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CN113142292A - Method for prolonging preservation period of cold fresh meat by using antibacterial lactobacillus composite preservation solution - Google Patents

Method for prolonging preservation period of cold fresh meat by using antibacterial lactobacillus composite preservation solution Download PDF

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Publication number
CN113142292A
CN113142292A CN202011588896.1A CN202011588896A CN113142292A CN 113142292 A CN113142292 A CN 113142292A CN 202011588896 A CN202011588896 A CN 202011588896A CN 113142292 A CN113142292 A CN 113142292A
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China
Prior art keywords
meat
lactobacillus plantarum
fresh
liquid
lactobacillus
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沈佳敏
孙小梅
曾小群
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Huzhou Zhanwang Food Technology Co ltd
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Huzhou Zhanwang Food Technology Co ltd
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
    • A23B4/00—General methods for preserving meat, sausages, fish or fish products
    • A23B4/14—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12
    • A23B4/18—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12 in the form of liquids or solids
    • A23B4/20—Organic compounds; Microorganisms; Enzymes
    • A23B4/22—Microorganisms; Enzymes; Antibiotics
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
    • A23B4/00—General methods for preserving meat, sausages, fish or fish products
    • A23B4/06—Freezing; Subsequent thawing; Cooling
    • A23B4/066—Freezing; Subsequent thawing; Cooling the materials not being transported through or in the apparatus with or without shaping, e.g. in the form of powder, granules or flakes
    • A23B4/068—Freezing; Subsequent thawing; Cooling the materials not being transported through or in the apparatus with or without shaping, e.g. in the form of powder, granules or flakes with packages or with shaping in the form of blocks or portions
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
    • A23B4/00—General methods for preserving meat, sausages, fish or fish products
    • A23B4/10—Coating with a protective layer; Compositions or apparatus therefor
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
    • A23B4/00—General methods for preserving meat, sausages, fish or fish products
    • A23B4/14—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12
    • A23B4/18—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12 in the form of liquids or solids
    • A23B4/20—Organic compounds; Microorganisms; Enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/169—Plantarum

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  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Microbiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Food Preservation Except Freezing, Refrigeration, And Drying (AREA)

Abstract

The invention discloses a method for prolonging the fresh-keeping period of cold fresh meat by using bacteriostatic lactic acid bacteria, which comprises the following steps of (1) removing redundant fascia and fat in purchased meat to be used as meat raw materials; (2) activating and expanding culture of lactobacillus plantarum; (3) separating and purifying the expanded lactobacillus plantarum; (4) redissolving the purified lactobacillus plantarum to prepare lactobacillus plantarum solution; (5) obtaining lactobacillus plantarum suspension; (6) resuspending baicalin and lactoferrin in the bacterial suspension, finally adding chitosan with final concentration of 0.1-0.3%, and vortex mixing to obtain a composite fresh-keeping solution; spraying the lactobacillus plantarum composite preservative solution on the chilled meat; packaging the treated cold fresh meat with preservative film, and refrigerating and storing in a refrigerator at 4 + -0.5 deg.C. By using the method, the fresh-keeping period of the chilled meat is prolonged to 12 days, which is prolonged by 9 days compared with the untreated meat, and the effect is obvious.

Description

Method for prolonging preservation period of cold fresh meat by using antibacterial lactobacillus composite preservation solution
Technical Field
The invention relates to a method for prolonging the fresh-keeping period of cold fresh meat, in particular to a method for prolonging the fresh-keeping period of cold fresh meat by using the synergistic effect of lactobacillus plantarum, baicalin, lophatherum gracile extract, active lactoferrin and chitosan with broad-spectrum antibacterial effect.
Background
The cold fresh meat is also called cold fresh meat, and more accurately called 'cooling and acid-removing meat'. The method is characterized in that the carcass temperature (usually the center of the ham is a measuring point) of livestock such as cattle, sheep and pigs or poultry such as chicken, duck and goose is rapidly reduced to 0-4 ℃ within 24 hours after the poultry is slaughtered, and the fresh meat products are maintained in the temperature range in a series of processes such as subsequent processing, storage and sale. Because the cold fresh meat is subjected to processes of precooling, acid discharging and 'ripening' after being slaughtered, the cold fresh meat is more moist and elastic compared with other meat products, and is easier to taste in subsequent processing.
