CN107354144A - A kind of preparation method of the glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier - Google Patents
A kind of preparation method of the glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier Download PDFInfo
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- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
- C12N11/12—Cellulose or derivatives thereof
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- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/04—Enzymes or microbial cells immobilised on or in an organic carrier entrapped within the carrier, e.g. gel or hollow fibres
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
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- C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
- C12Y101/03—Oxidoreductases acting on the CH-OH group of donors (1.1) with a oxygen as acceptor (1.1.3)
- C12Y101/03004—Glucose oxidase (1.1.3.4)
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Abstract
The present invention provides a kind of preparation method of the glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier, and specific preparation method is：Glucose oxidase is added in bacterial cellulose solution, solvent flashing is vibrated, sealing, stand at low temperature, takes out, obtains adsorbing the bacterial cellulose solution of glucose oxidase；The bacterial cellulose solution of absorption glucose oxidase will be added in algal biomass, order input anionic polymer and cationic surfactant, or cationic polymer and anion surfactant, stirring at low speed, filtering retention obtain the glucose oxidase of algae flocks body cladding；The glucose oxidase that algae flocks body is fixed is added in solvent, adds auxiliary agent, is well mixed, obtains the glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier.Solidification glucose oxidase stability prepared by the present invention is good, and enzyme activity is high, good with fabric bonding force, and preparation method is easy, and controllability is strong.
Description
Technical field
The invention belongs to solidify enzyme field of material technology, and in particular to a kind of to coagulate body as carrier using bacteria cellulose and algae
The preparation method of the glucose oxidase of solidification.
Background technology
As the organic matter caused by living cells with catalytic action, have compared with other catalyst to replace biology enzyme
The advantage in generation, such as nontoxic, environmentally friendly, treatment conditions are gentle, and effect is single-minded, and dosage is few, and the enzyme for reacting release also may be used
To continue to be catalyzed another reaction, the features such as easily administering is polluted, the weaving of biology enzyme and application are more and more extensive, especially exist
Field of textile industry is widely used.
In field of textiles, the biological treatment of biological enzyme formulation is mainly applied to yam surface with specific enzyme preparation, with
Reach clean textile surface, reduce surface fine hair, prevent fabric pilling, and the fabric after processing is weightless small, fiber damage
Hinder the small purpose of degree.At present, the use of relatively broad enzyme preparation is mainly cellulase, protease, shallow lake in field of textiles
Powder enzyme, pectase, lipase, catalase, laccase and glucose oxidase, wherein glucose oxidase belong to redox
Old processing is done in enzyme, the bleaching for being mainly used in fabric portion or whole fabrics, and the fibre damage degree of the fabric after processing is low, knits
Thing has that feel is abundant, any surface finish, it is soft plentiful the characteristics of.
Curing technology refers to strong cell microorganism or electrodes method using chemically or physically means in the sky of restriction
Between region and its is kept a kind of active and repeatedly used basic fundamental, immobilised enzymes is compared with dissociating not, immobilised enzymes
While keeping its efficient, single-minded and gentle processing is specific, storage stability height, separation and recovery are also showed easily, can
Repeatedly use, the advantages that operation continuous controllable, simple process.A kind of application is solid disclosed in Chinese patent CN101302716A
Surely the method that laccase carries out complete cotton knitted fabric dyeing post processing is changed, using sodium alginate, gelatina polyethylene glycol enzyme pearl to laccase
Embedded, calcium chloride solidification and glutaraldehyde cross-linking being fixed laccase, immobilization laccase is handled to dyeing at 45-55 DEG C
After fabric, processing may be recovered.The disclosed immobilized lipases using red halloysite as carrier of Chinese patent CN103146675B
Preparation method, processing red halloysite is added in lipase after hydroxylating and silane post processing, and low temperature vibration is fixed, and freezing is dry
It is dry, then coated with acid sodium solution, calcium chloride solidification, the lipase for being fixed.From above-mentioned prior art, at present
The technique for fixing of field of textiles biology enzyme have it is a variety of, but for glucose oxidase rapid solidification techniques in terms of research
It is actually rare.
