CN108374033A - A kind of extracting method of nisin - Google Patents
A kind of extracting method of nisin Download PDFInfo
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- CN108374033A CN108374033A CN201810150346.8A CN201810150346A CN108374033A CN 108374033 A CN108374033 A CN 108374033A CN 201810150346 A CN201810150346 A CN 201810150346A CN 108374033 A CN108374033 A CN 108374033A
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- nisin
- extracting method
- thalline
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- streptococcus
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/315—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Streptococcus (G), e.g. Enterococci
Abstract
The invention discloses a kind of extracting methods of nisin, are related to a kind of method extracted from microorganism and nisin is made.The extracting method of the nisin of the present invention includes the following steps：Fermentation lactic acid streptococcus obtains nisin fermented liquid；The pH value of zymotic fluid is adjusted to 4~5, concussion is adsorbed；After centrifuging thalline, with sodium radio-phosphate,P-32 solution washing thalline；Concussion elution, isolated supernatant；Concentrated supernatant；Removal of impurities.The extracting method of the nisin of the present invention is easy to operate, extraction obtains nisin purity is high, stability is strong, is consequently adapted to large-scale industrial production, has very high economic benefit.The invention belongs to molecule microorganisms technical fields.
Description
Technical field
The present invention relates to molecule microorganisms technical fields, and in particular to one kind is extracted from microorganism and streptococcus lactis is made
The method of element.
Background technology
Bacteriocin is certain bacteriums has bacteriostatic activity in metabolic process by one kind that Ribosome biogenesis mechanism generates
Polypeptide or Precursor Peptide.Narrow inhibition spectrum is presented to nearly edge bacterial strain of the same race, by perforating on target cell, inhibiting peptide glycan
Synthesis inhibits protein to synthesize with ribosomes or tRNA interactions, and directly degrade target cell DNA, to play fungistatic effect.
Bacteriocin is by bacterium or archeobacteria gene code, a kind of sterilization protein or polypeptide of Ribosome biogenesis, bacteriocin
Production bacterium there is immunity to the bacteriocin itself secreted.Bacteriocin is the one kind being present in the natural surroundings of bacterium life
The research of antibacterial material, early stage thinks that bacteriocin only works to of the same race or with Characterization bacterium, however, getting in recent years
Show that a kind of bacteriocin may also play inhibitory or killing effect to other various bacterias come more results of study.
Nineteen twenty-five, the uncommon Asia of A Grays report for the first time V plant of Escherichia coli generate it is a kind of have to Φ plant of Escherichia coli sterilize work
Substance, this substance are referred to as colicine.Find that many Escherichia coli can generate colicine again later, and
And col factor can be transferred to recipient cell from donorcells.
After the 1970s, molecular biology level is entered to the research of bacteriocin, has carried out the change to bacteriocin
Learn the exploration of property, structure, biosynthesis, release and mode of action etc..It is combined using bacteriocin or with phagocytosis body method,
The parting of certain bacteriums can be effectively performed and the popular of pathogen checks.
The generation of most of bacteriocin and host cell are controlled the immunity of bacteriocin by plasmid.Bacterium generates bacteriocin
It is the lethal processes of cell.Lethal substance can be divided into two classes by its property, and one kind is the protein or peptide of low molecular weight, it is difficult to
Observed under electron microscope to this substance structure, it is how unstable to trypsase；Another kind of is the egg for having labyrinth
White matter particle has bacteriophage portion forms structure, is easy to observe under an electron microscope, to trypsinase stability.
It is that Buddhist nun is pungent that nisin (Nisin), which is also known as nisin or transliteration, is the one of streptococcus lactis generation
Kind peptide material, is made of, molecular weight is about 3500Da 34 amino acid residues.Due to nisin can inhibit it is most of
Gram-positive bacterium, and have strong inhibiting effect to the spore of bacillus, therefore answered extensively as food preservative
For food service industry.It is hydrolyzed into amino acid quickly under physiological pH condition and α-chymotrypsin protein enzyme effect of human body after edible,
Normal flora in human body intestinal canal will not be changed and generate the resistance problem occurred such as other antibiotic, will not more be resisted with other
There is cross tolerance in rhzomorph, is a kind of efficient, nontoxic, safe, without side-effects antiseptics for natural food.