However, the preservation condition of the cold fresh meat cannot completely inhibit the growth and propagation of microorganisms, and certain pathogenic bacteria can still grow at 0-4 ℃, thereby causing the spoilage of meat. For a long time, how to extend the originally short shelf life of chilled meat has been an important concern for many food related researchers. Prolonging the shelf life of the meat can not only produce great economic benefit, but also greatly improve the popularization of the chilled meat. In the existing research, chemical preservatives such as organic acids and salts thereof (such as formic acid, lactic acid, acetic acid, propionic acid, citric acid, ascorbic acid, sorbic acid and potassium salts thereof) and ozone, inorganic salts (such as mixed phosphate) and vitamin E are mainly used singly or in a compound way, generally, the compound use effect is better than that of the single use, and some natural preservatives such as extracts of onion, ginger, garlic or spices are used for preservation, but the report of using biological preservatives such as lactic acid bacteria to cold fresh meat is not found.
According to the patent application, fermentation liquor obtained by culturing lactobacillus plantarum in an MRS culture medium is centrifuged to collect supernatant, 0.1-0.3% of chitosan, 0.1-0.35% of natamycin and 0-50% of sterile water are added, and the mixture is uniformly mixed to obtain the lactobacillus fermentation liquor preservative, but the lactobacillus fermentation liquor is used, and the culture medium contains various matrix components, which may affect the quality, particularly the flavor, of cold fresh meat. In the patent of lactobacillus plantarum antistaling agent for low-salt meat products (patent number: 201910080097.4), lactobacillus plantarum viable bacteria are sprayed on the surface of low-salt meat products (sausages and bacon), and then are quickly packaged in vacuum and stored in a cold storage at 4 ℃; this patent is a preservation of processed meat products and requires vacuum packaging. No report that the biological preservative lactobacillus viable bacteria, the natural preservative baicalin and the lophatherum gracile extract and the animal-derived preservative are applied to the fresh-keeping aspect of the fresh meat by combining the synergistic effect of the biological preservative lactobacillus viable bacteria, the natural preservative baicalin and the lophatherum gracile extract and the coating agent is available.
Biological preservation is the use of naturally or artificially controlled microorganisms and their produced metabolites to extend the shelf life of food and improve the safety of food. The lactobacillus is a biological preservative, can normally grow on the surface of the meat product, and can not generate adverse effects on the tissue state of the cold fresh meat product and the like under the condition of low-temperature refrigeration storage. The lactobacillus secretes antibacterial substance nisin during the growth and development process, which has broad-spectrum and high-efficiency antibacterial activity, and in addition, the lactobacillus can indirectly inhibit the growth of pathogenic bacteria due to the antagonistic action among strains during the growth process. The lactobacillus is added to effectively inhibit the growth of putrefying microorganisms in the meat and prolong the shelf life.
Baicalin (baicailin) is a flavonoid compound extracted and separated from dried root of Scutellaria baicalensis Georgi (Scutellaria baicalensis Georgi) belonging to family Labiatae. Has remarkable biological activity, has the functions of bacteriostasis, diuresis, anti-inflammation, cholesterol reduction, thrombosis resistance, asthma relief, fire purging, detoxification, hemostasis, miscarriage prevention, allergy resistance and spasmolysis, is a specific inhibitor of liver sialidase of mammals, has the function of regulating certain diseases, and also has stronger physiological efficiency of anticancer reaction.
The functional factors contained in the lophatherum gracile are mainly as follows: flavone, phenolic acid compounds, amino acids, manganese, zinc and other trace elements. Experiments show that the effective components have good antibacterial activity and can clear active oxygen free radicals in vivo; the preservative prepared from Lophatherum gracile serving as a raw material has strong inhibitory effect on bacteria, mold and yeast, and can be added into sterilized food.
Scientific research on lactoferrin, namely lactoferrin, mainly provides an iron binding protein extracted from cow milk, is a safe and reliable natural substance, and has good inhibitory effect on pathogenic bacteria and putrefying bacteria.
0.25 percent of chitosan CS0811 has good inhibition effect on pathogenic bacteria, putrefying bacteria and the like such as staphylococcus aureus, escherichia coli, bacillus subtilis, pseudomonas and the like.
At present, the research on prolonging the fresh-keeping period of the chilled meat by utilizing biological preservation is little at home and abroad, and the synergistic effect of the lactobacillus plantarum, the plant extract, the lactoferrin and the chitosan added to the meat can prolong the fresh-keeping period of the chilled meat and ensure the diet safety of people.
