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Method for preparing high-purity nisin

A nisin, high-purity technology, applied in the separation and purification of nisin, the field of separation and purification of nisin, can solve the problems of complicated steps and high cost, and achieve the effect of safe process

Inactive Publication Date: 2014-05-07
SHANGHAI INST OF PHARMA IND CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The steps of this method are complicated, the cost is too high, and it is not suitable for industrial scale-up

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] a. Pretreatment and solid-liquid separation:

[0024] Fermentation technology culture pretreatment: take Lactococcus lactis subsp. lactic acid CGMCC1.1936 (purchased by China General Microorganism Culture Collection Management Center, and the titer is increased to about 6000IU / mL by conventional physical and chemical mutagenesis methods and medium optimization) for fermentation The resulting fermented liquid containing nisin had a titer of 6380 IU / mL, adjusted the pH to 2.0 with hydrochloric acid, heated the fermentation technology culture to 80° C., and maintained it for 30 minutes to obtain a nisin pretreatment liquid.

[0025] Solid-liquid separation of the fermentation technology culture pretreatment liquid: the pretreatment liquid is centrifuged to remove solids such as bacterial fragments in the fermentation technology culture to obtain a clarified filtrate, and take 1L of the filtrate for extraction.

[0026] b. Extraction:

[0027] Add 460 mL of n-butanol to th...

Embodiment 2

[0031] a. Nisin product pretreatment:

[0032] The nisin product is a commercially available nisin product containing about 2.5% of nisin. Its source: Zhengzhou Qihong Biotechnology Co., Ltd., the product name is nisin, and the potency is 1094IU / mg.

[0033] Weigh 5.0 g of nisin product and dissolve it with 1 L of pH 2.0 hydrochloric acid aqueous solution to obtain nisin product pretreatment solution with a titer of 5470 IU / mL.

[0034] b. Extraction:

[0035] Add the n-butanol of 460mL of product pretreatment liquid volume to product pretreatment liquid, after fully mixing, adjust pH to 7.0 with sodium hydroxide solution, leave standstill layering, nisin is transferred from product pretreatment liquid to in the extract phase. Take the extract phase, wash it three times with 460mL n-butanol-saturated aqueous solution, back-extract part of the impurities in the extract phase into n-butanol-saturated aqueous solution, then add 460mL n-butanol-saturated aqueous solution, mix we...

Embodiment 3

[0039] The method used is the same as in Example 1, the difference is that the fermentation technology culture is nisin fermentation concentrate, the concentration of the concentrate is 5160IU / mL, and the powdered nisin with a purity of 74.8% is obtained, that is, through the agar diffusion method The titer determined was 29923IU / mg.

[0040] a. Pretreatment and solid-liquid separation:

[0041] Fermentation technology culture pretreatment: Lactococcus lactis CGMCC1.2030 (purchased from the China General Microorganism Culture Collection Management Center, and the titer was increased to about 1000IU / mL by conventional physical and chemical mutagenesis methods) produced by fermentation containing lactic acid streptococcus The fermented liquid of bacterin has a titer of 1030IU / mL, and the fermented liquid is concentrated to a titer of 5160IU / mL, adjusted to pH 2.0 with hydrochloric acid, heated to 80°C for 30 minutes to obtain nisin Pretreatment solution.

[0042] Solid-liquid se...

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Abstract

The invention discloses a method for preparing high-purity nisin, which comprises the following steps of: pretreating a fermentation technology culture and performing solid-liquid separation to obtain filtrate or nisin product pretreatment fluid; adding an extracting agent in a volume which is 40 to 80 percent of the volume of the filtrate or nisin product pretreatment fluid into the filtrate or nisin product pretreatment fluid, fully mixing uniformly, regulating the pH to be 6.0-8.0 by using alkali, and standing for demixing to obtain an extract phase; washing the extract phase by using saturated aqueous solution of the extracting agent with the same volume as the extract phase for three times, adding saturated aqueous solution of the extracting agent with the same volume as the extract phase, fully mixing uniformly, regulating the pH to be 2.0-3.0 by using acid, and standing for demixing to obtain an aqueous phase; taking the aqueous phase as feed liquid and repeating the extraction circle at least once; and concentrating the aqueous solution of the nisin subjected to extraction, and performing spray drying on concentrated solution to obtain the nisin with the purity of over 70 percent. The method is simple and convenient, and low in time consumption; the organic solvent can be recycled; and the method is suitable for the large-scale industrial production process.

Description

technical field [0001] The present invention relates to a method for separation and purification of microbial products, in particular to a method for separation and purification of nisin, in particular to a method of extracting with an organic solvent from the fermentation technology culture of nisin and nisin products Method for purifying nisin. Background technique [0002] Nisin is a polypeptide with strong antibacterial effect synthesized and secreted by some Lactococcus lactis in the metabolic process, with a molecular weight of about 3510Da. Because it has a strong inhibitory effect on many Gram-positive bacteria, especially Bacillus, Clostridium, Staphylococcus, Listeria, etc. It is a toxic natural food preservative and has been widely used as a preservative for dairy products, canned foods, meat products and beverages. [0003] The product of nisin is obtained from the fermented liquid obtained by fermentation of Lactococcus lactis which can produce nisin, and then...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/315C07K1/14
Inventor 严宇媛赵波赵文杰那可魏晓东顾敏
Owner SHANGHAI INST OF PHARMA IND CO LTD