Renewable chromatographic system and method for removing nisin pigment and peculiar smell

A nisin and chromatographic system technology, applied in chemical instruments and methods, separation methods, solid adsorbent liquid separation, etc., can solve the problems of nisin loss of activity, loss of its own activity, low activity recovery rate, etc. Achieve the effect of shortening the separation process and preparation cycle, high separation efficiency and recovery rate, fast and efficient separation

Pending Publication Date: 2021-06-15
宁波博睿瀚达生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] But on the other hand, there is still the problem of low recovery rate of peptide compounds after separation and purification. Generally, peptides or proteins are easily affected by factors such as temperature, pH value, buffer ionic strength, solvent, etc. during the purification process, resulting in the loss of their own activity.
For example, the patent document CN100554276C establishes the purification of nisin from the culture supernatant of nisin-containing streptococcus nisin. Although the purity and yield are improved, the total recovery rate of the active polypeptide can only reach About 57%, it can be seen that its nisin lost part of its activity in the purification operation
However, the traditional process of removing nisin pigment and odor also has many shortcomings, such as the low activity recovery rate of the foam adsorption method, the solvent extraction method is more polluting to the environment and is prone to solvent residues, and the removal of impurities by the salting out method is incomplete. There are still a lot of miscellaneous proteins and pigments remaining

Method used

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  • Renewable chromatographic system and method for removing nisin pigment and peculiar smell
  • Renewable chromatographic system and method for removing nisin pigment and peculiar smell
  • Renewable chromatographic system and method for removing nisin pigment and peculiar smell

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Embodiment 1

[0042] This embodiment provides a renewable chromatographic system for removing lactic acid chain bacteria pigment and odor, such as figure 1 As shown, the liquid system 1, the high pressure infusion pump, the mixer (not shown), the filter protection column 2, the column 3, the detector 4, the liquid system 5, wherein the liquid system 1 includes injection. Tube 11, inlet valve 101, balance liquid tube 12, balance liquid valve 102, regenerative liquid tube 13 and regenerative roll valve 103, the liquid system 5 includes a sample tube 51, a sample valve 501, a balanced liquid tube 52, a balance liquid Valve 502, regenerated liquid tube 53 and regenerative roll valve 503; medium of the column use a phenyl bonding silica gel (particle diameter 5 μm, aperture 120 angstrina), the chromatographic column size is: a diameter of 100 mm × length 1000 mm.

[0043] The medium packing of the column of the column of this example is produced by Ningbo Bocrupture Biotechnology Co., Ltd.

Embodiment 2

[0045] This embodiment provides a method of removing lactic acid chain bacteria pigment and odor, using the chromatographic system of Example 1 to separate the lactic acid Socolioma blended liquid, including the steps of:

[0046] Forward treatment: First adjust the pH of the lactic acid Socolioplasm to 3.5, then centrifuged, discard the precipitate, and take the supernatant.

[0047] Injective: Automatically open the balanced liquid tube valve in the inlet system in the liquid system, and the flow rate of 300 ml / min is injected;

[0048] Collection: Set the UV detector wavelength of 220 nm, when the detector is displayed to rise, the liquid system automatically turns off the balanced liquid valve, and the sample valve in the outlet system opens, collecting Nisin samples from the sample tube, After the injection, close the injective valve, the in-liquid system automatically opens the balanced valve, eluted with sodium acetate-acetic acid buffer having a concentration of 50 mmol ...

Embodiment 3

[0055] This embodiment provides a method of removing lactic acid chain bacteria pigment and odor, using the chromatographic system of Example 1 to separate the lactic acid Socolioma blended liquid, including the steps of:

[0056] Forward treatment: First adjust the pH of the lactic acid Socolioplasm to 3.5, then centrifuged, discard the precipitate, and take the supernatant.

[0057] Injective: Automatically open the balanced liquid pipe valve in the inlet system in the in-liquid system, and the flow rate of 200 ml / min is sampled;

[0058] Collection: Set the UV detector wavelength of 220 nm, when the detector is displayed to rise, the liquid system automatically turns off the balanced liquid valve, and the sample valve in the outlet system opens, collecting Nisin samples from the sample tube, After the injection, close the injective valve, the in-liquid system automatically opens the balanced valve, eluted with sodium acetate-acetic acid buffer having a concentration of 50 mmo...

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Abstract

The invention relates to the technical field of separation of natural food preservatives, in particular to a reproducible chromatographic system and method for removing nisin pigment and peculiar smell. The chromatographic system comprises a liquid inlet system, a high-pressure liquid conveying pump, a mixer, a filtering protection column, a chromatographic column, a detector and a liquid outlet system. Separation is realized on the basis of different acting forces between different components and hydrophobic groups in lactic streptococci fermentation liquor, the lactic streptococci fermentation liquor with low ionic strength is adjusted to be acidic, Nisin with weak adsorption acting force rapidly flows out of a chromatographic column by adopting high-flow-rate power, pigment and odor substances are adsorbed on the surface of a medium, and dissociation is carried out through elution of regeneration liquid. The method provided by the invention overcomes the defects of the traditional process, improves the taste and quality of the food-grade nisin, can realize rapid and efficient separation, has no organic solvent residue, is green and environment-friendly, and has long service life of the chromatographic column and low production cost.

Description

Technical field [0001] The present invention relates to the separation technology of natural food preservatives, and specific relates to regenerative chromatography systems and methods of removing lactic acid-chain bacteria pigment and odor. Background technique [0002] Lactic acid celase (Nisin, Nitern) is also known as lactic acid celase peptide, is a polypeptide compound synthesized and secreted in the metabolic process, consisting of 34 amino acid residues, and the molecular terminal has an amino group and a carboxyl group, amino group. The end is isoleucine, the end of the carboxyl group is lysine, and the molecular weight is approximately 3500 Da. Lactic acid Societium has stable, efficient antibacterial activity, can be absorbed by human body without altering the normal flora in the intestine of the human body, and does not produce drug resistance or cross resistance such as other antibiotics. Lactic acid cemetin can effectively inhibit a variety of Gram-positive bacteria...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01D15/26
CPCB01D15/265
Inventor 朱文瑾李浛君陈平李浛民
Owner 宁波博睿瀚达生物科技有限公司
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