CN112369510A - Biological bacteriostatic composition and preparation method and application thereof - Google Patents
Biological bacteriostatic composition and preparation method and application thereof Download PDFInfo
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- CN112369510A CN112369510A CN202011230168.3A CN202011230168A CN112369510A CN 112369510 A CN112369510 A CN 112369510A CN 202011230168 A CN202011230168 A CN 202011230168A CN 112369510 A CN112369510 A CN 112369510A
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K20/00—Accessory food factors for animal feeding-stuffs
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- A23K20/163—Sugars; Polysaccharides
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/174—Vitamins
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
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Abstract
The invention belongs to the technical field of biology, and particularly relates to a biological antibacterial composition, and a preparation method and application thereof. The biological bacteriostatic composition provided by the invention comprises multiple functional components such as composite antibacterial peptide, soybean protein powder, maltose, type II collagen, vitamin C, glycerol and the like. The biological bacteriostatic composition provided by the invention has a certain digestion promoting function besides a good bacteriostatic function.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a biological antibacterial composition, and a preparation method and application thereof.
Background
In recent years, the breeding industry gradually becomes the dominant industry of agricultural strategies, the proportion of the breeding industry in developed countries reaches more than 85%, and the proportion of the breeding industry in agriculture is lower, so that the breeding industry has a larger space for promotion. By 2030 years, the ratio of the output value of the domestic aquaculture industry to the total agricultural output value is further improved and is expected to reach 53.5 percent; however, after 2030 years, the GDP in the breeding industry and the GDP in the planting industry do not change too much. This group of data tells us that the farming industry is still on a climbing road and that market space is constantly being released.
The pig is the main breeding animal in China, the pork is also the main meat food of residents in China, and the pork demand rises every year. At present, the factors influencing the development of the pig industry in China mainly include the immaturity of the pig raising technology and small scale investment. The immature pig raising technology is a difficult technical problem to overcome, and specific technology screening is carried out according to biological characteristics, varieties, types and the like of pigs in the pig raising process. In the prior art, the feed contains too many additives or bacteria in the feed in the breeding process of pigs, so that the immunity of the pigs is reduced, various diseases occur, and substances added in the feed stimulate too many substances to influence the digestion and absorption capacity of the pigs, and finally the growth and development of the pigs are influenced.
Based on this, chinese patent application CN108341882A discloses a preparation and application of a fused pig antibacterial peptide FPAP recombinant yeast preparation, which discloses that FPAP can promote the proliferation of lymphocytes, erythrocytes and leukocytes, promote the secretion of nonspecific antibodies and disease specific antibodies, and improve the immunocompetence and survival rate of animals, however, the antibacterial peptide is added with various amino acids and proteins, which easily affects the digestion of pigs and destroys their appetite, and the proteins themselves are the hotbeds of bacteria, which easily cause bacterial infection, and can also infect other diseases while improving the immunity of animals.
Meanwhile, Chinese patent CN108558998B discloses preparation and application of a recombinant yeast preparation co-expressed by pig interleukin 4/6 and fused pig antibacterial peptide, and the application discloses that FPAPIL46 can promote the proliferation of lymphocytes, white blood cells and red blood cells, inhibit the growth of pathogenic microorganisms and improve the immunocompetence and survival rate of animals, but the antibacterial peptide still influences the digestive ability of pigs to cause the appetite of the pigs to be reduced, and on the other hand, the immunocompetence of the pigs is reduced.
In conclusion, the acceptance additives in the prior art generally have the defects that the acceptance additives are easily infected by bacteria, the immunity of pigs is reduced, and the appetite of the pigs is influenced.
Disclosure of Invention
Aiming at the defects generally existing in the prior art, the invention creatively provides a biological bacteriostatic composition and a preparation method and application thereof. The biological bacteriostatic composition provided by the invention has a good bacteriostatic function, improves the immunity of a human body, has a certain digestion promoting function, and is beneficial to improving the appetite of animals.
In order to achieve the purpose, the invention adopts the technical scheme that:
a biological bacteriostatic composition comprises the following components in parts by weight: 10-20 parts of composite antibacterial peptide, 3-5 parts of soybean protein powder, 6-10 parts of maltose, 5-8 parts of II type collagen, 1-3 parts of vitamin C, 1-2 parts of glycerol and 40-50 parts of water.
Preferably, the biological bacteriostatic composition comprises the following components in parts by weight: 15 parts of composite antibacterial peptide, 4 parts of soybean protein powder, 8 parts of maltose, 7 parts of II type collagen, 2 parts of vitamin C, 1.5 parts of glycerol and 45 parts of water.
Preferably, the composite antibacterial peptide is prepared from cecropin, pig beta-defensin and nisin according to the weight ratio of 1-2: 8-10: 2 to 5.
Preferably, the composite antibacterial peptide is prepared from cecropin, pig beta-defensin and nisin according to the weight ratio of 1.5: 9: 4.
