CN109762671A - It is a kind of for stablizing the compound stabilizer of protease in liquid detergent - Google Patents
It is a kind of for stablizing the compound stabilizer of protease in liquid detergent Download PDFInfo
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- CN109762671A CN109762671A CN201910095742.XA CN201910095742A CN109762671A CN 109762671 A CN109762671 A CN 109762671A CN 201910095742 A CN201910095742 A CN 201910095742A CN 109762671 A CN109762671 A CN 109762671A
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Abstract
The invention discloses a kind of for stablizing the compound stabilizer of protease in liquid detergent, belongs to detergent applications.The present invention provides a kind of for stablizing the compound stabilizer of protease in liquid detergent, which is formed with ammonium citrate calcium, APG0810 and APG1214.Compared to borax, compound stabilizer provided by the invention can significantly improve enzyme activity conservation rate of the protease in laundry liquid under the conditions of room temperature and 40 DEG C.
Description
Technical field
The present invention relates to a kind of for stablizing the compound stabilizer of protease in liquid detergent, belongs to detergent applications.
Background technique
Protease is most widely used enzyme preparation in detergent industry, but the stability in liquid detergent is well below it
Stability in washing powder.Borax was once widely used in liquid enzymatic liquid detergent, but borax is forbidden by Europe at present
It is used in enzyme detergent；Therefore it is highly desirable to the substitute for seeking borax or improves protease by other approach and washing
Stability in clothing liquid.
The factor for influencing stability of the protease in liquid detergent, in addition to for example some nonionics of the active constituent in liquid detergent
Surfactant, the zymologic property of most important or protease itself.Currently, protease is helped to keep certain in detergent
The approach of stability include: transformation production protease strain gene, by albumen enzyme immobilization and addition stabilizer.Egg is transformed
White enzyme gene and enzyme is immobilized, all there are problems that technology complexity and higher cost.Stabilizer is added to keep enzyme
Method living is relatively simple, but stability it is good stabilizer or compound stabilizer it is less.Some stabilizers need to synthesize and close
It maintains secrecy at technique；The effect of some stabilizers, which belongs to, generally acknowledges preferable such as borax, and is limited by a variety of causes and be even forbidden to use.
Protease and detergent powder, that is, surfactant coexist or are easy in pole dilute aqueous solution (such as wash environment)
Stablized by stabilizer.But the surfactant of a certain concentration range is easy to inactivate enzyme.Therefore enzyme liquid detergent is as added
It is always a the problem of perplexing industry that the stabilizer of protease, which is chosen, in enzyme liquid detergent.Educational circles usually only notices that single surface is living
Property agent perhaps influence of the low concentration surfactant to enzyme activity or the testing conditions chosen be enzyme best enzyme activity condition without
It is liquid detergent (stablizing effect of such borax or other stabilizers can all be got well naturally)；In fact, detergent must be placed in enzyme
In, under detergent concentration, using borax as positive control, choosing stablizing effect is more than or the stabilizer close with borax,
The stabilizer of selection protease can really be facilitated.
From the affinity analysis to protein, possible stabilizer has dicarboxylate, polyalcohol, polyacid and some nothings
Machine salt.In inorganic salts, the effect of borate family stabilizer such as borax is relatively preferable, can keep adding at room temperature to a certain extent
The enzyme activity of enzyme liquid detergent was once widely used in liquid enzymatic liquid detergent；But current borax is forbidden washing enzyme by Europe
It washs in agent and uses.It is disclosed in the patent application of 107815354 A of Publication No. CN and utilizes polyhydric alcohol amine, polyalcohol, halogenation
The active retention agent that calcium, polyacid sodium, sulfonate, compatibility of citric acid obtain can provide a life steady in a long-term for protease
Dis environment；But the stabilizer that the method for the patent application publication uses simultaneously is too many, to the compatibility band of surfactant in formula
Uncertainty is carried out；And need the pH value of strict control reaction system that could make effect of the active retention agent to protease
Perform to maximum.CN201810585252.3 discloses a kind of Superconcentrated washing liquid and preparation method thereof, the Superconcentrated washing
Each raw material and its parts by weight of liquid match are as follows: 20-40 parts of water, 10-20 parts of coconut oil potassium, nonionic surfactant 40-70
Part, DE1-5 parts of coconut oleoyl amine, 1-5 parts of propylene glycol .1-1 parts of proteinase-10,0.1-1 parts of cellulase, EDT0.1-1 parts, lemon
0.1-1 parts sour, 0.1-1 parts of preservative, 0.1-1 parts of essence；The stability of protease is not referred to.Publication No. CN
It is stabilizer come storage time and the shortening for guaranteeing multienzyme that 105154253 A, which are disclosed using citric acid, propylene glycol, sodium hydroxide,
The time of multienzyme decomposing organic matter and greasy dirt, no and surfactant compound are used as liquid detergent.