Disclosure of Invention
The invention aims to solve the technical problem of providing a method for prolonging the fresh-keeping period of cold fresh meat by adding lactobacillus plantarum, baicalin, lophatherum gracile extract, active lactoferrin and chitosan into the cold fresh meat for synergistic action.
The technical scheme adopted by the invention for solving the technical problems is as follows: a method for prolonging the preservation period of cold fresh meat comprises the following steps:
(1) treatment of meat
The chopping board and the knife are sterilized in a sterilizing pot and wiped with sterilized alcohol before use. Under the aseptic condition of an aseptic table, removing redundant fascia and fat in purchased meat from fresh meat which is slaughtered by a meat complex on the day, and cutting the meat into meat blocks with the weight of about 100-1000 g for later use;
(2) expanded culture of lactobacillus
Inoculating lactobacillus plantarum into a liquid MRS culture medium in a sterile manner, and culturing in a thermostat at 37 ℃ for about 12 hours to fully activate the lactobacillus plantarum; then inoculating the activated strain into a large conical bottle filled with a liquid MRS culture medium, and expanding in a shaking table at the constant temperature of 37 ℃ for about 12 hours for later use;
(3) collection of Lactobacillus thallus
Pouring the cultured lactobacillus bacterial liquid into a centrifugal tube in a super-clean workbench, then putting the centrifugal tube into a centrifugal machine, centrifuging for 10min at 8000r/min, removing the upper layer liquid, adding physiological saline with the same volume as the bacterial liquid added at the beginning into the centrifugal tube, homogenizing, putting the centrifugal machine into the centrifugal machine, centrifuging for 10min at 8000r/min, removing the upper layer clear liquid, adding the physiological saline with the same volume, repeating the operation, taking the precipitate, and obtaining lactobacillus plantarum thallus precipitate;
(4) preparation of lophatherum gracile extract
Adding 100g of herba Lophatheri into 1000mL of water, boiling at a material-to-liquid ratio of 1:10g/mL, keeping the boiling state for 20min, performing ultrasonic extraction for 20min at an ultrasonic power of 300W to fully extract effective components of the herba Lophatheri, filtering the supernatant, and storing in a sterilized glass ware.
(5) Preparation of composite fresh-keeping liquid
And adding the cooled lophatherum gracile extract into a centrifugal tube containing the thallus precipitate according to the mass-volume ratio of the thallus to the lophatherum gracile extract of 1-3%, and blowing or whirling the thallus precipitate by using a pipette to uniformly mix so as to resuspend the lactobacillus plantarum precipitate to obtain lactobacillus plantarum suspension. Adding baicalin with final concentration of 15-30mg/L and active lactoferrin with final concentration of 10-30mg/L to make baicalin and lactoferrin be suspended in the bacterial suspension, finally adding chitosan with final concentration of 0.1-0.3%, and vortex and mixing uniformly to obtain a composite fresh-keeping solution for later use;
(6) spraying preservation liquid
According to the compound fresh-keeping liquid: uniformly spraying the chilled fresh meat with a small sprayer to a meat sample in a volume mass fraction (V/W) of 1.0-2.0%, standing for 1min, and draining for later use;
(7) packaging and preserving
And (3) coating the meat blocks sprayed with the lactobacillus plantarum suspension by using a food-grade polyethylene preservative film, and refrigerating and storing in a refrigerator at the temperature of (4 +/-0.5 ℃).
The preparation method of the MRS liquid culture medium comprises the following steps: 10g of peptone, 10g of beef extract, 5g of yeast extract, 2g of dipotassium phosphate, 2g of diammonium citrate, 5g of sodium acetate, 10g of glucose, 801 mL of tween-801, 0.5g of magnesium sulfate heptahydrate and 0.25g of manganese sulfate heptahydrate are dissolved in 1000mL of distilled water, the pH value is adjusted to 6.8, and sterilization is carried out for 15min at 121 ℃.
