CN101418287B - Aspergillus niger liquid state fermentation pectic enzyme and control to colloidal matter in white water and paper stuff - Google Patents
Aspergillus niger liquid state fermentation pectic enzyme and control to colloidal matter in white water and paper stuff Download PDFInfo
- Publication number
- CN101418287B CN101418287B CN2008101536434A CN200810153643A CN101418287B CN 101418287 B CN101418287 B CN 101418287B CN 2008101536434 A CN2008101536434 A CN 2008101536434A CN 200810153643 A CN200810153643 A CN 200810153643A CN 101418287 B CN101418287 B CN 101418287B
- Authority
- CN
- China
- Prior art keywords
- polygalacturonase
- liquid
- aspergillus niger
- fermentation
- paper
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Enzymes And Modification Thereof (AREA)
- Paper (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a liquid-state fermentation pectinase of aspergillus niger and a method for controlling the dissolving and colloidal substances in white water and paper pulp. The method determines the optimal combination of the culture medium through optimizing a culture medium of aspergillus niger strains for producing pectinase to so as to increase the yield and the stability of the pectinase, and the method is used for processing the paper making white paper and controlling the dissolving and the colloidal substances (DCS) respectively. The demand amount of cations in the white water is reduced by 25 to 30 percent, the particle size is decreased, and the conductivity is increased after the treatment. The colloidal substances in the paper pulp are degraded. The invention has the advantages of simple operation of the treatment process, low cost, easy control and the like.
Description
Technical field
The present invention relates to a kind of Aspergillus niger liquid state fermentation polygalacturonase, relate in particular to the Aspergillus niger liquid state fermentation polygalacturonase that is applied to mthod of white water from paper making and paper pulp.
Background technology
Along with the continuous development of mthod of white water from paper making cyclically utilizing technology, the paper grade (stock) water yield has had minimizing significantly at present.But along with the raising of whitewater closure circulation degree and the continuous increase of cycle index, objectionable impurities also can accumulate gradually in the plain boiled water.In these objectionable impuritiess, to copy paper and paper making wet part chemical affect maximum, the most unmanageable be dissolving and the colloidalmaterial (DCS) of being called in the plain boiled water " anionic trash ".The continuous enrichment of colloidalmaterial, and then changed the specific conductivity balance, and increase cationic demand.Effects such as these anionic trashes are easier to neutralize with cationic paper making additive, absorption, thereby the effect or the inefficacy of reduction auxiliary agent.And then can cause l Water Paper page or leaf intensity and tensile strength to reduce, and paper machine broken end increases, and operating performance reduces.
Traditional plain boiled water treatment process mainly concentrates in the processing to suspended solids (SS), by mechanical effect such as disk filter, flotation and precipitation settler etc., or carry out biochemical treatment, adopt active sludge or anaerobic-aerobic bio-reactor, but these methods are very ineffective to the removal of DCS in the waste water, can only be as a kind of initial treatment measure.(Dissolved Air Flotation DAF) is present Newsprint Mill water-purification plant commonly used in dissolved air floatation.DAF is effectively for the removal of suspended solid material, but also bad to the removal effect of DCS in the plain boiled water.
In the modern paper system, often adopt chemical process that DCS is controlled.Wherein, the effective means of influence that reduces anionic trash in the papermaking systems is with anionic trash trapping agent (ATC) or is deciding agent (fixative) slurry is carried out pre-treatment.But, want strict control consumption when adding ATC, consumption can not effectively be controlled the anionic trash problem very little, and consumption too conference caused cationization and influenced follow-up sheet forming, influence the additive effect of retention filtration adjuvant, addition should be determined by the titration of dissolved charge and colloid surface electric charge.
The dissolving pectin that in not floating the TMP slurry, contains about slurry 2kg per ton.
There are some researches show that the dissolving of bleached mechanical pulp and colloidalmaterial (DCS) material mainly is created in H
2O
2In the bleaching process, pectic acid is the significant contribution person of the positively charged ion requirement of DCS in the bleached mechanical pulp.Pectin is a kind of saccharan of mainly being made up of galacturonic acid, and demethylation pectin is commonly referred to as polygalacturonic acid or pectic acid.In the defibrination process of machinery pulping, because high temperature and high-intensity mechanical effect, the part pectin substance dissociates out from fiber.
