CN109938245B - Physical method for reducing N-glycolylneuraminic acid in red meat - Google Patents
Physical method for reducing N-glycolylneuraminic acid in red meat Download PDFInfo
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Abstract
The invention discloses a method for reducing N-glycolylneuraminic acid in red meat, which comprises the following steps: taking beef, stirring, weighing 1 part of minced meat, adding 10 parts of purified water into a homogenate bottle, grinding uniformly, directly placing an electrode into the minced beef, and electrically stimulating for 10-50s by using a 30-120V direct current stabilized power supply; or taking the beef block, using a 100-grade 300V direct current stabilized power supply to directly contact the electrodes with the two sides of the beef block, and under the premise of ensuring that the beef block cannot be burnt, intermittently electrically stimulating for 60s, wherein the time of each electrical stimulation is 1-5 s. The invention can safely and efficiently reduce the Neu5Gc content and improve the tenderness of the red meat.
Description
Technical Field
The invention belongs to the technical field of biochemistry, and particularly relates to a physical method for reducing N-glycolylneuraminic acid in red meat.
Background
Red meat, which at present generally refers to the muscle of unprocessed mammals, such as pork, mutton, beef, and the like. The red meat can provide high-quality animal protein, a plurality of mineral substances, B-group vitamins and the like for people. However, in 2015, 10 months, the international cancer research institute classified red meat and processed meat products as more likely carcinogens and carcinogens respectively, and attracted extensive social attention. Many epidemiological investigations have shown that red meat intake is closely related to the occurrence of several diseases, such as colorectal, lung, pancreatic, prostate, etc. The risk substances present in red meat are generally considered to be saturated fatty acids, N-nitroso compounds, heterocyclic amines, polycyclic aromatic hydrocarbons, hematin and the like, but these substances are not specific to red meat. Therefore, sialic acid-N-glycolylneuraminic acid of non-human origin proposed by the professor group of Varki is a marker risk substance for red meat carcinogenesis.
N-glycolylneuraminic acid (Neu5Gc), a member of the sialic acid family, is a derivative of a 9-carbon monosaccharide specific for mammals, acidic in red meat and containing a total of 11 carbon atoms, and is ubiquitous in the non-neural tissues and fluids of most posterior animals, including echinoderms and vertebratesAnimals are component sugars such as animal cell membranes or secretion glycoproteins, glycolipids, or bacterial capsular substances. Neu5Gc is N-acetylneuraminic acid (Neu5Ac) biosynthesized by CMP-Neu5Ac hydroxylase, but about 2-3 million years ago, a human gene encoding CMP-Neu5Ac hydroxylase is mutated, so that Neu5Gc cannot be synthesized in a human body, and Neu5Gc is a non-human sialic acid. At present, Neu5Gc in human body is mainly from the intake of animal food, the main sialic acid form in human body is Neu5Ac, and the molecular formula of Neu5Gc is C11H19NO10The molecular weight is 325.27, and the structure is shown in the following formula.
Many researchers adopt different immunochemical techniques to detect Neu5Gc from different visceral canceration tissues such as liver cancer tissues, stomach cancer tissues, rectal cancer tissues and the like. Ingestion of Neu5Gc produces antibodies in the body that recognize Neu5Gc, which promotes the development of chronic inflammation and thus stimulates tumor growth. Thus, Neu5Gc attached to the surface of tumor cells actually indirectly promoted the growth of cancer. This raises concerns about the consumer buying edible red meat products and the risk of health hazards to the consumer, and it is therefore highly desirable to be able to reduce the risk of materials in the red meat by effective means.