Compared with the prior art, the invention has the advantages that: the invention relates to a method for prolonging the fresh-keeping period of cold fresh meat, which comprises the steps of taking fresh meat slaughtered by a meat company factory on the day, spraying a lactobacillus plantarum compound fresh-keeping liquid on the fresh meat, and refrigerating and storing the fresh meat in a refrigerator at the temperature of (4 +/-0.5) DEG C to prolong the fresh-keeping period of the cold fresh meat. The lophatherum gracile extract and the baicalin have the effects of inhibiting the growth of the putrefying bacteria of the chilled fresh meat, simultaneously avoiding the excessive growth of lactic acid bacteria, avoiding the excessive acid production of the lactic acid bacteria to become new putrefying bacteria, and controlling the acidity and the microbial quantity within an effective range. Under the condition of not adding chemical substances and using chemical preservatives, the method for safely and conveniently prolonging the fresh-keeping period of the chilled fresh meat is provided. As the demand for chilled fresh meat increases, chilled fresh meat is becoming a mainstream trend for consumption of meat products. The short preservation period and shelf life of the cold fresh meat become main problems restricting the development of the cold fresh meat, and meanwhile, the cold fresh meat is often rancid during preservation and is rotten and mildewed due to spoilage microorganisms, so that the nutritional characteristics and the quality of the cold fresh meat are influenced. The cold fresh meat is treated by the lactobacillus plantarum suspension, so that the cold fresh meat is prevented from being decayed, the preservation period is prolonged, and the health and safety problems caused by the traditional chemical preservative method can be solved. The lactobacillus plantarum growth process is a complex process. After lactobacillus plantarum is added into the cold fresh meat, the lactobacillus can secrete a plurality of antibacterial substances in the growth process to inhibit the growth and reproduction of spoilage microorganisms; in addition, the lactobacillus plantarum added in a large amount is the dominant strain in the meat, and the growth and the propagation of putrefying microorganisms are inhibited through competitive inhibition, namely growth antagonism. Meanwhile, the main bacteriostatic substance lactein secreted by lactobacillus is quicker and more effective than the traditional chemical fresh-keeping reagent, and is digested and absorbed by human body after entering the human body, so that the normal flora of intestinal tract is not influenced, and the allergy phenomenon is avoided. Meanwhile, the lactobacillus preferentially utilizes glucose, and can not participate in decomposing protein amino acid unless the glucose is exhausted, and other nutritional ingredients in the meat can not be damaged under the normal preservation condition; meanwhile, the antibacterial action of the lactobacillus inhibits other microorganisms from damaging the meat, thereby reducing the putrefaction of the meat. Therefore, the quality such as nutritive value characteristic and the like of the chilled fresh meat treated under the condition is kept, and the chilled fresh meat does not have adverse effect on the health of human bodies. The lactoferrin can effectively protect the bright red color and quality of meat, and is a good antioxidant, can inhibit the oxidation speed of oxymyoglobin, protect the reduction system of the methemoglobin and inhibit the generation of malondialdehyde and thiobarbituric acid. The chitosan is also a good bacteriostatic agent and antioxidant, and forms a protective layer on the surface of the chilled fresh pork, so that the shelf life of the chilled fresh pork can be prolonged.
The cold fresh meat preserved after being treated by the method of the invention is evaluated by sensory and physicochemical indexes, and the result shows that: the reduction range and speed of the sensory quality of the meat sample are greatly inhibited, the elasticity is good, the color is ruddy, and no bad smell such as rancidness is generated; when the meat is preserved for 12 days, the loss rate of the meat juice is averagely reduced by 0.3-0.5 percent; the pH value is not more than 6.2 and is the standard of fresh meat specified in GB/T9695.5-2008 'determination of pH value of meat and meat products'; the total number of bacterial colonies and the volatile basic nitrogen value rise slowly; globulin precipitation test solutions were light blue in color with only a small amount of white precipitate. Compared with the cold fresh meat stored at the same temperature for preservation, the quality guarantee period of the cold fresh meat treated by the method is prolonged by 9 days, and the quality characteristics and safety of the cold fresh meat food are ensured.
Detailed Description
The present invention will be described in further detail with reference to examples.
