CN105017446A - Deproteinised natural rubber and preparation method thereof - Google Patents
Deproteinised natural rubber and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a deproteinised natural rubber preparation method including the following steps: 1) adding a stabilizer in a fresh natural rubber latex or a concentrated natural rubber latex for stabilizing; 2) adding deionized water to dilute the rubber latex, carrying out centrifugal separation to remove impurities, and collecting an upper rubber phase; 3) diluting the upper rubber phase obtained in the step 2) with deionized water, adding a compound enzyme preparation under stirring and weak alkali conditions, and degrading proteins in the rubber latex under certain reaction conditions; and 4) carrying out centrifugal separation with a centrifugal machine to remove impurities obtained after the proteins are degraded, carrying out repeated degradation to remove the proteins contained in the latex, and thus obtaining deproteinised natural rubber. In addition, the invention also discloses the deproteinised natural rubber prepared by the method. The content of protein amino acids of the deproteinised natural rubber can reach less than a 0.67 ppm reagent detection limit value, and the deproteinised natural rubber can be used in production of various medical latex products and can avoid generation of allergy caused by natural rubber.
Description
Technical field
The present invention relates to a kind of DPNR and preparation method thereof, relate to a kind of allergy avoiding being caused by natural rubber further and occur.
Background technology
Natural rubber is a kind of macromolecular material naturally synthesized by Brazilian para ruber, based on cis-polyisoprene, emulsion liquid containing tens kinds of components, there is the character that such as elongation is large, elasticity is high, film toughness is good, be widely used in the fields such as industrial or agricultural, space flight and aviation, health care.Because natural rubber latex has good biocompatibility and process operation, therefore, up to the present, at medical assistance apparatuses such as the widely used surgical gloves of medical field, medical catheter, tube for blood transfusion,latex, inspection Check gloves, child care articles for use, and family planning articles for use condom etc., the main Nong of employing Shrink natural rubber latex is produced.
Along with widely using of medical emulsion products, the safety issue of medical emulsion products is paid close attention to all the more.Some doctor patients even contact in the process of medical emulsion products in use, and the instant type (I type) of expiratory dyspnea or allergic conditions (such as angioedema, measles or cyanosis) may be caused irritated.There will be the anaphylaxis such as fash, nose eye conjunctivitis, asthma, doctor patient time serious, may be caused to occur shock, even threat to life safety.
The U.S. reports recently, uses the medicine equipment such as surgical gloves of natural rubber, and various conduit and narcotic face shield cause expiratory dyspnea or hypersensitive symptom (such as, angioedema, urticaria, collapse, cyanosis).Also report female patients to ache from hand owing to using family expenses natural rubber gloves, collapse and the supersensitivity impaired memory around ocular vascular oedema.
Result of study shows, the anaphylaxis of medical emulsion products is mainly present in that the soluble protein in natural rubber latex Ruzhong causes.Now from fresh latex, isolated the quite high protein of two kinds of content, one is called alpha globulin, is made up of 17 seed amino acids.This protein is insoluble to pure water, but be soluble in neutral salt solution, basic solution and pH value lower than 3 acidic solution.The iso-electric point of alpha globulin is 4.55, close with the iso-electric point of rubber particles, is one of protein forming fresh latex particle protective layer, and is the principal element determining rubber particles colloidal stability; Another kind is called hevein, is made up of 14 seed amino acids, and its molecular weight is very low, generally only 10000 ± 500.Hevein can dissolve under any pH value, also can not be precipitated out from solution when boiling.Because the surfactivity of this protein is extremely low, seem remarkably influenced to occur to the colloidal property of fresh latex.
Test-results shows, when soluble protein content mark residual in natural rubber products is less than 110ppm, substantially protein allergy problem does not occur.Therefore, people's examination for a long time
figurefully protein is shifted out from natural rubber, reduce the protein content in natural rubber latex Ruzhong, to reduce medical emulsion products supersensitivity risk from starting material angle, expand natural rubber application category, preparation, to the unresponsive health-care medical material of tissue and utensil, is the research emphasis of this area always.Numerous investigator, conduct in-depth research shifting out the natural rubber performance after albumen, result shows, not only can not produce too large negative impact after deproteinated, greatly can promote the many performances of natural rubber on the contrary, as water-absorbent, electroconductibility, biocompatibility etc.