The invention also provides a preparation method of the biological bacteriostatic composition, which comprises the following steps:
s1, heating water to 40-50 ℃, adding maltose, vitamin C and glycerol, and mixing and stirring uniformly to obtain a mixture I;
s2, cooling the mixture I to 20-25 ℃, adding the composite antibacterial peptide, and mixing and stirring uniformly to prepare a mixture II;
s3, adding the soybean protein powder and the II-type collagen into the mixture II obtained in the step S2, and uniformly mixing and stirring to prepare a mixture III;
s4, freeze-drying the mixture III prepared in the step S3, and collecting dried powder to obtain the compound III.
Preferably, the mixing and stirring process in steps S1, S2 and S3 is to stir at a rotation speed of 100-200 rpm for 30-40 min.
Preferably, the specific operation process of freeze-drying in step S4 is as follows: and (3) placing the mixture III under a vacuum condition, and freezing at a temperature of between 15 ℃ below zero and 20 ℃ below zero for 20 to 30min to obtain the compound III.
The invention also provides application of the biological bacteriostatic composition in preparation of a feed additive.
Preferably, the addition ratio of the biological antibacterial composition to commercial feed is 6-8: 92-94.
Preferably, the addition ratio of the biological bacteriostatic composition to the commercial feed is 7: 93 of the composition.
In the invention, the composite antibacterial peptide is added, so that the antibacterial performance of the composition is effectively improved, the mixed bacteria pollution is prevented, and the immunity is improved, wherein in the added soybean protein powder, the protein content is more than 55 percent, the total fiber content is less than 4 percent, and the fat content is less than 1 percent, so that various nutrient elements can be supplemented, and the immunity of animals is further improved; type II collagen, maltose and vitamin C can be used for further supplementing different proteins and saccharides for animals.
In addition, the invention discovers in the actual screening process that the collagen II and the glycerol are added simultaneously according to a certain proportion, so that the immunity of animals is improved, the digestion and absorption functions of the animals can be improved to a certain extent when the collagen II and the glycerol are taken as feed additives and eaten by the animals, and the composite antibacterial peptide is added, so that the antibacterial property is improved, the digestion and absorption functions are also improved, and the diet of the animals is promoted.
Compared with the prior art, the biological bacteriostatic composition provided by the invention has the following advantages:
(1) the biological antibacterial composition provided by the invention can be used as a feed additive to supplement various nutrient substances for animals, balance the nutrition of the animals and improve the immunity of the animals;
(2) according to the biological antibacterial composition provided by the invention, antibacterial peptide consisting of cecropin, pig beta-defensin and nisin according to a certain weight ratio is added, so that the antibacterial performance of the feed is effectively improved, and the immunity of animals is further improved;
(3) the biological bacteriostatic composition provided by the invention can improve the digestion and absorption functions of animals, has few components in each formula, is common in all components, is simple in preparation process, effectively reduces the production cost, and is beneficial to further popularization and nutrition in the feed industry.
Detailed Description
The present invention is further explained with reference to the following specific examples, but it should be noted that the following examples are only illustrative of the present invention and should not be construed as limiting the present invention, and all technical solutions similar or equivalent to the present invention are within the scope of the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are commercially available products.
The cecropin can be purchased from hekavas bioengineering, ltd; the porcine beta-defensin is available from shanghai mengxi biotechnology limited; the nisin is available from Shanghai eosin industries GmbH under the product name nisin with the model number of 1414-45-5; the type II collagen is available from shanghai fusheng industries ltd; the soybean protein powder can be purchased from Nanjing Tongying Biotechnology Co., Ltd and the commercially available pig feed can be purchased from Jindingyi puffed feed Co., Ltd, Binzhou city.
Example 1A Biostatic composition
The biological bacteriostatic composition comprises the following components in parts by weight: 10 parts of composite antibacterial peptide, 3 parts of soybean protein powder, 6 parts of maltose, 5 parts of II type collagen, 1 parts of vitamin C, 1 part of glycerol and 40 parts of water; the composite antibacterial peptide is prepared from cecropin, pig beta-defensin and nisin according to the weight ratio of 1: 8: 2.
The preparation method of the biological bacteriostatic composition comprises the following steps:
s1, heating water to 40 ℃, adding maltose, vitamin C and glycerol, mixing and stirring for 30min at the rotating speed of 100rpm, and uniformly mixing to obtain a mixture I;
s2, cooling the mixture I to 20 ℃, adding the composite antibacterial peptide, mixing and stirring for 30min at the rotating speed of 100rpm, and uniformly mixing to prepare a mixture II;
s3, adding the soybean protein powder and the II-type collagen into the mixture II obtained in the step S2, mixing and stirring for 30min at the rotating speed of 100rpm, and uniformly mixing to prepare a mixture III;
s4, freeze-drying the mixture III prepared in the step S3, and collecting dried powder to obtain the compound III; the freeze drying process comprises the following steps: and (3) placing the mixture III under vacuum condition, and freezing at-15 ℃ for 20min to obtain the product.
