阅读说明：本技术 一种肉制品生物保鲜菌剂及其制备方法 (Biological fresh-keeping microbial agent for meat products and preparation method thereof ) 是由 胡传炯 朱颍琨 李姗 陈桂萍 徐杰 张康 于 2021-09-16 设计创作，主要内容包括：本发明公开了一种肉制品生物保鲜菌剂及其制备方法,包含乳酸菌培养基、乳酸菌及乳酸菌代谢物,所述乳酸菌培养基的配方为：蛋白胨8-12％、酵母提取液4-8％、牛肉膏10-15％、葡萄糖13-20％、乙酸钠1-6％、柠檬酸钠1.5-2.5％、淡竹叶10-15％、肉桂粉20-30％、MgSO-(4)·7H-(2)O 0.5-0.6％、MnSO-(4)·4H-(2)O 0.01-0.05％、K-(2)HPO-(4) 1-3％和CaCO-(3) 5-15％,调整pH值为5.5。本发明提供的肉制品生物保鲜菌剂,通过改变pH和营养竞争的方式抑制腐败菌的生长和挥发性盐基氮的产生,从而延长冷鲜肉保质期,改善保质期中的品质。(The invention discloses a biological fresh-keeping microbial agent for meat products and a preparation method thereof, wherein the biological fresh-keeping microbial agent comprises a lactic acid bacteria culture medium, lactic acid bacteria and lactic acid bacteria metabolites, and the formula of the lactic acid bacteria culture medium is as follows: 8-12% of peptone, 4-8% of yeast extract, 10-15% of beef extract, 13-20% of glucose, 1-6% of sodium acetate, 1.5-2.5% of sodium citrate, 10-15% of lophatherum gracile, 20-30% of cinnamon powder and MgSO (MgSO) water 4 ·7H 2 O 0.5‑0.6％、MnSO 4 ·4H 2 O 0.01‑0.05％、K 2 HPO 4 1-3% and CaCO 3 5-15%, and adjusting the pH value to 5.5. The biological fresh-keeping microbial agent for meat products provided by the invention can inhibit the growth of putrefying bacteria and the generation of volatile basic nitrogen by changing the pH and nutrition competition modes, thereby prolonging the quality guarantee period of cold fresh meat and improving the quality in the quality guarantee period.)

1. The biological fresh-keeping microbial agent for the meat products is characterized by comprising a lactic acid bacteria culture medium, lactic acid bacteria and lactic acid bacteria metabolites, wherein the formula of the lactic acid bacteria culture medium is as follows: 8-12% of peptone, 4-8% of yeast extract, 10-15% of beef extract, 13-20% of glucose, 1-6% of sodium acetate, 1.5-2.5% of sodium citrate, 10-15% of lophatherum gracile, 20-30% of cinnamon powder and MgSO (MgSO) water₄·7H₂O 0.5-0.6％、MnSO₄·4H₂O 0.01-0.05％、K₂HPO₄1-3% and CaCO₃5-15%, and adjusting the pH value to 5.5.

2. The biological fresh-keeping agent for meat products as claimed in claim 1, wherein the lactic acid bacteria strain is obtained by screening fermented sausage.

3. The preparation method of the biological fresh-keeping microbial agent for meat products as claimed in claim 1, characterized by comprising the following steps:

step 1: obtaining of anti-putrefactive lactic acid bacteria: taking fermented sausage, cutting sausage casing after surface sterilization, grinding the content of the fermented sausage in a sterile grinder into homogenate, and separating and purifying the homogenate in an MRS culture medium after gradient dilution;

step 2: screening of anti-putrefactive lactic acid bacteria: culturing the purified lactobacillus in a liquid MRS culture medium for 24h, centrifuging at 6000rpm for 5min, taking the supernatant, adding the supernatant into a putrefying bacteria liquid culture system at different concentrations, and testing the bacteriostatic effect;

and step 3: and (3) metabolite detection: culturing the strain screened in the step 2 in a liquid MRS culture medium at 37 ℃ for 24 hours, centrifuging and taking a supernatant to detect the components of the strain by a college gas chromatography;

and 4, step 4: preparing a biological fresh-keeping microbial agent: the strains obtained by screening are cultured by strain rejuvenation, primary strains, secondary strains, a seeding tank and a fermentation tank, and then the supernatant is centrifugally collected to obtain the biological fresh-keeping microbial inoculum.