Domestic and international many scientific research personnel, large quantity research has been carried out to shifting out method of protein in natural rubber, relatively the method for accreditation has to use and comprises proteolytic enzyme and tensio-active agent, for the method for the deproteinated agent process natural rubber latex of natural rubber latex, totally say and can be summarized as by (1) the repeatedly centrifugal soluble protein removing natural rubber latex Ruzhong; (2) soluble protein (CN 1246485A, CN 1865292A, CN 1376720A) in hydrolysis by novo removing natural rubber latex Ruzhong; (3) mode of Bian urea complexation removes the soluble protein (CN 1930191A) in natural rubber latex Ruzhong.Wherein, the method for external hydrolysis by novo deproteination matter is for the production of low protein natural rubber latex.Above method respectively has its advantages and disadvantages part, as centrifuging, need to use heavy equipment, and centrifugally can only slough soluble proteins, the albumen be attached on micelle then can not be removed, this part albumen can come off out gradually with the change of solvent and cause allergy, Zong must thoroughly remove or make it to be less than in danger threshold containing protein content.
Protease hydrolysis is the method for comparative maturity and accreditation.But, use in conventional art in the consistency of the method for proteolytic enzyme between enzyme and latex and have problems.Wyler's process is a kind of new technology of Japanese scientific research personnel invention in 2006, Japanese Patent Publication (Kokai) No.8-253506A (1996) summary of the invention is to provide a kind of production in a large number at low cost on industrial level substantially containing the method for the deproteinized natural rubber latex of the protein and peptide that cause allergy, but its removing result actual nitrogen content minimum also still up to 0.015% (protein content is 0.015X6.25=0.093%), and there is not yet the report of practical application.These sides
method altogetherbe only reduce protein content with defect, in rubber, protein content is still very high, and the possibility with the change stripping again of solvent is high, and use is absolutely unsafe, and the soluble protein content far not reaching international endorsement is less than the minimum requirements of 110ppm.
Therefore, urgent need will develop a kind of preparation method of stable deproteinized natural rubber latex to address these problems.Reduce the content of non-rubber component in natural rubber contribute to reducing the water specific absorption of natural rubber and contribute to improving the electrical property (such as, electrical insulating property) of Natural Rubber Products.By almost removing non-rubber component completely, can provide a kind of useful material for the production of rubber item, it there almost is not power loss and has excellent mechanical property, the crepe rubber performance of improvement and the ageing resistance of improvement.One of natural rubber critical defect is that material character changes according to production area and production time, and this is the characteristic issues of natural product.Therefore, the non-rubber component of this problem of removing generation just makes curability stablize.Such natural rubber just becomes a kind of elastomeric material with stability compared with synthetic rubber, improves the mechanical precision of Natural Rubber Products.
Summary of the invention
One of the technical problem to be solved in the present invention is to provide a kind of preparation method with the DPNR of good stability.
Two of the technical problem to be solved in the present invention is to provide a kind of DPNR adopting aforesaid method obtained.
Three of the technical problem to be solved in the present invention is to provide a kind of important new strains for the preparation of DPNR.
Specifically, the invention provides a kind of method being prepared deproteinized natural rubber latex by complex enzyme degradation DNA techniques, after Nong Shrink natural rubber latex is suitably stablized with stablizer, under agitation add microbial compound enzyme preparation successively, proteolytic degradation in natural rubber is made under certain reaction conditions, by centrifugation, the fragment after proteolytic degradation and other impurity are removed again, adopt Lowry micromethod, and Xylene Brilliant Cyanine G, the methods such as triketohydrindene hydrate developer detect, the DPNR that protein content is less than detection limit 0.67ppm can be obtained finally by centrifugal concentrating.