It can effectively inhibit many gram-positive bacteriums for causing food spoilage, such as clostridium botulinum, Staphylococcus aureus
Bacterium, Streptococcus hemolyticus, Listera, the growth and breeding of stearothermophilus bud pole bacterium, especially to producing sporogenic gram
Positive bacteria has special efficacy.The antibacterial action of nisin is to cause cell membrane by the normal function of interference cell v films
Infiltration, nutrient loss and film potential decline, so as to cause the death of pathogenic bacteria and spoilage organisms cell.It is a kind of nontoxic natural
Preservative has no adverse effects to color, the mouthfeel etc. of food.It is existing that oneself is widely used in dairy products, tin product, fish system
In product and alcoholic beverage.
It is purified into various bacteria element at present, the method for purification of bacterial element is varied, mainly has：Organic solvent deposit
Method, absorption method, heating denaturalization, ion exchange, gel chromatography, high performance liquid chromatography etc..Due to bacteriocin activity easily by
To the influence of extraneous physical and chemical condition, prodigious difficulty is brought for the extraction of bacteriocin.Existing nisin system
Preparation Method is complicated for operation, prepares difficulty, and product purity is low, is unfavorable for industrialized production.
Invention content
The purpose of the present invention is to provide a kind of nisin purity that easy to operate, extraction obtains, and high, antibacterial is lived
Property high nisin extracting method, it is complicated for operation to solve existing nisin preparation method, prepare
Difficulty, and product purity is low, the problem of being unfavorable for industrialized production.
To achieve the above object, the technical scheme is that a kind of extracting method of nisin, the extraction
Method includes the following steps：
(1) fermentation lactic acid streptococcus obtains nisin fermented liquid；
(2) pH value of zymotic fluid is adjusted to 4~5, and 0.5~5h is shaken at 10~25 DEG C, nisin is made to be adsorbed on
On streptococcus lactis thalline；
(3) streptococcus lactis thalline is centrifuged, the sodium radio-phosphate,P-32 solution washing thalline 1~3 time for being 6.0 with pH value, then
Thalline is suspended in the NaCl solution that pH is 2.0, vibrates centrifugation removal thalline after 0.5~4h, obtain supernatant；
(4) pH value of supernatant is adjusted to 6.0, is concentrated into the 5%~10% of original volume, is obtained concentrate；
(5) use Sephadex G-25 chromatographic columns removal concentrate in salt and impurity protein, collect eluent to get
To the nisin of the present invention.
Preferably, the culture medium that fermentation lactic acid streptococcus uses in the step (1) for：Sucrose 4.22g/100mL, soybean
Peptone 1.0g/100mL, yeast extract 0.5g/100mL, Na2HPO4For 0.8g/100mL, MgSO4·7H2O 0.02g/100mL,
Remaining is water.
Preferably, in the step (1) the streptococcic fermentation temperature of fermentation lactic acid be 28~30 DEG C, fermentation period be 10~
24h。
Preferably, in the step (3) sodium radio-phosphate,P-32 solution a concentration of 5mmol/L.
Preferably, in the step (3) NaCl solution a concentration of 100mmol/L.
Preferably, the mode concentrated in the step (4) is rotary evaporation.
Preferably, the elution requirement of chromatographic column is in the step (5)：Flow velocity is 0.5~2mL/min, uses deionized water
It is eluted.
Preferably, the step (5) also carries out following operation after collecting eluent：Rotary evaporation is concentrated into effluent volume
20%~80%.
The invention has the advantages that：
1, the extracting method of nisin of the invention is easy to operate, extracts obtained nisin purity
Height, stability are strong, are consequently adapted to large-scale industrial production, have very high economic benefit.
2, the nisin antibacterial activity that method culture using the present invention and extraction obtain is high, can effectively inhibit
With a variety of harmful bacterias of killing, lay a good foundation for its application in food preservative field.
Specific implementation mode
The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..