The content of the invention
Solidified the technical problem to be solved in the present invention is to provide a kind of as carrier using bacteria cellulose and algae flocks body
The preparation method of glucose oxidase, glucose oxidase is adsorbed first with bacteria cellulose, is then added to algal biomass
In, using the polymer and surfactant of different electric charges, the rapid flocculation and cladding of algal biomass are realized, is finally added
Product is prepared in auxiliary agent.Solidification glucose oxidase stability prepared by the present invention is good, and enzyme activity is high, with fabric bonding force
Good, preparation method is easy, and controllability is strong, green.
In order to solve the above technical problems, the technical scheme is that：
A kind of preparation method of the glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier, its feature
It is：Comprise the following steps：
(1) glucose oxidase is added in bacterial cellulose solution, vibrated while solvent flashing, sealing, put
4-6h is stood in 4 DEG C of refrigerators, is taken out, obtains adsorbing the bacterial cellulose solution of glucose oxidase；
(2) it is the bacterium that absorption glucose oxidase prepared by step (1) is added in 0.1-4g/L algal biomass is fine
Tie up plain solution, after being well mixed, order puts into anionic polymer and cationic surfactant, or cationic polymer and
Anion surfactant, stirring at low speed, cotton-shaped particle is formed, retain to obtain the Portugal of algae flocks body cladding through coarse filtration membrane filtration
Grape carbohydrate oxidase；
(3) glucose oxidase that the algae flocks body for preparing step (2) is fixed is added in solvent, adds auxiliary agent,
It is well mixed, obtain the glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier.
As the preferred of above-mentioned technical proposal, in the step (1), the raw material of bacteria cellulose in bacterial cellulose solution
For coconut palm fruit bacteria cellulose, the content of coconut palm fruit bacteria cellulose is 2-8wt%.
As the preferred of above-mentioned technical proposal, in the step (1), the mass ratio of glucose oxidase and bacteria cellulose
For 1:3-5.
As the preferred of above-mentioned technical proposal, in the step (1), the bacterial cellulose solution of glucose oxidase is adsorbed
Solid content be not less than 25%.
As the preferred of above-mentioned technical proposal, in the step (2), anionic polymer is polyacrylic acid PAA, cation
Surfactant is Cetylpyridinium Chloride CPC, and cationic polymer is polyethyleneimine PEI, and anion surfactant is dodecane
Base sodium sulphate SDS.
As the preferred of above-mentioned technical proposal, in the step (2), the amount ratio of polymer and surfactant is 90-
100ppm/mM。
As the preferred of above-mentioned technical proposal, in the step (2), the glucose oxidase of algae flocks body cladding contains
Water rate is less than 65%.
As the preferred of above-mentioned technical proposal, in the step (2), algae in the glucose oxidase of algae flocks body cladding
The mass fraction of class flocks body is 15-30%.
As the preferred of above-mentioned technical proposal, in the step (3), auxiliary agent is gelling agent, thickener and surfactant,
The gelling agent is calcium carbonate, and the thickener is sodium cellulose glycolate, and the surfactant is lived for non-ionic surface
Property agent.
As the preferred of above-mentioned technical proposal, in the step (3), the dosage of auxiliary agent is 0.1-5%.
Compared with prior art, the invention has the advantages that：
(1) glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier prepared by the present invention is by grape
Carbohydrate oxidase is formed by bacteria cellulose absorption and algae flocks body cladding, and the crystallinity of the molecule of bacteria cellulose is high, point
The sub- degree of orientation is good, and mechanical strength is more than plant cellulose, intramolecular and it is intermolecular hydrogen bond action be present, there is high-selenium corn characteristic
And with reference to outlet capacity, glucose oxidase can be adsorbed onto between inside and the bacteria cellulose of bacteria cellulose, and pass through
The solidification for combining to form microballoon, being more beneficial for glucose oxidase between concentration technique promotion bacteria cellulose is vibrated, afterwards
Using the flocks performance of algal biomass, one layer of algae flocks material is wrapped up on the surface of bacteria cellulose and glucose oxidase
Material, using the effect of the polymer and surfactant of different electric charges, quick flocks and the sedimentation of algal biomass are realized, and it is aobvious
Writing reduces the water content of algae flocks material, and therefore, glucose oxidase coats through double-deck three-dimensional network material, and stability is good,
Impact resistance is good, is advantageous to keep the activity of enzyme for a long time, and covering material and the compatibility of fabric are good, are more beneficial for afterwards
Use in textile finishing.