For solving the problems of the technologies described above, the present invention is achieved through the following technical solutions:
In one aspect of the invention, provide a kind of preparation method of DPNR, the method comprises the steps:
(1) in fresh natural rubber latex or concentrated natural rubber latex, add stablizer stablize;
(2) add deionized water to be diluted by rubber latex, impurity is removed in centrifugation, collects upper rubber phase;
(3) to the deionized water dilution of step (2) gained upper rubber, under stirring and weak basic condition, add compound enzymic preparation, under certain reaction conditions, make protein degradation in rubber latex;
(4) be separated the impurity after removing protein degradation by centrifuge, make protein removal contained by latex by repeatedly degrading, obtaining DPNR.
In step (1), described fresh natural rubber latex or concentrated natural rubber latex, can be selected from the fresh natural rubber breast of Brazilian para ruber, also can select through the natural rubber latex from the dense Shrink of the heart.
As the preferred technical scheme of the present invention, in step (1), described stablizer, can be selected from Surfactant SDS, the moon potassium silicate, saccharan, potassium hydroxide, fat alcohol polyethylene ether sodium sulfate (AES), the stablizers such as alkyl polyoxyethylene ether, be preferably Surfactant SDS, the weight ratio of the sodium lauryl sulphate added and fresh natural rubber latex or concentrated natural rubber latex is (0.1-1): 100.
As the preferred technical scheme of the present invention, in step (1), described in add stablizer after, with acidity regulator, the pH value of rubber latex is adjusted to 8-9, the preferred potassium hydroxide of described acidity regulator (KOH) and aqueous formic acid.
As the preferred technical scheme of the present invention, in step (3), describedly under certain reaction conditions, make protein degradation in rubber latex, concrete reaction conditions is: under temperature 30 DEG C of conditions, the reaction times is 20 minutes-24 hours.Have no particular limits the time that latex processes with prozyme and tensio-active agent, it can according to the content of protein in the factor such as latex of process, and the kind of the quality of prozyme and consumption and tensio-active agent and consumption are determined.But this process is preferably carried out 20 minutes to about 24 hours from adding prozyme.In this case, if needed, the performance level of deproteinization reaction is by measuring the content of protein in latex to the sampling of latex and determined.
As the preferred technical scheme of the present invention, in step (3), described compound enzymic preparation, is selected from the microbial compound enzyme preparation of Haitobacter sufflavus Tih-w37, azotobacter vinelandii Azotobacter vinelandii, bacillus pumilus Bacillus pumilus, subtilis Bacillus subtilis fermentative production; The prozyme configuration proportion of Haitobacter sufflavus Tih-w37, azotobacter vinelandii Azotobacter vinelandii, bacillus pumilus Bacillus pumilus, subtilis Bacillus subtilis is 4:1:0.5:0.5, or 2:2:1:1.
As the preferred technical scheme of the present invention, in step (3), under stirring and weak basic condition, add compound enzymic preparation be specially: first add Haitobacter sufflavus Tih-w37 and the reaction of azotobacter vinelandii Azotobacter vinelandii compound enzymic preparation; And then add Haitobacter sufflavus Tih-w37, bacillus pumilus Bacillus pumilus, the reaction of subtilis Bacillus subtilis compound enzymic preparation.
Usually known natural rubber is molecular weight is 1,000,000 to 2,500, the high molecular weight component of 000 and 100, the mixture of 000 to 200,000 lower-molecular-weight component.Be 100 when an amino acid molecular (namely a nucleidic mass is the nitrogen-atoms of 14) is attached to molecular weight, on a molecule of the low molecular weight rubber of 000, it gets on to the eye is formed by intrinsic biosynthesizing, and the content of nitrogen is 0.014%, i.e. protein content 0.0875%.Therefore, when natural molecular weight rubber quantity is 1, also not containing an Amino acid score period of the day from 11 p.m. to 1 a.m in 500,000 mixture, namely nitrogen content is less than 0.001%, and when namely protein content is less than 0.0063%, this natural rubber just should be called DPNR.
As the preferred technical scheme of the present invention, in step (4), described in the protein content of DPNR that obtains can reach lower than 0.67ppm detectability.
