CN114868914B - A prebiotics and its preparation method - Google Patents

A prebiotics and its preparation method Download PDF

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CN114868914B
CN114868914B CN202210366346.8A CN202210366346A CN114868914B CN 114868914 B CN114868914 B CN 114868914B CN 202210366346 A CN202210366346 A CN 202210366346A CN 114868914 B CN114868914 B CN 114868914B
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彭孝雄
尹修权
徐彩虹
旷文丰
陈晓霞
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Guangzhou Zhengming Houshengyuan Technology Co ltd
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Abstract

The invention discloses a metazoan and a preparation method thereof, belonging to the technical field of biology. The preparation method of the metancholia comprises the following steps: s1, inoculating lactobacillus acidophilus into a lactobacillus culture solution for culturing, culturing at 35-38 ℃ to obtain a first fermentation solution, adding citric acid and hydrolyzed protein into the first fermentation solution, continuously culturing at 25-30 ℃ to obtain a second fermentation solution, and culturing the second fermentation solution at 35-38 ℃ to obtain a third fermentation solution; s2, filtering the third fermentation liquor to obtain a supernatant, and concentrating the supernatant to obtain a concentrated solution; s3, adding gelatin into the concentrated solution, and then refrigerating at 5-10 ℃ to obtain a fourth fermentation liquid; and S4, inactivating the fourth fermentation liquor. The invention also comprises the metazoan prepared by the preparation method. The obtained Lactobacillus acidophilus has high content and the metazoan has high content of antibacterial peptide.

Description

A prebiotics and its preparation method
Technical Field
The invention relates to the technical field of biology, in particular to a metazoan and a preparation method thereof.
Background
The metazoan is a general name of the metabolite components of the probiotics after the probiotics are processed and treated, and comprises thalli and metabolites. Recent studies have found that inactivated probiotics (non-viable probiotics) and their lysates (lysates), extracts or isolates (known as postbiotics) have similar efficacy to probiotics and are more acid tolerant and easier to preserve than probiotics.
Lactobacillus acidophilus, one of lactic acid bacteria, produces many antibacterial substances in the metabolic process, mainly including acidic substances, antibacterial peptides, etc., and can effectively resist food deterioration and food-borne pathogenic microorganisms. The antibacterial peptide has broad-spectrum antibacterial activity and strong killing effect on bacteria, and particularly the killing effect on certain drug-resistant pathogenic bacteria draws more attention from people; the method is a technical problem which needs to be solved urgently in the prior art, the prior art promotes the production of the antibacterial peptide by adding salmonella into the lactic acid bacteria as a competitive relationship, but the survival rate of the lactic acid bacteria can be influenced in the competitive process, and further the yield of the antibacterial peptide can be influenced, and how to improve the yield of the antibacterial peptide while ensuring the number of lactobacillus acidophilus is a technical problem which needs to be solved urgently in the prior art.
Disclosure of Invention
The invention aims to overcome the technical defects, provides a metazoan and a preparation method thereof, and solves the technical problem of how to ensure the number of lactobacillus acidophilus and improve the yield of antibacterial peptide in the prior art.
In order to achieve the technical purpose, the technical scheme of the invention provides a preparation method of a metazoan, which comprises the following steps:
s1, inoculating lactobacillus acidophilus into a lactobacillus culture solution for culturing, culturing at 35-38 ℃ to obtain a first fermentation solution, adding citric acid and hydrolyzed protein into the first fermentation solution, continuously culturing at 25-30 ℃ to obtain a second fermentation solution, and culturing the second fermentation solution at 35-38 ℃ to obtain a third fermentation solution;
s2, filtering the third fermentation liquor to obtain a supernatant, and concentrating the supernatant to obtain a concentrated solution;
s3, adding gelatin into the concentrated solution, and then refrigerating at 5-10 ℃ to obtain a fourth fermentation liquid;
and S4, inactivating the fourth fermentation liquor.
Further, in step S1, the addition amount of the citric acid is 0.1-0.5% of the volume of the first fermentation solution, and the concentration of the citric acid is 0.3-0.5 mol/L.
Further, in step S1, the amount of the hydrolyzed protein added is 0.1 to 0.5% by mass of the first fermentation solution.
Further, in step S4, the adding amount of the gelatin is 0.5-1% of the mass of the concentrated solution.
