CN103740690A - Method for concentrating, stabilizing and separating liquid enzyme - Google Patents
Method for concentrating, stabilizing and separating liquid enzyme Download PDFInfo
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- CN103740690A CN103740690A CN201410000994.7A CN201410000994A CN103740690A CN 103740690 A CN103740690 A CN 103740690A CN 201410000994 A CN201410000994 A CN 201410000994A CN 103740690 A CN103740690 A CN 103740690A
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- 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
- C12N9/96—Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
Abstract
The invention provides a method for concentrating, stabilizing and separating liquid enzyme. The liquid enzyme comprises the following components by weight percent: 1.0-20.0% of macrocyclic molecular synthetic receptor, 0.1-50% of nonionic surfactant, 0.25-5% of enzyme stabilizer, 0.01-80% of biological enzyme, and the balance of polar solvent. The method comprises the following steps: mixing the components, and reacting for 1-24 hours under the conditions that the pH is 3-12 and the temperature is 0-80 DEG C; forming supramolecular microspheres containing the biological enzyme in the solution, wherein the particle size of the supramolecular microspheres is 10-1000nm. The selected materials are friendly to the environment, harmless, and easy to degrade. Above all, the method does not damage the activity of enzyme, the enzyme has high activity within an extended period, and the activity of the enzyme coated in the supramolecular microspheres is improved by 20% in comparison with the activity of common liquid enzyme. The activity loss of the biological enzyme is smaller than 10% four weeks after the liquid enzyme is applied to the formula of a textile emulsifier or a liquid detergent.
Description
Technical field
The present invention relates to liquid enzyme formulation, specifically belong to method concentrated, stable, separating liquid zymin.
Background technology
Liquid enzyme formulation is a kind of easy to use, environmental protection, the raw material that biological degradability is good.Single-minded catalysis characteristics and gentle reaction conditions, more and more used and paid attention in industry member.But, liquid enzyme formulation concentrated, stable and to separate be a difficult problem that is perplexing people always.
Certainly, there are many people to solve relevant liquid enzyme formulation in trial searching way concentrated, the problem of stablizing and separating.And propose some feasible technology, have interpolation enzyme stabilizers, membrane separation technique, microwave vacuum method of enrichment, reagent is concentrated, and temperature-sensitive hydrogel method etc.
Adding enzyme stabilizers method is more traditional stablizing, the method for concentrated enzyme preparation.In US20070134375A1, the people such as Andreas Habich relate to a kind of stable solid-state or liquid enzyme preparation in invention, it comprises at least one enzyme and at least one stablizer that is selected from gum arabic, at least one vegetable-protein and composition thereof, has improved storage and the stability of zymin.
Membrane separation technique is an isolation technique of rising the sixties in 20th century, it is by means of the promotion of outside energy or chemical potential difference, by the osmosis of certain films, realize that gas or liquid that two components or polycomponent are mixed separate, the technology of classification, purification and enrichment.Conventional mould material has cellulose acetate (CA), polyacrylonitrile (PAN), poly-alum (PSF), polymeric amide (PA), polyvinylidene difluoride (PVDF) (PVDF) etc.Common membrane separation technique: (1) micro-filtration (Micro filtration, MF); (2) ultrafiltration (Ultrafiltration, UF); (3) nanofiltration (Nanofiltration, NF); (4) reverse osmosis (Reverse osmosis, RO).At present, in Enzymes Industry, applying more is micro-filtration and ultra-filtration technique, especially ultra-filtration technique, and ultra-filtration technique is employed more and more widely with the use of combining of other isolation technique in recent years.
The propositions such as nineteen sixty-five Blatt are carried out the concentrated of microorganism by membrane separation technique, and test.The all external pressure tubular membrane of using of Wang Ping are by concentration extraction bromeline in pineapple juice, and enzyme work can concentrate nearly 4 times, enzyme rejection 95% in concentrated solution, and the rate of recovery 79.8%, enzyme powder yield exceeds 47% than traditional method.In CN201110389274, the people such as Zhou Xing have carried out a series of processing with ultrafiltration and nanofiltration to the thick enzyme of nitrilase, finally prepare the nitrilase product that concentration is higher.In patent CN103122343A, the people such as Wei Xingye concentrate broken glue enzyme compound bacteria-fermented liquid with the ultra-filtration membrane of tubular type or tubular fibre formula, and result obtains the enzyme product that purity is higher.
