CN105851063B - A kind of production method of the antibacterial spraying of Luo Yishi soft white hands machine - Google Patents
A kind of production method of the antibacterial spraying of Luo Yishi soft white hands machine Download PDFInfo
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- CN105851063B CN105851063B CN201610276532.7A CN201610276532A CN105851063B CN 105851063 B CN105851063 B CN 105851063B CN 201610276532 A CN201610276532 A CN 201610276532A CN 105851063 B CN105851063 B CN 105851063B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/10—Animals; Substances produced thereby or obtained therefrom
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/02—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing liquids as carriers, diluents or solvents
- A01N25/04—Dispersions, emulsions, suspoemulsions, suspension concentrates or gels
- A01N25/06—Aerosols
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- Wood Science & Technology (AREA)
- Dispersion Chemistry (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Chemical & Material Sciences (AREA)
- Environmental Sciences (AREA)
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- Agronomy & Crop Science (AREA)
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Abstract
A kind of production method of the antibacterial spraying of Luo Yishi soft white hands machine, belongs to natural bacteriostatic agent processing technique field.It includes：1）Strain selects；2）L. reuteri Lactic acid bacteria activates；3）L. reuteriLactic acid bacteria is cultivated；4）L. reuteriThe eccentric cleaning of lactic acid bacteria；5) coenzyme B12The preparation of reaction solution；6）reuterinProduction；7）reuterinExtraction；8）It is prepared by microemulsion；9）Product compounds；10）Product packaging.The antibacterial spraying of Luo Yishi soft white hands machines that the present invention obtains not only has very high antibacterial efficiency, but also production cost is low.
Description
Technical field
The invention belongs to natural bacteriostatic agent processing technique fields, and in particular to a kind of life of the antibacterial spraying of Luo Yishi soft white hands machine
Production method.
Background technology
With the exploitation of smart mobile phone and mobile phone A pp and universal, the time of people's mobile phone is more and more.Especially university
Raw, investigation display, the average every university student in China one day is 4.5 hours the time with mobile phone, it is seen that mobile phone to university student very
It is attractive.Handrail is tested with the presence of superbacteria in Guangzhou Underground for the previous period, it is seen that people's on hand may contamination
Very harmful bacterium, therefore a kind of new product of urgent need is antibacterial to mobile phone, to protect the health of oneself.
It is found after investigating existing bactericidal product on the market, most of antibacterial sprayings are all chemistry currently on the market
Material not environmentally has health potentially hazardous, may cause secondary pollution.
ε-poly-D-lysine is the microbiology class food preservative with superior antiseptic property in current natural antiseptic agent.It
It is to be polymerized by 25~30 lysine residues, there is strong bacteriostasis, it can be as preservative for the fresh-keeping of food.
ε-poly-D-lysine antimicrobial spectrum is wide, in acid and slightly acidic environment to gram-positive bacteria, Gram-negative bacteria, saccharomycete,
Mould has certain fungistatic effect, the gram-negative large intestine that ε-poly-D-lysine is not easy to inhibit to other natural antiseptic agents
Bacillus, salmonella fungistatic effect are very good, and it also has inhibiting effect to heat resistance bacillus and some viruses.
The present invention combinesreuterinWith natural, healthy, the efficient antibacterial feature of ε-poly-D-lysine, swashed again using proenzyme
Mode living promotesreuterinFermenting and producing, using food-grade ε-poly-D-lysine microemulsion as the mode of carrying system,
The antibacterial spraying of mobile phone that both they compounding obtains is had into efficient bacteriostasis, is carried out for the microbe species on mobile phone
The antibacterial spraying of mobile phone of best fungistatic effect is obtained after the allotment of antipathogenic composition.
Invention content
In view of the problems of the existing technology, it is an object of the invention to design to provide a kind of antibacterial spray of Luo Yishi soft white hands machine
The technical solution of the production method of mist.
