Embodiment
Below in conjunction with embodiment, the invention will be further described.
Embodiment 1:
A kind of production method of chemical-mechanical pulping complex enzyme liquid, 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; Described alkaline pectase be CCTCCNO:M2010004 bacterial classification through spawn culture, shake flask fermentation, seeding tank is prepared after fermentor tank, filtration, degerming, rotproofing; Described zytase be CGMCCNO:5466 bacterial classification through spawn culture, shake flask fermentation, seeding tank is prepared after fermentor tank, filtration, degerming, rotproofing; Described mannase be CGMCCNO:6226 bacterial classification through spawn culture, shake flask fermentation, seeding tank is prepared after fermentor tank, filtration, degerming, rotproofing; Described lignoenzyme be CGMCCNO:6225 bacterial classification through spawn culture, shake flask fermentation, seeding tank is prepared after fermentor tank, filtration, degerming, rotproofing; The vigor of described complex enzyme liquid neutral and alkali polygalacturonase is 800u/ml ~ 1500u/ml, and the vigor of zytase is 1000u/ml ~ 1800u/ml, and the vigor of mannase is 300u/ml ~ 1000u/ml, and the vigor of lignoenzyme is 100u/ml ~ 180u/ml.
CCTCCNO:M2010004 bacterial classification of the present invention is by naturally gathering bacterium sample in Ningxia, China soil, two strain starting strains are obtained through shaking flask screening from nearly hundred strain bacterial strains, and under cell levels, adopt nitrosoguanidine and ultraviolet radiation mutagenesis to obtain nearly ten thousand strain bacterial strains, the bacterial strain that a strain laboratory is numbered MAPLE61 is obtained through shaking flask screening, i.e. heat-resisting Bacillus subtillis (Heat ~ resistantBacillussubtilis), its biological characteristics is: thalline is shaft-like, thalline two ends are more smooth, individual cells (0.7 ~ 0.75) * (2.5 ~ 2.9) micron, without pod membrane, peritrichous, bacterium colony oyster white, Gram-positive, aerophil, liquid culture thalline in vegetative period majority in chain row example and with three disjunctors or tetrad in the majority, form brood cell, gemma form ellipse is to shaft-like, and (0.6 ~ 0.8) * (1.0 ~ 1.4) micron, is positioned at thalline central authorities or slightly inclined.This bacterial classification submits China typical culture collection center preservation (Wuhan City, Hubei Province Wuhan University) on January 11st, 2010, and preserving number is: CCTCCNO:M2010004, and have submitted preservation survival proof on January 15th, 2010.
The slant culture based formulas of this bacterial classification is: in 1000mL distilled water, adds extractum carnis 5g ~ 10g, yeast extract paste 5g ~ 10g, peptone 10g ~ 15g, glucose 5g ~ 10g, sodium-chlor 5g ~ 10g, sodium carbonate 0.3 ~ 0.5g, agar powder 20g, adjust pH 6.9 ~ 7.1,0.1MPa sterilizing 20 minutes ~ 30 minutes, make test tube slant, inoculate this bacterial classification, 33 DEG C ~ 35 DEG C, cultivate 18 hours ~ 24 hours.
After this slant strains is qualified by shake flask fermentation inspection bacterial classification enzymatic productivity, can spreads cultivation as seed and use or enzymatic production use.
The culture condition of shake flask fermentation is: in mass, culture medium prescription (﹪) wheat bran 6 ~ 8, Semen Maydis powder 0.5 ~ 1, corn steep liquor (nitrogen content 40 ﹪) 1 ~ 2, sodium-chlor 0.5 ~ 0.8, sodium carbonate 0.2 ~ 0.3, cellulosic powder 1 ~ 2, water 85.0 ~ 91.0, wherein said each component sum is 100 ﹪, before sterilizing, adjust pH is 8.0 ~ 8.5,0.1MPa sterilizing 25 minutes ~ 35 minutes, culture condition: 33 DEG C ~ 35 DEG C, rotating speed 260r/ minute ~ 280r/ minute, cultivate 18 hours ~ 22 hours.
