Natural preservative industrialized purification process

A process and chromatographic column technology, which is applied in the field of natural preservative purification process, can solve the problems of low purification yield and achieve the effect of improving yield

Active Publication Date: 2006-12-13
SHANGHAI HI TECH BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved in the present invention is to disclose a nisin purification proces

Method used

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  • Natural preservative industrialized purification process
  • Natural preservative industrialized purification process
  • Natural preservative industrialized purification process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Ion exchange chromatography:

[0028] 10 ml of strong cationic agarose gel with a particle size of 100-300 μm (produced by Amersia Biotechnology Co., Ltd., a chromatography column with a trade name of SP Sepharose BigBead medium, with a diameter of 2.5 cm.

[0029] Sample: 100ml culture supernatant of Lactobacillus containing nisin; containing 85300 units of nisin.

[0030] Chromatography equipment: AKTA explorer 100 produced by Amersya Biotechnology Co., Ltd.;

[0031] Equilibrium and washing buffer: 15mmol / L acetate buffer with pH 4.5;

[0032] Elution buffer: 15mmol / L pH 4.5 acetate buffer containing 1mol / L NaCl.

[0033] Equilibration: Use 15mmol / L acetic acid buffer solution with a pH of 4.5 to equilibrate the chromatographic column containing 10ml volume of SPSepharose BigBead medium, equilibrate 8 column volumes at a flow rate of 120cm per hour, and ensure that the chromatographic column has been equilibrated by ultraviolet detection to baseline;

[0034] Sam...

Embodiment 2

[0044] reverse HPLC purification

[0045] The medium used in said HPLC column is a C18 silica gel filler (provided by Fuji Corporation) with a particle size of 10 μm and a pore diameter of 100 A, and the diameter of the chromatographic column is 2.5 centimeters.

[0046] Sample: Add an equal volume of 15mmol / L acetate buffer solution with pH 4.5, containing nisin 70372.5U, to the above-mentioned chromatographic eluent;

[0047] Chromatography equipment: SD-1 produced by Varian;

[0048] Equilibrium and washing buffer: 0.1% TFA aqueous solution;

[0049] Elution buffer: 0.1% TFA in water containing 50% acetonitrile.

[0050]Elution: After washing to the baseline, use 50% acetonitrile in 0.1% TFA aqueous solution to elute the polypeptide from the chromatographic column at a flow rate of 250 cm per hour, and collect the elution peaks. The eluent of ion exchange chromatography can remove pigments and other peptides very well through reverse high performance liquid chromatograph...

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Abstract

The invention discloses the purification technology of streptococcus lactis. The method comprises the ion exchange and backward high efficiency liquid chromatography. The recovery ratio is above 50%, and the purity displayed by HPLC is 98%.

Description

technical field [0001] The invention relates to a natural preservative purification process, in particular to an industrial purification process of nisin. Background technique [0002] Nisin, also known as lactin or nisin, is called Nisin in English. It is a small peptide with strong bactericidal effect synthesized and secreted by certain lactococcus in N-type serum during the metabolic process. Nisin can inhibit the growth and reproduction of most Gram-positive bacteria and their spores, such as Staphylococcus, Streptococcus, Clostridium and Bacillus, especially Staphylococcus aureus and Streptococcus hemolyticus , Botox effect is obvious. In addition, Nisin can also work with some complexing agents (such as EDTA or citric acid), but only some Gram-negative bacteria are sensitive to it. Nisin is used in food preservation and cosmetics, but its purity is low. If it is to be used further as medicine, the problem of its purity must be solved. [0003] J.Meghrous et al. hav...

Claims

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Application Information

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IPC IPC(8): C07K1/16C07K2/00
Inventor 黄青山李国栋
Owner SHANGHAI HI TECH BIOENG
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