The production method of a kind of antibacterial spraying of Luo Yishi soft white hands machine, it is characterised in that include the following steps：
1）Take lactobacillus reuteri（L. reuteri）After bacterial strain thaws, it is incubated on solid MRS tablets and is activated；
2）It takes the single bacterium colony after activation to be placed in stationary culture in sterilized liquid MRS culture mediums, is passed through nitrogen, keep
It is cultivated in the environment of anaerobism；
3）It takes culture gained supernatant to be centrifuged, centrifugation gained supernatant liquor is gone, is cultivated after washing bacterium；
4）Prepare coenzyme B12Reaction solution, per 1mL coenzyme B12Containing 0.01~0.1 mol/L KCl in reaction solution, 0.05~
0.5 mol/L 1,2-PD, 10~20umol/L coenzyme B12, 0.01~0.05mol/L buffer solution of potassium phosphate, pH3~4；
5）Take step 3）Obtained thalline is added glycerine and glucose, ferments under anaerobic；
6）Take step 5）Obtained zymotic fluid, centrifugation, takes supernatant zymotic fluid, obtains lactobacillus reuteri thalline；
7）Take step 6）Step 4 is added in obtained lactobacillus reuteri thalline）Obtained coenzyme B12In reaction solution, according to step
Rapid 5）Condition anaerobic fermentation after, it is Luo Yishi element extracting solutions to take supernatant zymotic fluid, and the Luo Yishi reactivated is replaced after 48h
Lactobacillus（L.reuteri）；
8）Take ε-poly-D-lysine as solute, as solvent, the ε-poly for being configured to 10~12mg/L concentration relies glycerine
Propylhomoserin microemulsion；
9）Step 7 is added in ε-poly-D-lysine microemulsion）Obtained Luo Yishi element extracting solutions, mix well；
10）Solution after compounding is added in spray bottle, the antibacterial spray product of mobile phone is obtained.
A kind of production method of antibacterial spraying of Luo Yishi soft white hands machine, it is characterised in that the step 1）Middle Roy
Family name's lactobacillus（L. reuteri）Activation condition：37 DEG C of 10~12h of culture.
A kind of production method of antibacterial spraying of Luo Yishi soft white hands machine, it is characterised in that the step 2）Middle Roy
Family name's lactobacillus（L. reuteri）Anaerobic culture conditions：37 DEG C of 15~16h of culture.
A kind of production method of antibacterial spraying of Luo Yishi soft white hands machine, it is characterised in that the step 3）Middle centrifugation
Condition：3000 rpm of rotating speed, 20~25 DEG C of centrifuging temperature, 10~15min of centrifugation time；Wash bacterium condition：0.85% sodium chloride solution
Wash bacterium twice；Condition of culture：0.5~1h is cultivated at 37 DEG C of temperature in 0.85% sodium chloride solution.
A kind of production method of antibacterial spraying of Luo Yishi soft white hands machine, it is characterised in that the step 5）Middle fermentation
Condition：37 DEG C of 2~2.5h of fermentation.
A kind of production method of antibacterial spraying of Luo Yishi soft white hands machine, it is characterised in that the step 5）Middle glycerine
Weight ratio with glucose is 3:1, addition is 10 times of thalline weight.
A kind of production method of antibacterial spraying of Luo Yishi soft white hands machine, it is characterised in that the step 6）Middle centrifugation
Condition is：10000 rpm of rotating speed, 4 DEG C of centrifuging temperature, 10~12min of centrifugation time.
A kind of production method of antibacterial spraying of Luo Yishi soft white hands machine, it is characterised in that the step 9）Middle Roy
Family name's element extracting solution it is a concentration of：7~10mL/L.
Compared with prior art, the invention has the advantages that：
1. its main antipathogenic composition ---reuterinFor natural bacteriostatic agent, there is wider antimicrobial spectrum.
2.reuterinAs the bacteriostatic agent of food-grade, health will not be damaged.
3. being eaten with ε-poly-D-lysine glyceritereuterinCarrier systems improve the stability of antibacterial spraying, and
Mobile phone screen will not be caused to damage.
4. ε-poly-D-lysine withreuterinIt is used in compounding, not only there is very high antibacterial efficiency, but also production cost
It is low.
