CN1836599A - Preparation method of chitosan immobilized hydrogen peroxidase for cigarette filter tip additive - Google Patents
Preparation method of chitosan immobilized hydrogen peroxidase for cigarette filter tip additive Download PDFInfo
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- CN1836599A CN1836599A CN 200610039838 CN200610039838A CN1836599A CN 1836599 A CN1836599 A CN 1836599A CN 200610039838 CN200610039838 CN 200610039838 CN 200610039838 A CN200610039838 A CN 200610039838A CN 1836599 A CN1836599 A CN 1836599A
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- hydrogen peroxidase
- filter tip
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- cigarette filter
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Abstract
The preparation process of chitosan immobilized hydrogen peroxidase as cigarette filter tip additive relates to the production of cigarette modifying diacetate fiber tow and belongs to the field of cigarette filter tip preparing technology. The chitosan immobilized hydrogen peroxidase is prepared with chitosan as carrier and ethanedial and sodium pyrophosphate as cross-linking agent and through adsorption and cross-linking of hydrogen peroxidase, filtering, cleaning, stoving and grinding. The chitosan immobilized hydrogen peroxidase is added into filter tip through a dry process in the amount of 10 mg each cigarette. The filter tip with added chitosan immobilized hydrogen peroxidase has obvious eliminating effect on freedom radical in main cigarette smoke, including gaseous freedom radical eliminating rate of 25.07 % and solid freedom radical eliminating rate of 21.34 %.
Description
Technical field
A kind of preparation method of chitosan immobilized hydrogen peroxidase for cigarette filter tip additive relates to the production method of modification cigarette with the diacetate fiber tow, belongs to the preparing technical field of cigarette filter.
Background technology
Shitosan has another name called chitin, chitin, and formal name used at school is poly--2-acetylaminohydroxyphenylarsonic acid 2-deoxidation-D-β-1,4-glucose.Chitin is very abundant in the occurring in nature resource, is mainly derived from the organisms such as arthropod, mollusk, annelid, protozoan, coelenterate, marine alga and fungi, and be one of important derivative of occurring in nature.In recent years, shitosan has caused a lot of scholars' attention as the research of fixation support, because the amino in the chitosan molecule is very high to the affinity of range protein, can be effectively applied in the immobilization technology of enzyme.
Catalase (Catalase, be called for short CAT), claiming catalase again, is a kind of efficient, safety, nontoxic biocatalyst, white is to the unformed powder or the liquid of light brown, and molecular weight is about 240,000, and is water-soluble, the aqueous solution generally is light brown to brown, is dissolved in organic solvents such as ethanol, chloroform and ether hardly.This enzyme is an iron porphin purine enzyme, contains 4 iron atoms in the molecule, and wherein acting on group is ferroheme, can the quick catalysis decomposing hydrogen dioxide solution be water and oxygen.Catalase is distributed widely in the animal vegetable tissue, and content is also more in the bacteriums such as micrococcus lysodeikticus.This enzyme just is widely used in some assay determination in biology, medicine, the clinical and food from nineteen fifty; In food and dairy processing industry, be used to remove the hydrogen peroxide of remnants after sterilization and the sterilization; Residual hydrogen peroxide when in textile industry, being used to remove oxygen and floating.
Summary of the invention
The preparation method who the purpose of this invention is to provide a kind of chitosan immobilized hydrogen peroxidase for cigarette filter tip additive, be further to expand the using value of shitosan, improve catalatic actual application value simultaneously, develop a kind of chitosan immobilized hydrogen peroxidase for cigarette filter tip additive.
Technical scheme of the present invention: with shitosan is carrier immobilized catalase.The ratio of shitosan and 1% acetum is that 3: 1000 (g/ml) is prepared into the shitosan acidic sol, and the enzyme work of catalase liquid is 10,000CIU/ml, and the volume ratio of enzyme liquid consumption and shitosan acetic acid colloidal sol is 6: 20.The shitosan mixed liquor of enzyme under 0 ℃ condition, adsorption of immobilization 2-2.5 hour.The preparation of crosslinking agent, at first the glyoxal solution with the 4%-6% mass concentration activates 10 minutes in 80 ℃ of water-baths, mixes with the sodium pyrophosphate solution equal-volume of 15% mass concentration, and crosslinking agent is cooled to 0 ℃.Under stirring condition, lentamente catalase-shitosan mixed liquor is joined in the crosslinking agent, the volume ratio of catalase and crosslinking agent is 6-8: 100-120, under 0 ℃ of condition, crosslinked immobilization 0.5 hour.Filter, use pH=7.0,0.1mol/L KH
2PO
4/ Na
2HPO
4Flushing liquor washing three times is removed glyoxal wherein, sodium pyrophosphate etc.Last under 60 ℃ condition, oven dry is ground, and crosses 85 mesh sieves, makes chitosan immobilized hydrogen peroxidase for cigarette filter tip additive.