4. The method for preparing the biological fresh-keeping microbial inoculum for the meat products as claimed in claim 3, wherein the biological fresh-keeping microbial inoculum is stored for later use after being cooled to 4 ℃.

5. The method for preparing biological fresh-keeping agent for meat products as claimed in claim 3, wherein the whole process of step 4 is carried out for 55-65 hours, the culture solution is centrifuged at 6000rpm for 5 minutes, and the supernatant fermentation broth is collected.

6. The preparation method of the biological fresh-keeping microbial inoculum for the meat products as claimed in claim 3, wherein the use method of the prepared biological fresh-keeping microbial inoculum comprises the following steps: and spraying alcohol to sterilize the pork split into the white streaky pork, spraying the biological fresh-keeping agent, and entering a freezer.

7. The method for preparing biological preservative for meat products as claimed in claim 6, wherein after the pork fillet is further divided into separate pieces, alcohol and the biological preservative are sprayed again before packaging and transportation.

Technical Field

The invention relates to the technical field of microbial fermentation, in particular to a biological fresh-keeping microbial inoculum for meat products and a preparation method thereof.

Background

In cold chain transportation of cold fresh meat, especially in terminal retail shelf life, due to the incomplete disinfection means and unstable refrigeration temperature, the terminal shelf life of the current cold fresh meat is short, and the transportation cost and range of the cold fresh meat are influenced. Therefore, it is a real need to increase the shelf life of chilled meat and improve the shelf life quality of chilled meat. Lactic acid bacteria can produce various metabolites mainly comprising organic acids during the growth process, and inhibit the growth of putrefying bacteria and the generation of volatile basic nitrogen in a mode of changing pH and nutrition competition. However, metabolite compositions of different lactobacillus strains are greatly different, so that the separated strain with good inhibition capability on common pollution bacteria has important significance for preparing the biological fresh-keeping microbial inoculum.

Disclosure of Invention

The biological fresh-keeping microbial agent for the meat products provided by the invention can be used for inhibiting the growth of putrefying bacteria and the generation of volatile basic nitrogen in a pH and nutrition competition changing manner, so that the quality guarantee period of cold fresh meat is prolonged, the quality in the quality guarantee period is improved, and the problems in the background art are solved.

In order to achieve the purpose, the invention provides the following technical scheme:

a biological fresh-keeping microbial agent for meat products comprises a lactic acid bacteria culture medium, lactic acid bacteria and lactic acid bacteria metabolites, wherein the formula of the lactic acid bacteria culture medium is as follows: 8-12% of peptone, 4-8% of yeast extract, 10-15% of beef extract, 13-20% of glucose, 1-6% of sodium acetate, 1.5-2.5% of sodium citrate, 10-15% of lophatherum gracile, 20-30% of cinnamon powder and MgSO (MgSO) water₄·7H₂O 0.5-0.6％、MnSO₄·4H₂O 0.01-0.05％、K₂HPO₄1-3% and CaCO₃5-15%, and adjusting the pH value to 5.5.

Further, the strain of the lactic acid bacteria is obtained by screening fermented sausages.

The invention provides another technical scheme: a preparation method of biological fresh-keeping microbial inoculum for meat products comprises the following steps:

step 1: obtaining of anti-putrefactive lactic acid bacteria: taking fermented sausage, cutting sausage casing after surface sterilization, grinding the content of the fermented sausage in a sterile grinder into homogenate, and separating and purifying the homogenate in an MRS culture medium after gradient dilution;

step 2: screening of anti-putrefactive lactic acid bacteria: culturing the purified lactobacillus in a liquid MRS culture medium for 24h, centrifuging at 6000rpm for 5min, taking the supernatant, adding the supernatant into a putrefying bacteria liquid culture system at different concentrations, and testing the bacteriostatic effect;

and step 3: and (3) metabolite detection: culturing the strain screened in the step 2 in a liquid MRS culture medium at 37 ℃ for 24 hours, centrifuging and taking a supernatant to detect the components of the strain by a college gas chromatography;

and 4, step 4: preparing a biological fresh-keeping microbial agent: the strains obtained by screening are cultured by strain rejuvenation, primary strains, secondary strains, a seeding tank and a fermentation tank, and then the supernatant is centrifugally collected to obtain the biological fresh-keeping microbial inoculum.