Further, after step S4, the method further includes: and S5, adding chitosan into the inactivated fourth fermentation liquor, and then carrying out spray drying to obtain the metagens.
Further, in step S5, the amount of the chitosan added is 1-3% of the mass of the inactivated fourth fermentation broth.
Further, in step S3, the volume of the concentrated solution is 0.5 to 0.6 times the volume of the supernatant.
Further, in step S3, the refrigerating time is 30-60min.
Further, in step S1, culturing at 35-38 deg.C for 2-3 days to obtain a first fermentation solution; culturing at 25-30 deg.C for 3-5h to obtain second fermentation broth, and culturing at 35-38 deg.C for 1-2 days to obtain third fermentation broth.
In addition, the invention also provides a metazoan prepared by the preparation method.
Compared with the prior art, the invention has the beneficial effects that: the lactobacillus acidophilus is inoculated into a lactobacillus culture solution for culturing, the lactobacillus acidophilus is cultured at a higher temperature of 35-38 ℃ to be favorable for the growth of the lactobacillus acidophilus, citric acid and hydrolyzed protein are added into a first fermentation solution to be continuously cultured at a lower temperature of 25-30 ℃ to obtain a second fermentation solution, the citric acid and the hydrolyzed protein are added into the first fermentation solution to be continuously cultured at a lower temperature of 35-38 ℃ to obtain a third fermentation solution, the hydrolyzed protein provides amino acid required by the generation of antibacterial peptide, the citric acid improves the stability of the lactobacillus acidophilus, the low-temperature and high-temperature culture is favorable for promoting the growth of the lactobacillus acidophilus again and promoting the generation of the antibacterial peptide, then the third fermentation solution is filtered and concentrated to obtain a concentrated solution, gelatin is added into the concentrated solution, and is refrigerated at a temperature of 5-10 ℃ to obtain a fourth fermentation solution, the gelatin has a binding effect, the position of the lactobacillus acidophilus can be stabilized at the lower temperature, the inactivation treatment can improve the precision of the lactobacillus acidophilus after further inactivation treatment, the efficiency of inactivation can be improved, the lactobacillus acidophilus can be realized only at a temperature of 30-60min at 80-85 ℃, and the high content of the inactivated lactobacillus acidophilus can be completely, and the high content of the lactobacillus acidophilus can be obtained.
Detailed Description
The specific embodiment provides a preparation method of a metazoan, which comprises the following steps:
s1, inoculating lactobacillus acidophilus into a lactobacillus culture solution for culturing, culturing for 1-2 days at 35-38 ℃ to obtain a first fermentation solution, adding citric acid and hydrolyzed protein into the first fermentation solution, continuously culturing for 3-5 hours at 25-30 ℃ to obtain a second fermentation solution, and culturing the second fermentation solution at 35-38 ℃ for 1-2 days to obtain a third fermentation solution; the hydrolyzed protein is preferably wheat hydrolyzed protein; the addition amount of the citric acid is 0.1-0.5% of the volume of the first fermentation liquid, and the concentration of the citric acid is 0.3-0.5 mol/L; the addition amount of the hydrolyzed protein is 0.1-0.5% of the mass of the first fermentation solution;
s2, filtering the third fermentation liquor to obtain a supernatant, and concentrating the supernatant to obtain a concentrated solution; the volume of the concentrated solution is 0.5-0.6 times of the volume of the supernatant;
s3, adding gelatin into the concentrated solution, and then refrigerating at 5-10 ℃ for 30-60min to obtain a fourth fermentation liquid; the adding amount of the gelatin is 0.5-1% of the mass of the third fermentation liquor;
s4, inactivating the fourth fermentation liquor at 80-85 ℃ for 30-60min;
s5, adding chitosan into the inactivated fourth fermentation broth, and then performing spray drying to obtain a secondary biomass; the addition amount of the chitosan is 1-3% of the mass of the inactivated fourth fermentation broth. Chitosan enables the anagen to form microcapsules, the microcapsules can start to release the anagen when entering the intestinal tract, and the adhesion force with the intestinal tract is improved under the action of gelatin in the anagen, so that the action time of the anagen is prolonged, and the treatment effect on the intestinal inflammation is favorably improved.
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
In the following examples, the lactic acid bacteria culture medium was composed of 1000ml of water, 10g of beef extract, 20g of peptone, 10g of yeast extract, 200g of tomato juice, 10g of glucose, 0.5ml of tween, 17g of calcium carbonate, 0.1g of bromocresol green, and 15g of agar. The Lactobacillus acidophilus in the following examples is a known strain from the prior art collected by itself.