Enzyme liquid is prepared as by eggplant bottle inclined-plane to seeding tank to fermentor tank, second order fermentation, preparation standard enzyme liquid.In mass, its culture medium prescription is (﹪) wheat bran 6 ~ 8, Semen Maydis powder 0.5 ~ 1, corn steep liquor (nitrogen content 40 ﹪) 1 ~ 2, sodium-chlor 0.5 ~ 0.8, sodium carbonate 0.2 ~ 0.3, cellulosic powder 1 ~ 2, water 85.0 ~ 91.0, wherein said each component sum is 100 ﹪, before sterilizing, adjust pH is 8.0 ~ 8.5,0.1MPa sterilizing 25 minutes ~ 35 minutes, tinning cumulative volume is no more than 70%; The culture condition of seeding tank: temperature (DEG C): 36 DEG C ~ 38 DEG C, mixing speed (r/ minute): 220, during air flow (v/v) 0 ~ 5 time, 1:0.40 ~ 0.46,5 time after 1:0.50 ~ 0.56; The culture condition of fermentor tank: temperature (DEG C): 0 hour ~ 4 hours, 36 ~ 38,4 hours later 33 ~ 35, mixing speed (r/min): 0 hour ~ 4 hours, 180,4 hours ~ 6 hours, 200,6 hours later 220, air flow (v/v): 0 hour ~ 4 hours, 1:0.25 ~ 0.27,4 hours ~ 6 hours, 1:0.33 ~ 0.36,1:0.5 ~ 0.55 after 6 hours.
After the preparation of enzyme liquid, preparation standard alkaline pectase liquid, fermentation liquid is degerming through Plate Filtration, diatomite, and obtain under three months conditions are deposited in 15 DEG C of sealings through rotproofing, enzyme activity conservation rate is the standard alkaline type pectase liquid of 95 more than ﹪.
The enzyme activity determination of alkaline pectase of the present invention is that spectrophotometric colo method records with DNS colour developing.Its condition determination is: with the pectin of 1 ﹪ for substrate was pH9.6,60 DEG C of water-baths 10 minutes, add the colour developing of DNS boiling water bath, 550nm colorimetric.Enzyme activity unit is defined as: under condition determination, and the enzyme amount that hydrolysis of pectin per hour produces needed for 1mg reducing sugar (in galacturonic acid) is defined as an enzyme activity unit.
Above content the patent No. be 201010238630.4, denomination of invention is describe in detail in the patent application of a kind of alkaline pectinase production method and the application in paper-making pulping, its preservation survival proves also to submit to Patent Office of the People's Republic of China with application.
CGMCCNO:5466 bacterial classification of the present invention is in Ningxia, China collection, in strain improvement screening process, have employed the method for test tube liquid culture radical reaction DNS colour developing initial characterization screening and shaking flask product enzyme quantitative screening, filter out the superior strain that a strain laboratory is numbered KERR89, according to major physiological biochemical character and 16SrRNA gene order, the experimental datas such as gyrB gene order are comprehensively analyzed and are accredited as husky good fortune genus bacillus (Bacillussafensis), its biological characteristics is: G+ bacteria, produce gemma, thalline is rod-short, unicellular diameter is less than 1um, breed in binary fission mode, on substratum of the present invention, bacterium colony is rounded, white, smooth surface, homogeneous, and neat in edge, slightly gloss are sticky.This bacterial classification submits China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation (No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City) on November 17th, 2011, preserving number: CGMCCNO:5466, and proves in have submitted preservation survival on the same day.
The slant culture based formulas of this bacterial classification is: peptone 0.5 ﹪ ~ 1 ﹪, extractum carnis 0.3 ﹪ ~ 0.5 ﹪, yeast extract paste 0.3 ﹪ ~ 0.5 ﹪, glucose 0.3 ﹪ ~ 0.5 ﹪, sodium-chlor 0.3 ﹪ ~ 0.5 ﹪, agar 1.8 ﹪ ~ 2.2 ﹪, water 94.8 ﹪ ~ 96.5 ﹪, wherein said each component sum is 100 ﹪, adjust pH 6.9 ~ 7.1,0.1MPa sterilizing 20 minutes ~ 30 minutes, make test tube slant, inoculate this bacterial classification, 32 DEG C ~ 35 DEG C, cultivate 24 hours ~ 26 hours.
After this slant strains is qualified by shake flask fermentation inspection bacterial classification enzymatic productivity, can spreads cultivation as seed and use or enzymatic production use.