Using membrane separating technology concentrates and refines enzyme, and operating process is simple, has reduced the chance of living contaminants and enzyme deactivation, has improved the rate of recovery of enzyme and has improved the quality of product.
Also someone attempts by microwave vacuum method of enrichment, zymin being concentrated at present, and has obtained success.In CN102899303A, the people such as Li Bing disclose a kind of microwave vacuum concentration method of papoid solution, and they concentrate melon proteolytic enzyme with microwave vacuum concentration method, have obtained papoid concentrated solution.This method concentration process temperature is low, and concentrated speed is fast, and the papoid concentrated solution enzyme rate of recovery alive obtaining is high.Meanwhile, equipment is less scaling, and energy consumption is low.
Reagent for method of enrichment polyoxyethylene glycol (PEG) absorb the moisture in enzyme liquid, this method is easy, efficiency is high, but reagent price is more expensive, as suitability for industrialized production, does not consider because of Cost Problems.
Reduction vaporization method of enrichment adopts reliever that enzyme liquid to be concentrated is carried out at certain vacuum tightness and temperature, and to heat-sensitive substances such as enzyme liquid, reliever adopts centrifugal-film evaporator and scraper-type vaporizer more, but this method energy consumption is larger.
Temperature-sensitive hydrogel also can concentrate with Fen Li enzyme, and it is according to being concentrated the size of separate substance and cross-linking density or the monomer structure of character design gel.But gel during to the separation efficiency low temperature of albumen and enzyme (lower than phase transition temperature) separation efficiency very high, during high temperature, (higher than phase transition temperature) separation efficiency reduces, near separation efficiency generation hop phase transition temperature.(Wang Jin hall Zhong Hui Hongjun ZHU Zhang Weiguang, Nanjing University of Chemical Technology's journal, 1998,20,2)
At present, also have some technology to mention and carry out enclosing liquid zymin with supramolecule, but because the feature of biological enzyme three-dimensional structure, in supramolecule technology in various degree all run into enzyme deactivation and the too low problem of encapsulation biological enzyme content.
Summary of the invention:
The object of the invention is to the problem for the ubiquitous enzyme deactivation of zymin, utilize the technology of supermolecule nano microballoon wrapping biological enzyme, a kind of method of concentrated, separation, stabilising liq enzyme is provided.
The method of a kind of concentrated, separation provided by the invention, stabilising liq enzyme,
Components based on weight percentage: macrocycle molecule synthesis of receptor 1.0-20.0%, nonionic surface active agent 0.1%-50%, enzyme stabilizers 0.25%-5%, biological enzyme 0.01%-80%, polar solvent surplus;
By said components proportioning, mix, control pH3-12, temperature 0-80 ℃, reaction 1-24h.In solution, formed the supramolecule microballoon containing biological enzyme, size is between 10nm-1000nm.
Preferred component proportion is above: macrocycle molecule synthesis of receptor 2.0-18.0%, nonionic surface active agent 0.5%-40%, enzyme stabilizers 0.5%-4.5%, biological enzyme 0.5%-70%, polar solvent surplus;
Between the preferred 5-70 of described temperature of reaction ℃, more preferably between 10-50 ℃.
The preferred 5-11 of described pH value.
Described macrocycle molecule synthesis of receptor is crown ether, ring kind, cyclodextrin, Cucurbituril, calixarene or post aromatic hydrocarbons etc.
Described crown ether is 15-crown ether-5,15-crown ether-6 or hexaoxacyclooctadecane-6-6 etc.;
Ring kind be to encircle greatly kind or little ring kind etc.;
Cyclodextrin is alpha-cylodextrin, beta-cyclodextrin, γ-cyclodextrin etc.;
Calixarene is cup [4] aromatic hydrocarbons or cup [5] aromatic hydrocarbons etc.;
Post aromatic hydrocarbons is post [5] aromatic hydrocarbons or post [6] aromatic hydrocarbons etc.
Described polar solvent is water or straight chain alcohol or ketone.
Described enzyme stabilizers can be dibasic alcohol or trivalent alcohol; Dibasic alcohol can be ethylene glycol, 1,2-PD, 1,3-PD, 1,2-butyleneglycol, 1,3 butylene glycol, BDO or 1,5-PD; Trivalent alcohol can be glycerol, BT or 1,2,5-, penta triol.
