoside (APG), methyl glucamine (AGA) 18 parts, 12 parts of N- dodecyl ethylenediamine triacetic acid sodium (ED3A), isomery alcohol ether carboxylate AEC-110712 parts, bay 9 parts of Amide Ether Carboxylates (LAEC), peregal c-1254 parts, 4 parts of JFC；

The activator is uniformly mixed by the inorganic salts of following quality component：17 parts of sodium sulphate, 7 parts of zinc chloride, 6 parts of magnesium chloride, 4 parts of calcium chloride；

The protective agent is made up of the raw material of following parts by weight:40 parts of trehalose, 15 parts of GL-B, 12 parts of NaCl, (NH₄)₂SO₄7 parts, 4 parts of cysteine；

The antioxidant is grape pip procyanidin, Rosmarinus officinalis extract and apricot leaf extract in mass ratio 4:2:1.5 Uniformly mixing；

The preparation method of above-mentioned bleaching complex enzyme, comprises the following steps：The protective agent, activator are distinguished into ultra micro first Crush, uniformly mixing, immediately sequentially adds nonionic surfactant, bacillus alcalophilus culture uniformly mixes 25min, it is equal then to sequentially add alkalescent xylanase, laccase, dextranase, seminase, lipase, tannase, pectase Even mixing, finally sequentially add double phosphorus derivants (BBHPE), the antioxygen of 1- hydroxy benzo triazoles, EDTA, trihydroxy methyl phosphoric acid Agent, packed after well mixed and produce bleaching complex enzyme.

Embodiment 6

10 parts of alkalescent xylanase, 8 parts of bacillus alcalophilus culture, 8 parts of laccase, 5 parts of dextranase, 1- hydroxy benzos 3 parts of triazole, 3 parts of EDTA, 4 parts of protective agent, 4 parts of activator, 4 parts of seminase, 3 parts of lipase, 3 parts of tannase, pectase 3 parts, 2 parts of trihydroxy methyl phosphoric acid (THP), 2 parts of nonionic surfactant, 1 part of antioxidant；

The quality component of the nonionic surfactant is：20 parts of alkyl polyglycoside (APG), methyl glucamine (AGA) 30 parts, 15 parts of N- dodecyl ethylenediamine triacetic acid sodium (ED3A), isomery alcohol ether carboxylate AEC-110710 parts, bay 8 parts of Amide Ether Carboxylates (LAEC), peregal c-1253 parts, 3 parts of JFC；

The activator is uniformly mixed by the inorganic salts of following quality component：15 parts of sodium sulphate, 6 parts of zinc chloride, 5 parts of magnesium chloride, 3 parts of calcium chloride；

The protective agent is made up of the raw material of following parts by weight:30 parts of trehalose, 12 parts of GL-B, 10 parts of NaCl, (NH₄)₂SO₄5 parts, 3 parts of cysteine；

The antioxidant is grape pip procyanidin, Rosmarinus officinalis extract and apricot leaf extract in mass ratio 3:1:1 is equal Even mixing；

The preparation method of above-mentioned bleaching complex enzyme is the same as embodiment 2.

Embodiment 7

20 parts of alkalescent xylanase, 10 parts of bacillus alcalophilus culture, 10 parts of laccase, 8 parts of dextranase, 1- hydroxy benzenes And 8 parts of triazole, 8 parts of EDTA, 7 parts of protective agent, 7 parts of activator, 6 parts of seminase, 5 parts of lipase, 5 parts of tannase, pectin 5 parts of enzyme, 5 parts of whiteness stabilizer, 4 parts of nonionic surfactant, 3 parts of antioxidant；

The whiteness stabilizer is pressed for double phosphorus derivants (BBHPE) of trihydroxy methyl phosphoric acid (THP) and trihydroxy methyl phosphoric acid Mass ratio 1:2 uniformly mixing；

The quality component of the nonionic surfactant is：40 parts of alkyl polyglycoside (APG), methyl glucamine (AGA) 30 parts, 20 parts of N- dodecyl ethylenediamine triacetic acid sodium (ED3A), isomery alcohol ether carboxylate AEC-110715 parts, bay 10 parts of Amide Ether Carboxylates (LAEC), peregal c-1255 parts, 5 parts of JFC；

The activator is uniformly mixed by the inorganic salts of following quality component：18 parts of sodium sulphate, 8 parts of zinc chloride, 8 parts of magnesium chloride, 5 parts of calcium chloride；

The protective agent is made up of the raw material of following parts by weight:50 parts of trehalose, 18 parts of GL-B, 15 parts of NaCl, (NH₄)₂SO₄9 parts, 5 parts of cysteine；