Embodiment 1
The extracting method of the nisin of the present embodiment carries out according to the following steps：
(1) nisin fermented liquid is prepared：
1. preparing fermentation medium：Sucrose 4.22g/100mL, soy peptone 1.0g/100mL, yeast extract 0.5g/
100mL, Na2HPO4For 0.8g/100mL, MgSO4·7H2O 0.02g/100mL, remaining is water；
2. accessing streptococcus lactis strain into the fermentation medium after sterilizing；
3. at 28 DEG C, shake culture fermentation 12h makes biomass in fermentation medium reach 108Cfu/mL is to get to lactic acid
Streptococcal fermentation liquid；
(2) pH value of zymotic fluid is adjusted to 4, and 1h is shaken at 10 DEG C, nisin is made to be adsorbed on streptococcus lactis bacterium
On body；
(3) streptococcus lactis thalline is centrifuged, the sodium radio-phosphate,P-32 solution washing thalline 1 time for being 6.0 with pH value, then by bacterium
Body is suspended in the NaCl solution that pH is 2.0, is vibrated centrifugation removal thalline after 3h, is obtained supernatant；The concentration of sodium radio-phosphate,P-32 solution
For 5mmol/L；A concentration of 100mmol/L of NaCl solution；
(4) pH value of supernatant is adjusted to 6.0, and rotary evaporation is concentrated into the 5%~10% of original volume, obtains concentrate；
(5) Sephadex G-25 chromatographic columns are used to remove salt and impurity protein in concentrate, elution requirement is：Flow velocity
It for 1mL/min, is eluted with deionized water, according to the elution time of nisin standard items, collection has nisin
The eluent of rhzomorph absorption peak；Eluent is collected to get to the nisin of the present embodiment, after measured the breast of the present embodiment
A concentration of the 2% of acid streptococci element.
Embodiment 2
The extracting method of the nisin of the present embodiment carries out according to the following steps：
(1) nisin fermented liquid is prepared：
1. preparing fermentation medium：Sucrose 4.22g/100mL, soy peptone 1.0g/100mL, yeast extract 0.5g/
100mL, Na2HPO4For 0.8g/100mL, MgSO4·7H2O 0.02g/100mL, remaining is water；
2. accessing streptococcus lactis strain into the fermentation medium after sterilizing；
3. at 30 DEG C, shake culture fermentation 10h makes biomass in fermentation medium reach 109Cfu/mL is to get to lactic acid
Streptococcal fermentation liquid；
(2) pH value of zymotic fluid is adjusted to 4, and 2h is shaken at 25 DEG C, nisin is made to be adsorbed on streptococcus lactis bacterium
On body；
(3) streptococcus lactis thalline is centrifuged, the sodium radio-phosphate,P-32 solution washing thalline 3 times for being 6.0 with pH value, then by bacterium
Body is suspended in the NaCl solution that pH is 2.0, is vibrated centrifugation removal thalline after 1h, is obtained supernatant；The concentration of sodium radio-phosphate,P-32 solution
For 5mmol/L；A concentration of 100mmol/L of NaCl solution；
(4) pH value of supernatant is adjusted to 6.0, and rotary evaporation is concentrated into the 5%~10% of original volume, obtains concentrate；
(5) Sephadex G-25 chromatographic columns are used to remove salt and impurity protein in concentrate, elution requirement is：Flow velocity
It for 1.5mL/min, is eluted with deionized water, according to the elution time of nisin standard items, collection has lactic acid chain
The eluent of coccus element absorption peak；
(6) eluent is collected, rotary evaporation is concentrated into the 40% of effluent volume to get to the nisin of the present embodiment
Rhzomorph, after measured a concentration of the 2.5% of the nisin of the present embodiment.