Further, the biological fresh-keeping microbial inoculum is cooled to 4 ℃ and then stored for later use.

Further, the whole process of step 4 is cultured for 55-65 hours, the culture solution is centrifuged for 5 minutes at 6000rpm, and the supernatant fermentation broth is collected.

Further, the use method of the prepared biological fresh-keeping microbial inoculum comprises the following steps: and spraying alcohol to sterilize the pork split into the white streaky pork, spraying the biological fresh-keeping agent, and entering a freezer.

Further, after the pork white strip is further divided into divided meat, alcohol and the biological fresh-keeping agent are sprayed again before packaging and transportation.

Compared with the prior art, the invention has the beneficial effects that:

the invention can biologically inhibit putrefying bacteria existing in the environment by a small amount of live lactobacillus, lactobacillus metabolites, bamboo leaf flavone, cinnamaldehyde and the like, thereby reducing the growth probability of microorganisms on the cold fresh meat and prolonging the storage time of the cold fresh meat. The invention prolongs the fresh-keeping period by the competitive inhibition of the probiotics, can be compatible with the commonly used alcohol disinfection method at present, and has application value.

Drawings

FIG. 1 is a flow chart of the preparation of the biological fresh-keeping agent of the present invention.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1

A biological fresh-keeping microbial agent for meat products comprises a lactic acid bacteria culture medium, lactic acid bacteria and lactic acid bacteria metabolites, wherein the formula of the lactic acid bacteria culture medium is as follows: peptone 8%, yeast extract 8%, beef extract 15%, glucose 20%, sodium acetate 5.99%, sodium citrate 1.5%, lophatherum gracile 15%, cinnamon powder 20%, MgSO 2₄·7H₂O0.5％、MnSO₄·4H₂O 0.01％、K₂HPO₄1% and CaCO₃5%, and adjusting the pH value to 5.5.

The preparation process is shown in figure 1, and the anti-rancidity lactic acid bacteria strain is obtained: taking fermented sausage, cutting sausage casing after surface sterilization, grinding the content of the fermented sausage in a sterile grinder into homogenate, and separating and purifying the homogenate in an MRS culture medium after gradient dilution; screening of anti-putrefactive lactic acid bacteria: culturing the purified lactobacillus in a liquid MRS culture medium for 24h, centrifuging at 6000rpm for 5min, taking the supernatant, adding the supernatant into a putrefying bacteria liquid culture system at different concentrations, and testing the bacteriostatic effect; and (3) metabolite detection: culturing the strain screened in the step 2 in a liquid MRS culture medium at 37 ℃ for 24 hours, centrifuging and taking a supernatant to detect the components of the strain by a college gas chromatography; preparing a biological fresh-keeping microbial agent: and (3) rejuvenating the strains obtained by screening, culturing the first-stage strains, the second-stage strains, a seeding tank and a fermentation tank, wherein the total flow culture time is 60 hours, centrifuging the culture solution at 6000rpm for 5 minutes, collecting supernatant clear fermentation liquor to obtain a biological fresh-keeping microbial inoculum, and cooling the biological fresh-keeping microbial inoculum to 4 ℃ for storage for later use.

Example 2

A biological fresh-keeping microbial agent for meat products comprises a lactic acid bacteria culture medium, lactic acid bacteria and lactic acid bacteria metabolites, wherein the formula of the lactic acid bacteria culture medium is as follows: 10% of peptone, 6% of yeast extract, 12.4% of beef extract, 16% of glucose, 4% of sodium acetate, 2% of sodium citrate, 12% of lophatherum gracile, 25% of cinnamon powder and MgSO (MgSO) s₄·7H₂O0.55％、MnSO₄·4H₂O 0.05％、K₂HPO₄2% and CaCO₃10%, and adjusting the pH value to 5.5.