Method for detecting protein content of the present invention can be adopted with the following method: 1. Xylene Brilliant Cyanine G is to the detection of protein content; 2. Lowry micromethod detection by quantitative that protein is carried out; 3. triketohydrindene hydrate is to the test of protein, peptide and aminoacids content.
Lowry micromethod is the classical way of mensuration trace protein conventional both at home and abroad, and the concrete testing method that present method provides with reference to Chinese Pharmacopoeia 2010 editions is implemented.
Coomassie Brilliant Blue and triketohydrindene hydrate are all colorimetrys that laboratory is commonly used.Coomassie brilliant G-250 measures the one that protein content belongs to dye binding method.In red-brown under unbound state, maximum light absorption is at 488nm; When becoming cyanic colours behind it and protein bound, protein-pigment binding substances has maximum light absorption under 595nm wavelength.Its absorbance value is directly proportional to protein content, measures protein content by color standards curve.
Triketohydrindene hydrate is except proline(Pro), oxyproline and ninhydrin reaction generate yellow substance, all a-amino acids and all protein can generate bluish voilet material with ninhydrin reaction, according to reacting the depth of bluish voilet material generated, under 570nm wavelength, carry out colorimetric just can the content of Argine Monohydrochloride in working sample.
Three kinds of methods respectively have relative merits, and in order to avoid mushing error, detected value of the present invention is the mutual the result of method not of the same race.
In another aspect of this invention, a kind of DPNR adopting aforesaid method obtained is provided.
In another aspect of this invention, provide a kind of bacterial strain for the preparation of DPNR, this Strain Designation is Haitobacter sufflavus Tih-w37, on September 1st, 2004
preservationat Chinese Typical Representative culture
preservationcenter (referred to as CCTCC), its
preservation is compilednumber be CCTCC No.M204056,
preservationplace: China, Wuhan.
Beneficial effect of the present invention is: the invention provides a kind of method being prepared deproteinized natural rubber latex by complex enzyme degradation technology, it is characterized in that concentrated natural rubber latex stablizer is stable after, deionized water and prozyme is added successively under stirring and alkaline condition, protein degradation in rubber latex is made under certain reaction conditions, then, the impurity such as the amino acid after removing proteolytic degradation are separated by centrifuge, protein removal contained by latex is made by complex enzyme degradation, the DPNR that protein content is less than below detection limit 0.67ppm can be obtained.By the way, protein wherein has been examined not measure and has been called deproteinized natural rubber latex, can be used as the raw material of Industrial materials and all kinds of rubber product.
Adopt deproteinized natural rubber latex prepared by the present invention, histone amino acid content can reach the degree of reagent detection limit, is less than 0.67ppm, can adopt the sulfuration of vulcanization method, and produce various medical emulsion products by conventional production process.Deproteinized natural rubber latex prepared by the present invention has satisfactory stability, reduces the water specific absorption of natural rubber and contributes to improving the electrical property of Natural Rubber Products, improve the mechanical precision of Natural Rubber Products.In addition, the DPNR adopting the present invention to prepare can also avoid the allergy caused by natural rubber to occur.
Embodiment
In following examples, the raw material related to, reagent etc. if not otherwise specified, are then commercially produced product.
Below with reference to embodiment, the present invention is described in more detail, but technical scope of the present invention is not limited.The present invention will be described hereinafter to adopt nonlimiting examples.
The preparation of embodiment 1 compound enzymic preparation
Compound enzymic preparation used in the present invention comes from Haitobacter sufflavus Tih-w37, A.vinelandii (azotobacter vinelandii), B.pumilus (bacillus pumilus), B.subtilis (subtilis) fermentation obtain, bacterial classification source and acquisition methods as follows:
1. Haitobacter sufflavus Tih-w37 is independent intellectual
property rightmicroorganism, is stored in Chinese Typical Representative culture by present inventor Hao Wei
preservationcenter (CCTCC), is selected in as new species
countrynature scientific and technological resources platform.Haitobacter sufflavus Tih-w37 is on September 1st, 2004
preservationat Chinese Typical Representative culture
preservationcenter (referred to as CCTCC),
preservation is compilednumber: CCTCC NO.:M204056,
preservationplace: wuchang, wuhan district Bayi Road No. 299 Wuhan Universitys of Hubei China province in the school.
countrynature scientific and technological resources platform is numbered: 1542C0001WHM204056.