Example 1
This example presents a metazoan prepared by the following steps:
s1, mixing 10 4 Inoculating lactobacillus acidophilus into a lactobacillus culture solution for culturing, culturing for 2 days at 35 ℃ to obtain a first fermentation solution, adding citric acid and hydrolyzed protein into the first fermentation solution, continuously culturing for 3 hours at 25 ℃ to obtain a second fermentation solution, and culturing the second fermentation solution at 35 ℃ for 1 day to obtain a third fermentation solution; detecting the third fermentation liquid, the number of Lactobacillus acidophilus is 3.5 × 10 10 CFU/mL; the addition amount of the citric acid is 0.3 percent of the volume of the first fermentation liquid, and the concentration of the citric acid is 0.3mol/L; the addition amount of the hydrolyzed protein is 0.4 percent of the mass of the first fermentation solution;
s2, filtering the third fermentation liquor to obtain a supernatant, and concentrating the supernatant to obtain a concentrated solution; the volume of the concentrated solution is 0.5 times of the volume of the supernatant;
s3, adding gelatin into the concentrated solution, and then refrigerating at 5 ℃ for 30min to obtain a fourth fermentation liquid; the adding amount of the gelatin is 0.8 percent of the mass of the third fermentation liquid;
s4, inactivating the fourth fermentation liquor at 80 ℃ for 60min;
s5, adding chitosan into the inactivated fourth fermentation broth, then carrying out spray drying to obtain 162g of metazoan, and repeating the method in the embodiment by adopting a dialysis concentration method in the prior art to obtain 0.923g of antibacterial peptide; the addition amount of the chitosan is 1% of the mass of the inactivated fourth fermentation broth.
The yield of antimicrobial peptide was calculated to be 0.923 g/(162 g) = 100% =0.57%.
Example 2
This example presents a prebiotic made by the following steps:
s1, mixing 10 4 Inoculating lactobacillus acidophilus into a lactobacillus culture solution for culturing, culturing at 37 ℃ for 1 day to obtain a first fermentation solution, adding citric acid and hydrolyzed protein into the first fermentation solution, continuously culturing at 30 ℃ for 5 hours to obtain a second fermentation solution, and culturing the second fermentation solution at 37 ℃ for 2 days to obtain a third fermentation solution; detecting the third fermentation liquid, the number of Lactobacillus acidophilus is 3.2 × 10 10 CFU/mL; the addition amount of the citric acid is 0.1% of the volume of the first fermentation liquid, and the concentration of the citric acid is 0.5mol/L; the addition amount of the hydrolyzed protein is 0.2 percent of the mass of the first fermentation solution;
s2, filtering the third fermentation liquor to obtain a supernatant, and concentrating the supernatant to obtain a concentrated solution; the volume of the concentrated solution is 0.6 times of the volume of the supernatant;
s3, adding gelatin into the concentrated solution, and then refrigerating at 10 ℃ for 60min to obtain a fourth fermentation liquid; the adding amount of the gelatin is 0.5 percent of the mass of the third fermentation liquor;
s4, inactivating the fourth fermentation liquor at 85 ℃ for 45min;
s5, adding chitosan into the inactivated fourth fermentation broth, and then performing spray drying to obtain 158g of secondary prebiotics; the mass of the obtained antibacterial peptide obtained by repeating the method of the embodiment by adopting a dialysis concentration method of the prior art is 0.884g; the addition amount of the chitosan is 2% of the mass of the inactivated fourth fermentation liquid.
The yield of antimicrobial peptide was calculated to be 0.884 g/(158 g) × 100% =0.559%.