Polygalacturonase is the general name of the plurality of enzymes of decompose pectin matter, all has major application to be worth at aspects such as food-processing, feed processing, weaving, environment protection, inducing plant are disease-resistant.Microorganism because of have fast growth, growth conditions is simple, metabolic process is special and the characteristics such as wide that distribute become the important source of polygalacturonase.In recent years, many scholars are applied to polygalacturonase in the pulp and paper industry.
U.S. Patent number 5487172 has disclosed polygalacturonase and has handled and can reduce in the thermomechanical pulp (TMP) 40% cationic demand.
People such as Reid are published in " Enzyme and Microbial Technology " 2000 the 26th volumes the 2nd phase the 115th～123 page of " Pectinase in papermaking:solving retentionproblems in mechanical pulps bleached with hydrogen peroxide " literary composition and have reported that in plant experiment the cationic demand of thermomechanical pulp (TMP) recirculated water that the process polygalacturonase is handled can reduce by 28%～60%.
Li Zongquan, Zhan Huaiyu are published in studies show that in the 23rd～26 page of " Chinese papermaking " 2006 the 25th the 8th phase of volume " among the polygalacturonase treatments B CTMP DCS material and to the influence of cationic auxiliary action effect " literary composition: polygalacturonase can be degraded and be produced the essential substance pectic acid of anionic trash in solute and the colloidalmaterial (DCS).DCS is after polygalacturonase is handled, and the positive charge requirement obviously reduces, and compares with handling without polygalacturonase, and under identical cationic retention aid agent add-on, the retention of fiber fines obviously improves.Therefore, use polygalacturonase processing machine slurry or mthod of white water from paper making, can reduce anionic trash wherein, reduce the add-on of cationic polymers, improve the retention of fiber fines.
But because scholar before all adopts the commodity polygalacturonase to handle, it is higher to cause handling the DCS material cost with polygalacturonase.Therefore, China's papermaking circle demands urgently developing to replace the low polygalacturonase of cost of commodity polygalacturonase, in order to handle dissolving and colloidalmaterial (DCS) material in mthod of white water from paper making and the control paper pulp.
Summary of the invention
The object of the present invention is to provide a kind of zymotechnique of Aspergillus niger liquid state fermentation polygalacturonase, the polygalacturonase that ferments by this technology has enzyme height alive and good thermal stability.
Another object of the present invention is to provide a kind of above-mentioned polygalacturonase is applied to handle the dissolving in mthod of white water from paper making and the control paper pulp and the technology of colloidalmaterial, this technology dissolving and colloidalmaterial in the dissolving and colloidalmaterial and control paper pulp in the plain boiled water of degrading effectively reduces cationic demand.
The present invention is for achieving the above object the technical scheme that is adopted:
A kind of Aspergillus niger liquid state fermentation polygalacturonase, it is characterized in that: the production bacterial strain that uses is aspergillus niger (Aspergillus niger), depositary institution is China Committee for Culture Collection of Microorganisms common micro-organisms center, the address is the Datun Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, preservation date on November 25th, 2008, preserving number CGMCC No.2784, produce polygalacturonase by Aspergillus niger liquid state fermentation, its fermentation step comprises:
1) preparation of substratum
Seed culture medium: shake-flask seed substratum: wheat bran 10～30g/L; Glucose 20～30g/L; (NH
4)
2SO
410～20g/L; KH
2PO
41～3g/L; MgSO
47H
2O 0.5～1g/L, pH value 4.0～6.0;
Fermention medium: shake flask fermentation substratum: wheat bran 40～50g/L; (NH
4)
2SO
410～30g/L; NaCl 1～4g/L; MgSO
47H
2O 0.5～3g/L; The pH value is 4.0～6.0;
2) fermentation process
Aspergillus niger test tube slant bacterial classification is inserted in the liquid seeds fermentor tank, at 30 ℃ of temperature, stirring velocity 120r/min, ventilation 0.2m
3Condition bottom fermentation 36h～72h of/h obtains needed liquid seeds; According to 3%～5% inoculum size seed is inserted the fermentation of Aspergillus niger jar then, at 30 ℃～50 ℃ of temperature, stirring velocity 120r/min, ventilation 0.2m
3Condition bottom fermentation 36～72h of/h makes required pectase liquid through centrifugation then, and it is stored in the polygalacturonase storage tank with standby;
3) pectase liquid is purified
With the ultra-filtration membrane of molecular weight cut-off 10000 to fermentation broth sample carry out ultrafiltration and concentration to original volume 1/10～1/2 after, it is standby to collect concentrated crude enzyme liquid, carry out the ammonium sulfate precipitation preliminary experiment, the concentration of determining ammonium sulfate precipitation is 40%～100%, with the centrifugal removal thalline of crude enzyme liquid, in supernatant liquor, add ammonium sulfate to 40% saturation ratio, place 24h for 4 ℃, through 4 ℃, behind the centrifugal 10min of 8000r/min, supernatant liquor continues to add ammonium sulfate to 100% saturation ratio, places 24h for 4 ℃, through 4 ℃, behind the centrifugal 30min of 8000r/min, collecting precipitation is dissolved in it in acetate-sodium acetate buffer of pH6.0, dialyses with dialysis tubing, pectase liquid after obtaining purifying, its zymologic property: 30～60 ℃ of optimal reactive temperatures, optimum pH 4～6, residual enzyme work is higher than 90% behind 30～60 ℃ of following insulation 2h.