(2) the glucose oxidase preparation side solidified using bacteria cellulose and algae flocks body as carrier prepared by the present invention
Method is simple, and controllability is strong, and the solidification raw material of selection is all green natural materials, and cost is low, and technique is simple, the solidification of preparation
Glucose oxidase enzymatic activity it is strong, stability is good, and the rate of recovery is good, and impact resistance is good, good with the adhesion strength of fabric, more favorably
Acted in glucose oxidase.
Embodiment
The present invention is described in detail below in conjunction with specific embodiment, herein illustrative examples and explanation of the invention
For explaining the present invention, but it is not as a limitation of the invention.
Embodiment 1：
(1) it is 1 according to the mass ratio of glucose oxidase and bacteria cellulose:3, glucose oxidase is added to
In 2wt% coconut palm fruit bacterial cellulose solution, vibrate while solvent flashing, sealing, are placed in 4 DEG C of refrigerators and stand 4h, take
Go out, obtain the bacterial cellulose solution for the absorption glucose oxidase that solid content is 25%.
(2) bacterial cellulose solution for adsorbing glucose oxidase will be added in 0.1g/L algal biomass, mixing is equal
After even, order puts into 45ppm anionic polymer polyacrylic acid PAA and 0.5mM cationic surfactant Cetylpyridinium Chloride CPC,
The stirring at low speed 20min under 100r/min speed, the cotton-shaped particle that moisture content is 64.7% is formed, is retained through coarse filtration membrane filtration
Obtain the glucose oxidase of algae flocks body cladding, wherein algae flocks body in the glucose oxidase of algae flocks body cladding
Mass fraction be 15%.
(3) glucose oxidase that algae flocks body is fixed is added in deionized water, it is 1 to add mass ratio:1:1
Calcium carbonate gelling agent, sodium cellulose glycolate thickener and aliphatic acid polyethenoxy ether nonionic surface active agent, mixing are equal
It is even, the glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier is obtained, the wherein total dosage of auxiliary agent is
0.1%.
Embodiment 2：
(1) it is 1 according to the mass ratio of glucose oxidase and bacteria cellulose:5, glucose oxidase is added to
In 8wt% coconut palm fruit bacterial cellulose solution, vibrate while solvent flashing, sealing, are placed in 4 DEG C of refrigerators and stand 6h, take
Go out, obtain the bacterial cellulose solution for the absorption glucose oxidase that solid content is 27%.
(2) bacterial cellulose solution for adsorbing glucose oxidase will be added in 4g/L algal biomass, be well mixed
Afterwards, order input 100ppm anionic polymer polyacrylic acid PAA and 1mM cationic surfactant Cetylpyridinium Chloride CPC,
Stirring at low speed 30min under 150r/min speed, the cotton-shaped particle that moisture content is 63.7% is formed, is retained through coarse filtration membrane filtration
To the glucose oxidase of algae flocks body cladding, algae flocks body wherein in the glucose oxidase of algae flocks body cladding
Mass fraction is 30%.
(3) glucose oxidase that algae flocks body is fixed is added in deionized water, it is 1 to add mass ratio:1:1
Calcium carbonate gelling agent, sodium cellulose glycolate thickener and aliphatic acid polyethenoxy ether nonionic surface active agent, mixing are equal
It is even, the glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier is obtained, the wherein total dosage of auxiliary agent is
5%.
Embodiment 3：
(1) it is 1 according to the mass ratio of glucose oxidase and bacteria cellulose:4, glucose oxidase is added to
In 5wt% coconut palm fruit bacterial cellulose solution, vibrate while solvent flashing, sealing, are placed in 4 DEG C of refrigerators and stand 5h, take
Go out, obtain the bacterial cellulose solution for the absorption glucose oxidase that solid content is 27%.
(2) bacterial cellulose solution for adsorbing glucose oxidase will be added in 2g/L algal biomass, be well mixed
Afterwards, order input 76ppm cationic polymer polyethyleneimine PEI and 0.8mM anion surfactant dodecyl sulphates
Sodium SDS, the stirring at low speed 25min under 120r/min speed, the cotton-shaped particle that moisture content is 64.6% is formed, through coarse filtration film mistake
Filter retention obtains the glucose oxidase of algae flocks body cladding, wherein algae in the glucose oxidase of algae flocks body cladding
The mass fraction of flocks body is 20%.