Therefore, the influence of stabilizer, surfactant and environmental factor to enzyme activity is studied extensively, and is provided
The scheme of stability of the effective raising protease in liquid detergent has important practical significance.
Summary of the invention
[technical problem]
The technical problem to be solved in the present invention is to provide a kind of for stablizing the compound stabilizer of protease in liquid detergent.
[technical solution]
The present invention provides a kind of for stablizing the compound stabilizer of protease in liquid detergent, including with liquid detergent gross mass
0.5%-2% ammonium citrate calcium, 0.5%-1%APG0810,0.5%-2%APG1214 of meter.The ammonium citrate calcium be by
Ammonium citrate and calcium citrate are (25-20) according to molar ratio: what 1 ratio compounded.The stable liquid detergent protease,
Refer under 25~40 DEG C of preservation condition, compound stabilizer can reduce the loss of enzyme activity.
For more detergent use protease is used in the market, inventor has selected several stable in liquid detergent
Property weaker protease is captured.
Inventor has found ammonium citrate calcium and calcium citrate to the enzyme activity conservation rate of the protease in liquid detergent in an experiment
With significant facilitation, the specific stable work for mixing citrate than borax to several commercial detergent special protein enzymes
With stronger, this may be that prothetic group due to calcium ion as enzyme can make in proteinase activity in conjunction with the activated centre of enzyme
Hydrophobic structure allows the molecular structure of enzyme to become the raising for more closely and being conducive to the enzyme activity of protease to protease interior shifting.
For calcium citrate, though effect is good, it is difficult to be directly added into；Come using then can solve this in the form of ammonium citrate calcium
A problem.
It is compared with other ionic surfactants, nonionic surfactant is minimum to the damage of enzyme activity.In view of APG
Series can be used in liquid detergent as the nonionic surfactant for having certain dirt-removing power as detergent simultaneously.Inventor
Ammonium citrate calcium and eurytopic alkyl glucose polyglycoside surfactants APG (i.e. the mixture of different carbon chain product) are compounded
Afterwards, a kind of excellent ammonium citrate calcium-APG0810-APG1214 tri compound stabilizer is obtained, shown protease is being washed
The stabilization of ammonium citrate calcium and borax is had more than in clothing liquid.
In one embodiment of the invention, when the protease is Purafect 4000L, the compound stabilizer
Including 2% ammonium citrate calcium, 1%APG0810,2%APG1214.
In one embodiment of the invention, it is described to be used for when the protease is Purafect Prime 4000L
The protease composite stabilizer of liquid detergent, including 2% ammonium citrate calcium, 0.5%APG0810,0.5%APG1214.
In one embodiment of the invention, described for stablize in the compound stabilizer of liquid detergent protease can be with
Add AEO.Such as AEO7、AEO9。
It is described for stablizing the method for the compound stabilizer of protease in liquid detergent the present invention also provides using, it is described multiple
The mass ratio for closing stabilizer and enzyme is (0.5-5): 1.
[beneficial effect]
Under 25~40 DEG C of preservation condition, compound stabilizer can borax more good than generally acknowledged stabilization preferably subtract
The loss of few enzyme activity.For example, being stabilizer, Purafect 4000L and Purafect Prime with borax at pH7 and 37 DEG C
3 weeks enzyme activity conservation rates of 4000L are respectively 2.87% and 13.5%；And protease composite stabilizer provided by the invention is applied,
3 weeks enzyme activity conservation rates under similarity condition are respectively 47.1% and 42.0%.
At pH7 and 25 DEG C, protease composite stabilizer provided by the invention can make respectively Purafect 4000L and
3 weeks enzyme activity conservation rates of Purafect Prime 4000L reach 84.3% and 71.5%, still higher than borax under similarity condition
Caused 65.7% and 68.4%.