The characteristic contrast accompanying drawing of chitosan immobilized hydrogen peroxidase is illustrated.Accompanying drawing 1 is the influence that adsorption time is lived to immobilised enzymes.This figure explanation, under the situation that other conditions are fixed, the work of immobilised enzymes enzyme increased with the adsorption of immobilization time, and when reaching 2-2.5 hour, enzyme work reaches maximum.Accompanying drawing 2 is influences that crosslinking time is lived to immobilised enzymes.Illustrate, along with the increase of crosslinking time, immobilised enzymes is lived and is reduced on the contrary.But contain sodium pyrophosphate in the crosslinking agent, it mainly acts on and makes chitosan gel rubberization, generates chitosan microball, separates from reaction system.Need the certain reaction time and form the gel solid particle, though therefore Fig. 2 demonstration is along with the prolongation of crosslinking time, the enzyme of immobilised enzymes is lived and is descended, and for guaranteeing the complete of gelation, determines that crosslinking time is 0.5h.Accompanying drawing 3 is influences that glyoxal concentration is lived to immobilised enzymes, and the result shows that when the concentration of glyoxal was 4%-6%, immobilised enzymes was lived higher.The influence that accompanying drawing 4 dosage of crosslinking agent are lived to immobilised enzymes is under the condition of 100-120ml at crosslinking agent, and the enzyme of immobilized catalase is lived the highest.Accompanying drawing 5 is influences that catalatic addition is lived to immobilised enzymes, and when the enzyme addition was 6-8ml, enzyme was lived higher.Accompanying drawing 6 is that the pH stability of resolvase and immobilised enzymes compares.As can be seen, the enzyme work when pH=7 of immobilised enzymes and resolvase all is the highest.Resolvase is when pH=3, and resolvase is inactivation almost completely; Comparatively speaking, chitosan immobilized hydrogen peroxidase diminishes to the sensitiveness that pH changes, and is very favourable for the practical application of immobilized catalase.Accompanying drawing 7 and accompanying drawing 8 are immobilized catalase removal effects to free radical in the cigarette main flume.Mainly in the mode of dry method interpolation, addition is that 10mg/ props up.Measured the removing effect of additive-chitosan immobilized hydrogen peroxidase to cigarette main flume free radical, clearance is: gaseous phase free radical 25.07%, solid phase free radical 21.34%.
Beneficial effect of the present invention: the present invention has further expanded the using value of shitosan, has improved catalatic actual application value simultaneously, develops a kind of chitosan immobilized hydrogen peroxidase for cigarette filter tip additive.In the mode that dry method is added, addition is that 10mg/ props up, and has added chitosan immobilized hydrogen peroxidase, and obvious to the removing effect of the main flume free radical of cigarette, clearance is: gaseous phase free radical 25.07%, solid phase free radical 21.34%.
Description of drawings
The influence that Fig. 1 adsorption time is lived to immobilised enzymes.
The influence that Fig. 2 crosslinking time is lived to immobilised enzymes.
The influence that Fig. 3 glyoxal concentration is lived to immobilised enzymes.
The influence that Fig. 4 dosage of crosslinking agent is lived to immobilised enzymes.
The influence that Fig. 5 enzyme concentration is lived to immobilised enzymes.
The pH stability of Fig. 6 resolvase and immobilised enzymes relatively.
Fig. 7 immobilised enzymes is to the scavenging action (the ESR collection of illustrative plates relatively) of gaseous phase free radical in the main flume.
Fig. 8 immobilised enzymes is to solid phase measured by esr technique in the main flume (the ESR collection of illustrative plates relatively).
The specific embodiment
The preparation of shitosan acidic sol: take by weighing the 3.0g shitosan and be dissolved in 1000ml, in 1.00% the acetum, be prepared into 0.3% shitosan acetic acid colloidal sol.
The preparation of crosslinking agent: 50ml 6% glyoxal solution is in 80 ℃ of waters bath with thermostatic control, and insulation activates 10min.Mix with isopyknic 15% sodium pyrophosphate, be prepared into crosslinking agent.
After immobilization is finished, filter, remove residual liquid.Potassium dihydrogen phosphate-sodium hydrogen phosphate buffer solution with 0.1mol/L pH=7.0 washs immobilized enzyme particle three times again, removes residual component.Behind the suck dry moisture, under 60 ℃ of conditions, oven dry.Grind 85 mesh sieves, promptly be prepared into chitosan immobilized hydrogen peroxidase for cigarette filter tip additive.With dry method chitosan immobilized hydrogen peroxidase is added in the cigaratte filter diacetate fiber tow, addition is that 10mg/ props up.
The preparation of shitosan acidic sol: take by weighing the 3.0g shitosan and be dissolved in 1000ml, in 1.00% the acetum, be prepared into 0.3% shitosan acetic acid colloidal sol.
The preparation of crosslinking agent: 60ml 4% glyoxal solution is in 80 ℃ of waters bath with thermostatic control, and insulation activates 10min.Mix with isopyknic 15% sodium pyrophosphate, be prepared into crosslinking agent.