First, experimental determination method
1. Sensory evaluation
And performing sensory evaluation on the preserved meat sample according to national standard GB/T22210-2008 sensory evaluation Specification for meat and meat products. Setting a full score of 5, wherein the score is from low to high and represents that the meat quality is from bad to good: the score of 5 is the fresest, the score of 4 is the fresher, the score of 3 is general, the score of 2 is poor, and the score of 1 is very poor. Meat samples were evaluated organoleptically by food professionals in the laboratory in terms of color, smell, texture, etc., averaged and recorded. 2. Determination of peroxide number
2. Determination of volatile basic Nitrogen (TVB-N)
Mincing cold fresh meat, uniformly weighing 10g, placing in a conical flask, adding 100mL water, shaking (or shaking with an electromagnetic stirrer), soaking for 30min, filtering with filter paper, and placing the filtrate in a refrigerator for use. Placing a small conical bottle containing 10mL of boric acid absorption liquid and 5-6 drops of mixed indicating liquid at the lower end of a condensation tube (the lower end of the condensation tube is inserted below the liquid level of the absorption liquid), accurately sucking 5.0mL of the filtrate into a reaction chamber of a distiller, adding 5.0mL of magnesium oxide suspension (10g/L), quickly covering and plugging, adding water to prevent air leakage, introducing steam for distillation, stopping distillation for 5min, titrating the absorption liquid with a hydrochloric acid standard solution, and ending the point to blue-purple. And simultaneously, carrying out a reagent blank test.
According to the provisions of GB/T5009.44-2003 analytical methods for the hygienic Standard of meat and meat products. The TVB-N value of the meat and meat products is primary freshness when the TVB-N value is less than or equal to 15mg/100g, secondary freshness when the TVB-N value is not more than 25mg/100g, and deteriorated meat when the TVB-N value is more than 25mg/100 g.
3. Determination of pH
Taking 5g of meat sample in a large centrifuge tube, adding sterile water to the large centrifuge tube 2/3, homogenizing the meat sample by using a homogenizer, adding ice on the periphery of the centrifuge tube in the homogenizing process to prevent the temperature of the test solution from being too high, and paying attention to the liquid level height of the test solution. And calibrating the pH meter, after the calibration is finished, carrying out pH measurement on the test solution by using the pH meter, flushing a measuring head of the pH meter every time of measurement, carrying out next measurement, and taking an average number and recording.
Fresh meat with pH of 5.8-6.2 according to the regulation of GB/T9695.5-2008 'determination of pH value of meat and meat products'; the meat with pH of 6.3-6.6 is sub-fresh meat; deteriorated meat having a pH of 6.7 or more.
4. Determination of juice loss Rate
Firstly, weighing the total mass (M1) of the meat sample without the preservative film and the preservative film, then removing the preservative film of the meat sample, carefully leaching out juice on the surface of the meat sample, weighing the total mass (M2) of the juice and the preservative film, and finally weighing the mass (M3) of the dried preservative film, so as to calculate the juice loss rate W, wherein the calculation formula is as follows:
W＝(M2-M3)/(M1-M3)。
5. determination of the Total number of colonies
About 25g of each meat sample was cut into pieces, placed in different small beakers, and 225mL of sterile water was added to each beaker, and then the sample solution was homogenized using a homogenizer. And counting the homogenized sample solution according to the total number of bacterial colonies determined by the food safety national standard food microbiology inspection of GB 4789.2-2016 according to a conventional method, and recording data.
6. Detection of globulin precipitate
Taking meat samples of different groups, taking about 5g of samples at different parts, shearing the samples into pieces, and placing the pieces in a small beaker. An equal amount of sterile water is added to the beaker, preferably with the water level submerged in the meat sample (about 50 mL). Shaking up by shaking, fully soaking the meat sample for about 30min, filtering, measuring 2mL of filtrate, dripping about 10 drops of copper sulfate solution with the mass fraction of 10%, standing, and observing the whole color change of the test solution after 5 min.