The culture condition of shake flask fermentation is: by weight, culture medium prescription is wheat bran 5 ﹪ ~ 8 ﹪, cellulosic powder 1.5 ﹪ ~ 2 ﹪, glucose 0.3 ﹪ ~ 0.5 ﹪, ammonium sulfate 0.3 ﹪ ~ 0.5 ﹪, corn steep liquor 1 ﹪ ~ 2 ﹪, dipotassium hydrogen phosphate 0.1 ﹪ ~ 0.2 ﹪, sodium-chlor 0.3 ﹪ ~ 0.5 ﹪, water 86.3 ﹪ ~ 91.5 ﹪, wherein said each component sum is 100 ﹪, before sterilizing, adjust pH is 8.0 ~ 8.5,0.1MPa sterilizing 20 minutes ~ 35 minutes, culture condition: 33 DEG C ~ 35 DEG C, shaking speed 260r/min ~ 280r/min, cultivates 48 hours ~ 52 hours.
Enzyme liquid is prepared as by eggplant bottle inclined-plane to seeding tank to fermentor tank, second order fermentation, preparation standard enzyme liquid.
Seed culture based formulas is: by weight, wheat bran 5 ﹪ ~ 8 ﹪, Semen Maydis powder 1.5 ﹪ ~ 2 ﹪, ammonium sulfate 0.3 ﹪ ~ 0.5 ﹪, corn steep liquor 1 ﹪ ~ 2 ﹪, dipotassium hydrogen phosphate 0.1 ﹪ ~ 0.2 ﹪, sodium-chlor 0.3 ﹪ ~ 0.5 ﹪, water 86.8 ﹪ ~ 91.8 ﹪, wherein said each component sum is 100 ﹪, before sterilizing, adjust pH is 6.5 ~ 7.0,0.1MPa sterilizing 30 minutes ~ 35 minutes, seeding tank charging cumulative volume is no more than 70%; Culture condition: temperature 33 DEG C ~ 38 DEG C, mixing speed 180r/min ~ 220r/min, air flow was in v/v: 0 hour ~ 5 hours, 1:0.40 ~ 0.46, after 5 hours, 1:0.50 ~ 0.56, tank pressure 0.06MPa ~ 0.07MPa, culture cycle is 7 hours ~ 9 hours.
The culture medium prescription producing enzymic fermentation tank is: by weight, wheat bran 5 ﹪ ~ 8 ﹪, cellulosic powder 1.5 ﹪ ~ 2 ﹪, glucose 0.3 ~ 0.5 ﹪, ammonium sulfate 0.3 ﹪ ~ 0.5 ﹪, corn steep liquor 1 ﹪ ~ 2 ﹪, dipotassium hydrogen phosphate 0.1 ﹪ ~ 0.2 ﹪, sodium-chlor 0.3 ﹪ ~ 0.5 ﹪, water 86.3 ﹪ ~ 91.5 ﹪, wherein said each component sum is 100 ﹪, before sterilizing, adjust pH is 8.0 ~ 8.5,0.1MPa sterilizing 30 minutes ~ 35 minutes, charging cumulative volume is no more than 70 ﹪; Culture condition: temperature 33 DEG C ~ 38 DEG C, mixing speed 180r/min ~ 220r/min, air flow in v/v: 0 hour ~ 2 hours, 1:0.40 ~ 0.42,2 hours ~ 4 hours, 1:0.50 ~ 0.52,4 hours ~ 6 hours, 1:0.55 ~ 0.68,1:0.60 ~ 0.62 after 6 hours, tank pressure 0.06MPa ~ 0.07MPa, fermentation period is 48 hours ~ 52 hours.
After the preparation of enzyme liquid, preparation standard zytase liquid.By degerming through Plate Filtration, diatomite for the fermentation liquid producing enzymic fermentation tank, obtain under three months conditions are deposited in 15 DEG C of sealings through rotproofing, enzyme activity conservation rate is the standard xylanases liquid of 95 more than ﹪.