2. A.vinelandii (azotobacter vinelandii), molecular cell biological study institute of Tokyo University cell function polymer integrating center provides, ATCC 478.
3. B.subtilis (subtilis), Chinese agriculture microorganism health-protection administrative center provides, ACCC 10619.
4. B.pumilus (bacillus pumilus), Chinese agriculture microorganism health-protection administrative center provides, ACCC 10113.
Substratum: bacterial classification is put down
platform is publicbe furnished with respective substratum.In distilled water 1000ml, add respective substratum in proportion; Adjusted to ph to 7.0, in Autoclave 120 DEG C, as the substratum of respective fermentative production prozyme after sterilizing in 20 minutes.
The preparation of compound enzymic preparation: respectively get above-mentioned 200ml substratum, pour in the triangular flask of 500ml, add corresponding bacteria culture fluid 1ml again, then under 30 DEG C of conditions, shaking table is cultivated after 24 hours, obtain the corresponding mixed enzyme fermentation liquid of respective bacterial classification, centrifugation (3000rpm*15min) can be carried out further, obtain respective compound enzymic preparation.
Then, by 1., 2., 3. and 4. obtained prozyme according to I) 4:1:0.5:0.5; II) population proportion of 2:2:1:1 carries out composite use, but will carry out composite in use again, 1. and 2., and 1., 3. and 4. composite use respectively.These compound enzymic preparations can be placed in 4 DEG C of refrigerators separately and save backup.
Embodiment 2
Take 100g total solids level be 60% without An Nong Shrink natural rubber latex, add 0.1g sodium lauryl sulphate (SDS), use the potassium hydroxide aqueous solution adjusted to ph to 9 of 10% subsequently, make latex keep stable; Under agitation add 140g deionized water, the total solids level of reaction system remains on 25%.After abundant stirring, by separating centrifuge after 10000 turns/min, 30min centrifugation, remove lower floor's glue clear, leave and take upper strata concentrating colloidal particles layer, then, under agitation add 240g deionized water, make the total solids level of reaction system remain on about 25%; Then, at 30 DEG C, slowly under agitation condition, add the proportions of compound enzymic preparation (1., 2., 3. and 4. according to I) 4:1:0.5:0.5 prepared by embodiment 1), first will 1. and 2. compound preparation 8g, then, react 5 hours with this understanding.After reaction terminates, by separating centrifuge after 7000 turns/min, 20min centrifugation, remove lower floor's glue clear, leave and take the dense latex granulosa in upper strata; Then, then 140g deionized water is added to the dense breast of centrifugal gained upper rubber grain, keep solid content at 20-25%.Then, under agitation again add 1., 3. and 4. compound preparation 2g (first time 1. and 2. compound preparation 8g, with second time 1., and 4. 3. 1. the population proportion of compound preparation 2g meets, 2., 3. with 4. according to I) proportions of 4:1:0.5:0.5), under 30 DEG C of conditions, limit is slowly stirred after limit reacts 5 hours, after reaction terminates, reuse separating centrifuge and centrifugation is carried out to it, slough the impurity such as the amino acid under enzymolysis, get the rubber grain that upper strata is pure, allotment is to the latex rubber of required solid content (60%), namely the DPNR of protein content lower than 0.67ppm is obtained.
