CN106223096A - A kind of high temperature esterase is utilized to remove the method for gluing thing in secondary stock - Google Patents
A kind of high temperature esterase is utilized to remove the method for gluing thing in secondary stock Download PDFInfo
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- CN106223096A CN106223096A CN201610594944.5A CN201610594944A CN106223096A CN 106223096 A CN106223096 A CN 106223096A CN 201610594944 A CN201610594944 A CN 201610594944A CN 106223096 A CN106223096 A CN 106223096A
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- D—TEXTILES; PAPER
- D21—PAPER-MAKING; PRODUCTION OF CELLULOSE
- D21C—PRODUCTION OF CELLULOSE BY REMOVING NON-CELLULOSE SUBSTANCES FROM CELLULOSE-CONTAINING MATERIALS; REGENERATION OF PULPING LIQUORS; APPARATUS THEREFOR
- D21C5/00—Other processes for obtaining cellulose, e.g. cooking cotton linters ; Processes characterised by the choice of cellulose-containing starting materials
- D21C5/005—Treatment of cellulose-containing material with microorganisms or enzymes
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
- C12N9/18—Carboxylic ester hydrolases (3.1.1)
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y301/00—Hydrolases acting on ester bonds (3.1)
- C12Y301/01—Carboxylic ester hydrolases (3.1.1)
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- D—TEXTILES; PAPER
- D21—PAPER-MAKING; PRODUCTION OF CELLULOSE
- D21C—PRODUCTION OF CELLULOSE BY REMOVING NON-CELLULOSE SUBSTANCES FROM CELLULOSE-CONTAINING MATERIALS; REGENERATION OF PULPING LIQUORS; APPARATUS THEREFOR
- D21C5/00—Other processes for obtaining cellulose, e.g. cooking cotton linters ; Processes characterised by the choice of cellulose-containing starting materials
- D21C5/02—Working-up waste paper
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/50—Reuse, recycling or recovery technologies
- Y02W30/64—Paper recycling
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Abstract
The invention discloses and a kind of utilize high temperature esterase to remove the method for gluing thing in secondary stock, described esterase is esterase EstE1, the PDB ID:2C7B deriving from grand genomic library.Specifically including following steps: add the esterase of 0.1U～1.0U/g (relative to oven dry stock quality) in secondary stock, process the time 45～120min, temperature 40～80 DEG C, pH is 5.0～9.0, mixing speed 100～150rpm.Finally according to gluing thing content in TAPPIT 277 standard technique detection secondary stock.Result confirms that stickies removal rate reaches 54.1%～66.5%, even if the most still having preferable removal effect.The present invention utilizes high temperature esterase EST1 that the gluing thing in secondary stock carries out efficient degradation, and operating process is easy and without additional machinery equipment, and stickies removal effect is obvious, and production cost is low, pollution-free.
Description
Technical field
The present invention relates to a kind of utilize high temperature esterase to remove the method for gluing thing in secondary stock.
Background technology
In recent years, secondary stock in global pulp total quantity consumed accounting more than 60%.Waste paper is as China's pulping and paper-making
A kind of important source material of industry, the recycling of waste paper is possible not only to save a large amount of plant fiber material, reduce production cost, and
, alleviation environmental pollution energy-saving and cost-reducing for realization has profound significance.But the formation of gluing thing and control problem are in reality
Waste paper recycle during day by day highlight, gluing thing during referring to waste paper recycling various sources different and have permanent or
The interim stickiness of person also can cause problem and all substances causing product quality to decline in paper-making process.Gluing thing main source
Hot-melt object in waste paper, pressure-sensitive thing, sizing agent, coating gluing thing and ink residue thing etc., these materials are mainly by polypropylene
Acid esters, polyvinyl acetate etc. mix with other impurity, and the basic structure component of gluing thing is connected by they by ester bond
Together.Owing to gluing thing presents elecrtonegativity in slurry system, thus various cationic substance can be adsorbed, therefore in slurrying
Paper-making process is easy for result in blockage mesh, paper cavity occurs, produce anionic trash, cause disconnected paper and paper machine to shut down
Etc. problem.
At present, remove and control the method for gluing thing in slurry and mainly use Mechanical Method and chemical method.Mechanical Method includes sieve
Select, purify, wash, the physical method such as flotation and dissipation of heat, be mainly used to remove the big gluing thing in secondary stock.Chemistry rule
By adding stickies control machine, utilize the methods such as chemisorbed, modification, dispersion, surface passivation to remove microstickies.This
A little methods are the stickies control method that regenerated papermaking enterprise is conventional, although have certain removal effect, however exist highly energy-consuming,
Efficiency is low, be easily generated the problems such as a large amount of industrial wastewaters, and removal efficiency also will be by factors such as the qualities of equipment performance
Impact.