After immobilization is finished, filter, remove residual liquid.Potassium dihydrogen phosphate-sodium hydrogen phosphate buffer solution with 0.1mol/L pH=7.0 washs immobilized enzyme particle three times again, removes residual component.Behind the suck dry moisture, under 60 ℃ of conditions, oven dry.Grind 85 mesh sieves, promptly be prepared into chitosan immobilized hydrogen peroxidase for cigarette filter tip additive.With dry method chitosan immobilized hydrogen peroxidase is added in the cigaratte filter diacetate fiber tow, addition is that 10mg/ props up.
Claims (5)
1, a kind of preparation method of chitosan immobilized hydrogen peroxidase for cigarette filter tip additive, it is characterized by: shitosan is dissolved preparation shitosan acidic sol fully with 1% acetum, with catalase liquid, under stirring condition, join lentamente in the shitosan acidic sol, place for 0 ℃ and carried out the adsorption of immobilization reaction in 2-2.5 hour, to be hybridly prepared into crosslinking agent at the glyoxal solution of 10 minutes 4%-6% mass concentration of 80 ℃ of activation and the sodium pyrophosphate solution equal-volume of 15% mass concentration, under the condition of ice bath, join catalase-shitosan colloidal sol in the crosslinking agent lentamente, carried out crosslinked immobilization in 0.5 hour 0 ℃ of stirring, after filtration, washing makes the wet granular of chitosan immobilized hydrogen peroxidase, dry under 60 ℃ of conditions again, grind 85 mesh sieves and obtained chitosan immobilized hydrogen peroxidase for cigarette filter tip additive.
2, the preparation method of chitosan immobilized hydrogen peroxidase for cigarette filter tip additive according to claim 1 is characterized in that the chitosan mass volumetric concentration of shitosan acidic sol counts 0.3% with g/ml.
3, the preparation method of chitosan immobilized hydrogen peroxidase for cigarette filter tip additive according to claim 1, the volume ratio that it is characterized in that catalase enzyme liquid and shitosan acidic sol is 6: 20.
4, the preparation method of chitosan immobilized hydrogen peroxidase for cigarette filter tip additive according to claim 1, the volume ratio that it is characterized in that catalase enzyme liquid and crosslinking agent is 6-8: 100-120.
5, the preparation method of chitosan immobilized hydrogen peroxidase for cigarette filter tip additive according to claim 1, the enzyme work that it is characterized in that catalase liquid is 10,000CIU/ml.
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| CN 200610039838 CN1836599A (en) | 2006-04-19 | 2006-04-19 | Preparation method of chitosan immobilized hydrogen peroxidase for cigarette filter tip additive |
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| CN 200610039838 CN1836599A (en) | 2006-04-19 | 2006-04-19 | Preparation method of chitosan immobilized hydrogen peroxidase for cigarette filter tip additive |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102793271A (en) * | 2012-08-31 | 2012-11-28 | 福建中烟工业有限责任公司 | Application of cross-linked chitosan porous microspheres in cigarette filter tips |
| CN103535852A (en) * | 2013-10-18 | 2014-01-29 | 贵州中烟工业有限责任公司 | Immobilized catalase tobacco additive and preparation method and application thereof |
| CN106706911A (en) * | 2016-11-25 | 2017-05-24 | 云南中烟工业有限责任公司 | Method for detecting influence of oral smoking products on enzyme activity of catalase in cells |
-
2006
- 2006-04-19 CN CN 200610039838 patent/CN1836599A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102793271A (en) * | 2012-08-31 | 2012-11-28 | 福建中烟工业有限责任公司 | Application of cross-linked chitosan porous microspheres in cigarette filter tips |
| CN102793271B (en) * | 2012-08-31 | 2015-04-22 | 福建中烟工业有限责任公司 | Application of cross-linked chitosan porous microspheres in cigarette filter tips |
| CN103535852A (en) * | 2013-10-18 | 2014-01-29 | 贵州中烟工业有限责任公司 | Immobilized catalase tobacco additive and preparation method and application thereof |
| CN103535852B (en) * | 2013-10-18 | 2015-11-25 | 贵州中烟工业有限责任公司 | A kind of immobilized catalase cigarette additive and its preparation method and application |
| CN105146745A (en) * | 2013-10-18 | 2015-12-16 | 贵州中烟工业有限责任公司 | Immobilized catalase additive for tobacco as well as preparation method and application of additive |
| CN106706911A (en) * | 2016-11-25 | 2017-05-24 | 云南中烟工业有限责任公司 | Method for detecting influence of oral smoking products on enzyme activity of catalase in cells |
| CN106706911B (en) * | 2016-11-25 | 2019-01-25 | 云南中烟工业有限责任公司 | A method of detection mouth containing tobacco product influences cellular superoxide hydrogen enzyme enzyme activity |
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Open date: 20060927 |