The vigour-testing method of zytase of the present invention is, is that 2 ﹪ xylans (sigma comes from birch product) solution prepared by solvent with glycine-sodium hydrate buffer solution that pH value is 9.0,0.1mol/L.Draw 0.9ml substrate in 15ml scale test tube, requiring that temperature water bath balances 3min, add the suitable diluted enzyme solution of 0.1ml, reaction 10min, add 3ml3,5 ~ edlefsen's reagent, develop the color in boiling water bath 15min, 15ml is settled to distilled water, spectrophotometer 550nm wavelength light-metering absorption value after cooling.Enzyme activity is defined as, and under condition determination, hydrolysis substrate per hour produces the required enzyme amount of 1mg reducing sugar (in wood sugar) and is defined as an enzyme activity unit (u).
CGMCCNO:6226 bacterial classification of the present invention is in Ningxia, China collection, in strain improvement screening process, filter out the superior strain that a strain laboratory is numbered PARK9639, according to major physiological biochemical character and 16SrRNA gene order, the experimental datas such as gyrB gene order are comprehensively analyzed and are accredited as subtilis (Bacillussubtilis), this bacterial classification submitted China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation (No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City) on 06 15th, 2012, preserving number: CGMCCNO:6226, and prove in have submitted preservation survival on the same day.
The slant culture based formulas of this bacterial classification is: in 1000mL distilled water, add extractum carnis 3g ~ 8g, yeast extract paste 3g ~ 8g, peptone 8g ~ 12g, glucose 3g ~ 5g, sodium-chlor 3g ~ 5g, adjust pH 6.9 ~ 7.1,0.1MPa sterilizing 28 minutes ~ 32 minutes, make test tube slant, inoculate this bacterial classification, 34 DEG C ~ 36 DEG C, cultivate 20 hours ~ 24 hours.
After this slant strains is qualified by shake flask fermentation inspection bacterial classification enzymatic productivity, can spreads cultivation as seed and use or enzymatic production use.
The culture condition of shake flask fermentation is: in mass, culture medium prescription is: Rhizoma amorphophalli powder 3 ﹪ ~ 4 ﹪, wheat bran 2 ﹪ ~ 4 ﹪, corn steep liquor 0.5 ﹪ ~ 1 ﹪ of nitrogen content 40%, yeast extract paste 0.5 ﹪ ~ 1 ﹪, Secondary ammonium phosphate 0.2 ﹪ ~ 0.3 ﹪, dipotassium hydrogen phosphate 0.02 ﹪ ~ 0.05 ﹪, peptone 0.1 ﹪ ~ 0.2 ﹪, magnesium sulfate 0.02 ﹪ ~ 0.05 ﹪, sodium carbonate 0.20 ﹪ ~ 0.25 ﹪, water 89.15 ﹪ ~ 93.46 ﹪, wherein said each component sum is 100 ﹪, before sterilizing, adjust pH is pH8.0 ~ pH8.5, 0.1MPa sterilizing 30 minutes ~ 35 minutes, culture condition: 32 DEG C ~ 37 DEG C, shaking speed 260r/min ~ 280r/min, cultivate 34 hours ~ 42 hours.
Enzyme liquid is prepared as by eggplant bottle inclined-plane to seeding tank to fermentor tank, second order fermentation, preparation standard enzyme liquid.
Seed culture based formulas is: in mass, corn steep liquor 1 ﹪ ~ 2 ﹪ of wheat bran 5 ﹪ ~ 6 ﹪, Semen Maydis powder 1 ﹪ ~ 2 ﹪, nitrogen content 40 ﹪, sodium-chlor 0.4 ﹪ ~ 0.6 ﹪, sodium carbonate 1 ﹪ ~ 2 ﹪, ammonium sulfate 0.3 ﹪ ~ 0.5 ﹪, water 86.9 ﹪ ~ 91.3 ﹪, wherein said each component sum is 100 ﹪, before sterilizing, adjust pH is 6.5 ~ 7.0,0.1MPa sterilizing 30 minutes ~ 35 minutes; Seeding tank charging cumulative volume is no more than 70 ﹪, culture condition is: temperature 33 DEG C ~ 38 DEG C, mixing speed 180r/min ~ 220r/min, air flow in v/v: 0 hour ~ 4 hours, 1:0.25 ~ 0.27,4 hours ~ 6 hours, 1:0.33 ~ 0.36, after 6 hours, 1:0.50 ~ 0.55, tank pressure 0.06MPa ~ 0.07MPa, culture cycle is 7 hours ~ 9 hours.