Enzyme has efficient, single-minded, a stable catalytic, and environmentally safe, therefore in pulping and paper-making field
Application of increased attention, many enterprises utilize biological enzyme to process the gluing in waste paper removal process in recent years
Thing problem.Along with the development of enzyme engineering technology, the catalytic efficiency of enzyme, substrate specificity and stability can be carried further
Height, is particularly suited for actual production.The biological enzyme formulation being currently used for paper industry has protease, cellulase, pectase, wood
Dextranase, lipase and esterase etc..Wherein lipase and esterase are class carboxylic ester hydrolases, have quickening emplastic ester bond
Fracture so that it is stick object and amass and diminish and stick the characteristic that efficiency weakens, can be used for the gluing thing controlling and removing in waste paper.
Ester bond once ruptures, and the solvent of gluing thing is difficult to regroup in systems, and enzyme can be degraded voluntarily, will not
Pollute environment.Wan Jinquan etc. [ten thousand Jin Quan etc., in de inked pulp, gluing thing biological enzyme processes] utilize lipase to de inked pulp
Middle gluing thing processes, and finds that different types of lipase treatment effect is different, but is difficult to real under common Pulping conditions
Now preferable treatment effect.The soughing of the wind in forest trees etc. [soughing of the wind in forest trees etc., the application in Waste Paper Handling of the stickies control enzyme] utilize enzyme
Optimyze525 processes waste paper gluing thing, and stickies removal rate reaches 29.9%.
Although common lipase and esterase have certain stickies removal effect, but secondary stock composition is complex,
Containing the chemical substance that enzyme can be made in a large number to inactivate, and the extreme environment such as high temperature is easily caused biological enzyme formulation in actual production
The efficiency of application is suppressed.
Summary of the invention
For highly energy-consuming in existing waste paper stickies removal technique, efficiency is low, be easily generated a large amount of industrial wastewater and general
Logical enzyme is the problem such as easy in inactivation in hot environment.The present invention provides a kind of high temperature esterase EST1 to remove gluing thing in secondary stock
Method, the present invention provide technique simple, effect is obvious, easily operated.
For achieving the above object, technical scheme is as follows for the present invention:
A kind of utilizing high temperature esterase to remove the method for gluing thing in secondary stock, described esterase is high temperature esterase EST1, comes
Come from esterase EstE1, the PDB ID:2C7B_B of grand genomic library.Comprise the steps:
(1) preparation of high temperature esterase EST1:
Esterase EST1 strain is inoculated into seed fluid medium shaken cultivation, then strain is inoculated into fermentation medium shakes
Swing cultivation, prepare high temperature esterase EST1 crude enzyme liquid, by high temperature esterase EST1 crude enzyme liquid purification, use paranitrophenol colorimetry
The enzyme measuring high temperature esterase EST1 is lived.
(2) utilize high temperature esterase EST1 process secondary stock:
In secondary stock add 0.1U～1.0U/g (relative to oven dry stock quality) esterase EST1, process the time 45～
120min, temperature 40～80 DEG C, pH 5.0～9.0, mixing speed 100～150rpm.
(3) by gluing thing content in TAPPIT-277 standard technique detection secondary stock:
Pulmac-MasterScreen pulp classifier is utilized to be sieved by slurry according to TAPPIT-277 gluing thing algoscopy
Choosing, will sift out afterwards gluing thing carry out dyeing, tabletting, finally be analyzed scanning with scanning software, and then record gluing thing and contain
Amount.
Preferably, described esterase EST1 addition is 0.3～0.8U/g (relative to oven dry stock quality).
It is further preferable that the addition of described esterase is 0.5U/g (relative to oven dry stock).
Preferably, described temperature is 40～60 DEG C.
Preferably, described pH is 7.0～8.0.
It is further preferable that described temperature is 55 DEG C.
Preferably, the described process time is 60min.
Preferably, described mixing speed is 120rpm.
Preferably, the enzyme of described esterase is lived as 125.0U/ml.
Preferably, the slurry of described secondary stock is dense is 4%～6%.
Compared with prior art, the beneficial effects of the present invention is:
(1) preparation of the high temperature esterase EST1 in the present invention is easily, can effectively hydrolyze polyacrylate in waste paper gluing thing, gather
The compositions such as vinylacetate；Remove waste paper gluing thing effect obvious, result confirm stickies removal rate reach 54.1%～
66.5%, even if the most still having preferable removal effect.