The polygalacturonase enzyme liquid that adopts preparation method of the present invention to make is applied to handle in the mthod of white water from paper making method of dissolving and colloidalmaterial, polygalacturonase handle dissolve and the technology of colloidalmaterial as follows:
Add pectase liquid to plain boiled water, treatment temp is 30～60 ℃, treatment time 20～60min.
Described plain boiled water is for being the paper machine plain boiled water of main slurry with mechanical pulp and chemimechanical pulp.
Described polygalacturonase consumption is 15U～50U/100mL plain boiled water.
The polygalacturonase that adopts preparation method of the present invention to make is applied to control dissolving and colloidalmaterial in the paper pulp, and polygalacturonase is made an addition in the paper pulp, is added in an amount of 3U～10U/g oven dry stock, and described paper pulp is mechanical pulp and chemimechanical pulp.
Beneficial effect of the present invention:
Do blank relatively with traditional mechanical process and chemical Treatment plain boiled water and with the plain boiled water that does not add pectase liquid, the present invention's pectic substance in the DCS material of degrading effectively, plain boiled water cationic demand (CD value) reduces 25%-30%, and grain diameter reduces, and specific conductivity increases.
Pectase liquid of the present invention makes an addition in the paper pulp, after pulp processing, and the colloidalmaterial in the paper pulp of having degraded effectively.
Embodiment
In order further to understand content of the present invention, characteristics, with specific embodiment the present invention is described below.
Embodiment
Polygalacturonase is produced in liquid state fermentation
Aspergillus niger test tube slant bacterial classification is inserted in the liquid seeds fermentor tank, and the seed fermentation substratum adopts: wheat bran 10g/L, glucose 20g/L, (NH
4)
2SO
420g/L, KH
2PO
42g/L, MgSO
47H
2O 1g/L, pH value 4.0; Be 4.0 in 30 ℃ of temperature, pH value, stirring velocity 120r/min, ventilation 0.2m
3The condition bottom fermentation 48h of/h obtains needed liquid seeds; According to 3% inoculum size seed is inserted the fermentation of Aspergillus niger jar then, fermention medium adopts: wheat bran 40g/L; (NH
4)
2SO
410g/L; NaCl 1g/L; MgSO
47H
2O 1g/L; PH value 4.0; At 30 ℃ of temperature, pH5, stirring velocity 120r/min, ventilation 0.2m
3The condition bottom fermentation 72h of/h.Filter and centrifugal fermented liquid, measure the polygalacturonase enzyme and live, absorbancy reaches 0.725.
The purifying of pectase liquid
Crude enzyme liquid is filtered centrifugal back film with molecular weight cut-off 10000 it is carried out ultrafiltration to 1/5 of original volume.Set sulfuric acid and press concentration, in enzyme liquid, add the ammonium sulfate powder and carry out well-regulated stirring, place 24h for 4 ℃, observe and measure the vigor of three kinds of polygalacturonases in the supernatant liquor from 0%～100%.The result shows that the concentration of the ammonium sulfate precipitation of polygalacturonase in the crude pectin enzyme liquid is 50%～100%.
Pectase liquid is handled dissolving and the colloidalmaterial technology in the mthod of white water from paper making: add the 20U pectase liquid in filling 100mL plain boiled water, handle 40min down at 40 ℃.