Embodiment 3
The extracting method of the nisin of the present embodiment carries out according to the following steps：
(1) nisin fermented liquid is prepared：
1. preparing fermentation medium：Sucrose 4.22g/100mL, soy peptone 1.0g/100mL, yeast extract 0.5g/
100mL, Na2HPO4For 0.8g/100mL, MgSO4·7H2O 0.02g/100mL, remaining is water；
2. accessing streptococcus lactis strain into the fermentation medium after sterilizing；
3. at 28 DEG C, shake culture fermentation 14h makes biomass in fermentation medium reach 108Cfu/mL is to get to lactic acid
Streptococcal fermentation liquid；
(2) pH value of zymotic fluid is adjusted to 5, and 5h is shaken at 15 DEG C, nisin is made to be adsorbed on streptococcus lactis bacterium
On body；
(3) streptococcus lactis thalline is centrifuged, the sodium radio-phosphate,P-32 solution washing thalline 3 times for being 6.0 with pH value, then by bacterium
Body is suspended in the NaCl solution that pH is 2.0, is vibrated centrifugation removal thalline after 0.5h, is obtained supernatant；Sodium radio-phosphate,P-32 solution it is dense
Degree is 5mmol/L；A concentration of 100mmol/L of NaCl solution；
(4) pH value of supernatant is adjusted to 6.0, and rotary evaporation is concentrated into the 5%~10% of original volume, obtains concentrate；
(5) Sephadex G-25 chromatographic columns are used to remove salt and impurity protein in concentrate, elution requirement is：Flow velocity
It for 0.5mL/min, is eluted with deionized water, according to the elution time of nisin standard items, collection has lactic acid chain
The eluent of coccus element absorption peak；
(6) eluent is collected, rotary evaporation is concentrated into the 60% of effluent volume to get to the nisin of the present embodiment
Rhzomorph, after measured a concentration of the 2.7% of the nisin of the present embodiment.
Embodiment 4
The extracting method of the nisin of the present embodiment carries out according to the following steps：
(1) nisin fermented liquid is prepared：
1. preparing fermentation medium：Sucrose 4.22g/100mL, soy peptone 1.0g/100mL, yeast extract 0.5g/
100mL, Na2HPO4For 0.8g/100mL, MgSO4·7H2O 0.02g/100mL, remaining is water；
2. accessing streptococcus lactis strain into the fermentation medium after sterilizing；
3. at 29 DEG C, shake culture is fermented for 24 hours, and biomass in fermentation medium is made to reach 109Cfu/mL is to get to lactic acid
Streptococcal fermentation liquid；
(2) pH value of zymotic fluid is adjusted to 4.5, and 3h is shaken at 20 DEG C, nisin is made to be adsorbed on streptococcus lactis
On thalline；
(3) streptococcus lactis thalline is centrifuged, the sodium radio-phosphate,P-32 solution washing thalline 3 times for being 6.0 with pH value, then by bacterium
Body is suspended in the NaCl solution that pH is 2.0, is vibrated centrifugation removal thalline after 2h, is obtained supernatant；The concentration of sodium radio-phosphate,P-32 solution
For 5mmol/L；A concentration of 100mmol/L of NaCl solution；
(4) pH value of supernatant is adjusted to 6.0, and rotary evaporation is concentrated into the 5%~10% of original volume, obtains concentrate；
(5) Sephadex G-25 chromatographic columns are used to remove salt and impurity protein in concentrate, elution requirement is：Flow velocity
It for 2mL/min, is eluted with deionized water, according to the elution time of nisin standard items, collection has nisin
The eluent of rhzomorph absorption peak；
(6) eluent is collected, rotary evaporation is concentrated into the 20% of effluent volume to get to the nisin of the present embodiment
Rhzomorph, after measured a concentration of the 3.1% of the nisin of the present embodiment.
Embodiment 5
The extracting method of the nisin of the present embodiment carries out according to the following steps：
(1) nisin fermented liquid is prepared：
1. preparing fermentation medium：Sucrose 4.22g/100mL, soy peptone 1.0g/100mL, yeast extract 0.5g/
100mL, Na2HPO4For 0.8g/100mL, MgSO4·7H2O 0.02g/100mL, remaining is water；
2. accessing streptococcus lactis strain into the fermentation medium after sterilizing；
3. at 30 DEG C, shake culture fermentation 10h makes biomass in fermentation medium reach 108Cfu/mL is to get to lactic acid
Streptococcal fermentation liquid；
(2) pH value of zymotic fluid is adjusted to 5, and 0.5h is shaken at 22 DEG C, nisin is made to be adsorbed on streptococcus lactis
On thalline；
(3) streptococcus lactis thalline is centrifuged, the sodium radio-phosphate,P-32 solution washing thalline 2 times for being 6.0 with pH value, then by bacterium
Body is suspended in the NaCl solution that pH is 2.0, is vibrated centrifugation removal thalline after 4h, is obtained supernatant；The concentration of sodium radio-phosphate,P-32 solution
For 5mmol/L；A concentration of 100mmol/L of NaCl solution；
(4) pH value of supernatant is adjusted to 6.0, and rotary evaporation is concentrated into the 5%~10% of original volume, obtains concentrate；
(5) Sephadex G-25 chromatographic columns are used to remove salt and impurity protein in concentrate, elution requirement is：Flow velocity
It for 0.5mL/min, is eluted with deionized water, according to the elution time of nisin standard items, collection has lactic acid chain
The eluent of coccus element absorption peak；
(6) eluent is collected, rotary evaporation is concentrated into the 80% of effluent volume to get to the nisin of the present embodiment
Rhzomorph, after measured a concentration of the 3.2% of the nisin of the present embodiment.
