CN104770827A - Nisin microemulsion - Google Patents
Nisin microemulsion Download PDFInfo
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- CN104770827A CN104770827A CN201410016022.7A CN201410016022A CN104770827A CN 104770827 A CN104770827 A CN 104770827A CN 201410016022 A CN201410016022 A CN 201410016022A CN 104770827 A CN104770827 A CN 104770827A
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Abstract
The invention discloses a nisin microemulsion, and a preparation method and an application scope thereof, and belongs to the food fresh keeping technical field. The microemulsion contains nisin, blend oil, Triton X-100, a 95% ethanol, Tween 80, cetyl trimethyl ammonium bromide, purified water and other components; after ultrasonic treatment by ultrasonic waves and high-pressure homogenization treatment, the nisin solubility is enhanced, and the microemulsion having relatively good stability is obtained; the microemulsion has good performance of inhibiting bacillus subtilis, also can inhibit Gram-negative bacteria such as escherichia coli and staphylococcus aureus.
Description
Technical field
The present invention relates to food antiseptic technical field of preservation of fresh, particularly relate to a kind of have press down food-borne pathogens containing the microemulsion of nisin.
Background technology
Nisin is the peptide species class antibacterial substance that streptococcus lactis produces, and belong to natural safe biological preservative, also referred to as nisin, English name is NISIN, and be made up of 34 amino acid residues, molecular weight is about 3500Da.Nisin reason lactic acid bacteria produces, most of gram-positive bacterium can be suppressed, and have good inhibition to the spore of bacillus, can amino acid be degraded to and there is not safety problem in human body, being widely used in food service industry as anticorrisive agent at present.Nisin is the pressed powder of a kind of light brown to white, and the solubility in water is less, more soluble in acid solution, is dissolved in alkaline solution hardly.
Microemulsion has the features such as stability is high, solubilizing amount is large, interfacial tension is low, and be widely used in the fields such as food, medicine, daily-use chemical industry, application prospect is very wide.In field of food, the application of microemulsion technology in beverage production, heat sensitive material embedding, food antiseptic etc. increases gradually, becomes study hotspot at present.The preparation method of current microemulsion mainly comprises the method such as ultrasonic emulsification or high-pressure homogeneous emulsification, needs to add suitable surfactant in ultrasonic emulsification and homogenizing process.
Food origin disease is the major issue causing food security, and food origin disease is defined as by the World Health Organization: " every human body that to enter by ingesting makes human body suffer from infectious or toxic disease." tentative disease causes primarily of food-borne pathogens, cause diarrhoea, the various diseases such as poisoning, cause larger economic loss every year.Common food-borne pathogens comprises Escherichia coli, staphylococcus aureus etc.
Nisin has wider application as efficient security natural antiseptic agent, but its dissolubility is relatively low, and price is higher, by adding the compositions such as emulsifying agent, the microemulsion of preparation containing nisin, its effect can be improved, if current nisin is to suppress gram-positive bacteria, prepare in conjunction with the method that high-pressure homogeneous process combines the range of application that microemulsion likely widens nisin by ultrasonic emulsification.
Summary of the invention
1. in order to overcome above-mentioned the deficiencies in the prior art, the invention provides one (nisin microemulsion, this microemulsion contains nisin, ready-mixed oil, triton x-100, 95% ethanol, Tween 80, the composition such as softex kw and pure water, after ultrasonic wave and high-pressure homogeneous process, improve the dissolubility of nisin, obtain the microemulsion with good stability, this microemulsion has good suppression gram-positive bacteria as outside bacillus subtilis, Gram-negative bacteria can also be suppressed as Escherichia coli and staphylococcus aureus).
2. in application process, there is dissolubility low for nisin, to suppress gram-positive bacterium, and because of nisin be natural anticorrisive agent, price is relatively high, be widely used in the anticorrosion of the different food products such as meat products, mixed congee, beverage at present, but carrying out microemulsified process, to carry out antibacterial research report few.For the uniformity in raising nisin application process and effect, adopt ultrasonic emulsification and high pressure homogenization technique to process the solution containing nisin and emulsifying agent, obtain stable microemulsion.