Example 3
This example presents a metazoan prepared by the following steps:
s1, mixing 10 4 Inoculating lactobacillus acidophilus into a lactobacillus culture solution for culturing, culturing at 38 ℃ for 1.5 days to obtain a first fermentation solution, adding citric acid and hydrolyzed protein into the first fermentation solution, continuously culturing at 28 ℃ for 4 hours to obtain a second fermentation solution, and culturing the second fermentation solution at 38 ℃ for 1 day to obtain a third fermentation solution; detecting third fermentation broth, lactobacillus acidophilusThe number is 2.9 × 10 10 CFU/mL; the addition amount of the citric acid is 0.5% of the volume of the first fermentation liquid, and the concentration of the citric acid is 0.4mol/L; the addition amount of the hydrolyzed protein is 0.1 percent of the mass of the first fermentation solution;
s2, filtering the third fermentation liquor to obtain a supernatant, and concentrating the supernatant to obtain a concentrated solution; the volume of the concentrated solution is 0.6 times of the volume of the supernatant;
s3, adding gelatin into the concentrated solution, and then refrigerating at 8 ℃ for 50min to obtain a fourth fermentation liquid; the adding amount of the gelatin is 1% of the mass of the third fermentation liquor;
s4, inactivating the fourth fermentation liquor at 85 ℃ for 40min;
s5, adding chitosan into the inactivated fourth fermentation broth, and then performing spray drying to obtain 162g of secondary prebiotics; the mass of the antibacterial peptide prepared by repeating the method of the embodiment by adopting a dialysis concentration method of the prior art is 0.824g; the addition amount of the chitosan is 3% of the mass of the inactivated fourth fermentation liquid.
The yield of antimicrobial peptide was calculated to be 0.828 g/(162 g) × 100% =0.511%.
Example 4
This example presents a prebiotic made by the following steps:
s1, mixing 10 4 Inoculating lactobacillus acidophilus into a lactobacillus culture solution for culturing, culturing at 36 ℃ for 1 day to obtain a first fermentation solution, adding citric acid and hydrolyzed protein into the first fermentation solution, continuously culturing at 27 ℃ for 5 hours to obtain a second fermentation solution, and culturing the second fermentation solution at 36 ℃ for 2 days to obtain a third fermentation solution; detecting the third fermentation liquid, the number of Lactobacillus acidophilus is 3.1 × 10 10 CFU/mL; the addition amount of the citric acid is 0.4% of the volume of the first fermentation liquid, and the concentration of the citric acid is 0.4mol/L; the addition amount of the hydrolyzed protein is 0.3 percent of the mass of the first fermentation liquid;
s2, filtering the third fermentation liquor to obtain a supernatant, and concentrating the supernatant to obtain a concentrated solution; the volume of the concentrated solution is 0.5 times of the volume of the supernatant;
s3, adding gelatin into the concentrated solution, and then refrigerating at 7 ℃ for 40min to obtain a fourth fermentation liquid; the adding amount of the gelatin is 0.8 percent of the mass of the third fermentation liquid;
s4, inactivating the fourth fermentation liquor at 80 ℃ for 30min;
s5, adding chitosan into the inactivated fourth fermentation broth, and then performing spray drying to obtain 159g of secondary prebiotics; the mass of the antibacterial peptide prepared by repeating the method of the embodiment by adopting a dialysis concentration method in the prior art is 0.817g; the adding amount of the chitosan is 3% of the mass of the inactivated fourth fermentation liquor.
The yield of antimicrobial peptide was calculated to be 0.817 g/(159 g) = 100% =0.514%.
Example 5
This example presents a prebiotic made by the following steps:
s1, adding 10 4 Inoculating lactobacillus acidophilus into a lactobacillus culture solution for culturing, culturing at 37 ℃ for 1 day to obtain a first fermentation solution, adding citric acid and hydrolyzed protein into the first fermentation solution, continuously culturing at 25 ℃ for 4 hours to obtain a second fermentation solution, and culturing the second fermentation solution at 37 ℃ for 2 days to obtain a third fermentation solution; detecting the third fermentation liquid, the number of Lactobacillus acidophilus is 3.2 × 10 10 CFU/mL; the addition amount of the citric acid is 0.5% of the volume of the first fermentation liquid, and the concentration of the citric acid is 0.3mol/L; the addition amount of the hydrolyzed protein is 0.1 percent of the mass of the first fermentation liquid;
s2, filtering the third fermentation liquor to obtain a supernatant, and concentrating the supernatant to obtain a concentrated solution; the volume of the concentrated solution is 0.5 times of the volume of the supernatant;
s3, adding gelatin into the concentrated solution, and then refrigerating at 5 ℃ for 30min to obtain a fourth fermentation liquid; the adding amount of the gelatin is 0.6 percent of the mass of the third fermentation liquid;
s4, inactivating the fourth fermentation liquor at 80 ℃ for 50min;
s5, adding chitosan into the inactivated fourth fermentation broth, then carrying out spray drying to obtain 155g of postbiotic, and repeating the method of the embodiment by adopting a dialysis concentration method in the prior art to obtain 0.851g of antibacterial peptide; the adding amount of the chitosan is 2% of the mass of the inactivated fourth fermentation liquor.