The antibacterial activity of nisin produced by the present invention is measured in the following ways：
Test strain：Staphylococcus aureus, listeria spp, Escherichia coli, salmonella typhimurium, Freund shiga
Bacterium, pseudomonas aeruginosa, Lactobacillus casei.
Bacteriostatic activity analysis method：The agar (2%) that 10mL heating and meltings are first poured into sterilized petri dishes waits for that it is fully cold
But after solidifying, it is several to be put into sterilized Oxford cup, and marshalling in a certain order.It is respectively that cultured test strain is dilute
It releases to 106~107The 15mL solid MRS culture mediums to have sterilized are cooled to 50 DEG C or so by cfu/mL, and test bacterium solution 1mL is added,
It is rapid to be uniformly mixed, it pours into and is placed in the plate of Oxford cup, Oxford cup is taken out with aseptic nipper after cooling.It will be in embodiment 1-5
After nisin obtained dilutes 10 times, then the 100 μ L of nisin after dilution is taken to be added to Oxford cup formation
In aperture, 37 DEG C are incubated overnight, and antibacterial circle diameter is measured with vernier caliper.
Specific inhibition zone result is as shown in table 1-5.
The bacteriostatic activity of product is made in 1 embodiment 1 of table
| Indicator bacteria | Antibacterial circle diameter (mm) |
| Staphylococcus aureus | 12.6 |
| Listeria spp | 17.5 |
| Escherichia coli | 14.8 |
| Shigella flexneri | 15.9 |
| Lactobacillus casei | 11.2 |
| Salmonella typhimurium | 16.4 |
| Pseudomonas aeruginosa | 17.6 |
The bacteriostatic activity of product is made in 2 embodiment 2 of table
| Indicator bacteria | Antibacterial circle diameter (mm) |
| Staphylococcus aureus | 13.5 |
| Listeria spp | 18.2 |
| Escherichia coli | 14.5 |
| Shigella flexneri | 15.7 |
| Lactobacillus casei | 11.4 |
| Salmonella typhimurium | 16.8 |
| Pseudomonas aeruginosa | 17.9 |
The bacteriostatic activity of product is made in 3 embodiment 3 of table
| Indicator bacteria | Antibacterial circle diameter (mm) |
| Staphylococcus aureus | 12.1 |
| Listeria spp | 17.9 |
| Escherichia coli | 15.1 |
| Shigella flexneri | 16.2 |
| Lactobacillus casei | 11.4 |
| Salmonella typhimurium | 16.4 |
| Pseudomonas aeruginosa | 17.3 |
The bacteriostatic activity of product is made in 4 embodiment 4 of table
| Indicator bacteria | Antibacterial circle diameter (mm) |
| Staphylococcus aureus | 12.4 |
| Listeria spp | 17.8 |
| Escherichia coli | 14.4 |
| Shigella flexneri | 15.7 |
| Lactobacillus casei | 11.5 |
| Salmonella typhimurium | 16.6 |
| Pseudomonas aeruginosa | 17.5 |
The bacteriostatic activity of product is made in 5 embodiment 5 of table
| Indicator bacteria | Antibacterial circle diameter (mm) |
| Staphylococcus aureus | 12.7 |
| Listeria spp | 17.5 |
| Escherichia coli | 14.6 |
| Shigella flexneri | 16.1 |
| Lactobacillus casei | 10.8 |
| Salmonella typhimurium | 16.6 |
| Pseudomonas aeruginosa | 17.3 |
By experimental result in table 1-5 it is found that the nisin antimicrobial spectrum of the present invention is wider, it can not only inhibit leather blue
Family name's positive bacteria also has inhibition to Gram-negative bacteria, can divide bacillus with suppressing portion.