The culture medium prescription producing enzymic fermentation tank is: in mass, corn steep liquor 0.5 ﹪ ~ 1 ﹪ of Rhizoma amorphophalli powder 3 ﹪ ~ 4 ﹪, wheat bran 2 ﹪ ~ 4 ﹪, nitrogen content 40%, yeast extract paste 0.5 ﹪ ~ 1 ﹪, Secondary ammonium phosphate 0.2 ﹪ ~ 0.3 ﹪, dipotassium hydrogen phosphate 0.02 ﹪ ~ 0.05 ﹪, peptone 0.1 ﹪ ~ 0.2 ﹪, magnesium sulfate 0.02 ﹪ ~ 0.05 ﹪, sodium carbonate 0.20 ﹪ ~ 0.25 ﹪, water 89.15 ﹪ ~ 93.46 ﹪, wherein said each component sum is 100 ﹪, before sterilizing, adjust pH is 8.0 ~ 8.5,0.1MPa sterilizing 30 minutes ~ 35 minutes, charging cumulative volume is no more than 70 ﹪; Culture condition: temperature 33 DEG C ~ 38 DEG C, mixing speed 180r/min ~ 220r/min, air flow was in v/v: 0 hour ~ 4 hours, 1:0.30 ~ 0.32,4 hours ~ 6 hours, 1:0.40 ~ 0.42,1:0.50 ~ 0.55 after 6 hours, tank pressure 0.06MPa ~ 0.07MPa, fermentation period is 34 hours ~ 42 hours.
After the preparation of enzyme liquid, preparation standard mannase liquid.By degerming through Plate Filtration, diatomite for the fermentation liquid producing enzymic fermentation tank, obtain under three months conditions are deposited in 15 DEG C of sealings through rotproofing, enzyme activity conservation rate is the standard mannase liquid of 95 more than ﹪.
The enzyme activity determination method of mannase is, utilizes the disodium hydrogen phosphate dodecahydrate of 0.05mol/LpH6.0 and the locust bean gum solution of monohydrate potassium buffer 0.5%.In this substrate solution of 0.9ml, add the enzyme solution that 0.1ml suitably dilutes, 50 DEG C of accurate clock reaction 10min, in 550nm light-metering absorption value.Its enzyme activity is defined as, and under condition determination, hydrolysis locust bean gum per hour produces the required enzyme amount of 1mg reducing sugar (in seminose) and is defined as an enzyme activity unit (u).
CGMCCNO:6225 bacterial classification of the present invention is in Ningxia, China collection, in strain improvement screening process, filter out the superior strain that a strain laboratory is numbered KERR7989, according to major physiological biochemical character and 16SrRNA gene order, the experimental datas such as gyrB gene order are comprehensively analyzed and are accredited as me and carry genus bacillus (Bacillusaltitudinis), this bacterial classification submitted China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation (No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City) on 06 15th, 2012, preserving number: CGMCCNO:6225, and prove in have submitted preservation survival on the same day.
The slant culture based formulas of this bacterial classification is: in 1000mL distilled water, add extractum carnis 5g ~ 10g, yeast extract paste 5g ~ 10g, peptone 10g ~ 15g, glucose 5g ~ 10g, sodium-chlor 3g ~ 8g, adjust pH 6.9 ~ 7.1,0.1MPa sterilizing 20 minutes ~ 30 minutes, make test tube slant, inoculate this bacterial classification, 33 DEG C ~ 35 DEG C, cultivate 18 hours ~ 24 hours.
After this slant strains is qualified by shake flask fermentation inspection bacterial classification enzymatic productivity, can spreads cultivation as seed and use or enzymatic production use.
The culture condition of shake flask fermentation is: in mass, culture medium prescription is: wheat bran 6 ﹪ ~ 8 ﹪, cellulosic powder 1 ﹪ ~ 2 ﹪, corn steep liquor 1 ﹪ ~ 2 ﹪ of nitrogen content 40 ﹪, ammonium sulfate 0.3 ﹪ ~ 0.5 ﹪, dipotassium hydrogen phosphate 0.1 ﹪ ~ 0.2 ﹪, peptone 0.5 ﹪ ~ 1 ﹪, magnesium sulfate 0.03 ﹪ ~ 0.05 ﹪, sodium carbonate 0.2 ﹪ ~ 0.3 ﹪, water 85.95 ﹪ ~ 90.7 ﹪, wherein said each component sum is 100 ﹪, before sterilizing, adjust pH is 8.0 ~ 8.5, 0.1MPa sterilizing 25 minutes ~ 35 minutes, culture condition: 33 DEG C ~ 35 DEG C, shaking speed 260r/ minute ~ 280r/ minute, cultivate 48 hours ~ 52 hours.