3. compared with prior art, the present invention constructs a kind of microemulsion containing nisin, has widened range of application and the action effect of nisin, has reduced its actual application cost, in food-borne pathogens control, show good effect.
4. the technical solution adopted in the present invention is:
(1) a certain amount of nisin of accurate weighing, with 0.02M aqueous hydrochloric acid solution for solvent is mixed with solution containing nisin,
(2) certain streptococcus lactis cellulose solution is drawn in Boiling tube, add 95% ethanol, salad oil, triton x-100, Tween 80, softex kw successively, and be supplemented to 100mL with deionized water, each step all fully shakes test tube above, in vitro solution is mixed
(3) solution mixed is placed in ultrasonic cleaner and carries out ultrasonic emulsification process, adopt 200W Power Processing a few minutes, subsequently emulsion is placed in high pressure homogenizer (ATS engineering INC) and carries out high-pressure homogeneous process, wherein pressure 200-600bar, homogenizing time 2-10min
(4) to obtained microemulsion, laser fineness gage is adopted to carry out grain size analysis,
(5) microemulsion obtained mixed in right amount or coat on beef extract-peptone flat board, to mix or to be coated with the flat board of sterilized water for contrast, then being coated with certain density microorganism suspension respectively, the antibacterial situation of contrast nisin.
At present, have in the application process of nisin that dissolubility is bad, cost is higher and the problem such as antimicrobial spectrum is not wide, and rare nisin is carried out research prepared by microemulsified as antipathogenic composition, in order to improve the effect in nisin application process, adopt ultrasonic emulsification and high pressure homogenization technique to carry out emulsification and homogeneous process to the solution containing compositions such as nisin, ethanol, salad oil, triton x-100, Tween 80, softex kws, obtain stable microemulsion.Compare with technology with the application process of existing nisin, ultrasonic wave and the homogeneous process of nisin improve its range of application, obtain the microemulsion with good stability, in the suppression of gram-positive bacterium and food-borne pathogenic negative bacteria, there is good effect.
Accompanying drawing explanation
Fig. 1 is nisin microemulsion grain size distribution.
Fig. 2 is 100 μ L0.5% (m/v) nisin microemulsion fungistatic effect figure.
Detailed description of the invention
Embodiment 1
In the present embodiment, the formula of microemulsion contains nisin, 95% ethanol, ready-mixed oil, triton x-100, Tween 80, softex kw and pure water.First with the streptococcus lactis cellulose solution of the hydrochloric acid solution of 0.02M preparation 5%, streptococcus lactis cellulose solution 0.5mL is drawn by liquid-transfering gun, pipette etc., add 95% ethanol 0.3mL successively, ready-mixed oil 0.1mL, triton x-100 0.1mL, Tween 80 0.1mL, softex kw (10% aqueous solution) 0.2mL is in Boiling tube, mix with vortex oscillator in dropping process, and repeatedly add pure water on a small quantity gradually, being supplemented to mixture solution total amount is 100mL.Then ultrasonic wave is adopted to carry out ultrasonic process to mixed solution, processing power 200W, processing time is 3min, homogeneous process is carried out again afterwards, homogenization pressure 300bar, homogenizing time 5min by high pressure homogenizer, the microemulsion of uniformity, after high-pressure homogeneous, the uniformity of emulsion significantly improves, and the stability of put procedure also has remarkable increase.After 50 μ L microemulsions are coated beef-protein medium, add 100 μ L and be diluted to 10
6the bacillus subtilis of concentration, Escherichia coli and staphylococcus glucose coccus suspension, first blank group is coated with sterilized water 50 μ L, add the above-mentioned bacterium liquid of 100 μ L again, counting after cultivating 24 hours at 37 DEG C, on the flat board dripping microemulsion, the quantity of bacillus subtilis, Escherichia coli and staphylococcus glucose ball three kinds of microorganisms is respectively 21.5% of blank, 22.3% and 4%, microemulsion has good fungistatic effect.