Obtaining of anti-putrefactive lactic acid bacteria: taking fermented sausage, cutting sausage casing after surface sterilization, grinding the content of the fermented sausage in a sterile grinder into homogenate, and separating and purifying the homogenate in an MRS culture medium after gradient dilution; screening of anti-putrefactive lactic acid bacteria: culturing the purified lactobacillus in a liquid MRS culture medium for 24h, centrifuging at 6000rpm for 5min, taking the supernatant, adding the supernatant into a putrefying bacteria liquid culture system at different concentrations, and testing the bacteriostatic effect; and (3) metabolite detection: culturing the strain screened in the step 2 in a liquid MRS culture medium at 37 ℃ for 24 hours, centrifuging and taking a supernatant to detect the components of the strain by a college gas chromatography; preparing a biological fresh-keeping microbial agent: and (3) rejuvenating the strains obtained by screening, culturing the primary strains, the secondary strains, a seeding tank and a fermentation tank, wherein the total flow culture time is 65 hours, centrifuging the culture solution at 6000rpm for 5 minutes, collecting supernatant clear fermentation liquor to obtain a biological fresh-keeping microbial inoculum, and cooling the biological fresh-keeping microbial inoculum to 4 ℃ and storing the biological fresh-keeping microbial inoculum for later use.

Example 3

A biological fresh-keeping microbial agent for meat products comprises a lactic acid bacteria culture medium, lactic acid bacteria and lactic acid bacteria metabolites, wherein the formula of the lactic acid bacteria culture medium is as follows: peptone 12%, yeast extract 4%, beef extract 10%, glucose 13%, sodium acetate 6%, sodium citrate 2.5%, lophatherum gracile 10%, cinnamon powder 28.87%, MgSO 2₄·7H₂O0.6％、MnSO₄·4H₂O 0.03％、K₂HPO₄3% and CaCO₃10%, and adjusting the pH value to 5.5.

Obtaining of anti-putrefactive lactic acid bacteria: taking fermented sausage, cutting sausage casing after surface sterilization, grinding the content of the fermented sausage in a sterile grinder into homogenate, and separating and purifying the homogenate in an MRS culture medium after gradient dilution; screening of anti-putrefactive lactic acid bacteria: culturing the purified lactobacillus in a liquid MRS culture medium for 24h, centrifuging at 6000rpm for 5min, taking the supernatant, adding the supernatant into a putrefying bacteria liquid culture system at different concentrations, and testing the bacteriostatic effect; and (3) metabolite detection: culturing the strain screened in the step 2 in a liquid MRS culture medium at 37 ℃ for 24 hours, centrifuging and taking a supernatant to detect the components of the strain by a college gas chromatography; preparing a biological fresh-keeping microbial agent: and (3) rejuvenating the strains obtained by screening, culturing the first-stage strains, the second-stage strains, a seeding tank and a fermentation tank, wherein the total flow culture time is 60 hours, centrifuging the culture solution at 6000rpm for 5 minutes, collecting supernatant clear fermentation liquor to obtain a biological fresh-keeping microbial inoculum, and cooling the biological fresh-keeping microbial inoculum to 4 ℃ for storage for later use.

Example 4

A biological fresh-keeping microbial agent for meat products comprises a lactic acid bacteria culture medium, lactic acid bacteria and lactic acid bacteria metabolites, wherein the formula of the lactic acid bacteria culture medium is as follows: 8% of peptone, 4% of yeast extract, 10% of beef extract, 15% of glucose, 4.95% of sodium acetate, 1.5% of sodium citrate, 10% of lophatherum gracile, 30% of cinnamon powder and MgSO (MgSO) s₄·7H₂O0.5％、MnSO₄·4H₂O 0.05％、K₂HPO₄1% and CaCO₃15%, and adjusting the pH value to 5.5.