Enzyme liquid is prepared as and obtains cellular liquid seed to seeding tank by eggplant bottle inclined-plane, then to producing enzymic fermentation tank, second order fermentation, preparation standard enzyme liquid.
Seed culture based formulas is: in mass, corn steep liquor 1 ﹪ ~ 2 ﹪ of wheat bran 5 ﹪ ~ 6 ﹪, Semen Maydis powder 1 ﹪ ~ 2 ﹪, nitrogen content 40 ﹪, sodium-chlor 0.4 ﹪ ~ 0.6 ﹪, sodium carbonate 1 ﹪ ~ 2 ﹪, ammonium sulfate 0.3 ﹪ ~ 0.5 ﹪, water 86.9 ﹪ ~ 91.3 ﹪, wherein said each component sum is 100 ﹪, before sterilizing, adjust pH is 6.5 ~ 7.0,0.1MPa sterilizing 25 minutes ~ 35 minutes; Seeding tank charging cumulative volume is no more than 70 ﹪, culture condition is: temperature 33 DEG C ~ 38 DEG C, mixing speed 180r/min ~ 220r/min, air flow in v/v: 0 hour ~ 4 hours, 1:0.25 ~ 0.27,4 hours ~ 6 hours, 1:0.33 ~ 0.36, after 6 hours, 1:0.50 ~ 0.55, tank pressure 0.06MPa ~ 0.08MPa, culture cycle is 7 hours ~ 9 hours.
The culture medium prescription producing enzymic fermentation tank is: in mass, corn steep liquor 1 ﹪ ~ 2 ﹪ of wheat bran 6 ﹪ ~ 8 ﹪, cellulosic powder 1 ﹪ ~ 2 ﹪, nitrogen content 40 ﹪, ammonium sulfate 0.3 ﹪ ~ 0.5 ﹪, dipotassium hydrogen phosphate 0.1 ﹪ ~ 0.2 ﹪, peptone 0.5 ﹪ ~ 1 ﹪, magnesium sulfate 0.03 ﹪ ~ 0.05 ﹪, sodium carbonate 0.2 ﹪ ~ 0.3 ﹪, water 85.95 ﹪ ~ 90.87 ﹪, wherein said each component sum is 100 ﹪, before sterilizing, adjust pH is 8.0 ~ 8.5,0.1MPa sterilizing 25 minutes ~ 35 minutes, charging cumulative volume is no more than 70 ﹪; Culture condition: temperature 33 DEG C ~ 38 DEG C, mixing speed 180r/min ~ 220r/min, air flow was in v/v: 0 hour ~ 4 hours, 1:0.30 ~ 0.32,4 hours ~ 6 hours, 1:0.40 ~ 0.42,1:0.50 ~ 0.55 after 6 hours, tank pressure 0.06MPa ~ 0.07MPa, fermentation period is 36 hours ~ 42 hours.
After the preparation of enzyme liquid, preparation standard lignoenzyme liquid.By degerming through Plate Filtration, diatomite for the fermentation liquid producing enzymic fermentation tank, obtain under three months conditions are deposited in 15 DEG C of sealings through rotproofing, enzyme activity conservation rate is the standard lignoenzyme liquid of 95 more than ﹪.
The measuring method of the enzyme activity of lignoenzyme: the syringaldazine solution of configuration 0.2mg/L to load in black agents bottle in Refrigerator store 5 days effectively.During use, the Tris damping fluid of 25mmol/LpH7.5 dilutes 10 times, and draw in this substrate solution of 2.5ml the enzyme solution adding 0.5ml and suitably dilute, 30 DEG C of accurate clock reaction 5min, in 530nm light-metering absorption value.Enzyme activity is defined as, and is defined as 1 enzyme activity unit (u) when enzyme substrate specificity per hour under condition determination makes light absorption value often increase by 1.0 units.