(2) the high temperature esterase EST1 in the present invention is best suitable for reaction temperature is 55 DEG C, and lives in 80 DEG C of insulations catalysis in 2 hours
Trying hard to keep, it is constant to hold, and this esterase shows good catalytic performance and structural stability, and that can tackle in waste paper recovery process is each
Plant high temperature and acid or alkali environment, remain under the high temperature conditions keep preferable stickies removal effect.
(3) method that the present invention removes waste paper gluing thing by high temperature esterase EST1 is carried out the most on a production line, need not
Increase any plant equipment, simple to operate, low cost, pollution-free, solve in regenerated papermaking field tight efficiently, steadily
Ghost image rings product quality gluing thing problem.
Detailed description of the invention
The enforcement of the present invention is introduced in more detail by embodiment.In the described embodiment, described enzyme concentration is relative to absolutely
Dry pulp Mass Calculation, experimental result is taken the mean the most in triplicate.Esterase used in the present invention, its protein sequence number is by egg
White matter warehouse publication (http://www.rcsb.org/pdb/home/home.do)；Derive from the esterase of grand genomic library
(EstE1, PDB ID:2C7B), its aminoacid sequence as shown in SEQ ID NO:1, nucleotide sequence such as SEQ ID NO:2 institute
Show.
The preparation process of esterase EST1: utilize the encoding gene of the EST1 esterase that full genome synthetic method obtains, and clone
Recombinant expression carrier is obtained on expression vector pET23a-CBD carrier.Utilize CaCl2Conversion method, will encode containing EST1 esterase
The expression plasmid of gene proceeds in BL21 (DE3) strain, it is thus achieved that genetic engineering bacterium.The strain of esterase EST1 is received containing Amp
(100ug/ml) amplification culture in LB seed culture medium, when OD value reaches 0.6～0.8, by bacterium solution according to the ratio of 1:100
Be inoculated in the LB fermentation medium containing Amp (100ug/ml), shake to OD value be 0.6～0.8 time add IPTG (20mmol/L)
Derivant carries out abduction delivering, receives bacterium after 18～24h.Bacterium solution is collected tube wall after centrifugal 10min under the conditions of 12000r/min
Cell carries out ultrasonication, the most again to take supernatant after 12000r/min pelleted by centrifugation 10min, i.e. obtains crude enzyme liquid.Existing
Research report utilizes cellulose to carry out specific binding with esterase, carries out enzyme action with HRV 3CP the most again thus obtains pure enzyme.
1L bacterium solution correspondence 1g cellulose, is combined 30min by crude enzyme liquid with cellulose under room temperature, stirs in good time.High speed centrifugation after in conjunction with
10min, abandoning supernatant, adhesion protein PBS is washed twice.HRV 3CP the most centrifugal, supernatant discarded,
PBS is utilized to clean 2 times.In adhesion protein, add appropriate PBS, mix in the ratio of 1L bacterium solution correspondence 1ml HRV 3CP
Even rear enzyme action 4h or overnight.High speed centrifugation after enzyme action, supernatant is pure enzyme.Utilize paranitrophenol colorimetric method for determining system afterwards
The enzyme of standby high temperature esterase EST1 is lived, and records enzyme and lives as 125.0U/ml.
Lipase CALB is commercial lipases, records enzyme and lives as 150.0U/ml.
Embodiment 1
Weighing 100g OCC oven dry stock, reduction paste is dense to 4%, and esterase EST1 addition is that 0.2U/g is (relative to over dry
Slurry), treatment temperature is 40 DEG C, and pH is 5.0, processes time 45min, and speed of agitator is 100rpm, and embodiment is according to TAPPIT-
277 gluing thing algoscopys utilize Pulmac-MasterScreen pulp classifier to be screened by slurry, the gluing thing that will sift out afterwards
Carry out dyeing, tabletting, be finally analyzed scanning with scanning software, and then record the gluing thing content after ferment treatment.It is being not added with
In the case of enzyme, processing secondary stock with remaining the same terms, measuring result is blank group gluing thing content.