(3) glucose oxidase that algae flocks body is fixed is added in deionized water, it is 1 to add mass ratio:1:1
Calcium carbonate gelling agent, sodium cellulose glycolate thickener and aliphatic acid polyethenoxy ether nonionic surface active agent, mixing are equal
It is even, the glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier is obtained, the wherein total dosage of auxiliary agent is
0.5%.
Embodiment 4：
(1) it is 1 according to the mass ratio of glucose oxidase and bacteria cellulose:3.5, glucose oxidase is added to
In 5wt% coconut palm fruit bacterial cellulose solution, vibrate while solvent flashing, sealing, be placed in 4 DEG C of refrigerators and stand 5.5h,
Take out, obtain the bacterial cellulose solution for the absorption glucose oxidase that solid content is 26%.
(2) bacterial cellulose solution for adsorbing glucose oxidase will be added in 1.35g/L algal biomass, mixing is equal
After even, order puts into 100ppm cationic polymer polyethyleneimine PEI and 1mM anion surfactant dodecyl sulphates
Sodium SDS, the stirring at low speed 20min under 150r/min speed, the cotton-shaped particle that moisture content is 63.7% is formed, through coarse filtration film mistake
Filter retention obtains the glucose oxidase of algae flocks body cladding, wherein algae in the glucose oxidase of algae flocks body cladding
The mass fraction of flocks body is 25%.
(3) glucose oxidase that algae flocks body is fixed is added in deionized water, it is 1 to add mass ratio:1:1
Calcium carbonate gelling agent, sodium cellulose glycolate thickener and aliphatic acid polyethenoxy ether nonionic surface active agent, mixing are equal
It is even, the glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier is obtained, the wherein total dosage of auxiliary agent is
1%.
Embodiment 5：
(1) it is 1 according to the mass ratio of glucose oxidase and bacteria cellulose:4.5, glucose oxidase is added to
In 7wt% coconut palm fruit bacterial cellulose solution, vibrate while solvent flashing, sealing, are placed in 4 DEG C of refrigerators and stand 6h, take
Go out, obtain the bacterial cellulose solution for the absorption glucose oxidase that solid content is 28%.
(2) bacterial cellulose solution for adsorbing glucose oxidase will be added in 0.9g/L algal biomass, mixing is equal
After even, order puts into 70ppm anionic polymer polyacrylic acid PAA and 0.7mM cationic surfactant Cetylpyridinium Chloride CPC,
The stirring at low speed 25min under 110r/min speed, the cotton-shaped particle that moisture content is 64.1% is formed, is retained through coarse filtration membrane filtration
Obtain the glucose oxidase of algae flocks body cladding, wherein algae flocks body in the glucose oxidase of algae flocks body cladding
Mass fraction be 18%.
(3) glucose oxidase that algae flocks body is fixed is added in deionized water, it is 1 to add mass ratio:1:1
Calcium carbonate gelling agent, sodium cellulose glycolate thickener and aliphatic acid polyethenoxy ether nonionic surface active agent, mixing are equal
It is even, the glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier is obtained, the wherein total dosage of auxiliary agent is
3%.
Embodiment 6：
(1) it is 1 according to the mass ratio of glucose oxidase and bacteria cellulose:5, glucose oxidase is added to
In 2wt% coconut palm fruit bacterial cellulose solution, vibrate while solvent flashing, sealing, are placed in 4 DEG C of refrigerators and stand 6h, take
Go out, obtain the bacterial cellulose solution for the absorption glucose oxidase that solid content is 26%.
(2) bacterial cellulose solution for adsorbing glucose oxidase will be added in 3g/L algal biomass, be well mixed
Afterwards, order input 90ppm cationic polymer polyethyleneimine PEI and 1mM anionics
SDS, the stirring at low speed 25min under 120r/min speed, the cotton-shaped particle that moisture content is 64.4% is formed, through coarse filtration membrane filtration
Retention obtains the glucose oxidase of algae flocks body cladding, and wherein algae coagulates in the glucose oxidase of algae flocks body cladding
The mass fraction of flco is 27%.