The patent of Wako et al (U.S. Pat. No. 3, 06/22282) mentions that non-human sialic acid Neu5Gc is metabolically incorporated into cultured cells and human tissues, respectively, to contaminate biotherapeutic products and humans. In the first case, the contamination comes from the animal-derived components in the culture medium and the animal cell line used; and in the second case, the contamination comes from the dietary intake from food (e.g. red meat). Administration of an effective Neu5Ac, derivative, analog, or precursor to an animal comprising the Neu5Gc enzymatic pathway is effective to reduce or eliminate Neu5 Gc. Jiangyu and the like develop research on reducing the content of Neu5Gc in red meat, and the aim is to dissociate part of Neu5Gc by pretreatment before cooking, namely pork and beef are treated by water boiling, microwave oven heating, organic acid pickling and galactosidase hydrolysis, and non-human sialic acid dissociation modes are searched, and the results show that the pretreatment modes before cooking of the red meat have different degrees of effects on reducing the content of Neu5Gc in products. The content of Neu5Gc in red meat is reduced by frying, cooking and the like, but the content of Neu5Gc is reduced when the frying temperature reaches 150 ℃; with the prolonged cooking time, the Neu5Gc content in the broth is obviously increased, and Neu5Gc in the beef is dissociated, which shows that the cooking can effectively reduce the Neu5Gc content of the meat. However, none of the methods adopted at present can achieve the effect of rapidly and nondestructively reducing the Neu5Gc content in the red meat on the basis of keeping the original quality of the red meat, and the method is not strong in practicability for industrial production.
Electrical stimulation is a technology which effectively prevents cold contraction from hardening, and electrical stimulation can accelerate glycolysis and reduce pH to prevent cold contraction, destroy the myofibrillar structure and further improve the tenderness of meat; the electrical stimulation obviously accelerates the degradation of troponin-T and shortens the maturation time of the slaughtered beef, and the electrical stimulation is mainly used for improving the tenderness of the beef in the carcasses of animals such as cattle, sheep and the like at present.
Disclosure of Invention
The invention aims to overcome the defects and provide a physical method for reducing N-glycolylneuraminic acid in red meat, which is safe and efficient, improves the tenderness of red meat and simultaneously reduces the content of Neu5 Gc.
The purpose of the invention and the main technical problem of solving the invention are realized by adopting the following technical scheme:
the invention relates to a method for reducing N-glycolylneuraminic acid in red meat, which comprises the following steps:
(1) electrically stimulating the minced beef: taking beef, stirring, weighing 1 part of minced meat, adding 10 parts of purified water into a homogenate bottle, grinding uniformly, directly placing an electrode into the minced beef, and electrically stimulating for 10-50s by using a 30-120V direct current stabilized power supply;
(2) electrically stimulating the beef blocks: taking beef blocks, using a 100-plus-300V direct current stabilized power supply to directly contact the electrodes with the two sides of the beef blocks, and under the premise of ensuring that the beef blocks cannot be burnt, intermittently electrically stimulating for 60s, wherein the time of each electrical stimulation is 1-5 s.
Compared with the prior art, the beef provided by the invention has obvious beneficial effects, and the beef selected by the invention is higher in Nue5Gc content compared with other types of red meat, so that the change of Neu5Gc before and after treatment can be observed. The collocation of the voltage and the time of the invention is obtained by repeatedly screening the electrical stimulation parameters, and the minced beef and the blocky beef are electrically stimulated under the condition. The high performance liquid chromatography fluorescence detector is adopted to detect the Neu5Gc content in the beef before and after the electric stimulation, which proves that the Neu5Gc content in the red beef can be reduced after the electric stimulation is carried out on the beef by the method, and meanwhile, the tenderness of the beef is improved.
Detailed Description
Example 1
A physical method for reducing the N-glycolylneuraminic acid content in beef emulsion, comprising the following steps:
the beef is taken and smashed by a smashing machine, 1g of minced meat is weighed and placed in a homogenate bottle, 10ml of purified water is added and evenly ground, an electrode is directly placed in the minced beef, and the minced beef is electrically stimulated for 10s by a 90V direct current stabilized voltage power supply.
Example 2
A physical method for reducing the N-glycolylneuraminic acid content in beef emulsion, comprising the following steps:
the beef is taken and smashed by a smashing machine, 1g of minced meat is weighed and placed in a homogenate bottle, 10ml of purified water is added and evenly ground, an electrode is directly placed in the minced beef, and the minced beef is electrically stimulated for 30s by a 90V direct current stabilized voltage power supply.