Embodiment 3
Take 100g total solids level be 60% without An Nong Shrink natural rubber latex, add 0.5g sodium lauryl sulphate (SDS), use the potassium hydroxide aqueous solution adjusted to ph to 9 of 10% subsequently, make latex keep stable; Add 140g deionized water successively subsequently, the total solids level of reaction system remains on 25%.After abundant stirring, by separating centrifuge after 10000 turns/min, 30min centrifugation, remove lower floor's glue clear, leave and take upper strata concentrating colloidal particles layer, then, under agitation add 240g deionized water, make the total solids level of reaction system remain on about 20%; Then, at 30 DEG C, slowly under agitation condition, add the proportions of compound enzymic preparation (1., 2., 3. and 4. according to I) 4:1:0.5:0.5 prepared by embodiment 1), first will 1. and 2. compound preparation 8g, react 10 hours; Then, directly add 1. while stirring, 3. and 4. compound preparation 2g (first time 1. and 2. compound preparation 8g, meet 1., 2., 3. and 4. according to I with the population proportion of second time 1., 3. and 4. compound preparation 2g) proportions of 4:1:0.5:0.5), under 30 DEG C of conditions, limit is slowly stirred after limit reacts 10 hours, after reaction terminates, use separating centrifuge 7000 turns/min, 20min carry out centrifugation to it, slough the impurity such as the amino acid under enzymolysis, get the rubber grain that upper strata is pure.Then, be adjusted to required solid content (60%) by centrifugal concentrating, namely obtain the DPNR of protein content lower than 0.67ppm.
Embodiment 4
Take 100g total solids level be 60% fresh natural rubber revertex (be selected from the fresh natural rubber breast of Brazilian para ruber, the solid content of fresh natural rubber breast is not fixed, may only have 30-40% not), add 1g sodium lauryl sulphate (SDS), use the potassium hydroxide aqueous solution adjusted to ph to 8 of 10% subsequently, make latex keep stable; Add 140g deionized water successively subsequently, the total solids level of reaction system remains on 25%.After abundant stirring, by separating centrifuge after 10000 turns/min, 30min centrifugation, remove lower floor's glue clear, leave and take upper strata concentrating colloidal particles layer, then, under agitation add 240g deionized water, make the total solids level of reaction system remain on about 23%; Then, at 30 DEG C, slowly under agitation condition, add the proportions of compound enzymic preparation (1., 2., 3. and 4. according to I) 4:1:0.5:0.5 prepared by embodiment 1), first will 1. and 2. compound preparation 6g, react 8 hours.After reaction terminates, by separating centrifuge after 7000 turns/min, 20min centrifugation, remove lower floor's glue clear, leave and take the dense latex granulosa in upper strata; Then, then 140g deionized water is added to the dense breast of centrifugal gained upper rubber grain, keep solid content at 20-25%.Then, under agitation again add 1., 3. and 4. compound preparation 4g (first time 1. and 2. compound preparation 6g, meet 1., 2., 3. and 4. according to I with the population proportion of second time 1., 3. and 4. compound preparation 4g) proportions of 4:1:0.5:0.5), under 30 DEG C of conditions, limit is slowly stirred limit and is reacted 8 hours.After reaction terminates, reuse separating centrifuge 7000 turns/min, 20min carry out centrifugation to it, slough the impurity such as the amino acid under enzymolysis, get the rubber grain that upper strata is pure, allotment, to required drc (60%), obtains the DPNR that protein content is less than 0.67ppm.
Embodiment 5
Take 100g total solids level be 60% without An Nong Shrink natural rubber latex, add 0.2g month potassium silicate, use the potassium hydroxide aqueous solution adjusted to ph to 9 of 10% subsequently, make latex keep stable; Under agitation add 140g deionized water, the total solids level of reaction system remains on 25%.After abundant stirring, by separating centrifuge after 10000 turns/min, 30min centrifugation, remove lower floor's glue clear, leave and take upper strata concentrating colloidal particles layer, then, under agitation add 240g deionized water, make the total solids level of reaction system remain on 25%; Then, at 30 DEG C, slowly under agitation condition, add the proportions of compound enzymic preparation (1., 2., 3. and 4. according to I) 4:1:0.5:0.5 prepared by embodiment 1), first will 1. and 2. compound preparation 8g, then, react 6 hours with this understanding.After reaction terminates, by separating centrifuge after 7000 turns/min, 20min centrifugation, remove lower floor's glue clear, leave and take the dense latex granulosa in upper strata; Then, then 140g deionized water is added to the dense breast of centrifugal gained upper rubber grain, keep solid content at 20-25%.Then, under agitation again add 1., 3. and 4. compound preparation 2g (first time 1. and 2. compound preparation 8g, with second time 1., and 4. 3. 1. the population proportion of compound preparation 2g meets, 2., 3. with 4. according to I) proportions of 4:1:0.5:0.5), under 30 DEG C of conditions, limit is slowly stirred after limit reacts 6 hours, after reaction terminates, reuse separating centrifuge and centrifugation is carried out to it, slough the impurity such as the amino acid under enzymolysis, get the rubber grain that upper strata is pure, allotment is to the latex rubber of required solid content (60%), namely the DPNR of protein content lower than 0.67ppm is obtained.