Do the blank example with the plain boiled water that does not add pectase liquid.Compare with the blank example, handle through pectase liquid, the cationic demand after plain boiled water is handled reduces 25%-30%, and grain diameter reduces, the specific conductivity increase
Pectase liquid is applied to control the technology of colloidalmaterial DCS in the paper pulp:
Pectase liquid is made an addition in the paper pulp, be added in an amount of 3U～10U/g oven dry stock.Described paper pulp is mechanical pulp and chemimechanical pulp, by the processing of polygalacturonase to paper pulp, and the colloidalmaterial in the paper pulp of having degraded.
It is simple that the present invention has process operations, and cost is low and be easy to the advantages such as control.
Claims (3)
1. Aspergillus niger liquid state fermentation polygalacturonase preparation method, it is characterized in that: the production bacterial strain that uses is aspergillus niger (Aspergillus niger), depositary institution is China Committee for Culture Collection of Microorganisms common micro-organisms center, the address is the Datun Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, preservation date on November 25th, 2008, preserving number CGMCC No.2784, produce polygalacturonase by Aspergillus niger liquid state fermentation, fermentation step comprises:
Fermentation process:
1) preparation of substratum
Seed culture medium: shake-flask seed substratum: wheat bran 10～30g/L; Glucose 20～30g/L; (NH
4)
2SO
410～20g/L; KH
2PO
41～3g/L; MgSO
47H
2O 0.5～1g/L, pH value 4.0～6.0;
Fermention medium: shake flask fermentation substratum: wheat bran 40～50g/L; (NH
4)
2SO
410～30g/L; NaCl 1～4g/L; MgSO
47H
2O 0.5～3g/L; PH value 4.0～6.0;
2) fermentation culture
Aspergillus niger test tube slant bacterial classification is inserted in the liquid seeds fermentor tank, at 30 ℃ of temperature, stirring velocity 120r/min, ventilation 0.2m
3Condition bottom fermentation 36h～72h of/h obtains needed liquid seeds; According to 3%～5% inoculum size seed is inserted the fermentation of Aspergillus niger jar then, at 30 ℃～50 ℃ of temperature, stirring velocity 120r/min, ventilation 0.2m
3Condition bottom fermentation 36～72h of/h makes required pectase liquid through centrifugation then, and it is stored in the polygalacturonase storage tank with standby;
3) pectase liquid is purified
With the ultra-filtration membrane of molecular weight cut-off 10000 to fermentation broth sample carry out ultrafiltration and concentration to original volume 1/10～1/2 after, it is standby to collect concentrated crude enzyme liquid, carry out the ammonium sulfate precipitation preliminary experiment, the concentration of determining ammonium sulfate precipitation is 40%～100%, with the centrifugal removal thalline of crude enzyme liquid, in supernatant liquor, add ammonium sulfate to 40% saturation ratio, place 24h for 4 ℃, through 4 ℃, behind the centrifugal 10min of 8000r/min, supernatant liquor continues to add ammonium sulfate to 100% saturation ratio, places 24h for 4 ℃, through 4 ℃, behind the centrifugal 30min of 8000r/min, collecting precipitation is dissolved in it in acetate-sodium acetate buffer of pH6.0, dialyse with dialysis tubing, pectase liquid after obtaining purifying, the zymologic property of pectase liquid are 30～60 ℃ of optimal reactive temperatures, optimum pH 4～6, residual enzyme work is higher than 90% behind 30～60 ℃ of following insulation 2h.
2. one kind is adopted the polygalacturonase of claim 1 preparation to be applied to handle dissolving and colloidalmaterial in the mthod of white water from paper making, it is characterized in that: add pectase liquid to plain boiled water, treatment temp is 30～60 ℃, treatment time 20～60min, described plain boiled water is for being the paper machine plain boiled water of main slurry with mechanical pulp and chemimechanical pulp, and the consumption that is used to handle the polygalacturonase enzyme liquid of plain boiled water is 15U～50U/100mL plain boiled water.