Detailed description of the invention
Fig. 1 ammonium citrate calcium and other stabilizers compare the stabilization of protease, a:Purafect 4000L, b:
EFFECTENZ P150, c:Purafect Prime 4000L.
Influence of Fig. 2 surfactant to proteinase activity, a:Purafect 4000L, b:Purafect Prime
4000L。
Influence of Fig. 3 mixed surfactant to Purafect 4000L and Purafect Prime 4000L enzyme activity.
Specific embodiment
The method for measuring proteinase activity: Folin method.Specifically, 1% casein solution 1mL is placed in test tube, in
The enzyme solution of 1mL dilution certain multiple is added in constant temperature 15min in 40 DEG C of waters bath with thermostatic control.After reacting 15min, 2mL trichloroacetic acid is added
(0.4mol/L) terminates reaction.Continue to be placed in constant temperature 15min in water-bath, filters.Filtrate 1mL is pipetted, it is molten that 5mL sodium carbonate is added
Liquid (0.4mol/L) and the Folin reagent 1mL diluted continue to be placed in 20min in 40 DEG C of thermostatic water baths after shaking up, restore to room
Absorbance after temperature at ultraviolet specrophotometer measurement wavelength 660nm.The 1 μ g junket of caseinhydrolysate generation per minute at 40 DEG C
Enzyme amount required for propylhomoserin is defined as 1 proteinase activity unit.Enzyme-activity unit is U/mL.
In formula, A, the absorbance measured by sample compares with standard curve and corresponding tyrosine quality (μ g) can be obtained；4,
The measurement of 1mL solution is taken out from 4mL reaction；T, reaction time, min；N, enzyme extension rate.
In addition to error line clearly indicates, the relative error of all data is less than 5%.
The preparation of enzyme liquid detergent and the method for measuring enzyme activity conservation rate:
Typically (middle data are shown in Table when orthogonal experiment), (enzyme used in the present embodiment is by stabilizer and 0.5 gram of protease
Solid powder, liquid enzymes can dry basis) it is added sequentially in the sample bottle that capacity is 200mL, it is mixed with eddy oscillating device
Even, stabilizer and enzyme mass ratio are respectively as follows: magnesium citrate: enzyme=1: 10, calcium citrate: and enzyme=1: 4000, it is other
Stabilizer: enzyme=1: 1.Enzymatic mixture is added in liquid detergent (liquid detergent) again later, solution gross mass is 100g.It stirs
Mixing is mixed, the enzyme activity measured at this time is initial enzyme activity, and this solution is placed at 37 DEG C or 25 DEG C and is saved, and is taken in different time
Sample, in duplicate, using proteinase activity in the measurement of Folin method at this time system after dilution.Protease in system under experimental period
Enzyme activity is the enzyme activity conservation rate of corresponding time to the percentage of proteinase activity in initial system.
Laundry liquid formulation (mass percent, each component is in terms of butt): AES, 21.5%；AEO7, 2.5%；AEO9,
2.5%；Trisodium citrate, 3.5%；Polyether antifoam agent, 0.05%；Citric acid, 0.1%；NaCl, 0.5%；Fluorescent whitening agent,
0.14%；Shredding, 0.05%；Lemon extract, 0.1%；Surplus is deionized water.Experimental water is deionized water.Wherein, alkyl
Polyglycoside surfactants APG0810, alkyl glucoside surfactant APG1214 are purchased from Shanghai Fakai Chemical Co., Ltd.；Lemon
Sour (food-grade), fatty alcohol polyoxyethylene ether AEO7(organic matter mass concentration is 99%), fatty alcohol polyoxyethylene ether AEO9(have
Machine amount of substance concentration be 99%), sodium sulfate of polyethenoxy ether of fatty alcohol AES (organic matter mass concentration is 70%) be purchased from Shandong
You Suo Chemical Industry Science Co., Ltd.
1 citrate of embodiment and other stabilizers compare the stabilization of protease
Stabilizer: magnesium citrate, calcium citrate, ammonium citrate calcium, citric acid monoglyceride, citric acid, ammonium citrate,
EDTA-2Na, sorbierite, APG0810, APG1214, glycerol, polysorbas20, polysorbate40, polysorbate60, Tween 80, Span 20.Wherein,
Ammonium citrate calcium is after ammonium citrate and calcium citrate are dissolved in water according to the ratio that molar ratio is 20:1, to use as stock solution
Or it is spare after drying.