By the alkaline pectase liquid prepared, zytase liquid, mannase liquid and lignoenzyme liquid are through weighing, composite tank mixed preparing, anticorrosion, inspection, deposit in 5 DEG C of storehouses for subsequent use after packaging, the vigor of described complex enzyme liquid neutral and alkali polygalacturonase is 800u/ml ~ 1500u/ml(the present embodiment is 1400u/ml), the vigor of zytase is 1000u/ml ~ 1800u/ml(the present embodiment is 1600u/ml), the vigor of mannase is 300u/ml ~ 1000u/ml(the present embodiment is 600u/ml), the vigor of lignoenzyme is 100u/ml ~ 180u/ml(the present embodiment is 180u/ml).
The one that paper making raw material of the present invention is wheat straw or straw or reed or Di Lu or bagasse or raises in wood chip or Eucalyptus sheet or Herba Poae Sphondylodis.The present embodiment is Eucalyptus sheet.
Chemical-mechanical pulping of the present invention is the one in SCMP sulphonation chemistry machinery pulping or the thermomechanical slurrying of BCTMP bleached chemical or APP alkaline peroxide chemical machinery pulping or APMP alkaline hydrogen peroxide chemical-mechanical pulping or the slurrying of P-RCAPMP alkaline hydrogen peroxide thermomechanical.The present embodiment is APMP chemical mechanical pulp-making method.
The application of above-mentioned complex enzyme liquid in chemical-mechanical pulping, before to paper making raw material decatize, complex enzyme liquid is added paper making raw material by volume pump, make it mix with paper making raw material and carry out enzyme digestion reaction, the addition of complex enzyme liquid and the mass percent of over dry paper making raw material are 1.0 ﹪ ~ 1.5 ﹪, ferment treatment condition is: temperature 40 DEG C ~ 60 DEG C, pH6.0 ~ 8.5, enzymolysis time 30 minutes ~ 240 minutes, after enzymolysis completes, its subsequent technique carries out routinely.
The addition of the present embodiment complex enzyme liquid and the mass percent of over dry paper making raw material are 1.5 ﹪, and ferment treatment condition is: temperature 50 C, pH8.0, enzymolysis time 240 minutes, after enzymolysis completes, its subsequent technique carries out routinely.Namely Eucalyptus sheet through enzymolysis, gas steam storehouse, one section add DTPA and Na
2sO
3extruding preimpregnation, two sections add NaOH and H
2o
2extruding preimpregnation, three sections add NaOH and H
2o
2extruding preimpregnation, chemical reaction, one section of defibrination, two sections of defibrinations, latent, the screening purification of disappearing, concentrated after pulping.
Raw material, after complex enzyme liquid pre-treatment, can reduce industrial chemicals consumption 22 ﹪ such as alkali, and reduce refining energy consumption 25 ﹪, pulp brightness improves 2 ﹪, and yellowing reversion of pulp phenomenon obviously reduces.
The present invention is before to paper making raw material decatize, complex enzyme liquid is added paper making raw material by volume pump, make it mix with paper making raw material and carry out enzyme digestion reaction, its effect one is for destroying containing phenols, the isostructural chromophoric group of carbonyl in paper making raw material, to reduce brightness reversion.Two is pectin, pectin substance, xylan, xylogen that complex enzyme liquid acts in the mesogloea of adhesion flanking cell wall and primary wall, secondary wall, brings out fibre swelling, and it is estranged with cell or be separated, to reduce refining energy consumption to be easier to cell.Three be make primary wall simultaneously, secondary wall bears crack, pore quantity increases, and is easier to chemical liquid and enters, and reduces Chemical Pretreatment chemical industry material consumption.
Pectin substance in alkaline pectase effect raw material, the xylan in zytase effect raw material, mannase decomposes the mixed polysaccharide such as the mannosans in raw material, decreases the consumption of the mixed polysaccharide such as chemical industry material effect pectin, xylan and mannosans such as alkali; By the hydrolysis to the mixed polysaccharide such as pectin substance and xylan, make the pectin substance in middle lamella, the adhesion materials such as xylan reduce, make cell and cell estranged or separate, pore increases increase, is beneficial to the infiltration of the chemical industry material such as alkali, increase their accessibility, improve its functioning efficiency.Fortifying fibre water-absorbent, brings out fibre swelling, is beneficial to the dispersion of fibrous bundle, thus reduces refining intensity minimizing refining energy consumption; Interpolation lignoenzyme directly acts on the xylogen in raw material, makes it decompose or comes off, and the potential chromophoric group and the auxochrome group that contain a large amount of lignin and part extract in slurry are effectively removed, and reduce yellowing reversion of pulp phenomenon, improve pulp quality.