Embodiment 6
Take 100g total solids level be 60% without An Nong Shrink natural rubber latex, add 0.1g fat alcohol polyethylene ether sodium sulfate (AES), use the potassium hydroxide aqueous solution adjusted to ph to 9 of 10% subsequently, make latex keep stable; Under agitation add 140g deionized water, the total solids level of reaction system remains on 25%.After abundant stirring, by separating centrifuge after 10000 turns/min, 30min centrifugation, remove lower floor's glue clear, leave and take upper strata concentrating colloidal particles layer, then, under agitation add 240g deionized water, make the total solids level of reaction system remain on about 25%; Then, at 30 DEG C, slowly under agitation condition, add the proportions of compound enzymic preparation (1., 2., 3. and 4. according to II) 2:2:1:1 prepared by embodiment 1), first will 1. and 2. compound preparation 8g, then, react 12 hours with this understanding.After reaction terminates, by separating centrifuge after 7000 turns/min, 20min centrifugation, remove lower floor's glue clear, leave and take the dense latex granulosa in upper strata; Then, then 140g deionized water is added to the dense breast of centrifugal gained upper rubber grain, keep solid content at 20-25%.Then, under agitation again add 1., 3. and 4. compound preparation 2g (first time 1. and 2. compound preparation 8g, with second time 1., and 4. 3. 1. the population proportion of compound preparation 2g meets, 2., 3. with 4. according to II) proportions of 2:2:1:1), under 30 DEG C of conditions, limit is slowly stirred after limit reacts 12 hours, after reaction terminates, reuse separating centrifuge and centrifugation is carried out to it, slough the impurity such as the amino acid under enzymolysis, get the rubber grain that upper strata is pure, allotment is to the latex rubber of required solid content (60%), namely the DPNR of protein content lower than 0.67ppm is obtained.
Test example 1
Above embodiment, through verification experimental verification, all can avoid the allergy caused by natural rubber to occur.
Below be only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (10)
1. a preparation method for DPNR, is characterized in that, the method comprises the steps:
(1) in fresh natural rubber latex or concentrated natural rubber latex, add stablizer stablize;
(2) add deionized water to be diluted by rubber latex, impurity is removed in centrifugation, collects upper rubber phase;
(3) to the deionized water dilution of step (2) gained upper rubber, under stirring and weak basic condition, add compound enzymic preparation, under certain reaction conditions, make protein degradation in rubber latex;
(4) be separated the impurity after removing protein degradation by centrifuge, make protein removal contained by latex by repeatedly degrading, obtaining DPNR.
2. the preparation method of DPNR according to claim 1, it is characterized in that, in step (1), described stablizer, is selected from Surfactant SDS, the moon potassium silicate, saccharan, potassium hydroxide, fat alcohol polyethylene ether sodium sulfate, alkyl polyoxyethylene ether.
3. the preparation method of DPNR according to claim 2, it is characterized in that, in step (1), described stablizer is Surfactant SDS, and the weight ratio of the sodium lauryl sulphate added and fresh natural rubber latex or concentrated natural rubber latex is (0.1-1): 100.
4. the preparation method of DPNR according to claim 1, is characterized in that, in step (1), described in add stablizer after, with acidity regulator, the pH value of rubber latex is adjusted to 8-9.
5. the preparation method of DPNR according to claim 1, it is characterized in that, in step (3), describedly under certain reaction conditions, make protein degradation in rubber latex, concrete reaction conditions is: under temperature 30 DEG C of conditions, the reaction times is 20 minutes-24 hours.
