CN105440113A - Extraction method for nisin - Google Patents
Extraction method for nisin Download PDFInfo
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- CN105440113A CN105440113A CN201410516668.1A CN201410516668A CN105440113A CN 105440113 A CN105440113 A CN 105440113A CN 201410516668 A CN201410516668 A CN 201410516668A CN 105440113 A CN105440113 A CN 105440113A
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Abstract
The invention relates to an extraction method for nisin. The extraction method is characterized by comprising the following steps: (1) adjusting a pH value of a nisin fermentation concentrate to 1.7 to 3.0, and mixing the obtained concentrate with an organic solvent slightly soluble in water or an organic solvent insoluble in water, wherein the organic solvent is any one or a combination of any two selected from the group consisting of dichloromethane, n-butanol, ethyl acetate and tetrachloromethane; and (2) carrying out stirring and standing, allowing a solid phase to be separated so as to obtain a wet nisin cake, and carrying out pressure-reduced drying so as to obtain the nisin. The extraction method provided by the invention has the following main advantages: the organic solvent is used for precipitation of the nisin, so the disadvantages of high salt content and low product purity in the prior art are overcome; product purity of the nisin is higher than 50%; and the method is safe, simple, convenient, practical, and low in cost.
Description
Technical field
The present invention relates to a kind of preparation method of antiseptics for natural food, specifically a kind of method of separation and Extraction nisin from streptococcus acidi lactici or Lactococcus lactis Fermented Condensed liquid.
Background technology
Nisin is also known as Nisin or nisin, and be the polypeptide with very strong germicidal action produced in metabolic process by streptococcus acidi lactici or Lactococcus lactis, primary structure is made up of 34 amino acid, and molecular weight is about 3510Da.Nisin is a kind of antiseptics for natural food, can suppress most of G such as Staphylococcus, streptococcus and bacillus
+the Growth and reproduction of bacterium and gemma thereof, have germ resistance strong, to advantages such as human non-toxic, good water solubility, Heat stability is goods, be widely used in the anti-corrosive fresh-keeping of milk-product, tinned pre-, meat product and beverage.
Since nineteen sixties, the ways and means extracting nisin from fermented liquid is set up successively, and its removal process mainly comprises the steps: fermented liquid acidifying, heating, concentrates, saltout (or spraying dry), vacuum-drying.Concentration method mainly contains: cryogenic vacuum method of enrichment, ultrafiltration and concentration method, Foam concentration, Adsorption Concentration.Absorption method adds solid particulate adsorbent nisin in fermented liquid, is saltoutd or spraying dry by stripping liquid after desorb, makes Powdered nisin product; First membrane filter method adopts Plate Filtration to carry out pre-treatment to fermented liquid, removes the solid substances such as thalline, and after filtrate is concentrated with ultra-filtration membrane, concentrated solution is saltoutd or spraying dry, makes nisin powder.
Adopt the adsorbent such as diatomite, macroporous resin nisin, with spraying dry after the acidic aqueous solution desorption of polyoxyethylene glycol, although the cost of sorbent material is cheap, but not also the most desirable, and desorption agent polyoxyethylene glycol is difficult to remove, nisin finished product can be entered, affect product purity.
Adopt membrane filtering method, from fermented liquid, remove thalline and the solid equimolecular quantity material larger than nisin molecular weight by mineral membrane or tubular membrane; From fermented liquid, remove the little material of molecular weight ratio nisin molecular weight with rolled film ultrafiltration, the concentrate of fermentation liquid obtained, adds solid salt, and after spraying dry, salt content is 10 ~ 50%.
Adopt ion exchange layer analysis method to obtain high-purity nisin, the method due to cost of investment, running cost is higher does not also possess industrial utility value.
Employing ultrafiltration and concentration, Foam concentration two kinds of concentration techniques complete the concentrated of fermented liquid, finally adopt spray drying technology to obtain nisin product, still do not solve the low problem of product purity.
At present, nisin purifying technique mainly adopts saltouts and spray-dired mode, and in product, salts contg is high, and nisin purity is 20%, and extraction step is loaded down with trivial details, and purge process is complicated, there is the defects such as extraction cost is high.
Summary of the invention
Object of the present invention just high for salts contg existing in above-mentioned prior art, extraction process is complicated and the defect of product purity and the extracting method of a kind of new nisin developed specially.
The object of the invention is to be achieved through the following technical solutions:
The extracting method of nisin of the present invention comprises the following steps:
(1) by nisin fermentation concentrated solution, pH is adjusted to be 1.7 ~ 3.0 with hydrochloric acid, mix with water microsolubility organic solvent or water-insoluble organic solvents, the volume ratio of organic solvent used and nisin fermentation concentrated solution is 1:5 ~ 1:1, described organic solvent is any one in methylene dichloride, propyl carbinol, ethyl acetate, tetracol phenixin, or any two kinds of combinations; When organic solvent is two kinds of combinations, two kinds of solvent volume ratios are: 1:1 ~ 1:10
(2) stir and leave standstill, solid phase is separated, and obtains nisin and to wet cake, and in the baking oven of 40 DEG C ~ 60 DEG C, vacuum decompression is dry, obtains nisin product.
In the present invention, nisin fermentation concentrated solution is that the lactic bacteria streptococcus fermented element fermented liquid is adopted the concentrated solution that cryogenic vacuum concentrates, the method for ultrafiltration and concentration, Foam concentration, Adsorption Concentration is removed thalline and obtained.