Described nonionic surface active agent, its structural formula can be PEO
x-PPO
y-PEO
x, R-C
6h
4-O-(CH
2cH
2o)
n-H, R-CO-(OC
2h
4)
n-OH, RO-(CH
2cH
2o)
n-H or R-COO-(CH
2cH
2o)
n-H; And its molecular weight >10000, HLB value is between 5-60.
Described biological enzyme be in proteolytic enzyme, polygalacturonase, cellulase, lipase, hemicellulase, lipase, Novi's letter enzyme and subtilase enzymes any one or multiple.
Separation with prior art, method concentrated and stabilising liq zymin is compared, the present invention proposes to use macrocycle molecule synthesis of receptor, enzyme stabilizers, nonionic surface active agent and biological enzyme build supramolecule microballoon and reach separation, the object of concentrated and stabilising liq zymin, and selected materials of the present invention is environmental protection, without harm, the easily material of degraded.The most important thing is that this method do not damage the activity of enzyme, and make the activity of enzyme have higher activity in significant period of time, be wrapped in enzymic activity in supramolecule microballoon and improve 20% compared with the activity of common liq enzyme, in the formula of be applied to weave emulsifying agent or liquid washing agent, the loss of activity of surrounding artifact enzyme is less than 10%.
Accompanying drawing explanation:
Fig. 1 embodiment 1 dynamic light scattering (DLS) supramolecule microsphere particle size distribution figure
Fig. 2 embodiment 2 dynamic light scatterings (DLS) supramolecule microsphere particle size distribution figure
Fig. 3 embodiment 3 dynamic light scatterings (DLS) supramolecule microsphere particle size distribution figure
Fig. 4 embodiment 4 dynamic light scatterings (DLS) supramolecule microsphere particle size distribution figure
Fig. 5 embodiment 5 dynamic light scatterings (DLS) supramolecule microsphere particle size distribution figure
Fig. 6 embodiment 6 dynamic light scatterings (DLS) supramolecule microsphere particle size distribution figure
Fig. 7 embodiment 7 dynamic light scatterings (DLS) supramolecule microsphere particle size distribution figure
Fig. 8 embodiment 8 dynamic light scatterings (DLS) supramolecule microsphere particle size distribution figure
Fig. 9 embodiment 9 dynamic light scatterings (DLS) supramolecule microsphere particle size distribution figure
Figure 10 embodiment 10 dynamic light scatterings (DLS) supramolecule microsphere particle size distribution figure
Figure 11 embodiment 10 supramolecule microballoon electromicroscopic photographs
Embodiment:
Embodiment 1
By nonionic surface active agent EO
11pO
69eO
11(HLB value 6.0) 0.9g is dissolved in 50g water, then gets liquid protein zymin 1g and mixes with it, and mixture is added drop-wise in 0.8g alpha-cylodextrin, then adds 2.4g1, and 2-propylene glycol stablizer mixes.
By the above-mentioned liquid mixing, after sealing, at 20 ℃, stirring reaction 20h, determines that by GB GB/T23527-2009 the enzyme activity of liquid protein zymin sees the following form.
Dynamic light scattering Dynamic Light Scattering(DLS) nanostructure that is used for testing sample.Supramolecule microsphere particle size distribution (see figure 1), parcel enzyme particulate median size is in 270nm left and right.
Embodiment 2
By nonionic surface active agent EO
133pO
50eO
133(HLB value 26.0) 9g is dissolved in 50g ethanol, then gets liquid aliphatic zymin 38g and mixes with it, and mixture is added drop-wise in 1.5g beta-cyclodextrin, then adds 0.5g1, and ammediol stablizer mixes.
By the above-mentioned liquid mixing, after sealing, at 20 ℃, stirring reaction 22h, determines that by GB GB/T23527-2009 the enzyme activity of liquid aliphatic zymin sees the following form.
The nanostructure that dynamic light scattering Dynamic Light Scattering (DLS) is used for testing sample.Supramolecule microsphere particle size distribution (see figure 2), parcel enzyme particulate median size is in 298nm left and right.
Embodiment 3
By nonionic surface active agent EO
3pO
43eO
3(HLB value 6.0) 20g is dissolved in 100g ethyl ketone, then gets liquid hemicellulose zymin 18g and mixes with it, and mixture is added drop-wise in the little ring of 10g kind, then adds 0.55g1, and 2-butyleneglycol stablizer mixes.
By the above-mentioned liquid mixing, after sealing, at 30 ℃, stirring reaction 24h, determines that by GB GB/T23527-2009 the enzyme activity of liquid hemicellulose zymin sees the following form.