Second, the detailed description
Example 1
A method for prolonging the fresh-keeping period of cold fresh meat by using bacteriostatic lactic acid bacteria comprises the following steps:
(1) treatment of meat
The chopping board and the knife are sterilized in a sterilizing pot and wiped with sterilized alcohol before use. Under the aseptic condition of an aseptic table, removing redundant fascia and fat in purchased meat from fresh meat which is slaughtered by a meat complex on the day, and cutting the meat into meat blocks with the weight of about 100-1000 g for later use;
(2) expanded culture of lactobacillus
Inoculating lactobacillus plantarum into a liquid MRS culture medium in a sterile manner, and culturing in a thermostat at 37 ℃ for about 12 hours to fully activate the lactobacillus plantarum; then inoculating the activated strain into a large conical bottle filled with a liquid MRS culture medium, and expanding in a shaking table at the constant temperature of 37 ℃ for about 12 hours for later use;
(3) collection of Lactobacillus thallus
Pouring the cultured lactobacillus bacterial liquid into a centrifugal tube in a super-clean workbench, then putting the centrifugal tube into a centrifugal machine, centrifuging for 10min at 8000r/min, removing the upper layer liquid, adding physiological saline with the same volume as the bacterial liquid added at the beginning into the centrifugal tube, homogenizing, putting the centrifugal machine into the centrifugal machine, centrifuging for 10min at 8000r/min, removing the upper layer clear liquid, adding the physiological saline with the same volume, repeating the operation, taking the precipitate, and obtaining lactobacillus plantarum thallus precipitate;
(4) preparation of lophatherum gracile extract
Adding 100g of herba Lophatheri into 1000mL of water, boiling at a material-to-liquid ratio of 1:10g/mL, keeping the boiling state for 20min, performing ultrasonic extraction for 20min at an ultrasonic power of 300W to fully extract effective components of the herba Lophatheri, filtering the supernatant, and storing in a sterilized glass ware.
(5) Preparation of composite fresh-keeping liquid
And adding the cooled lophatherum gracile extract into a centrifugal tube containing the thallus precipitate according to the mass-volume ratio of the thallus to the lophatherum gracile extract of 1-3%, and blowing or whirling the thallus precipitate by using a pipette to uniformly mix so as to resuspend the lactobacillus plantarum precipitate to obtain lactobacillus plantarum suspension. Adding baicalin with final concentration of 15-30mg/L and active lactoferrin with final concentration of 10-30mg/L to make baicalin and lactoferrin be suspended in the bacterial suspension, finally adding chitosan with final concentration of 0.1-0.3%, and vortex and mixing uniformly to obtain a composite fresh-keeping solution for later use;
(6) spraying preservation liquid
According to the compound fresh-keeping liquid: uniformly spraying the chilled fresh meat with a small sprayer to a meat sample in a volume mass fraction (V/W) of 1.0-2.0%, standing for 1min, and draining for later use;
(7) packaging and preserving
And (3) coating the meat blocks sprayed with the lactobacillus plantarum suspension by using a food-grade polyethylene preservative film, and refrigerating and storing in a refrigerator at the temperature of (4 +/-0.5 ℃).
The preparation method of the MRS liquid culture medium comprises the following steps: 10g of peptone, 10g of beef extract, 5g of yeast extract, 2g of dipotassium phosphate, 2g of diammonium citrate, 5g of sodium acetate, 10g of glucose, 801 mL of tween-801, 0.5g of magnesium sulfate heptahydrate and 0.25g of manganese sulfate heptahydrate are dissolved in 1000mL of distilled water, the pH value is adjusted to 6.8, and sterilization is carried out for 15min at 121 ℃.
Example 2
The difference from the above example 1 is that:
and (5) in the preservation solution: spraying the lactobacillus plantarum suspension on the meat sample according to the volume fraction (V/W%) of 1.5%, standing for 1min, and draining for later use;
example 3
The difference from the above example 1 is that:
and (5) in the preservation solution: spraying the lactobacillus plantarum suspension to the meat sample according to the volume fraction (V/W%) of 2.0%, standing for 1min, and draining for later use;
example 4
The difference from the above example 1 is that:
and (5) in the preservation solution: spraying the lactobacillus plantarum suspension to the meat sample according to the volume fraction (V/W%) of 2.5%, standing for 1min, and draining for later use;
of course, the above description is not intended to limit the present invention, and the present invention is not limited to the above examples. Those skilled in the art should also realize that such changes, modifications, additions and substitutions are within the true spirit of the invention.