Service temperature in whole leaching process is 4 ~ 50 DEG C.
Major advantage of the present invention adopts organic solvent deposit nisin, and overcome the defect that existing technique salts contg is high, product purity is low, nisin product purity is higher than 50%.Method safety is easy, practical, cost is low.
Embodiment
Embodiment one:
In the present embodiment, nisin extracts according to the following steps:
(1) adjust pH: extracting lactic acid streptostacin concentrated broth (desorb after thalline absorption, obtained after filtering thalline, tiring is 51.8 ten thousand/ml) 1L, adjust pH to 2.0 with hydrochloric acid.
(2) precipitate: add 1L methylene dichloride in above-mentioned nisin concentrated solution, 20 DEG C are fully stirred, and isolate upper strata aqueous phase, mixed by aqueous phase, again concentrate with lower batch fermentation liquid after leaving standstill; Lower floor solid-liquid separation reclaims methylene dichloride, obtains nisin and to wet cake;
(3) dry: the cake vacuum decompression in 50 DEG C of baking ovens that wet by nisin is dry, and namely dry cake obtains the nisin product of food grade after pulverizing.
The biological assay of the present embodiment nisin product is tired as 20321IU/mg, and product purity is 50.80%.
Embodiment two:
(1) adjust pH: extracting lactic acid streptostacin concentrated broth (cryogenic vacuum is concentrated obtained, tires 58.4 ten thousand) 1L, adjust pH to 1.7 with hydrochloric acid.
(2) precipitate: add 0.5L propyl carbinol in above-mentioned nisin concentrated solution, 50 DEG C are fully stirred, and isolate lower floor's aqueous phase, mixed by aqueous phase, again concentrate with lower batch fermentation liquid after leaving standstill; Upper strata solid-liquid separation reclaims propyl carbinol, obtains nisin and to wet cake;
(3) dry: the cake vacuum decompression in 60 DEG C of baking ovens that wet by nisin is dry, and namely dry cake obtains the nisin product of food grade after pulverizing.
The biological assay of the present embodiment nisin product is tired as 26432IU/mg, and product purity is 66.08%.
Claims (7)
1. the extracting method of a nisin, it is characterized in that: this extracting method comprises the following steps: (1) is by nisin fermentation concentrated solution, pH is adjusted to be 1.7 ~ 3.0, mix with water microsolubility organic solvent or water-insoluble organic solvents, described organic solvent is any one in methylene dichloride, propyl carbinol, ethyl acetate, tetracol phenixin, or any two kinds of combinations; (2) stir and leave standstill, solid phase is separated, and obtains nisin and to wet cake, and vacuum decompression is dry, obtains nisin product.
2. the extracting method of the nisin according to claim l, is characterized in that: nisin fermentation concentrated solution is that the lactic bacteria streptococcus fermented element fermented liquid is adopted the concentrated solution that cryogenic vacuum concentrates, the method for ultrafiltration and concentration, Foam concentration, Adsorption Concentration is removed thalline and obtained.
3. the extracting method of the nisin according to claim l, is characterized in that: the volume ratio of organic solvent used and nisin fermentation concentrated solution is 1:5 ~ 1:1.
4. the extracting method of the nisin according to claim l, is characterized in that: when organic solvent is two kinds of combinations, two kinds of solvent volume ratios are: 1:1 ~ 1:10.
5. the extracting method of the nisin according to claim l, is characterized in that: the service temperature in leaching process is 4 ~ 50 DEG C.
6. the extracting method of the nisin according to claim l, is characterized in that: vacuum decompression drying process is carried out in the baking oven of 40 DEG C ~ 60 DEG C.
7. the extracting method of the nisin according to claim l, is characterized in that: the acid of adjust pH is hydrochloric acid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410516668.1A CN105440113A (en) | 2014-09-30 | 2014-09-30 | Extraction method for nisin |
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| CN201410516668.1A CN105440113A (en) | 2014-09-30 | 2014-09-30 | Extraction method for nisin |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106047966A (en) * | 2016-07-12 | 2016-10-26 | 汕头大学 | Antibacterial peptide Nisin and extraction method and application thereof |
| CN107286226A (en) * | 2017-06-28 | 2017-10-24 | 浙江新银象生物工程有限公司 | The preparation method of high clarity and the high-titer nisin aqueous solution |
| CN107347970A (en) * | 2017-07-06 | 2017-11-17 | 浦江县欧立生物技术有限公司 | The bio-preservative of shrimps |
| CN112159463A (en) * | 2020-09-23 | 2021-01-01 | 通辽市圣达生物工程有限公司 | Preparation method of nisin |
-
2014
- 2014-09-30 CN CN201410516668.1A patent/CN105440113A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106047966A (en) * | 2016-07-12 | 2016-10-26 | 汕头大学 | Antibacterial peptide Nisin and extraction method and application thereof |
| CN106047966B (en) * | 2016-07-12 | 2019-06-28 | 汕头大学 | A kind of antibacterial peptide Nisin and its extracting method and application |
| CN107286226A (en) * | 2017-06-28 | 2017-10-24 | 浙江新银象生物工程有限公司 | The preparation method of high clarity and the high-titer nisin aqueous solution |
| CN107347970A (en) * | 2017-07-06 | 2017-11-17 | 浦江县欧立生物技术有限公司 | The bio-preservative of shrimps |
| CN112159463A (en) * | 2020-09-23 | 2021-01-01 | 通辽市圣达生物工程有限公司 | Preparation method of nisin |
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Application publication date: 20160330 |