The nanostructure that dynamic light scattering Dynamic Light Scattering (DLS) is used for testing sample.Supramolecule microsphere particle size distribution is as figure below (see figure 3), and parcel enzyme particulate median size is in 806nm left and right.
Embodiment 4
By nonionic surface active agent EO
37pO
56eO
37(HLB value 14.0) 1.5g is dissolved in 50g acetone, then gets liquid cellulase preparation 10g and mixes with it, and mixture is added drop-wise in 5g Cucurbituril, then adds 1.2g1, and 3-butyleneglycol stablizer mixes.
By the above-mentioned liquid mixing, after sealing, at 36 ℃, stirring reaction 23h, determines that by GB GB/T23527-2009 the enzyme activity of liquid cellulase preparation sees the following form.
The nanostructure that dynamic light scattering Dynamic Light Scattering (DLS) is used for testing sample.Supramolecule microsphere particle size distribution is as figure below (see figure 4), and parcel enzyme particulate median size is in 353nm left and right.
Embodiment 5
Nonionic surface active agent polyoxyethylene nonylphenol ether 2.0g is dissolved in 30g propyl alcohol, then get liquid cellulase and hemicellulase composite preparation 8.0g mixes with it, mixture is added drop-wise in 0.7g γ-cyclodextrin, then adds 0.9g1, and 4-butyleneglycol stablizer mixes.
By the above-mentioned liquid mixing, after sealing, at 40 ℃, stirring reaction 24h determines that by GB GB/T23527-2009 the enzyme activity of liquid cellulase and hemicellulase composite preparation sees the following form.
The nanostructure that dynamic light scattering Dynamic Light Scattering (DLS) is used for testing sample.Supramolecule microsphere particle size distribution is as figure below (see figure 5), and parcel enzyme particulate median size is in 431nm left and right.
Embodiment 6
By nonionic surface active agent EO
103pO
39eO
103(HLB value 26.0) 3.0g is dissolved in 50g butanols, then gets liquid cellulase preparation 10g and mixes with it, and mixture is added drop-wise in 2.8g calixarene, then adds 1.5g1, and 5-pentanediol stablizer mixes.
By the above-mentioned liquid mixing, after sealing, at 25 ℃, stirring reaction 20h determines that by GB GB/T23527-2009 the enzyme activity of liquid cellulase preparation sees the following form.
The nanostructure that dynamic light scattering Dynamic Light Scattering (DLS) is used for testing sample.Supramolecule microsphere particle size distribution is as figure below (see figure 6), and parcel enzyme particulate median size is in 130nm left and right.
Embodiment 7
By nonionic surface active agent: lauric acid polyoxyethylene ester LAE-4,90.8g are dissolved in 40g ethyl ketone, then get liquid Bacillus subtilus zymin 0.6g and mix with it, mixture is added drop-wise in 0.6g beta-cyclodextrin, then adds 1.0g1,2,4-trihydroxybutane stablizer mixes.
By the above-mentioned liquid mixing, after sealing, at 20 ℃, stirring reaction 20h, determines that by GB GB/T23527-2009 the enzyme activity of liquid Bacillus subtilus zymin sees the following form.
The nanostructure that dynamic light scattering Dynamic Light Scattering (DLS) is used for testing sample.Supramolecule microsphere particle size distribution is as figure below (see figure 7), and parcel enzyme particulate median size is in 406nm left and right.
Embodiment 8
Nonionic surface active agent methacrylic Soxylat A 25-7 0.3g is dissolved in 10g acetone, then gets pectinase preparation 0.6g and mix with it, mixture is added drop-wise in 0.6g post [5] aromatic hydrocarbons, then adds 0.5g glycerol stablizer to mix.
By the above-mentioned liquid mixing, after sealing, at 20 ℃, stirring reaction 20h determines that by GB GB/T23527-2009 the enzyme activity of liquid lipase preparation sees the following form.
The nanostructure that dynamic light scattering Dynamic Light Scattering (DLS) is used for testing sample.Supramolecule microsphere particle size distribution is as figure below (see figure 8), and parcel enzyme particulate median size is in 676nm left and right.
Embodiment 9
By nonionic surface active agent EO
42pO
16eO
42(HLB value 26.0) 0.5g is dissolved in 30g propyl alcohol, then gets liquid protease and polygalacturonase composite preparation 0.5g mixes with it, and mixture is added drop-wise in 0.6g beta-cyclodextrin, then adds 1.0g1, and 3-butyleneglycol stablizer mixes.