Example 3
A physical method for reducing the N-glycolylneuraminic acid content in beef emulsion, comprising the following steps:
the beef is taken and smashed by a smashing machine, 1g of minced meat is weighed and placed in a homogenate bottle, 10ml of purified water is added and evenly ground, an electrode is directly placed in the minced beef, and the minced beef is electrically stimulated for 50s by a 30V direct current stabilized voltage power supply.
Example 4
A physical method for reducing the N-glycolylneuraminic acid content in beef emulsion, comprising the following steps:
the beef is taken and smashed by a smashing machine, 1g of minced meat is weighed and placed in a homogenate bottle, 10ml of purified water is added and evenly ground, an electrode is directly placed in the minced beef, and the minced beef is electrically stimulated for 50s by a 120V direct current stabilized voltage power supply.
Example 5
A physical method for reducing the content of N-glycolylneuraminic acid in beef cubes comprises the following steps:
taking beef blocks with weight of 400g, length, width and height of 120cm, 75cm and 48cm respectively, directly contacting electrodes with two sides of the beef blocks, and intermittently electrically stimulating for 60s by using a 100V direct current stabilized power supply, wherein the time of each electrical stimulation is 5 s.
Example 6
A physical method for reducing the content of N-glycolylneuraminic acid in beef cubes comprises the following steps:
taking beef blocks with weight of 400g, length, width and height of 120cm, 75cm and 48cm respectively, directly contacting electrodes with two sides of the beef blocks, and intermittently electrically stimulating for 60s by using a 300V direct current stabilized power supply, wherein the time of each electrical stimulation is 1 s.
Test example:
taking the beef paste treated in the embodiment 1-4, freeze-drying for 48h, adding 10ml of 30% saturated ammonium sulfate solution, standing at room temperature for 1h, and freezing;
taking the beef treated in the examples 5-6, mincing the beef by a mincing machine, weighing 1g of beef paste, adding 10ml of 30% saturated ammonium sulfate solution, grinding uniformly, standing for 1h at room temperature, and freezing;
freeze-drying the beef surimi treated by the conditions for 48h to obtain freeze-dried powder, adding 10mL of 2mol/L acetic acid into the freeze-dried powder, carrying out acidolysis for 3h in a water bath at 80 ℃, carrying out centrifugal filtration, carrying out freeze-drying for 48h again, adding 1mL of water and 0.2mL of 0.1mol/L NaOH into the freeze-dried powder, carrying out deacetylation for 30min in a water bath at 37 ℃, filtering through a 0.45 mu m filter head to obtain filtrate A, and carrying out derivatization on the filtrate A according to the following derivatization method for detection.
The blank group was not treated with electrical stimulation and the remaining steps were identical to those of the experimental group.
High performance liquid chromatography conditions:
the chromatographic column is a German Merck Li Chrosorb RP-18 column (150mm multiplied by 4.6mm, 5 mu m), the column temperature is 30 ℃, the fluorescence detector excitation wavelength is 373nm, the emission wavelength is 448nm, the mobile phase is ultrapure water-acetonitrile-methanol (85: 8: 7, V/V), the flow rate is 0.9mL/min, and the sample injection volume is 10 mu L.
Preparation of a derivative liquid and derivative conditions:
preparing a DMB derivative solution: 8mmol/L DMB, 1.5mol/L glacial acetic acid, 0.25mol/L sodium thiosulfate, 0.25mol/L sodium sulfite and 0.8 mmol/L2-mercaptoethanol.
The derivation conditions are as follows:
precisely sucking 900 μ L of the filtrate obtained after pretreatment, respectively adding 100 μ L of DMB derivative solution, performing light-shielding derivatization at 50 deg.C for 150min, cooling to room temperature, and detecting to obtain the results shown in tables 1 and 2.
TABLE 1 Change in Nue5Gc content in ground beef before and after Electrical stimulation treatment
As can be seen from table 1, the Neu5Gc content of the beef emulsion varied to varying degrees after treatment under the conditions of examples 1-4, but not every treatment group was effective in reducing the Neu5Gc content of the beef emulsion. Of these, example 4, which is the most effective, reduces by seventy percent the Neu5Gc in the beef emulsion. Thus, the proper electric stimulation condition can effectively reduce the content of Neu5Gc in the beef emulsion.