Compare stabilizer: borax.
Protease: Purafect 4000L, EFFECTENZ P150 or Purafect Prime 4000L.
Measure influence of the above-mentioned different stabilizer to the enzyme activity conservation rate of different protease.As shown in Figure 1, citric acid
The stablizing effect of salt especially ammonium citrate calcium is best.Compared to borax, ammonium citrate calcium make Purafect 4000L,
The conservation rate of enzyme activity for 24 hours of EFFECTENZ P150 and Purafect Prime 4000L has been respectively increased 19.75%, 66.09%
With 37.91%.
Embodiment 2 improves protease in liquid detergent using ammonium citrate calcium-APG0810-APG1214 compound stabilizer
Stability
The fundamental property of this 2 kinds of protease of Purafect 4000L and EFFECTENZ P150 relatively, therefore selects it
One implements for illustrating.
For the inactivation for accelerating enzyme, tested using (40 DEG C) of higher temperature.Under in different time points, chosen by orthogonal experiment
PH7, the proportion of the best stabilizer under 40 DEG C of preservation conditions.It is control stabilizer with borax.
Choose L934Orthogonal arrage, using the enzyme activity conservation rate of protease as index, 4 factors are respectively ammonium citrate calcium content
(w/w), APG0810 content (w/w), APG1214 content (w/w) and protease content (w/w).The factor and level of orthogonal test
(percentage composition refers in terms of the gross mass of liquid detergent in table 1) is shown in Table 1, and the test result of orthogonal experiment is shown in Table 2 under different time points
With 3.
The factor and level (40 DEG C) of 1 orthogonal experiment of table
2 Purafect 4000L enzyme activity stability orthogonal experiments (40 DEG C) of table*
* the relative error of all data is respectively less than 5%."-" indicates that enzyme activity conservation rate is lower than 60%, because without providing phase
Answer data.
3 Purafect Prime 4000L enzyme activity stability orthogonal experiments (40 DEG C) of table*
* the relative error of all data is respectively less than 5%."-" indicates that enzyme activity conservation rate is lower than 60%, because without providing phase
Answer data.
By table 2 and table 3 it is found that in compound stabilizer system, the 96h enzyme activity conservation rate of Purafect 4000L is superior to list
Solely using borax or single ammonium citrate calcium system.Although in compound system, the enzyme activity of Purafect Prime 4000L
Conservation rate is inferior to that ammonium citrate calcium system is used alone, but tests a number 7d enzyme activity conservation rate and lemon for the experimental group for being 5 and 9
Lemon acid ammonium calcium system approaches, also, in view of the attribute of APG and the use that can reduce ammonium citrate calcium, compound system is answered
This is a preferable selection.
By table 2 and table 3 it is found that the optimization formula of enzyme activity conservation rate be respectively as follows: 2% ammonium citrate calcium, 1%APG0810,
2%APG1214 and 0.5%Purafect 4000L；Alternatively, 2% ammonium citrate calcium, 0.5%APG0810,0.5%APG1214
With 0.5%Purafect Prime 4000L.In addition, after experiment number is stored 4 weeks at 40 DEG C for 5 and 9 experimental group, Purafect
4000L enzyme activity still keeps about 42.3%.
Using borax as positive control, the stable liquid detergent of above-mentioned 2 compound stabilizers will be used to be respectively placed in not equality of temperature
Degree is lower to be saved, and experimental result is as shown in table 4.
4 borax of table is compound stabilizer under positive control to proteinase activity stability*
As shown in Table 4, Purafect 4000L and Purafect Prime greatly improved in compound stabilizer used
The thermal stability of 4000L.At pH7,25 DEG C, the compound stabilizer of Purafect 4000L and Purafect Prime 4000L
3 weeks enzyme activity conservation rates of system respectively reach 84.3% and 71.5%；And correspondingly, 3 weeks enzyme activity conservation rates of borax system
Respectively 2.87% and 13.5%.Even if showing that under the stabilization of borax, this 2 kinds of enzymes are at 40 DEG C with corresponding data comparison in table 2
When liquid detergent in stability can sharply decline.