Embodiment 2
In the present embodiment, the formula of microemulsion contains nisin, 95% ethanol, ready-mixed oil, triton x-100, Tween 80, softex kw and pure water.First with the streptococcus lactis cellulose solution of the hydrochloric acid solution of 0.02M preparation 5%, streptococcus lactis cellulose solution 4mL is drawn by liquid-transfering gun, pipette etc., add 95% ethanol 3mL successively, ready-mixed oil 4mL, triton x-100 2mL, Tween 80 1mL, softex kw (10% aqueous solution) 3mL is in large beaker, mix with vortex oscillator in dropping process, and repeatedly add pure water on a small quantity gradually, being supplemented to mixture solution total amount is 1000mL.Then ultrasonic wave is adopted to carry out ultrasonic process to mixed solution, processing power 200W, processing time is 6min, homogeneous process is carried out again afterwards by high pressure homogenizer, homogenization pressure 500bar, homogenizing time 8min, the microemulsion of uniformity, the uniformity and the stability of high-pressure homogeneous rear emulsion significantly improve.After 100 μ L microemulsions are coated beef-protein medium, add 100 μ L and be diluted to 10
6the bacillus subtilis of concentration, Escherichia coli and staphylococcus glucose coccus suspension, first blank group is coated with sterilized water 100 μ L, add the above-mentioned bacterium liquid of 100 μ L again, counting after cultivating 36 hours at 37 DEG C, on the flat board dripping microemulsion, the quantity of bacillus subtilis, Escherichia coli and staphylococcus glucose ball three kinds of microorganisms is respectively 8.5% of blank, 14.3% and 2.1%, the microemulsion of gained shows good fungistatic effect.
Claims (2)
1. one kind (nisin microemulsion), it is characterized in that: (this microemulsion contains the compositions such as nisin, ready-mixed oil, triton x-100,95% ethanol, Tween 80, softex kw and pure water, after ultrasonic wave and high-pressure homogeneous process, improve the dissolubility of nisin, obtain the microemulsion with good stability, this microemulsion has good suppression gram-positive bacteria as outside bacillus subtilis, can also suppress Gram-negative bacteria as Escherichia coli and staphylococcus aureus).
2. (a kind of nisin microemulsion) according to claim 1, it is characterized in that:
(1) microemulsion containing nisin, this microemulsion contains the nisin with antipathogenic composition, and its content (m/v) is at 0.1-0.4%.
(2) a kind of microemulsion containing nisin, also containing ready-mixed oil in this microemulsion, triton x-100, 95% ethanol, Tween 80, the composition such as softex kw and water, wherein ready-mixed oil percentage (v/v) is 0.5-4%%, triton x-100 percentage (v/v) is 0.5%-3%, 95% ethanol percentage (v/v) is 1%-5%, Tween 80 percentage (v/v) is 1%-3%, softex kw (10% aqueous solution) percentage (v/v) is 0.05-4%, all the other are deionized water, emulsion total amount is 100 parts.
(3) microemulsion containing nisin, concrete grammar prepared by this microemulsion is as follows:
1. a certain amount of nisin of accurate weighing, is mixed with the nisin aqueous solution of 0.2-0.8% with the aqueous hydrochloric acid solution of 0.02M,
2. the streptococcus lactis cellulose solution of 5mL is drawn in Boiling tube, add concentration in 95% ethanol, salad oil, triton x-100, Tween 80, softex kw to (2) successively, and be supplemented to 100mL with deionized water, each step all fully shakes test tube above, in vitro solution is mixed
3. the solution mixed is placed in ultrasonic cleaner and carries out ultrasonic emulsification process, adopt 200W Power Processing 2-10min, subsequently emulsion is placed in high pressure homogenizer (ATS engineering INC) and carries out high-pressure homogeneous process, wherein pressure 200-600bar, homogenizing time 2-10min
4. a gained microemulsion part is used for grain size analysis, and adopt laser fineness gage (BT-9300H, Baite Instrument Co., Ltd., Dandong) to carry out grain size analysis mensuration, a part is used for bacteriostatic experiment.