The yield of the antimicrobial peptide was calculated to be 0.851 g/(155 g) × 100% =0.549%.
Comparative example 1
This comparative example differs from example 1 only in that: lactobacillus acidophilus was always cultured at 35 ℃ as follows:
this example presents a metazoan prepared by the following steps:
s1, adding 10 4 Inoculating lactobacillus acidophilus into a lactobacillus culture solution for culturing, culturing for 2 days at 35 ℃ to obtain a first fermentation solution, adding citric acid and hydrolyzed protein into the first fermentation solution, continuously culturing for 3 hours at 35 ℃ to obtain a second fermentation solution, and culturing the second fermentation solution at 35 ℃ for 1 day to obtain a third fermentation solution; detecting the third fermentation liquid, the number of Lactobacillus acidophilus is 3.5 × 10 7 CFU/mL; the addition amount of the citric acid is 0.3% of the volume of the first fermentation liquid, and the concentration of the citric acid is 0.3mol/L; the addition amount of the hydrolyzed protein is 0.4 percent of the mass of the first fermentation solution;
s2, filtering the third fermentation liquor to obtain a supernatant, and concentrating the supernatant to obtain a concentrated solution; the volume of the concentrated solution is 0.5 times of the volume of the supernatant;
s3, adding gelatin into the concentrated solution, and then refrigerating at 5 ℃ for 30min to obtain a fourth fermentation liquid; the adding amount of the gelatin is 0.8 percent of the mass of the third fermentation liquid;
s4, inactivating the fourth fermentation liquor at 80 ℃ for 60min;
s5, adding chitosan into the inactivated fourth fermentation broth, then performing spray drying to obtain 112g of metazoan, and repeating the method of the embodiment by adopting a dialysis concentration method in the prior art to obtain 0.512g of antibacterial peptide; the addition amount of the chitosan is 1% of the mass of the inactivated fourth fermentation broth.
The yield of antimicrobial peptide was calculated to be 0.512 g/(112 g) = 100% =0.457%.
Comparative example 2
The comparative example only differs from example 1 in that: citric acid and hydrolyzed protein were not added during the fermentation as follows:
this example presents a metazoan prepared by the following steps:
s1, adding 10 4 Inoculating Lactobacillus acidophilus into a lactic acid bacteria culture solution for culturing, culturing at 35 deg.C for 2 days to obtain a first fermentation solution, culturing at 25 deg.C for 3h to obtain a second fermentation solution, and culturing at 35 deg.C for 1 day to obtain a third fermentation solution; detecting the third fermentation liquid, the number of Lactobacillus acidophilus is 3.2 × 10 9 CFU/mL；
S2, filtering the third fermentation liquor to obtain a supernatant, and concentrating the supernatant to obtain a concentrated solution; the volume of the concentrated solution is 0.5 times of the volume of the supernatant;
s3, adding gelatin into the concentrated solution, and then refrigerating at 5 ℃ for 30min to obtain a fourth fermentation liquid; the adding amount of the gelatin is 0.8 percent of the mass of the third fermentation liquid;
s4, inactivating the fourth fermentation liquor at 80 ℃ for 60min;
s5, adding chitosan into the inactivated fourth fermentation broth, then performing spray drying to obtain 145g of metagens, and repeating the method of the embodiment by adopting a dialysis concentration method in the prior art to obtain 0.312g of antibacterial peptide; the adding amount of the chitosan is 1% of the mass of the inactivated fourth fermentation liquor.
The yield of the antimicrobial peptide was calculated to be 0.312 g/(145 g) × 100% =0.215%.
As can be seen from comparative example 1, the fermentation broth obtained without the preparation method of high and low temperature treatment has a small number of Lactobacillus acidophilus and a low yield of the antimicrobial peptide; it can also be seen from comparative example 2 that fermentation without the addition of citric acid and hydrolyzed protein had a smaller effect on the number of L.acidophilus but a larger effect on the yield of antimicrobial peptides.