Obtaining of anti-putrefactive lactic acid bacteria: taking fermented sausage, cutting sausage casing after surface sterilization, grinding the content of the fermented sausage in a sterile grinder into homogenate, and separating and purifying the homogenate in an MRS culture medium after gradient dilution; screening of anti-putrefactive lactic acid bacteria: culturing the purified lactobacillus in a liquid MRS culture medium for 24h, centrifuging at 6000rpm for 5min, taking the supernatant, adding the supernatant into a putrefying bacteria liquid culture system at different concentrations, and testing the bacteriostatic effect; and (3) metabolite detection: culturing the strain screened in the step 2 in a liquid MRS culture medium at 37 ℃ for 24 hours, centrifuging and taking a supernatant to detect the components of the strain by a college gas chromatography; preparing a biological fresh-keeping microbial agent: and (3) rejuvenating the strains obtained by screening, culturing the first-stage strains, the second-stage strains, a seeding tank and a fermentation tank, wherein the total flow culture time is 55 hours, centrifuging the culture solution at 6000rpm for 5 minutes, collecting supernatant fermentation liquor to obtain a biological fresh-keeping microbial inoculum, and cooling the biological fresh-keeping microbial inoculum to 4 ℃ for storage for later use.

Comparative example

Conventional biological fresh-keeping microbial inoculum

Taking 5 strips of white meat which have the same size and are cut from the same pig carcass, and numbering the 5 st group, the 2 nd group, the 3 rd group, the 4 th group and the 5 th group.

EXAMPLES 1 to 4

And (3) spraying alcohol on the strip meat of the group 1 for disinfection, then spraying the biological fresh-keeping agent, and entering a freezer. After the strip meat of the group 1 is further divided into cut meat, alcohol and the biological preservative are sprayed again before packaging and transportation.

And (3) spraying alcohol on the strip meat of the group 2 for disinfection, then spraying the biological fresh-keeping agent, and entering a freezer. After the strip meat of the group 2 is further divided into cut meat, alcohol and the biological preservative are sprayed again before packaging and transportation.

And spraying alcohol on the strip meat of the group 3 for disinfection, then spraying the biological fresh-keeping agent, and entering a freezer. After the white strip meat of the group 3 is further divided into cut meat, alcohol and the biological preservative are sprayed again before packaging and transportation.

And (4) spraying alcohol on the strip meat of the group 4 for disinfection, then spraying the biological fresh-keeping agent, and entering a freezer. After the strip meat of group 4 is further divided into cut meat, alcohol and the biological preservative are sprayed again before packaging and transportation.

After alcohol is sprayed on the strip meat of the group 5 for disinfection, a conventional biological fresh-keeping agent is sprayed on the strip meat, and the strip meat enters a freezer. After the strip meat of group 5 is further divided into cut meat, alcohol and conventional biological preservative agents are sprayed again before packaging and transportation.

The above 5 groups of the white-strip meat are transported by the same vehicle, and meat color identification is uniformly carried out after the white-strip meat arrives at the destination, and the identification results are as follows in the following table 1:

table 1: meat color discrimination

As can be seen from the above table 1, the color of the stripped meat treated by the biological fresh-keeping microbial agent in the examples 1-4 is bright red, and the color of the stripped meat treated by the conventional biological fresh-keeping microbial agent in the comparative example is dark red, which shows that the fresh-keeping effect of the biological fresh-keeping microbial agent on the meat product is superior to that of the conventional biological fresh-keeping microbial agent.

In addition, after identifying the meat color, the total number of colonies of the 5 groups of the white streaky meat was subjected to a microbiological examination, and the results are shown in the following table 2:

table 2: results of microbiological examination

It can be seen from table 2 that, in examples 1 to 4, the total number of the bacon albuginea treated by the biological preservative is lower than the limit value, and the bacon albuginea can be judged as fresh meat, while the total number of the bacon albuginea treated by the conventional biological preservative is obviously higher than the limit value, and the bacon albuginea can be judged as secondary fresh meat, which indicates that the effect of the biological preservative on the inhibition of common pathogenic or opportunistic pathogenic microorganisms in the environment of the meat product is better than the effect of the conventional biological preservative on the inhibition of common pathogenic or opportunistic pathogenic microorganisms in the environment.

Compared with the existing fresh-keeping mode, the invention can better reduce the drip loss, keep the meat color, inhibit the pollution of common pathogenic or opportunistic microorganisms in the environment to the surface of the meat and effectively improve the safety in the processes of cold chain transportation and shelf storage.

The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be able to cover the technical solutions and the inventive concepts of the present invention within the technical scope of the present invention.