Description of the drawings
Fig. 1 be plus the antibacterial spraying of mobile phone be not added under the conditions of antibacterial two kinds of the spraying of mobile phone to the shadow of salmonella total plate count
Ring figure；
Fig. 2 is the influence for adding the antibacterial spraying of mobile phone with being not added under the conditions of antibacterial two kinds of the spraying of mobile phone to salmonella OD values
Figure.
Specific implementation mode
It further illustrates the present invention with reference to embodiments.
Embodiment 1
1）Strain selects
It is selected to useL. reuteriBacterial strain.L. reuteriUsed culture medium is MRS broth medium.
AndL. reuteriPreservation be as antifreeze and to be stored in -80 so that 12.5% glycerite to be added in the mediumoC's
In refrigerator.
2）L. reuteriLactic acid bacteria activates
It willL. reuteriFrom -80oIt is taken out in C refrigerators, 4oAfter thawing under C, draws 100 microlitres of coatings and be incubated at solid
Make its activation on MRS tablets, 37oC cultivates 12h.
3）L. reuteriLactic acid bacteria is cultivated
Choose single bacterium colony（colony）It is placed in stationary culture in sterilized liquid MRS culture mediums, is passed through nitrogen 5 minutes
It keeps it in the environment of anaerobism, 37oC cultivates 16h.
4）L. reuteriThe eccentric cleaning of lactic acid bacteria
Culture gained supernatant 5mL is taken, is centrifuged in low-temperature and high-speed centrifuge, 3000 rpm of rotating speed, centrifuging temperature
20 DEG C, centrifugation time 10min.Then centrifugation gained supernatant is gone, bacterium is washed twice with 0.85% sodium chloride solution, and
Thalline, time 0.5h are cultivated at 37 DEG C in 0.85% sodium chloride solution.
5）Coenzyme B12The preparation of reaction solution
Contain 0.05 mol/L KCl, 0.2 mol/L 1,2-PD, 15 umol/L coenzyme B in 1mL reaction solutions12,0.035
Mol/L kaliumphosphate buffers（Ph3.6）.
6）reuterinProduction
The glycerine of 5mL is added in thalline after eccentric cleaning：Glucose（3:1）Solution（Glycerine and glucose solution add
Enter 10 times that amount is thalline weight）, under anaerobic 37oC fermentations 2h.
7）reuterinExtraction
Bacteria suspension is subjected to centrifugally operated, 10000 rpm of rotating speed, 4 DEG C, centrifugation time 10min of centrifuging temperature extracts supernatantreuterinZymotic fluid.
8）Proenzyme reactivation
By step 7）Separation obtainsL. reuteriMiddle addition 1mL actimide reaction solutions, according to step 6）Condition is sent out
After ferment, takes supernatant zymotic fluid to produce Luo Yishi element extracting solutions, the lactobacillus reuteri reactivated is replaced after 48h
（L.reuteri）.
9）It is prepared by microemulsion
Select ε-poly-D-lysine as solute, for glycerine as solvent, the ε-poly for being configured to 12 mg/L concentration relies ammonia
Acid solution 100mL.
10）Product matches
0.5mL is added in ε-poly-D-lysine microemulsionreuterinExtracting solution mixes well.
11）Product packaging
Solution after compounding is added in spray bottle, the antibacterial spray product of mobile phone is obtained.Aerosol container uses 25mL
Pet is bottled.
The antibacterial spraying of mobile phone that the embodiment is obtained carries out bacteriostatic test, fungistatic effect such as following table and attached drawing 1,2.
It is found that the antibacterial spraying of mobile phone that the present invention obtains has broad-spectrum antibacterial effect from table and attached drawing, and with very
High antibacterial efficiency.
Embodiment 2
1）Strain selects
It is selected to useL. reuteriBacterial strain.L. reuteriUsed culture medium is MRS broth medium.
AndL. reuteriPreservation be as antifreeze and to be stored in -80 so that 12.5% glycerite to be added in the mediumoC's
In refrigerator.
2）L. reuteriLactic acid bacteria activates
It willL. reuteriFrom -80oIt is taken out in C refrigerators, 4oAfter thawing under C, draws 100 microlitres of coatings and be incubated at solid
Make its activation on MRS tablets, 37oC cultivates 10h.