Embodiment 2
Weighing 100g OCC oven dry stock, reduction paste is dense to 5%, and esterase EST1 addition is that 0.5U/g is (relative to over dry
Slurry), treatment temperature is 55 DEG C, and pH is 5.0, processes time 60min, and speed of agitator is 120rpm, and embodiment is according to TAPPIT-
277 gluing thing algoscopys utilize Pulmac-MasterScreen pulp classifier to be screened by slurry, the gluing thing that will sift out afterwards
Carry out dyeing, tabletting, be finally analyzed scanning with scanning software, and then record the gluing thing content after ferment treatment.It is being not added with
In the case of enzyme, processing secondary stock with remaining the same terms, measuring result is blank group gluing thing content.
Embodiment 3
Weighing 100g OCC oven dry stock, reduction paste is dense to 5%, and esterase EST1 addition is that 0.5U/g is (relative to over dry
Slurry), treatment temperature is 55 DEG C, and pH is 7.0, processes time 60min, and speed of agitator is 120rpm, and embodiment is according to TAPPIT-
277 gluing thing algoscopys utilize Pulmac-MasterScreen pulp classifier to be screened by slurry, the gluing thing that will sift out afterwards
Carry out dyeing, tabletting, be finally analyzed scanning with scanning software, and then record the gluing thing content after ferment treatment.It is being not added with
In the case of enzyme, processing secondary stock with remaining the same terms, measuring result is blank group gluing thing content.
Embodiment 4
Weighing 100g OCC oven dry stock, reduction paste is dense to 5%, and esterase EST1 addition is that 0.5U/g is (relative to over dry
Slurry), treatment temperature is 55 DEG C, and pH is 9.0, processes time 60min, and speed of agitator is 120rpm, and embodiment is according to TAPPIT-
277 gluing thing algoscopys utilize Pulmac-MasterScreen pulp classifier to be screened by slurry, the gluing thing that will sift out afterwards
Carry out dyeing, tabletting, be finally analyzed scanning with scanning software, and then record the gluing thing content after ferment treatment.It is being not added with
In the case of enzyme, processing secondary stock with remaining the same terms, measuring result is blank group gluing thing content.
Embodiment 5
Weighing 100g OCC oven dry stock, reduction paste is dense to 6%, and esterase EST1 addition is that 1.0U/g is (relative to over dry
Slurry), treatment temperature is 80 DEG C, and pH is 7.0, processes time 120min, and speed of agitator is 150rpm, and embodiment is according to TAPPIT-
277 gluing thing algoscopys utilize Pulmac-Master Screen pulp classifier to be screened by slurry, the gluing that will sift out afterwards
Thing carries out dyeing, tabletting, is finally analyzed scanning with scanning software, and then records the gluing thing content after ferment treatment.Not
In the case of enzyme-added, processing secondary stock with remaining the same terms, measuring result is blank group gluing thing content.
Comparative example 1
Adding lipase CALB replaces esterase EST1 to process secondary stock, and other conditions are same as in Example 1, comparative example
Utilize Pulmac-Master Screen pulp classifier to be screened by slurry according to TAPPIT-277 gluing thing algoscopy, afterwards will
The gluing thing sifted out carries out dyeing, tabletting, is finally analyzed scanning with scanning software, and then records the gluing thing after ferment treatment
Content.In the case of the most enzyme-added, processing secondary stock with remaining the same terms, measuring result contains for blank group gluing thing
Amount.
Comparative example 2
Adding lipase CALB replaces esterase EST1 to process secondary stock, and other conditions are same as in Example 2, comparative example
Utilize Pulmac-Master Screen pulp classifier to be screened by slurry according to TAPPIT-277 gluing thing algoscopy, afterwards will
The gluing thing sifted out carries out dyeing, tabletting, is finally analyzed scanning with scanning software, and then records the gluing thing after ferment treatment
Content.In the case of the most enzyme-added, processing secondary stock with remaining the same terms, measuring result contains for blank group gluing thing
Amount.
Comparative example 3
Adding lipase CALB replaces esterase EST1 to process secondary stock, and other conditions are same as in Example 3, comparative example
Utilize Pulmac-Master Screen pulp classifier to be screened by slurry according to TAPPIT-277 gluing thing algoscopy, afterwards will
The gluing thing sifted out carries out dyeing, tabletting, is finally analyzed scanning with scanning software, and then records the gluing thing after ferment treatment
Content.In the case of the most enzyme-added, processing secondary stock with remaining the same terms, measuring result contains for blank group gluing thing
Amount.
Comparative example 4
Adding lipase CALB replaces esterase EST1 to process secondary stock, and other conditions are the same as in Example 4, comparative example
Utilize Pulmac-Master Screen pulp classifier to be screened by slurry according to TAPPIT-277 gluing thing algoscopy, afterwards will
The gluing thing sifted out carries out dyeing, tabletting, is finally analyzed scanning with scanning software, and then records the gluing thing after ferment treatment
Content.In the case of the most enzyme-added, processing secondary stock with remaining the same terms, measuring result contains for blank group gluing thing
Amount.
