Method of extracting myoglobin from beef

A myoglobin and extraction method technology, applied in hemoglobin/myoglobin, chemical instruments and methods, specific peptides, etc., can solve the problems of unfavorable sample storage application, difficult myoglobin, easy oxidation of myoglobin, etc. Achieve the effect of maintaining reduction and biological activity, improving purity and avoiding oxidation

Inactive Publication Date: 2019-08-09
甘肃省农业科学院农产品贮藏加工研究所
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0002] Among the extraction and purification methods of beef myoglobin, the Tris-HCl method is commonly used for crude myoglobin extraction, but the existing myoglobin extraction technology has complex muscle tissue components, and it is difficult for myoglobin to precipitate quickly and stably; And (oxygenated) myoglobin itself is easily oxidized, which is not conducive to the preservation and application of purified samples. It is necessary to improve the stability of myoglobin during the purification process and reduce the oxidation and denaturation loss of myoglobin.

Method used

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  • Method of extracting myoglobin from beef
  • Method of extracting myoglobin from beef
  • Method of extracting myoglobin from beef

Examples

Experimental program
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Embodiment 1

[0026] The method for extracting myoglobin in the beef of the present embodiment, the method may further comprise the steps:

[0027] Step 1, rough extraction: take beef longissimus dorsi, remove superficial fascia and connective tissue, chop to obtain beef meat samples, add 500 mL of Tris-HCl with a concentration of 10 mmol / L and a pH value of 8.2 to 100 g of beef meat samples The buffer solution was mixed, homogenized at 10000rpm for 1min, left to extract at 4°C for 1.5h, centrifuged at 9600rpm at 4°C for 10min and filtered to obtain Filtrate; The tris-HCl buffer solution a, the EDTA of 1mmol / L, the glycerol of 0.54mmol / L and the dithiothreitol that contain concentration in the Tris-HCl mixed buffer solution are 10mmol / L;

[0028] Step 2, salting out: in the filtrate that obtains in step 1, slowly and evenly add solid (NH 4 ) 2 SO 4 , to obtain the myoglobin salting-out solution, when the saturation of the myoglobin salting-out solution is 90%, let it stand for 30 minutes...

Embodiment 2

[0043] The method for extracting myoglobin in the beef of the present embodiment, the method may further comprise the steps:

[0044] Step 1, rough extraction: take the longissimus dorsi muscle of beef, remove the superficial fascia and connective tissue, chop to obtain a beef sample, add 500 mL of Tris-HCl with a concentration of 10 mmol / L and a pH value of 8.0 to 100 g of the beef sample Mix the buffer solution, then homogenize for 1 min under the condition of rotating speed of 10000 rpm, stand and extract for 1 h under the condition of temperature of 4 °C, then centrifuge for 10 min under the condition of rotating speed of 9600 rpm and temperature of 4 °C, and then filter to obtain the filtrate ; Containing the Tris-HCl damping solution a, the EDTA of 1mmol / L, the glycerol of 0.54mmol / L and the dithiothreitol of 1mmol / L in the Tris-HCl mixed buffer that concentration is;

[0045] Step 2, salting out: in the filtrate that obtains in step 1, slowly and evenly add solid (NH 4...

Embodiment 3

[0051] The method for extracting myoglobin in the beef of the present embodiment, the method may further comprise the steps:

[0052] Step 1, rough extraction: Take the longissimus dorsi muscle of beef, remove the superficial fascia and connective tissue, chop to obtain a beef sample, add 500 mL of Tris-HCl with a concentration of 10 mmol / L and a pH value of 8.4 to 100 g of the beef sample Mix the buffer solution, then homogenize at 10000rpm for 1min, stand and extract at 4°C for 2h, then centrifuge at 9600rpm and 4°C for 10min and filter to obtain the filtrate ; Containing the Tris-HCl damping solution a, the EDTA of 1mmol / L, the glycerol of 0.54mmol / L and the dithiothreitol of 1mmol / L in the Tris-HCl mixed buffer that concentration is;

[0053] Step 2, salting out: in the filtrate that obtains in step 1, slowly and evenly add solid (NH 4 ) 2 SO 4 , to obtain the myoglobin salting-out solution, when the saturation of the myoglobin salting-out solution is 95%, let it stand ...

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Abstract

The invention provides a method of extracting myoglobin from beef. The method includes the steps of adding a Tris-HCl mixed buffer solution to a beef meat sample, homogenizing, standing, centrifuging,adding (NH4)2SO4 and standing, centrifuging, adding a Tris-HCl buffer solution b, performing dialysis, and eluting with a NaCl mixed solution in a chromatography column to obtain the myoglobin in thebeef. According to the method, the pH value of the Tris-HCl mixed buffer solution, the extraction time and the saturation of the myoglobin salt precipitation solution are optimized to increase the extraction amount of myoglobin; the Tris-HCl mixed buffer solution contains glycerin and dithiothreitol, the glycerin maintains the structural stability of the myoglobin and facilitates rapid precipitation, and the dithiothreitol maintains the reductive and biological activity of the myoglobin; the solid (NH4)2SO4 is added for fractional salt precipitation to increase the purity of the myoglobin, the temperature is kept at 4 DEG C to prevent degeneration and oxidation of the myoglobin, residual salt ions and other small molecular proteins can be effectively removed through the dialysis to improve the purity of the myoglobin, and the separation by gel filtration chromatography carried out at last does not destroy the structure and function of the myoglobin.

Description

technical field [0001] The invention belongs to the technical field of protein extraction, and in particular relates to a method for extracting myoglobin in beef. Background technique [0002] Among the extraction and purification methods of beef myoglobin, the Tris-HCl method is commonly used for crude myoglobin extraction, but the existing myoglobin extraction technology has complex muscle tissue components, and it is difficult for myoglobin to precipitate quickly and stably; Moreover, (oxygenated) myoglobin itself is easily oxidized, which is not conducive to the preservation and application of purified samples. It is necessary to improve the stability of myoglobin during the purification process and reduce the oxidation and denaturation loss of myoglobin. Contents of the invention [0003] The technical problem to be solved by the present invention is to provide a method for extracting myoglobin in beef for the above-mentioned deficiencies in the prior art. The satura...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/805C07K1/14C07K1/16C07K1/30C07K1/36
CPCC07K14/805
Inventor 郑娅王晓璇史立学何元翔王佳妮
Owner 甘肃省农业科学院农产品贮藏加工研究所
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