TABLE 2 Change in Nue5Gc content in beef pieces before and after Electrical stimulation treatment
As can be seen from Table 2: respectively electrically stimulating the beef blocks with 100V and 300V voltages for 60s, wherein the content of Neu5Gc in the beef blocks is not obviously changed when the voltage of 100V is electrically stimulated; when the voltage is 300V for electric stimulation, the Neu5Gc content in 400g of beef is reduced, and is reduced by 21.02 percent compared with the control group (Table 2). Indicating that the proper electrical stimulation condition can reduce the content of Nue5Gc in the beef block.
Compared with the electrically stimulated minced beef, the electrically stimulated beef block has smaller reduction amplitude of the Neu5Gc content. The reasons are as follows: 1. although the voltage is increased, the meat-like mass increases and the volume increases, so that the effect of reduction is not significant as compared with beef emulsion. 2. The meat sample is enlarged, and the relative contact area is reduced when the meat sample is treated by electric stimulation. And 3, the electrical stimulation of the large meat sample causes voltage instability, intermittent electrical stimulation is required, and the electrical stimulation cannot be continuously performed.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention in any way, and any simple modification, equivalent change and modification made to the above embodiment according to the technical spirit of the present invention are within the scope of the present invention without departing from the technical spirit of the present invention.
Claims (2)
1. A method of reducing N-glycolylneuraminic acid in red meat comprising: the preparation method comprises the following steps of taking beef, smashing, weighing 1 part of minced meat, putting into a homogenate bottle, adding 10 parts of purified water, grinding uniformly, directly putting an electrode into the minced beef, and electrically stimulating for 10-50s by using a 30-120V direct current stabilized power supply.
2. A method of reducing N-glycolylneuraminic acid in red meat comprising: taking beef blocks, using a 100-plus-300V direct current stabilized power supply to directly contact the electrodes with the two sides of the beef blocks, and under the premise of ensuring that the beef blocks cannot be burnt, intermittently electrically stimulating for 60s, wherein the time of each electrical stimulation is 1-5 s.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| AU6611896A (en) * | 1995-07-07 | 1997-02-10 | Boehringer Mannheim Gmbh | Nucleic acid coding for cmp-n-acetyl-neuraminic acid hydroxylase and its use for the production of modified glycoproteins |
| WO2014194053A2 (en) * | 2013-05-29 | 2014-12-04 | Siamab Therapeutics, Inc. | Carbohydrate compounds for nutritional and therapeutic use |
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| EP2302390B1 (en) * | 2003-07-15 | 2013-06-05 | The Regents of The University of California | Methods for detecting and analyzing n-glycolylneuraminic acid (Neu5Gc) specific antibodies in biological materials |
| US20110195921A1 (en) * | 2008-09-09 | 2011-08-11 | Ajit Varki | Elimination of a contaminating non-human sialic acid by metabolic competition |
| WO2015048346A2 (en) * | 2013-09-25 | 2015-04-02 | Pronutria, Inc. | Compositions and formulations for prevention and treatment of diabetes and obesity, and methods of production and use thereof in glucose and caloric control |
| CN109223763A (en) * | 2018-09-28 | 2019-01-18 | 贵州大学 | It is a kind of effectively to inhibit NeuGc ALPHA2-3Gal synthetic method in meat products |
| CN109329713B (en) * | 2018-12-01 | 2022-04-22 | 西华大学 | Method for degrading N-glycolylnersialic acid in yak meat |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| AU6611896A (en) * | 1995-07-07 | 1997-02-10 | Boehringer Mannheim Gmbh | Nucleic acid coding for cmp-n-acetyl-neuraminic acid hydroxylase and its use for the production of modified glycoproteins |
| WO2014194053A2 (en) * | 2013-05-29 | 2014-12-04 | Siamab Therapeutics, Inc. | Carbohydrate compounds for nutritional and therapeutic use |
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