The absolute acid stability of nisin produced by the present invention is measured in the following ways：
By the 5mol/L HCl and 5mol/L NaOH of nisin obtained in embodiment 1-5 adjust pH be 2.0~
11.0.4h is incubated at 37 DEG C, then pH is adjusted to 6.0, measures its bacteriostatic activity respectively.The result shows that bacteriocins from streptococcus thermophilus
Activity keeps stablizing in the ranges of pH2.0~9.0, and activity reduces when pH ＞ 9.0.
Although above having used general explanation and specific embodiment, the present invention is described in detail, at this
On the basis of invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Therefore,
These modifications or improvements without departing from theon the basis of the spirit of the present invention belong to the scope of protection of present invention.
Claims (8)
1. a kind of extracting method of nisin, it is characterised in that：The extracting method includes the following steps：
(1) fermentation lactic acid streptococcus obtains nisin fermented liquid；
(2) pH value of zymotic fluid is adjusted to 4~5, and 0.5~5h is shaken at 10~25 DEG C, nisin is made to be adsorbed on lactic acid
On streptococcus thalline；
(3) streptococcus lactis thalline is centrifuged, the sodium radio-phosphate,P-32 solution washing thalline 1~3 time for being 6.0 with pH value, then by bacterium
Body is suspended in the NaCl solution that pH is 2.0, is vibrated centrifugation removal thalline after 0.5~4h, is obtained supernatant；
(4) pH value of supernatant is adjusted to 6.0, is concentrated into the 5%~10% of original volume, is obtained concentrate；
(5) salt and impurity protein in Sephadex G-25 chromatographic columns removal concentrate are used, collects eluent to get to originally
The nisin of invention.
2. the extracting method of nisin according to claim 1, it is characterised in that：Fermentation in the step (1)
The culture medium that streptococcus lactis uses for：Sucrose 4.22g/100mL, soy peptone 1.0g/100mL, yeast extract 0.5g/
100mL, Na2HPO4For 0.8g/100mL, MgSO4·7H2O 0.02g/100mL, remaining is water.
3. the extracting method of nisin according to claim 1, it is characterised in that：Fermentation in the step (1)
The fermentation temperature of streptococcus lactis be 28~30 DEG C, fermentation period be 10~for 24 hours.
4. the extracting method of nisin according to claim 1, it is characterised in that：Phosphoric acid in the step (3)
A concentration of 5mmol/L of sodium solution.
5. the extracting method of nisin according to claim 1, it is characterised in that：NaCl in the step (3)
A concentration of 100mmol/L of solution.
6. the extracting method of nisin according to claim 1, it is characterised in that：Concentration in the step (4)
Mode be rotary evaporation.
7. the extracting method of nisin according to claim 1, it is characterised in that：Chromatography in the step (5)
The elution requirement of column is：Flow velocity is 0.5~2mL/min, is eluted with deionized water.