3）L. reuteriLactic acid bacteria is cultivated
Choose single bacterium colony（colony）It is placed in stationary culture in sterilized liquid mrs culture mediums, is passed through nitrogen 5 minutes
It keeps it in the environment of anaerobism, 37oC cultivates 15h.
4）L. reuteriThe eccentric cleaning of lactic acid bacteria
Culture gained supernatant 3mL is taken, is centrifuged in low-temperature and high-speed centrifuge, 3000 rpm of rotating speed, centrifuging temperature
25 DEG C, centrifugation time 15min.Then centrifugation gained supernatant is gone, bacterium is washed twice with 0.85% sodium chloride solution, and
Thalline, time 1h are cultivated at 37 DEG C in 0.85% sodium chloride solution.
5）Coenzyme B12The preparation of reaction solution
Contain 0.01 mol/L KCl, 0.05 mol/L 1,2-PD, 10 umol/L coenzyme B in 1mL reaction solutions12,0.01
Mol/L kaliumphosphate buffers（Ph3）.
6）reuterinProduction
The glycerine of 5mL is added in thalline after eccentric cleaning：Glucose（3:1）Solution（Glycerine and glucose solution add
Enter 10 times that amount is thalline weight）, under anaerobic 37 DEG C fermentation 2.5h.
7）reuterinExtraction
Bacteria suspension is subjected to centrifugally operated, 10000 rpm of rotating speed, 4 DEG C, centrifugation time 12min of centrifuging temperature extracts supernatantreuterinZymotic fluid.
8）Proenzyme reactivation
By step 7）Separation obtainsL. reuteriMiddle addition 1mL coenzyme B12Reaction solution, according to step 6）Technology item
After part fermentation, takes supernatant zymotic fluid to produce Luo Yishi element extracting solutions, the lactobacillus reuteri reactivated is replaced after 48h（L. reuteri）.
9）It is prepared by microemulsion
Select ε-poly-D-lysine as solute, for glycerine as solvent, the ε-poly for being configured to 12 mg/L concentration relies ammonia
Acid solution 50mL.
10）Product matches
0.5mL is added in ε-poly-D-lysine microemulsionreuterinExtracting solution mixes well.
11）Product packaging
Solution after compounding is added in spray bottle, the antibacterial spray product of mobile phone is obtained.Aerosol container uses 25mL
Pet is bottled.
The antibacterial spraying of mobile phone that the embodiment obtains carries out such as 1 identical experiment of embodiment, finally can also reach as implemented
1 identical technique effect of example.
Claims (8)
1. a kind of production method of the antibacterial spraying of Luo Yishi soft white hands machine, it is characterised in that include the following steps：
1）Take lactobacillus reuteri（L. reuteri）After bacterial strain thaws, it is incubated on solid MRS tablets and is activated；
2）It takes the single bacterium colony after activation to be placed in stationary culture in sterilized liquid MRS culture mediums, is passed through nitrogen, is maintained at and detests
It is cultivated in the environment of oxygen；
3）It takes culture gained supernatant to be centrifuged, centrifugation gained supernatant liquor is gone, is cultivated after washing bacterium；
4）Prepare coenzyme B12Reaction solution, per 1mL coenzyme B12Containing 0.01~0.1 mol/L KCl in reaction solution, 0.05~0.5
Mol/L 1,2-PD, 10~20umol/L coenzyme B12, 0.01~0.05mol/L buffer solution of potassium phosphate, pH3~4；
5）Take step 3）Obtained thalline is added glycerine and glucose, ferments under anaerobic；
6）Take step 5）Obtained zymotic fluid, centrifugation, takes supernatant zymotic fluid, obtains lactobacillus reuteri thalline；
7）Take step 6）Step 4 is added in obtained lactobacillus reuteri thalline）Obtained coenzyme B12In reaction solution, according to step 5）
Condition anaerobic fermentation after, it is Luo Yishi element extracting solutions to take supernatant zymotic fluid, and the Luo Yishi breast bars reactivated are replaced after 48h
Bacterium（L.reuteri）；
8）Take ε-poly-D-lysine as solute, glycerine is configured to ε-poly-D-lysine of 10~12mg/L concentration as solvent
Microemulsion；
9）Step 7 is added in ε-poly-D-lysine microemulsion）Obtained Luo Yishi element extracting solutions, mix well；
10）Solution after compounding is added in spray bottle, the antibacterial spray product of mobile phone is obtained.