By the above-mentioned liquid mixing, after sealing, at 20 ℃, stirring reaction 20h determines that by GB GB/T23527-2009 the enzyme activity of liquid protease and polygalacturonase composite preparation sees the following form.
The nanostructure that dynamic light scattering Dynamic Light Scattering (DLS) is used for testing sample.Supramolecule microsphere particle size distribution is as figure below (see figure 9), and parcel enzyme particulate median size is in 588nm left and right.
Embodiment 10
By nonionic surface active agent EO
6pO
34eO
6(HLB value 5.0) 0.4g is dissolved in 30g ethanol, then gets pectinase preparation 0.5g and mixes with it, and mixture is added drop-wise in 0.6g γ-cyclodextrin, then adds 1.0g1, and 2,5-, penta triol stablizer mixes.
By the above-mentioned liquid mixing, after sealing, at 20 ℃, stirring reaction 20h, determines that by GB GB/T23527-2009 the enzyme activity of pectinase preparation sees the following form.
The nanostructure that dynamic light scattering Dynamic Light Scattering (DLS) is used for testing sample.Supramolecule microsphere particle size distribution is if figure below (see figure 10) parcel enzyme particulate median size is in 245nm left and right.
In addition, we also use transmission electron microscope (TEM) and scanning electron microscope (SEM) to observe molecule pattern, as figure below (seeing Figure 11), and the SEM figure that Figure 11 (A) is bag enzyme microballoon, as can be seen from Fig., the profile of this microballoon is the spheroidal of standard comparatively.From scheming, can read, the particle diameter of this microballoon is about 120nm.Figure 11 (B) is the TEM figure of bag enzyme microballoon, from scheming, can find out, this microballoon is solid microsphere, and this is the bag enzyme solid microsphere of standard, and particle diameter is approximately 120nm left and right.
Claims (15)
1. a method for concentrated, separation, stabilising liq enzyme, is characterized in that:
Components based on weight percentage: macrocycle molecule synthesis of receptor 1.0-20.0%, nonionic surface active agent 0.1%-50%, enzyme stabilizers 0.25%-5%, biological enzyme 0.01%-80%, polar solvent surplus;
By said components proportioning, mix, control pH3-12, temperature 0-80 ℃, reaction 1-24h.
2. the method for a kind of concentrated, separation as claimed in claim 1, stabilising liq enzyme, it is characterized in that: described components based on weight percentage: macrocycle molecule synthesis of receptor 2.0-18.0%, nonionic surface active agent 0.5%-40%, enzyme stabilizers 0.5%-4.5%, biological enzyme 0.5%-70%, polar solvent surplus.
3. the method for a kind of concentrated, separation as claimed in claim 1, stabilising liq enzyme, is characterized in that: described temperature of reaction is 5-70 ℃.
4. the method for a kind of concentrated, separation as claimed in claim 1, stabilising liq enzyme, is characterized in that: described pH value is 5-11.
5. the method for a kind of concentrated, separation as claimed in claim 1, stabilising liq enzyme, is characterized in that: described macrocycle molecule synthesis of receptor is crown ether, ring kind, cyclodextrin, Cucurbituril, calixarene or post aromatic hydrocarbons.
6. the method for a kind of concentrated, separation as claimed in claim 5, stabilising liq enzyme, is characterized in that: described crown ether is 15-crown ether-5,15-crown ether-6 or hexaoxacyclooctadecane-6-6.
7. the method for a kind of concentrated, separation as claimed in claim 5, stabilising liq enzyme, is characterized in that: described ring kind is large ring kind or little ring kind.
8. the method for a kind of concentrated, separation as claimed in claim 5, stabilising liq enzyme, is characterized in that: described cyclodextrin is alpha-cylodextrin, beta-cyclodextrin or γ-cyclodextrin.
9. the method for a kind of concentrated, separation as claimed in claim 5, stabilising liq enzyme, is characterized in that: described calixarene is cup [4] aromatic hydrocarbons or cup [5] aromatic hydrocarbons.
10. the method for a kind of concentrated, separation as claimed in claim 5, stabilising liq enzyme, is characterized in that: described post aromatic hydrocarbons is post [5] aromatic hydrocarbons or post [6] aromatic hydrocarbons.
The method of 11. a kind of concentrated, separation as claimed in claim 1, stabilising liq enzyme, is characterized in that: described polar solvent is water or straight chain alcohol or ketone.