The antioxidant is grape pip procyanidin, Rosmarinus officinalis extract and apricot leaf extract in mass ratio 5:3:2 is equal Even mixing；

Embodiment 8

20 parts of alkalescent xylanase, 10 parts of bacillus alcalophilus culture, 8 parts of laccase, 5 parts of dextranase, 1- hydroxy benzenes And 3 parts of triazole, 8 parts of EDTA, 7 parts of protective agent, 7 parts of activator, 4 parts of seminase, 5 parts of lipase, 5 parts of tannase, pectin 5 parts of enzyme, 5 parts of mercaptan, 2 parts of nonionic surfactant, 3 parts of grape pip procyanidin；

The quality component of the nonionic surfactant is：30 parts of alkyl polyglycoside (APG), methyl glucamine (AGA) 20 parts, 15 parts of N- dodecyl ethylenediamine triacetic acid sodium (ED3A), isomery alcohol ether carboxylate AEC-110710 parts, bay 8 parts of Amide Ether Carboxylates (LAEC), peregal c-1255 parts, 5 parts of JFC；

The activator is uniformly mixed by the inorganic salts of following quality component：15 parts of sodium sulphate, 8 parts of zinc chloride, 8 parts of magnesium chloride, 3 parts of calcium chloride；

The protective agent is made up of the raw material of following parts by weight:40 parts of trehalose, 12 parts of GL-B, 10 parts of NaCl, (NH₄)₂SO₄9 parts, 3 parts of cysteine；

Influence of the antioxidant of embodiment 9 to bleaching complex enzyme enzyme activity

The bleaching complex enzyme prepared using the embodiment of the present invention 5, other raw materials, material component, preparation method are identical, Unique difference is that raw material components are free of antioxidant, is contrasted example, stores 12 months, adopt under the conditions of 0 DEG C and 40 DEG C With《GB/T 23535-2009 lipase preparations》The enzyme activity of middle detection method measure lipase, calculates enzyme activity loss late, enzyme activity Loss late refers to that the difference of actually detected enzyme activity and product mark enzyme activity accounts for the percentage for marking enzyme activity, as a result such as table 2

Lipase activity power loss late in the Storage period complex enzyme of table 2

Result above shows, is stored 12 months under the conditions of 0 DEG C and 40 DEG C, and the lipase in embodiment 2 is than in comparative example Lipase activity loss respectively reduce by 18% and 65%, illustrate antioxidant science compounding, significantly improve in complex enzyme The vigor of each component enzyme, enzyme activity loss are greatly reduced, and especially effect is more notable under the high temperature conditions.

The present invention of embodiment 10 bleaching complex enzyme influences on the quality of the yellow slurry of different material

First, test site：Avenge beautiful paper industry in Hunan.

2nd, test period：- 2014 years on the 12nd January in 2014, on April 22, lasted 100 days.

3rd, plan design：1. experiment designs for single-factor, test group and control group, control group and test group are set Raw material, technique, equipment, operating personnel, environment and way to manage all same, difference be：In yellow slurry of the test group before chlorination Add bleaching complex enzyme prepared by the embodiment of the present invention 5, control group addition Market Pulp bleaching specific enzyme.2. test group and control Group is by the 0.15-0.3% addition bleaching complex enzymes of yellow slurry over dry weight.3. the yellow slurry test group of same raw material and control group are everywhere 10 batches are managed, calculate average value.Routinely 4. technique is carried out postorder chemical bleaching.5. the yellow slurry of pair different material is through bleaching system The paper pulp correlated quality index obtained carries out testing result such as table 3

Table 3 bleaches influence of the complex enzyme to the yellow slurry quality of different material

Result above shows, compared with Market Pulp bleaches specific enzyme, present invention bleaching complex enzyme is to leaf wood, needlebush Paper output and quality are remarkably improved with the yellow slurry of reed：Yield is respectively increased 8.94%, 8.80% and 10.05%；Alpha fibre Cellulose content is respectively increased 8.58%, 9.49% and 16.41%；Whiteness is respectively increased 10.94%, 11.97% and 11.10%；Return Huang value reduces by 39.84%, 37.31% and 24.32% respectively；Viscosity reduces by 1.20%, 1.18% and 1.43% respectively；Ash content point Not Jiang Di by 25%, 26% and 28%, improving alpha-cellulose content and while yield, especially substantially increasing pulp brightness, The YI yellow index of paper pulp is reduced, effectively prevents paper from yellowing.