2. a kind of production method of the antibacterial spraying of Luo Yishi soft white hands machine as described in claim 1, it is characterised in that the step
Rapid 1）Middle lactobacillus reuteri（L. reuteri）Activation condition：37 DEG C of 10~12h of culture.
3. a kind of production method of the antibacterial spraying of Luo Yishi soft white hands machine as described in claim 1, it is characterised in that the step
Rapid 2）Middle lactobacillus reuteri（L. reuteri）Anaerobic culture conditions：37 DEG C of 15~16h of culture.
4. a kind of production method of the antibacterial spraying of Luo Yishi soft white hands machine as described in claim 1, it is characterised in that the step
Rapid 3）Middle centrifugal condition：3000 rpm of rotating speed, 20~25 DEG C of centrifuging temperature, 10~15min of centrifugation time；Wash bacterium condition：0.85%
Sodium chloride solution washes bacterium twice；Condition of culture：0.5~1h is cultivated at 37 DEG C of temperature in 0.85% sodium chloride solution.
5. a kind of production method of the antibacterial spraying of Luo Yishi soft white hands machine as described in claim 1, it is characterised in that the step
Rapid 5）Middle fermentation condition：37 DEG C of 2~2.5h of fermentation.
6. a kind of production method of the antibacterial spraying of Luo Yishi soft white hands machine as described in claim 1, it is characterised in that the step
Rapid 5）The weight ratio of middle glycerine and glucose is 3:1, addition is 10 times of thalline weight.
7. a kind of production method of the antibacterial spraying of Luo Yishi soft white hands machine as described in claim 1, it is characterised in that the step
Rapid 6）Middle centrifugal condition is：10000 rpm of rotating speed, 4 DEG C of centrifuging temperature, 10~12min of centrifugation time.
8. a kind of production method of the antibacterial spraying of Luo Yishi soft white hands machine as described in claim 1, it is characterised in that the step
Rapid 9）Middle Luo Yishi elements extracting solution it is a concentration of：7~10mL/L.
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| CN108310441A (en) * | 2017-01-18 | 2018-07-24 | 南京理工大学 | A method of improving mobile phone screen surface flora |
| CN110269175A (en) * | 2019-06-05 | 2019-09-24 | 嘉兴浙华质检技术服务有限公司 | A kind of spraying micro-ecology preparation and preparation method thereof for feed anticorrosion |
Citations (2)
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| CN102026644A (en) * | 2008-06-10 | 2011-04-20 | 生命大地女神有限公司 | Controlled activation of the reuterin-production machinery of lactobacillus reuteri |
| CN103114051A (en) * | 2012-11-15 | 2013-05-22 | 上海交大昂立股份有限公司 | Roy bacterium lacticum and application thereof |
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- 2016-04-29 CN CN201610276532.7A patent/CN105851063B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102026644A (en) * | 2008-06-10 | 2011-04-20 | 生命大地女神有限公司 | Controlled activation of the reuterin-production machinery of lactobacillus reuteri |
| EP2303295B1 (en) * | 2008-06-10 | 2015-09-30 | Biogaia AB | Controlled activation of the reuterin-production machinery of lactobacillus reuteri |
| CN103114051A (en) * | 2012-11-15 | 2013-05-22 | 上海交大昂立股份有限公司 | Roy bacterium lacticum and application thereof |
Non-Patent Citations (2)
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| Purification and Characterization of Glycerol Dehydratase from Lactobacillus reuteri;TODD L. TALARICO et al;《Applied and Environmental Microbiology》;19900430;第56卷(第4期);第1195-1197页 * |
| 罗伊氏乳杆菌体外抑菌和免疫调节性能的研究;池海波 等;《中国乳品工业》;20151225;第43卷(第12期);第4-7页 * |
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