(3) glucose oxidase that algae flocks body is fixed is added in deionized water, it is 1 to add mass ratio:1:1
Calcium carbonate gelling agent, sodium cellulose glycolate thickener and aliphatic acid polyethenoxy ether nonionic surface active agent, mixing are equal
It is even, the glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier is obtained, the wherein total dosage of auxiliary agent is
3.5%.
After testing, what prepared by embodiment 1-6 is glycoxidative as the grape that carrier solidifies using bacteria cellulose and algae flocks body
The immobilized enzyme of the glucose oxidase of the solidification of enzyme and prior art, the result of repeat performance are as follows：
As seen from the above table, the present invention is prepared glycoxidative as the grape that carrier solidifies using bacteria cellulose and algae flocks body
The enzyme activity of enzyme improves, and stability is good, and the pH value and temperature that can be born improve, and illustrate that heat endurance and resistance to acids and bases also improve.
The above-described embodiments merely illustrate the principles and effects of the present invention, not for the limitation present invention.It is any ripe
Know the personage of this technology all can carry out modifications and changes under the spirit and scope without prejudice to the present invention to above-described embodiment.Cause
This, those of ordinary skill in the art is complete without departing from disclosed spirit and institute under technological thought such as
Into all equivalent modifications or change, should by the present invention claim be covered.
Claims (10)
1. a kind of preparation method of the glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier, its feature exist
In comprising the following steps：
(1) glucose oxidase is added in bacterial cellulose solution, vibrated while solvent flashing, sealing, are placed in 4 DEG C
4-6h is stood in refrigerator, is taken out, obtains adsorbing the bacterial cellulose solution of glucose oxidase；
(2) bacteria cellulose of absorption glucose oxidase prepared by step (1) will be added in 0.1-4g/L algal biomass
Solution, after being well mixed, order puts into anionic polymer and cationic surfactant, or cationic polymer and it is cloudy from
Sub- surfactant, stirring at low speed, cotton-shaped particle is formed, retain to obtain the glucose of algae flocks body cladding through coarse filtration membrane filtration
Oxidizing ferment；
(3) glucose oxidase that the algae flocks body for preparing step (2) is fixed is added in solvent, adds auxiliary agent, mixing
Uniformly, the glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier is obtained.
A kind of 2. glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier according to claim 1
Preparation method, it is characterised in that：In the step (1), the raw material of bacteria cellulose is that coconut palm fruit is thin in bacterial cellulose solution
Fungin, the content of coconut palm fruit bacteria cellulose is 2-8wt%.
A kind of 3. glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier according to claim 1
Preparation method, it is characterised in that：In the step (1), the mass ratio of glucose oxidase and bacteria cellulose is 1:3-5.
A kind of 4. glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier according to claim 1
Preparation method, it is characterised in that：In the step (1), the solid content of the bacterial cellulose solution of glucose oxidase is adsorbed
Not less than 25%.
A kind of 5. glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier according to claim 1
Preparation method, it is characterised in that：In the step (2), anionic polymer is polyacrylic acid PAA, cation surface activating
Agent is Cetylpyridinium Chloride CPC, and cationic polymer is polyethyleneimine PEI, and anion surfactant is lauryl sodium sulfate
SDS。
A kind of 6. glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier according to claim 1
Preparation method, it is characterised in that：In the step (2), the amount ratio of polymer and surfactant is 90-100ppm/mM.
A kind of 7. glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier according to claim 1
Preparation method, it is characterised in that：In the step (2), the moisture content of the glucose oxidase of algae flocks body cladding is less than
65%.
A kind of 8. glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier according to claim 1
Preparation method, it is characterised in that：In the step (2), algae flocks body in the glucose oxidase of algae flocks body cladding
Mass fraction be 15-30%.
A kind of 9. glucose oxidase solidified using bacteria cellulose and algae flocks body as carrier according to claim 1
Preparation method, it is characterised in that：In the step (3), auxiliary agent is gelling agent, thickener and surfactant, the gelling
Agent is calcium carbonate, and the thickener is sodium cellulose glycolate, and the surfactant is nonionic surface active agent.
It is 10. according to claim 1 a kind of glycoxidative as the grape that carrier solidifies using bacteria cellulose and algae flocks body
The preparation method of enzyme, it is characterised in that：In the step (3), the dosage of auxiliary agent is 0.1-5%.
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Application publication date: 20171117 |