Comparative example 5
Adding lipase CALB replaces esterase EST1 to process secondary stock, and other conditions are same as in Example 5, comparative example
Utilize Pulmac-Master Screen pulp classifier to be screened by slurry according to TAPPIT-277 gluing thing algoscopy, afterwards will
The gluing thing sifted out carries out dyeing, tabletting, is finally analyzed scanning with scanning software, and then records the gluing thing after ferment treatment
Content.In the case of the most enzyme-added, processing secondary stock with remaining the same terms, measuring result contains for blank group gluing thing
Amount.
Table 1
As shown in Table 1, under the conditions of 40～55 DEG C, add gluing thing content in the secondary stock after high temperature esterase EST1 processes
Significantly reduce, stickies removal rate up to 58.9%～66.5%, the removal effect of lipase CALB to be substantially better than.Even if
Under 80 DEG C of hot conditionss, the treatment effect after addition esterase EST1 is the most ideal, and the stickies removal rate in secondary stock reaches
54.1%, and lipase CALB inactivates the most substantially, it is difficult to remove gluing thing.In view of different in paper-making process from waste paper
The temperature of workshop section is not quite similar, and the effect that esterase EST1 removes gluing thing in waste paper reclaims different workshop section is the most ideal.
Claims (10)
1. one kind utilizes high temperature esterase to remove the method for gluing thing in secondary stock, it is characterised in that described esterase is for deriving from
Esterase EstE1, the PDB ID:2C7B of grand genomic library.
Method the most according to claim 1, it is characterised in that comprise the steps: in secondary stock add 0.1U～
The esterase of 1.0U/g (relative to oven dry stock quality), processes time 45～120min, temperature 40～80 DEG C, and pH is 5.0～9.0,
Mixing speed 100～150rpm.
Method the most according to claim 2, it is characterised in that described esterase addition is 0.3～0.8U/g (relative to absolutely
Dry pulp quality).
Method the most according to claim 3, it is characterised in that the addition of described esterase is that 0.5U/g is (relative to over dry
Slurry).
Method the most according to claim 2, it is characterised in that described temperature is 40～60 DEG C.
Method the most according to claim 5, it is characterised in that described temperature is 55 DEG C.
Method the most according to claim 2, it is characterised in that described pH is 7.0～8.0.
Method the most according to claim 2, it is characterised in that the described process time is 60min；Described mixing speed is
120rpm。
9. according to the method described in claim 1-8 any one, it is characterised in that the enzyme of described esterase is lived as 125.0U/ml.
10. according to the method for the esterase treatment secondary stock described in claim 1-8 any one, it is characterised in that described waste paper
The slurry of slurry is dense is 4%～6%.
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| CN201610594944.5A CN106223096B (en) | 2016-07-26 | 2016-07-26 | A method of utilizing gluing object in high temperature esterase removal secondary stock |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111197036A (en) * | 2020-01-08 | 2020-05-26 | 中南大学 | Esterase Est-24 and coding gene and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101220567A (en) * | 2008-01-24 | 2008-07-16 | 广东轻工职业技术学院 | Method for controlling with 100aste newsprint paper deinked pulp esterase |
| CN104781398A (en) * | 2012-10-26 | 2015-07-15 | 罗尔公司 | Novel esterases in the treatment of cellulosic and lignocellulosic material |
-
2016
- 2016-07-26 CN CN201610594944.5A patent/CN106223096B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101220567A (en) * | 2008-01-24 | 2008-07-16 | 广东轻工职业技术学院 | Method for controlling with 100aste newsprint paper deinked pulp esterase |
| CN104781398A (en) * | 2012-10-26 | 2015-07-15 | 罗尔公司 | Novel esterases in the treatment of cellulosic and lignocellulosic material |
Non-Patent Citations (1)
| Title |
|---|
| JUNG-SUE BYUN ETAL.: "Crystal structure of hyperthermophilic esterase EstE1 and the", <<BMC STRUCTURAL BIOLOGY>> * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111197036A (en) * | 2020-01-08 | 2020-05-26 | 中南大学 | Esterase Est-24 and coding gene and application thereof |
| CN111197036B (en) * | 2020-01-08 | 2022-07-05 | 中南大学 | Esterase Est-24 and coding gene and application thereof |
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