6. the preparation method of DPNR according to claim 1, it is characterized in that, in step (3), described compound enzymic preparation, is selected from the microbial compound enzyme preparation of Haitobacter sufflavus Tih-w37, azotobacter vinelandii Azotobacter vinelandii, bacillus pumilus Bacillus pumilus, subtilis Bacillus subtilis fermentative production; The prozyme configuration proportion of Haitobacter sufflavus Tih-w37, azotobacter vinelandii Azotobacter vinelandii, bacillus pumilus Bacilluspumilus, subtilis Bacillus subtilis is 4:1:0.5:0.5, or 2:2:1:1.
7. the preparation method of DPNR according to claim 6, it is characterized in that, in step (3), under stirring and weak basic condition, add compound enzymic preparation be specially: first add Haitobacter sufflavus Tih-w37 and the reaction of azotobacter vinelandii Azotobacter vinelandii compound enzymic preparation; And then add Haitobacter sufflavus Tih-w37, bacillus pumilus Bacillus pumilus, the reaction of subtilis Bacillus subtilis compound enzymic preparation.
8. the preparation method of DPNR according to claim 1, is characterized in that, in step (4), described in the protein content of DPNR that obtains lower than 0.67ppm detectability.
9. the DPNR adopting the method described in any one of claim 1-8 obtained.
10. for the preparation of a bacterial strain for DPNR, it is characterized in that, this Strain Designation is Haitobactersufflavus Tih-w37, and its deposit number is CCTCC No.M204056.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109456423A (en) * | 2018-09-23 | 2019-03-12 | 南通嘉得利安全用品有限公司 | A kind of low protein natural rubber gloves and its production method |
| CN111205379A (en) * | 2020-03-18 | 2020-05-29 | 田晓慧 | Method for processing natural latex by creaming |
| CN116020145A (en) * | 2021-10-25 | 2023-04-28 | 中国石油化工股份有限公司 | Apparatus and method for desolventizing polyisoprene latex |
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| CN1353116A (en) * | 2000-11-08 | 2002-06-12 | 住友橡胶工业株式会社 | Method of preparing low sensitivity natural rubber latex and deproteinized natural rubber latex and low sensitivity natural rubber and deproteinized natural rubber |
| CN101139622A (en) * | 2007-05-28 | 2008-03-12 | 上海居知园生物技术有限公司 | Organic compound and new biological material (JZIY-LC06) for protein resource conversion by detoxification of toxic waste liquor |
| CN102268106A (en) * | 2011-08-26 | 2011-12-07 | 中国热带农业科学院农产品加工研究所 | Preparation method of high-purity natural rubber |
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| CN1353116A (en) * | 2000-11-08 | 2002-06-12 | 住友橡胶工业株式会社 | Method of preparing low sensitivity natural rubber latex and deproteinized natural rubber latex and low sensitivity natural rubber and deproteinized natural rubber |
| CN101139622A (en) * | 2007-05-28 | 2008-03-12 | 上海居知园生物技术有限公司 | Organic compound and new biological material (JZIY-LC06) for protein resource conversion by detoxification of toxic waste liquor |
| CN102268106A (en) * | 2011-08-26 | 2011-12-07 | 中国热带农业科学院农产品加工研究所 | Preparation method of high-purity natural rubber |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109456423A (en) * | 2018-09-23 | 2019-03-12 | 南通嘉得利安全用品有限公司 | A kind of low protein natural rubber gloves and its production method |
| CN111205379A (en) * | 2020-03-18 | 2020-05-29 | 田晓慧 | Method for processing natural latex by creaming |
| WO2021184703A1 (en) * | 2020-03-18 | 2021-09-23 | 田晓慧 | Processing method for creaming natural latex |
| CN116020145A (en) * | 2021-10-25 | 2023-04-28 | 中国石油化工股份有限公司 | Apparatus and method for desolventizing polyisoprene latex |
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| CN105017446B (en) | 2017-01-18 |
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