Comparative example 3
This comparative example differs from example 1 in that no gelatin was added, as follows:
the comparative example proposes a prebiotic prepared by the steps of:
s1, adding 10 4 Inoculating lactobacillus acidophilus into a lactobacillus culture solution for culturing, culturing at 35 ℃ for 2 days to obtain a first fermentation solution, adding citric acid and hydrolyzed protein into the first fermentation solution, continuously culturing at 25 ℃ for 3 hours to obtain a second fermentation solution, and culturing the second fermentation solution at 35 ℃ for 1 day to obtain a third fermentation solution; detecting the third fermentation liquid, the number of Lactobacillus acidophilus is 3.5 × 10 10 CFU/mL; the addition amount of the citric acid is 0.3 percent of the volume of the first fermentation liquid, and the concentration of the citric acid is 0.3mol/L; the addition amount of the hydrolyzed protein is 0.4 percent of the mass of the first fermentation liquid;
s2, filtering the third fermentation liquor to obtain a supernatant, and concentrating the supernatant to obtain a concentrated solution; the volume of the concentrated solution is 0.5 times of the volume of the supernatant;
s3, refrigerating the concentrated solution at 5 ℃ for 30min to obtain a fourth fermentation liquid;
s4, inactivating the concentrated solution at 80 ℃ for 60min;
s5, adding chitosan into the inactivated concentrated solution, then carrying out spray drying to obtain 163g of metazoan, and repeating the method of the embodiment by adopting a dialysis concentration method in the prior art to obtain 0.911g of antibacterial peptide; the adding amount of the chitosan is 1% of the mass of the inactivated fourth fermentation liquor.
The yield of antimicrobial peptide was calculated to be 0.921 g/(163 g) × 100% =0.565%.
Comparative example 4
This comparative example differs from example 1 in that no refrigeration is performed, as follows:
the comparative example proposes a prebiotic prepared by the steps of:
s1, mixing 10 4 Inoculating Lactobacillus acidophilus into lactobacillus culture solution, culturing at 35 deg.C for 2 days to obtain first fermentation liquid, adding citric acid and hydrolyzed protein into the first fermentation liquid, and adding into the first fermentation liquidCulturing at 25 deg.C for 3h to obtain second fermentation broth, and culturing at 35 deg.C for 1 day to obtain third fermentation broth; detecting the third fermentation liquid, the number of Lactobacillus acidophilus is 3.5 × 10 10 CFU/mL; the addition amount of the citric acid is 0.3 percent of the volume of the first fermentation liquid, and the concentration of the citric acid is 0.3mol/L; the addition amount of the hydrolyzed protein is 0.4 percent of the mass of the first fermentation liquid;
s2, filtering the third fermentation liquor to obtain a supernatant, and concentrating the supernatant to obtain a concentrated solution; the volume of the concentrated solution is 0.5 times of the volume of the supernatant;
s3, adding gelatin into the concentrated solution to obtain a fourth fermentation liquid; the adding amount of the gelatin is 0.8 percent of the mass of the third fermentation liquor;
s4, inactivating the fourth fermentation liquor at 80 ℃ for 60min;
s5, adding chitosan into the inactivated fourth fermentation broth, then carrying out spray drying to obtain 163g of metazoan, and repeating the method in the embodiment by adopting a dialysis concentration method in the prior art to obtain 0.923g of antibacterial peptide; the addition amount of the chitosan is 1% of the mass of the inactivated fourth fermentation broth.
The yield of the antimicrobial peptide was calculated to be 0.923 g/(163 g) × 100% =0.566%.
The survival of Lactobacillus acidophilus in the fourth fermentation broth or concentrate after fire extinguishing in examples 1-5 and comparative examples 3 and 4 was examined and the results are shown in Table 1.
TABLE 1 survival of Lactobacillus acidophilus in fourth fermentation broth or concentrate after extinguishing fire in examples 1-5 and comparative example 3
Figure GDA0004066283970000121
As can be seen from Table 1, some of the Lactobacillus acidophilus bacteria were not completely inactivated after the inactivation of comparative example 3 without addition of gelatin, some of the Lactobacillus acidophilus bacteria were not completely inactivated after the inactivation of comparative example 4 without refrigeration, and all of the Lactobacillus acidophilus bacteria of examples 1 to 5 were inactivated.
The above-described embodiments of the present invention should not be construed as limiting the scope of the present invention. Any other corresponding changes and modifications made according to the technical idea of the present invention should be included in the protection scope of the claims of the present invention.