3 surfactant of embodiment influences the enzyme activity of Purafect 4000L and Purafect Prime 4000L
The primary surfactant AES, AEO for the laundry liquid formulation that the present invention chooses7And AEO9Be it is currently used yin, it is non-from
Sub- surfactant.3 kinds of surfactants (are respectively as follows: AES at its 25 DEG C in its cmc with the cmc that mass concentration is expressed
1.065g/L AEO70.048g/L, AEO90.056g/L) and Fig. 2 and Fig. 3 are shown in the influence under more low quality concentration to enzyme activity.Its
In, the testing liquid of Fig. 3 is the mixed surfactants solutions of AEO and AES, and the mass concentration of AEO is 4g/L (m (AEO7): m
(AEO9)=1:1).
Fig. 2 and Fig. 3 shows that anionic surfactant AES is affected to enzyme activity, and nonionic surfactant AEO7
And AEO9Influence it is smaller relative to AES.For mixed surfactant system, the mixing of AEO and AES can not reduce AES
Damage to enzyme activity.Therefore in the future it is contemplated that increasing the use of APG and AEO in liquid enzymatic detergents.
Although the present invention has been described by way of example and in terms of the preferred embodiments, it is not intended to limit the invention, any to be familiar with this skill
The people of art can do various change and modification, therefore protection model of the invention without departing from the spirit and scope of the present invention
Enclosing subject to the definition of the claims.
Claims (10)
1. a kind of for stablizing the compound stabilizer of the protease in liquid detergent, which is characterized in that include ammonium citrate calcium and alkane
Base polyglycoside surfactants, the ammonium citrate calcium are proportionally to be compounded by ammonium citrate with calcium citrate.
2. according to claim 1 a kind of for stablizing the compound stabilizer of the protease in liquid detergent, which is characterized in that
It according to molar ratio is (25-20) that the ammonium citrate calcium, which is by ammonium citrate and calcium citrate: 1 ratio compounds.
3. according to claim 1 a kind of for stablizing the compound stabilizer of the protease in liquid detergent, which is characterized in that
The ammonium citrate calcium is 1:1-1.5 to the mass ratio of alkyl glucoside surfactant.
4. according to claim 1 or 2 a kind of for stablizing the compound stabilizer of the protease in liquid detergent, feature exists
In the alkyl glucoside surfactant is C8-10Alkyl glucose glucosides APG0810 and C12-14Alkyl glucose glucosides
The mass ratio of the mixture of APG1214, APG0810 and APG1214 are 1:(0.25-4).
5. according to claim 1 or 2 or 4 is a kind of for stablizing the compound stabilizer of the protease in liquid detergent, special
Sign is that the mass ratio of the compound stabilizer and enzyme is (0.5-5): 1.
6. it is a kind of for stablizing the compound stabilizer of the protease in liquid detergent described according to claim 1 or 2 or 4 or 5,
It is characterized in that, the compound stabilizer includes 0.5%-2% ammonium citrate calcium, the 0.5%-1% in terms of liquid detergent gross mass
APG0810,0.5%-2%APG1214.
7. according to claim 6 a kind of for stablizing the compound stabilizer of the protease in liquid detergent, which is characterized in that
When the protease is Purafect 4000L, the compound stabilizer includes 2% ammonium citrate calcium, 1%APG0810,2%
APG1214。
8. according to claim 6 a kind of for stablizing the compound stabilizer of the protease in liquid detergent, which is characterized in that
It is described for stablizing the protease composite stabilizer in liquid detergent, packet when the protease is Purafect Prime 4000L
Include 2% ammonium citrate calcium, 0.5%APG0810,0.5%APG1214.
9. a kind of liquid detergent, which is characterized in that any described for stablizing the albumen in liquid detergent containing claim 1~8
The compound stabilizer of enzyme.
10. a kind of liquid detergent according to claim 9, which is characterized in that in terms of liquid detergent gross mass, contain 0.5%-
2% ammonium citrate calcium, 0.5%-1%APG0810,0.5%-2%APG1214.
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| CN111635832A (en) * | 2020-07-10 | 2020-09-08 | 华洁化工(新丰)有限公司 | Efficient clothes detergent and preparation method thereof |
| CN113004985A (en) * | 2021-03-15 | 2021-06-22 | 广州市家真惠实业有限公司 | Plant enzyme underwear laundry detergent |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113004985A (en) * | 2021-03-15 | 2021-06-22 | 广州市家真惠实业有限公司 | Plant enzyme underwear laundry detergent |
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