Claims (5)

1. A method for prolonging the fresh-keeping period of cold fresh meat by using bacteriostatic lactic acid bacteria is characterized by comprising the following steps:
(1) treatment of meat
The chopping board and the knife are sterilized in a sterilizing pot and wiped with sterilized alcohol before use. Under the aseptic condition of an aseptic table, removing redundant fascia and fat in purchased meat from fresh meat which is slaughtered by a meat complex on the day, and cutting the meat into meat blocks with the weight of about 100-1000 g for later use;
(2) expanded culture of lactobacillus
Inoculating lactobacillus plantarum into a liquid MRS culture medium in a sterile manner, and culturing in a thermostat at 37 ℃ for about 12 hours to fully activate the lactobacillus plantarum; then inoculating the activated strain into a large conical bottle filled with a liquid MRS culture medium, and expanding in a shaking table at the constant temperature of 37 ℃ for about 12 hours for later use;
(3) collection of Lactobacillus thallus
Pouring the cultured lactobacillus bacterial liquid into a centrifugal tube in a super-clean workbench, then putting the centrifugal tube into a centrifugal machine, centrifuging for 10min at 8000r/min, removing the upper layer liquid, adding physiological saline with the same volume as the bacterial liquid added at the beginning into the centrifugal tube, homogenizing, putting the centrifugal machine into the centrifugal machine, centrifuging for 10min at 8000r/min, removing the upper layer clear liquid, adding the physiological saline with the same volume, repeating the operation, taking the precipitate, and obtaining lactobacillus plantarum thallus precipitate;
(4) preparation of lophatherum gracile extract
Adding 100g of herba Lophatheri into 1000mL of water, boiling at a material-to-liquid ratio of 1:10g/mL, keeping the boiling state for 20min, performing ultrasonic extraction for 20min at an ultrasonic power of 300W to fully extract effective components of the herba Lophatheri, filtering the supernatant, and storing in a sterilized glass ware.
(5) Preparation of composite fresh-keeping liquid
And adding the cooled lophatherum gracile extract into a centrifugal tube containing the thallus precipitate according to the mass-volume ratio of the thallus to the lophatherum gracile extract of 1-3%, and blowing or whirling the thallus precipitate by using a pipette to uniformly mix so as to resuspend the lactobacillus plantarum precipitate to obtain lactobacillus plantarum suspension. Adding baicalin with final concentration of 15-30mg/L and active lactoferrin with final concentration of 10-30mg/L to make baicalin and lactoferrin be suspended in the bacterial suspension, finally adding chitosan with final concentration of 0.1-0.3%, and vortex and mixing uniformly to obtain a composite fresh-keeping solution for later use;
(6) spraying preservation liquid
According to the compound fresh-keeping liquid: uniformly spraying the chilled fresh meat with a small sprayer to a meat sample in a volume mass fraction (V/W) of 1.0-2.0%, standing for 1min, and draining for later use;
(7) packaging and preserving
And (3) coating the meat blocks sprayed with the lactobacillus plantarum suspension by using a food-grade polyethylene preservative film, and refrigerating and storing in a refrigerator at the temperature of (4 +/-0.5 ℃).
The preparation method of the MRS liquid culture medium comprises the following steps: peptone l0g, beef extract l0g, yeast extract 5g, dipotassium hydrogen phosphate 2g, diammonium citrate 2g, sodium acetate 5g, glucose 10g, Tween-80 l mL, magnesium sulfate heptahydrate 0.5g and manganese sulfate heptahydrate 0.25g are dissolved in 1000mL of distilled water, the pH is adjusted to 6.8, and sterilization is carried out at 121 ℃ for 15 min.
2. The method for prolonging the shelf life of cold fresh meat by using bacteriostatic lactic acid bacteria as claimed in claim 1, wherein the method comprises the following steps: and (3) enabling the whole process of the meat strips obtained in the step (1) to be in an aseptic condition, and applying the meat strips to the subsequent preservative solution spraying step.
3. The method for prolonging the shelf life of cold fresh meat by using bacteriostatic lactic acid bacteria as claimed in claim 1, wherein the method comprises the following steps: the lactobacillus used in the step (2) is lactobacillus plantarum.
4. The method for prolonging the shelf life of cold fresh meat by using bacteriostatic lactic acid bacteria according to claim 1, wherein the MRS liquid culture medium in the step (2) is prepared by the following steps: peptone l0g, beef extract l0g, yeast extract 5g, dipotassium hydrogen phosphate 2g, diammonium citrate 2g, sodium acetate 5g, glucose 10g, Tween-80 l mL, magnesium sulfate heptahydrate 0.5g and manganese sulfate heptahydrate 0.25g are dissolved in 1000mL of distilled water, the pH is adjusted to 6.8, and sterilization is carried out at 121 ℃ for 15 min.
5. The method for prolonging the shelf life of cold fresh meat by using bacteriostatic lactic acid bacteria as claimed in claim 1, wherein the method comprises the following steps: the fresh-keeping liquid in the step (4) is prepared by re-dissolving separated and purified lactobacillus plantarum in sterile water, wherein the concentration of the bacterial liquid is 107cfu/ml。
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