(4) a kind of nisin microemulsion, this microemulsion successively adopts ultrasonic emulsification and high pressure homogenizer homogeneous two kinds of methods to process, after certain hour process, the each composition of microemulsion of gained is uniformly dispersed unanimously, reach stable state, and decrease the consumption of nisin in actual application of comparing.
(5) a kind of nisin microemulsion, this microemulsion has the effect of good antagonism bacillus subtilis, making lactic acid streptostacin percentage composition (m/v) is respectively the microemulsion of 0.1%, 0.2%, 0.3%, 0.4%, respectively getting 50 μ L is mixed on beef-protein medium flat board, to drip the flat board of sterilized water for blank, measure microemulsion to the inhibiting rate of microorganism, gained nisin microemulsion has the effect significantly suppressing Gram-negative and positive bacteria.
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| CN201410016022.7A CN104770827A (en) | 2014-01-13 | 2014-01-13 | Nisin microemulsion |
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| CN201410016022.7A CN104770827A (en) | 2014-01-13 | 2014-01-13 | Nisin microemulsion |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105054217A (en) * | 2015-08-28 | 2015-11-18 | 佛山市立高食品有限公司 | Compound food preservative |
| CN105265994A (en) * | 2015-10-26 | 2016-01-27 | 天津科技大学 | Method effectively improving heat stability of Nisin |
| CN108410762A (en) * | 2018-03-07 | 2018-08-17 | 海南医学院 | A kind of streptococcus novel species HTS20 and its application |
| CN108485999A (en) * | 2018-03-07 | 2018-09-04 | 海南医学院 | A kind of streptococcus novel species HTS25 and its application |
| CN112075564A (en) * | 2020-08-31 | 2020-12-15 | 江苏大学 | Eucalyptus citriodora essential oil/nisin nano-emulsion and preparation method thereof |
| CN112826002A (en) * | 2021-01-22 | 2021-05-25 | 浙江工业大学 | Nisin compound and application thereof in preparation of antibacterial agent |
| CN113100376A (en) * | 2021-04-01 | 2021-07-13 | 浙江工业大学 | Nisin lipid microcapsule and preparation and application thereof |
-
2014
- 2014-01-13 CN CN201410016022.7A patent/CN104770827A/en active Pending
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105054217A (en) * | 2015-08-28 | 2015-11-18 | 佛山市立高食品有限公司 | Compound food preservative |
| CN105265994A (en) * | 2015-10-26 | 2016-01-27 | 天津科技大学 | Method effectively improving heat stability of Nisin |
| CN105265994B (en) * | 2015-10-26 | 2019-01-29 | 天津科技大学 | A method of effectively improving Nisin thermal stability |
| CN108410762A (en) * | 2018-03-07 | 2018-08-17 | 海南医学院 | A kind of streptococcus novel species HTS20 and its application |
| CN108485999A (en) * | 2018-03-07 | 2018-09-04 | 海南医学院 | A kind of streptococcus novel species HTS25 and its application |
| CN108410762B (en) * | 2018-03-07 | 2019-05-03 | 海南医学院 | A kind of streptococcus novel species HTS20 and its application |
| CN112075564A (en) * | 2020-08-31 | 2020-12-15 | 江苏大学 | Eucalyptus citriodora essential oil/nisin nano-emulsion and preparation method thereof |
| CN112826002A (en) * | 2021-01-22 | 2021-05-25 | 浙江工业大学 | Nisin compound and application thereof in preparation of antibacterial agent |
| CN112826002B (en) * | 2021-01-22 | 2023-10-20 | 浙江工业大学 | Nisin compound and application thereof in preparation of antibacterial agent |
| CN113100376A (en) * | 2021-04-01 | 2021-07-13 | 浙江工业大学 | Nisin lipid microcapsule and preparation and application thereof |
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