Claims

1. one kind bleaching complex enzyme, is prepared by the raw material of following parts by weight：Alkalescent xylanase 10-15 parts, thermophilic gemma bar Bacterium culture 8-9 parts, laccase 8-10 parts, dextranase 5-8 parts, 1- hydroxy benzo triazole 3-8 parts, EDTA 3-8 parts, protection Agent 4-7 parts, activator 4-7 parts, seminase 4-6 parts, lipase 3-5 parts, tannase 3-5 parts, pectase 3-5 parts, whiteness are steady Determine agent 2-5 parts, nonionic surfactant 2-4 parts, antioxidant 1-3 parts；

Alkalescent xylanase and the bacillus alcalophilus culture is by the thermophilic gemma bar for producing high temperature resistant alkalescent xylanase Bacterium CCTCC M 2013537 are starting strain, and are prepared by Optimal Medium composition and zymotechnique；

The activator is uniformly mixed by the inorganic salts of following quality component：Sodium sulphate 15-18 parts, zinc chloride 6-8 parts, Magnesium chloride 5-8 parts, calcium chloride 3-5 parts；

The protective agent is made up of the raw material of following parts by weight：Trehalose 30-50 parts, GL-B 12-18 parts, NaCl 10- 15 parts, (NH₄)₂SO₄5-9 parts, cysteine 3-5 parts；

The whiteness stabilizer is mercaptan, thioether, trihydroxy methyl phosphoric acid, tetra methylol phosphate, double phosphorus of trihydroxy methyl phosphoric acid One or more in derivative are uniformly mixed with arbitrary proportion；

The nonionic surfactant is environment-friendly type nonionic surfactant；

The quality component of the nonionic surfactant is：Alkyl polyglycoside 30-40 parts, methyl glucamine 15-20 parts, N- Dodecyl ethylenediamine triacetic acid sodium 10-15 parts, isomery alcohol ether carboxylate AEC-1107 10-15 parts, Lauric Acid Monoethanolamide Ether Carboxylate 8-10 parts, peregal c-125 3-5 parts, JFC 3-5 parts.

A kind of 2. bleaching complex enzyme as claimed in claim 1, it is characterised in that the preparation of the high temperature resistant alkalescent xylanase Method is as follows：By intact bacillus alcalophilus CCTCC M 2013537 slant strains it is activated and one, two, three Seed culture and first class seed pot culture obtain seed liquor, access fermentation tank with 6% inoculum concentration, 37-45 DEG C of cultivation temperature, stir Mix speed 200-700rpm, ventilation (V/V) 1:1-3, incubation time 25-30h；Then slowly dropped with 1-2 DEG C/h rate of temperature fall Temperature is to 15-20 DEG C, incubated 10-12h；Continue with 1-2 DEG C/h rate of temperature fall slow cooling to 8-10 DEG C, now, by one-level Seeding tank zymotic fluid adds access fermentation tank, incubated 7-12h with 4% inoculum concentration；It is finally slow with 1-2 DEG C/h heating rates 15-20 DEG C is warming up to, incubated 10-12h；Continue to be to slowly warm up to 37-45 DEG C with 1-2 DEG C/h heating rates, it is incubated 20-30h；Zymotic fluid is filtered, concentration, allotment, refined filtration, dry high temperature resistant alkalescent xylanase.

3. a kind of bleaching complex enzyme as claimed in claim 1, it is characterised in that the bacillus alcalophilus culture viable bacteria contains Measure as 7x10¹⁰-9x10¹⁰CFU/g。

4. a kind of bleaching complex enzyme as claimed in claim 1, it is characterised in that the antioxidant is grape pip original cyanine Any or several combination in element, Rosmarinus officinalis extract and apricot leaf extract.

5. the preparation method of a kind of bleaching complex enzyme as described in claim 1-4 is any, it is characterised in that including following step Suddenly：The protective agent, activator are distinguished into ultramicro grinding first, uniformly mixing, immediately sequentially add non-ionic surface work Property agent, bacillus alcalophilus culture uniformly mix 20-30min, then sequentially add alkalescent xylanase, laccase, glucan Enzyme, seminase, lipase, tannase, pectase uniformly mix, and finally sequentially add 1- hydroxy benzo triazoles, EDTA, white Stabilizer, antioxidant are spent, is packed after well mixed and produces bleaching complex enzyme.

6. a kind of preparation method for bleaching complex enzyme as claimed in claim 5, it is characterised in that the whiteness stabilizer is three Double phosphorus derivants of methylol phosphoric acid.

7. a kind of preparation method for bleaching complex enzyme as claimed in claim 5, it is characterised in that the antioxidant is grape Seed OPC, Rosmarinus officinalis extract and apricot leaf extract 3-5 in mass ratio:1-3:1-2 is uniformly mixed.