Embodiment 2:
Described in the present embodiment, the vigor of complex enzyme liquid neutral and alkali polygalacturonase is 1300u/ml, and the vigor of zytase is 1500u/ml, and the vigor of mannase is 800u/ml, and the vigor of lignoenzyme is 180u/ml.
Paper making raw material described in the present embodiment is for raising wood chip.
Chemical-mechanical pulping described in the present embodiment is APMP alkaline hydrogen peroxide chemical-mechanical pulping.
The addition of the present embodiment complex enzyme liquid and the mass percent of over dry paper making raw material are 1.5 ﹪, and ferment treatment condition is: temperature 50 C, pH8.0, enzymolysis time 230 minutes, after enzymolysis completes, its subsequent technique carries out routinely.Namely poplar sheet through enzymolysis, gas steam storehouse, one section add DTPA and Na
2sO
3extruding preimpregnation, two sections add NaOH and H
2o
2extruding preimpregnation, three sections add NaOH and H
2o
2extruding preimpregnation, chemical reaction, one section of defibrination, high dense reserving tower, squeezing machine, two sections of defibrinations, latent, the screening purification of disappearing, concentrated after pulping.
Raw material, after complex enzyme liquid pre-treatment, can reduce industrial chemicals consumption 23 ﹪ such as alkali, and reduce refining energy consumption 22 ﹪, pulp brightness improves 2 ﹪, and yellowing reversion of pulp phenomenon obviously reduces.
Remaining with embodiment 1.
Embodiment 3:
Described in the present embodiment, the vigor of complex enzyme liquid neutral and alkali polygalacturonase is 1200u/ml, and the vigor of zytase is 1300u/ml, and the vigor of mannase is 700u/ml, and the vigor of lignoenzyme is 130u/ml.
Paper making raw material described in the present embodiment is reed.
Chemical-mechanical pulping described in the present embodiment is the slurrying of BCTMP bleached chemical thermomechanical.
The addition of the present embodiment complex enzyme liquid and the mass percent of over dry paper making raw material are 1.5 ﹪, and ferment treatment condition is: temperature 55 DEG C, pH8.0, enzymolysis time 180 minutes, after enzymolysis completes, its subsequent technique carries out routinely.I.e. reed pulping after enzymolysis, gas steaming, chemical impregnation, preheating, pressurized refining, screening, bleaching.
Raw material, after complex enzyme liquid pre-treatment, can reduce industrial chemicals consumption 25 ﹪ such as alkali, and reduce refining energy consumption 21 ﹪, pulp brightness improves 3 ﹪, and yellowing reversion of pulp phenomenon obviously reduces.
Remaining with embodiment 1.
Embodiment 4:
Described in the present embodiment, the vigor of complex enzyme liquid neutral and alkali polygalacturonase is 1400u/ml, and the vigor of zytase is 1200u/ml, and the vigor of mannase is 800u/ml, and the vigor of lignoenzyme is 120u/ml.
Paper making raw material described in the present embodiment is wheat straw.
Chemical-mechanical pulping described in the present embodiment is SCMP sulphonation chemistry machinery pulping.
The addition of the present embodiment complex enzyme liquid and the mass percent of over dry paper making raw material are 1.5 ﹪, and ferment treatment condition is: temperature 55 DEG C, pH8.0, enzymolysis time 160 minutes, after enzymolysis completes, its subsequent technique carries out routinely.Namely wheat straw through enzyme leaching, storage, washing, pre-steamings, even steam, after steam, squeeze, one section of defibrination, two sections of defibrinations, disappear latent, selected after pulping.
Raw material, after complex enzyme liquid pre-treatment, can reduce industrial chemicals consumption 32 ﹪ such as alkali, and reduce refining energy consumption 23 ﹪, pulp brightness improves 3.5 ﹪, and yellowing reversion of pulp phenomenon obviously reduces.
Remaining with embodiment 1.