3. a polygalacturonase that adopts claim 1 preparation is applied to control the dissolving in the paper pulp and the method for colloidalmaterial, it is characterized in that: polygalacturonase is added in the paper pulp, be added in an amount of 3U～10U/g oven dry stock, described paper pulp slurry is planted and is mechanical pulp and chemimechanical pulp.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008101536434A CN101418287B (en) | 2008-11-28 | 2008-11-28 | Aspergillus niger liquid state fermentation pectic enzyme and control to colloidal matter in white water and paper stuff |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008101536434A CN101418287B (en) | 2008-11-28 | 2008-11-28 | Aspergillus niger liquid state fermentation pectic enzyme and control to colloidal matter in white water and paper stuff |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101418287A CN101418287A (en) | 2009-04-29 |
| CN101418287B true CN101418287B (en) | 2010-12-08 |
Family
ID=40629290
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2008101536434A Expired - Fee Related CN101418287B (en) | 2008-11-28 | 2008-11-28 | Aspergillus niger liquid state fermentation pectic enzyme and control to colloidal matter in white water and paper stuff |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101418287B (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102977184B (en) * | 2012-11-13 | 2014-07-09 | 天津耀宇生物技术有限公司 | Purification method of protein group of 30 kD in silkworm pupa |
| CN105040505B (en) * | 2015-06-30 | 2017-08-25 | 华南理工大学 | A kind of method that aspergillus oryzae whole-cell catalyst handles mthod of white water from paper making resin |
| CN105369633A (en) * | 2015-11-18 | 2016-03-02 | 德清县伊得利丝绸有限公司 | Cotton and linen bleaching and dyeing deoxygenization finishing agent |
| CN106248534A (en) * | 2016-10-11 | 2016-12-21 | 广西大学 | A kind of the change of temperature value in plain boiled water is utilized to carry out the system of early warning microstickies unstability |
| CN108585343A (en) * | 2017-08-01 | 2018-09-28 | 王建玲 | The recycle device and processing method of papermaking enriched water |
-
2008
- 2008-11-28 CN CN2008101536434A patent/CN101418287B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN101418287A (en) | 2009-04-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Shen et al. | The effect of selected heavy metals (Ni, Co and Fe) on anaerobic granules and their extracellular polymeric substance (EPS) | |
| AU2003250185B2 (en) | Production of a fermentation product | |
| CN101418287B (en) | Aspergillus niger liquid state fermentation pectic enzyme and control to colloidal matter in white water and paper stuff | |
| US9139857B2 (en) | Stepwise enzymatic hydrolysis process for converting cellulose to glucose | |
| JP4554314B2 (en) | Continuous saccharification method of lignocellulose | |
| MXPA05001407A (en) | Production of a fermentation product. | |
| AU2008322755A1 (en) | Method for producing lipid | |
| CN104558238B (en) | Process for extracting sodium alginate | |
| CN101418289B (en) | Trichoderma reesei liquid submerged fermentation cellulase and enzymatic beating process thereof | |
| CN110129380A (en) | Utilize the method for stalk production biomass fuel, corrugated paper, organic fertilizer | |
| CN1133775C (en) | Biological pretreatment process of preparing chemical mechanical straw pulp | |
| CN101899479B (en) | Production method for preparing xylitol by using agricultural and forestry waste | |
| CN106434759A (en) | Method of preparing biological flocculant from cornstalks | |
| CN107904266B (en) | Pretreatment method for efficiently and environmentally improving lignocellulose enzymolysis saccharification efficiency and application | |
| CN107177646B (en) | Method for strengthening acid pretreatment of waste biomass by using lignin-degrading bacteria | |
| CN103990441B (en) | A kind of adsorbent for heavy metal preparation method based on modified bacteria cellulose | |
| CN1053242C (en) | Straw pulp performance improving technology using xylanase | |
| CN100402658C (en) | Method and apparatus for preparing reducing sugar by enzymolysis of steam-exploded straw with membrane reactor | |
| CN107034255B (en) | Method for strengthening pretreatment of waste biomass sodium carbonate by using lignin-degrading bacteria | |
| WO2011022644A2 (en) | Fermentation and chemical treatment of pulp and paper mill sludge | |
| CN106467335A (en) | A kind of method of utilization cellulose base green flocculation material coagulating treatment ultrasound wave pulp-making waste-water | |
| CN104630909A (en) | Method for preparing flax fiber by ultrasonic combination of mucor circinelloides DK1 strain and hydrogen peroxide | |
| CN108103123A (en) | A kind of method that bacteria cellulose is produced using rice steeping water as raw material | |
| CN214183017U (en) | One set of device of preparation acid type sophorolipid | |
| KR102592850B1 (en) | Green Hydrogen Generator |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Assignee: Zhejiang Jingxing Paper Joint Stock Co., Ltd. Assignor: Tianjin University of Science & Technology Contract record no.: 2011330001098 Denomination of invention: Aspergillus niger liquid state fermentation pectic enzyme and control to colloidal matter in white water and paper stuff Granted publication date: 20101208 License type: Exclusive License Open date: 20090429 Record date: 20110824 |
|
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20101208 Termination date: 20171128 |