The method of 12. a kind of concentrated, separation as claimed in claim 1, stabilising liq enzyme, is characterized in that: described described enzyme stabilizers is dibasic alcohol or trivalent alcohol.
The method of 13. a kind of concentrated, separation as claimed in claim 12, stabilising liq enzyme, is characterized in that: described dibasic alcohol is ethylene glycol, 1,2-PD, 1,3-PD, 1,2-butyleneglycol, 1,3 butylene glycol, BDO or 1,5-PD; Described trivalent alcohol is glycerol, BT or 1,2,5-, penta triol.
The method of 14. a kind of concentrated, separation as claimed in claim 1, stabilising liq enzyme, is characterized in that: described nonionic surface active agent is that structural formula is PEO
x-PPO
y-PEO
x, R-C
6h
4-O-(CH
2cH
2o)
n-H, R-CO-(OC
2h
4)
n-OH, RO-(CH
2cH
2o)
n-H or R-COO-(CH
2cH
2o)
n-H, and its molecular weight >10000, the nonionic surface active agent of HLB value between 5-60.
The method of 15. a kind of concentrated, separation as claimed in claim 1, stabilising liq enzyme, is characterized in that: described biological enzyme be in proteolytic enzyme, polygalacturonase, cellulase, lipase, hemicellulase, lipase, Novi's letter enzyme and subtilase enzymes any one or multiple.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106893574A (en) * | 2017-02-22 | 2017-06-27 | 中国石油化工股份有限公司 | A kind of pressure break slow release type biological enzyme breaker and preparation method thereof |
| CN108559770A (en) * | 2018-01-16 | 2018-09-21 | 扬州大学 | Application of the Calixarene Derivatives in regulation and control bromelain and polyphenol oxidase activity |
| EP3540052A1 (en) | 2018-03-14 | 2019-09-18 | Indian Oil Corporation Limited | A stable lignocellulolytic enzyme composition |
| CN114929848A (en) * | 2019-12-20 | 2022-08-19 | 诺维信公司 | Stable liquid boron-free enzyme compositions |
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|---|---|---|---|---|
| CN102766540A (en) * | 2012-07-20 | 2012-11-07 | 山西大学 | Liquid enzyme stabilizing additive and preparation method and application thereof |
| CN103131557A (en) * | 2013-02-05 | 2013-06-05 | 山西勇宁记科技有限公司 | Enzymic preparation compound stabilizer used for liquid detergent |
| CN103131555A (en) * | 2013-02-05 | 2013-06-05 | 山西勇宁记科技有限公司 | Bio-enzyme preparation compound stabilizer for liquid detergent |
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2014
- 2014-01-02 CN CN201410000994.7A patent/CN103740690A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102766540A (en) * | 2012-07-20 | 2012-11-07 | 山西大学 | Liquid enzyme stabilizing additive and preparation method and application thereof |
| CN103131557A (en) * | 2013-02-05 | 2013-06-05 | 山西勇宁记科技有限公司 | Enzymic preparation compound stabilizer used for liquid detergent |
| CN103131555A (en) * | 2013-02-05 | 2013-06-05 | 山西勇宁记科技有限公司 | Bio-enzyme preparation compound stabilizer for liquid detergent |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106893574A (en) * | 2017-02-22 | 2017-06-27 | 中国石油化工股份有限公司 | A kind of pressure break slow release type biological enzyme breaker and preparation method thereof |
| CN106893574B (en) * | 2017-02-22 | 2019-08-16 | 中国石油化工股份有限公司 | A kind of pressure break slow release type biological enzyme breaker and preparation method thereof |
| CN108559770A (en) * | 2018-01-16 | 2018-09-21 | 扬州大学 | Application of the Calixarene Derivatives in regulation and control bromelain and polyphenol oxidase activity |
| CN108559770B (en) * | 2018-01-16 | 2021-07-09 | 扬州大学 | Application of calixarene derivative in regulation and control of activities of bromelain and polyphenol oxidase |
| EP3540052A1 (en) | 2018-03-14 | 2019-09-18 | Indian Oil Corporation Limited | A stable lignocellulolytic enzyme composition |
| US11028385B2 (en) | 2018-03-14 | 2021-06-08 | Indian Oil Corporation Limited | Stable lignocellulolytic enzyme composition |
| CN114929848A (en) * | 2019-12-20 | 2022-08-19 | 诺维信公司 | Stable liquid boron-free enzyme compositions |
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Application publication date: 20140423 |