Claims (5)

1. A preparation method of anagen is characterized by comprising the following steps:
s1, inoculating lactobacillus acidophilus into a lactobacillus culture solution for culturing, culturing for 1-2 days at 35-38 ℃ to obtain a first fermentation solution, adding citric acid and hydrolyzed protein into the first fermentation solution, continuously culturing for 3-5 hours at 25-30 ℃ to obtain a second fermentation solution, and culturing the second fermentation solution at 35-38 ℃ for 1-2 days to obtain a third fermentation solution; the addition amount of the citric acid is 0.1-0.5% of the volume of the first fermentation liquid, and the concentration of the citric acid is 0.3-0.5 mol/L; the addition amount of the hydrolyzed protein is 0.1-0.5% of the mass of the first fermentation solution;
s2, filtering the third fermentation liquor to obtain a supernatant, and concentrating the supernatant to obtain a concentrated solution;
s3, adding gelatin into the concentrated solution, and then refrigerating at 5-10 ℃ to obtain a fourth fermentation liquid; the addition amount of the gelatin is 0.5-1% of the mass of the concentrated solution;
s4, inactivating the fourth fermentation liquor; the inactivation treatment temperature is 80-85 deg.C, and the time is 30-60min;
and S5, adding chitosan into the inactivated fourth fermentation broth, and then performing spray drying to obtain the anagen.
2. The method for preparing metazoan according to claim 1, wherein the chitosan is added in an amount of 1-3% by mass of the fourth fermentation broth after inactivation in step S5.
3. The method for producing metazoan according to claim 1, wherein the volume of the concentrated solution is 0.5 to 0.6 times the volume of the supernatant in step S2.
4. The method for preparing metazoan according to claim 1, wherein the time for cold storage in step S3 is 30-60min.
5. A metazoan produced by the production method according to any one of claims 1 to 4.
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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101612169A (en) * 2008-06-27 2009-12-30 生物源生物技术（深圳）有限公司 Lactobacillus micro-ecological preparation of deactivation and preparation method thereof
CN103211143A (en) * 2013-03-26 2013-07-24 胡文锋 Inactivated lactobacillus effervescent tablets and preparation method thereof
CN105274044A (en) * 2015-11-05 2016-01-27 胡文锋 Inactivated lactic acid bacterium cell and application thereof to food
CN111304273A (en) * 2020-03-17 2020-06-19 广州正明生物科技有限公司 Method for promoting rapid secretion of antibacterial peptide by lactic acid bacteria
CN112841409A (en) * 2021-01-19 2021-05-28 山东泰山生力源集团股份有限公司 Method for producing postnatal prebiotics by semi-immobilized probiotics moist heat quenching

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103820323A (en) * 2014-03-20 2014-05-28 青岛根源生物技术集团有限公司 Storage method for increasing survival rate of liquid lactobacillus
CN112088978A (en) * 2020-09-16 2020-12-18 黄河三角洲京博化工研究院有限公司 Lactic acid bacteria stabilizer and application method thereof

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101612169A (en) * 2008-06-27 2009-12-30 生物源生物技术（深圳）有限公司 Lactobacillus micro-ecological preparation of deactivation and preparation method thereof
CN103211143A (en) * 2013-03-26 2013-07-24 胡文锋 Inactivated lactobacillus effervescent tablets and preparation method thereof
CN105274044A (en) * 2015-11-05 2016-01-27 胡文锋 Inactivated lactic acid bacterium cell and application thereof to food
CN111304273A (en) * 2020-03-17 2020-06-19 广州正明生物科技有限公司 Method for promoting rapid secretion of antibacterial peptide by lactic acid bacteria
CN112841409A (en) * 2021-01-19 2021-05-28 山东泰山生力源集团股份有限公司 Method for producing postnatal prebiotics by semi-immobilized probiotics moist heat quenching

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
不同发酵条件对O-2菌株代谢物抑制几种常见病原菌活性的影响;方祥等;《广州食品工业科技》;20031230;第19卷;第27-29页 *
柠檬酸的乳酸菌发酵降解途径及其应用;袁星星等;《食品研究与开发》;20170520;第38卷(第10期);第204-208页 *
热灭活嗜酸乳杆菌及其多肽类抗菌物质特性研究;胡文锋等;《饲料博览》;19991025;第11卷(第10期);第7-8页 *

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