8. the extracting method of nisin according to claim 1, it is characterised in that：The step (5) is collected and is washed
Following operation is also carried out after de- liquid：Rotary evaporation is concentrated into the 20%~80% of effluent volume.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112225784A (en) * | 2020-09-23 | 2021-01-15 | 通辽市圣达生物工程有限公司 | Method for extracting nisin |
| CN114732941A (en) * | 2022-03-22 | 2022-07-12 | 安徽徽科生物工程技术有限公司 | Preparation method of cleaning and antibacterial gel |
| CN115120550A (en) * | 2021-03-26 | 2022-09-30 | 株式会社现代百朗德 | Hair cosmetic composition comprising a culture of streptococci providing beneficial effects on the hair |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004000167A (en) * | 2002-03-22 | 2004-01-08 | Omugi Hakko Kenkyusho:Kk | Process for preparing fermentation product containing significant amount of nisin by using barley shochu distillation residual liquid as medium |
| JP3672259B2 (en) * | 2002-03-22 | 2005-07-20 | 株式会社大麦発酵研究所 | A method for producing a fermented product containing a significant amount of nisin using barley shochu distillation residue and rice shochu distillation residue as a medium |
| CN1724562A (en) * | 2004-07-22 | 2006-01-25 | 中国科学院生态环境研究中心 | The novel method of absorbing lacto streptococus peptide from fermented liquid |
| CN101608201A (en) * | 2009-07-13 | 2009-12-23 | 天津科技大学 | A kind of production method of novel streptococcus thermophilus bacteriocin |
| CN103798515A (en) * | 2012-11-09 | 2014-05-21 | 山东福瑞达生物科技有限公司 | Method for preparing feed additive by using nisin extraction waste liquid |
| CN104046585A (en) * | 2014-06-19 | 2014-09-17 | 北京工商大学 | Bifidobacterium animal bacteriocin, production method thereof and specific production strain |
| RU2585521C1 (en) * | 2015-03-10 | 2016-05-27 | Маргарита Анатольевна Иванова | Method and process line for production of nisin |
| CN105755074A (en) * | 2014-12-15 | 2016-07-13 | 瑞普（天津）生物药业有限公司 | Method for extracting Nisin |
-
2018
- 2018-02-13 CN CN201810150346.8A patent/CN108374033A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004000167A (en) * | 2002-03-22 | 2004-01-08 | Omugi Hakko Kenkyusho:Kk | Process for preparing fermentation product containing significant amount of nisin by using barley shochu distillation residual liquid as medium |
| JP3672259B2 (en) * | 2002-03-22 | 2005-07-20 | 株式会社大麦発酵研究所 | A method for producing a fermented product containing a significant amount of nisin using barley shochu distillation residue and rice shochu distillation residue as a medium |
| CN1724562A (en) * | 2004-07-22 | 2006-01-25 | 中国科学院生态环境研究中心 | The novel method of absorbing lacto streptococus peptide from fermented liquid |
| CN101608201A (en) * | 2009-07-13 | 2009-12-23 | 天津科技大学 | A kind of production method of novel streptococcus thermophilus bacteriocin |
| CN103798515A (en) * | 2012-11-09 | 2014-05-21 | 山东福瑞达生物科技有限公司 | Method for preparing feed additive by using nisin extraction waste liquid |
| CN104046585A (en) * | 2014-06-19 | 2014-09-17 | 北京工商大学 | Bifidobacterium animal bacteriocin, production method thereof and specific production strain |
| CN105755074A (en) * | 2014-12-15 | 2016-07-13 | 瑞普（天津）生物药业有限公司 | Method for extracting Nisin |
| RU2585521C1 (en) * | 2015-03-10 | 2016-05-27 | Маргарита Анатольевна Иванова | Method and process line for production of nisin |
Non-Patent Citations (4)
| Title |
|---|
| M.N. AMIALI等: "High nisin Z production by Lactococcus lactis UL719 in whey permeate with aeration", 《WORLD JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY》 * |
| MEENU CHOPRA等: "Surfactant assisted nisin loaded chitosan-carageenan nanocapsule synthesis for controlling food pathogens", 《FOOD CONTROL》 * |
| 叶嗣颖等: "乳链菌肽nisinZ的pH吸附法分离纯化技术研究", 《同济医科大学学报》 * |
| 田文利等: "乳酸链球菌素(Nisin)的研究进展", 《食品工业》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112225784A (en) * | 2020-09-23 | 2021-01-15 | 通辽市圣达生物工程有限公司 | Method for extracting nisin |
| CN115120550A (en) * | 2021-03-26 | 2022-09-30 | 株式会社现代百朗德 | Hair cosmetic composition comprising a culture of streptococci providing beneficial effects on the hair |
| CN114732941A (en) * | 2022-03-22 | 2022-07-12 | 安徽徽科生物工程技术有限公司 | Preparation method of cleaning and antibacterial gel |
| CN114732941B (en) * | 2022-03-22 | 2022-12-09 | 安徽徽科生物工程技术有限公司 | Preparation method of cleaning and antibacterial gel |
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