Example 2A Biostatic composition
The biological bacteriostatic composition comprises the following components in parts by weight: 20 parts of composite antibacterial peptide, 5 parts of soybean protein powder, 10 parts of maltose, 8 parts of II type collagen, 3 parts of vitamin C, 2 parts of glycerol and 50 parts of water; the composite antibacterial peptide is prepared from cecropin, pig beta-defensin and nisin according to the weight ratio of 2: 10: 5.
The preparation method of the biological bacteriostatic composition comprises the following steps:
s1, heating water to 50 ℃, adding maltose, vitamin C and glycerol, mixing and stirring for 40min at the rotating speed of 200rpm, and uniformly mixing to obtain a mixture I;
s2, cooling the mixture I to 25 ℃, adding the composite antibacterial peptide, mixing and stirring at the rotating speed of 200rpm for 40min, and uniformly mixing to prepare a mixture II;
s3, adding the soybean protein powder and the II-type collagen into the mixture II obtained in the step S2, mixing and stirring for 40min at the rotating speed of 200rpm, and uniformly mixing to prepare a mixture III;
s4, freeze-drying the mixture III prepared in the step S3, and collecting dried powder to obtain the compound III; the specific operation process of the freeze drying is as follows: and (3) placing the mixture III under vacuum condition, and freezing at-20 ℃ for 30min to obtain the product.
Example 3A Biostatic composition
The biological bacteriostatic composition comprises the following components in parts by weight: 15 parts of composite antibacterial peptide, 4 parts of soybean protein powder, 8 parts of maltose, 7 parts of II type collagen, 2 parts of vitamin C, 1.5 parts of glycerol and 45 parts of water; the composite antibacterial peptide is prepared from cecropin, pig beta-defensin and nisin according to the weight ratio of 1.5: 9: 4.
The preparation method of the biological bacteriostatic composition comprises the following steps:
s1, heating water to 45 ℃, adding maltose, vitamin C and glycerol, mixing and stirring for 35min at the rotating speed of 150rpm, and uniformly mixing to obtain a mixture I;
s2, cooling the mixture I to 23 ℃, adding the composite antibacterial peptide, mixing and stirring at the rotating speed of 150rpm for 35min, and uniformly mixing to prepare a mixture II;
s3, adding the soybean protein powder and the II type collagen into the mixture II obtained in the step S2, mixing and stirring for 35min at the rotating speed of 150rpm, and uniformly mixing to prepare a mixture III;
s4, freeze-drying the mixture III prepared in the step S3, and collecting dried powder to obtain the compound III; the specific operation process of the freeze drying is as follows: and (3) freezing the mixture III at-18 ℃ for 25min under vacuum condition to obtain the product.
Comparative example 1A Biobacteriostatic composition
The bacteriostatic biological composition is similar to example 3;
the difference from the example 3 is that the composite antibacterial peptide in the comparative example 1 is prepared by mixing cecropin, porcine beta-defensin and nisin in a weight ratio of 1: 1: 1.
Comparative example 2A Biobacteriostatic composition
The bacteriostatic biological composition is similar to example 3;
the difference from the example 3 is that the composite antibacterial peptide in the comparative example 2 is prepared by mixing cecropin and porcine beta-defensin according to the weight ratio of 1.5: 9.
Comparative example 3A Biobacteriostatic composition
The bacteriostatic biological composition is similar to example 3;
the difference from example 3 is that the composite antibacterial peptide in comparative example 3 is prepared from porcine beta-defensin and nisin in a weight ratio of 9: 4.
Comparative example 4A Biobacteriostatic composition
The bacteriostatic biological composition is similar to example 3;
the difference from example 3 is that comparative example 4 contains no type II collagen.
Comparative example 5A Biobacteriostatic composition
The bacteriostatic biological composition is similar to example 3;
the difference from example 3 is that comparative example 5 does not contain soy protein powder.
Test example 1 bacteriostatic test
1. Test samples: the biological bacteriostatic compositions prepared in examples 1 to 3 and comparative examples 1 to 3;
2. test strains: bacillus subtilis, staphylococcus aureus, pseudomonas aeruginosa of copper green;
3. the test method comprises the following steps: respectively adding purified water to dilute the test samples into 20 microgram/mL, detecting the bacteriostatic performance of the test samples by adopting a paper sheet method, soaking round paper sheets in a sterile environment, respectively dipping the round paper sheets into the test samples, respectively sticking the round paper sheets onto agar plates fully coated with bacillus subtilis, staphylococcus aureus and copper green pseudomonas, and observing and recording the size of the bacteriostatic circle diameter after overnight culture (the stronger the bacteriostatic effect, the larger the bacteriostatic circle diameter).
4. And (3) test results: the specific test results are shown in table 1.
TABLE 1 comparison of the bacteriostatic effects of different test samples
As can be seen from Table 1, the bacteriostatic effect of the biological bacteriostatic composition prepared in the examples 1-3 of the invention is significantly higher than that of the groups in the comparative examples 1-3, which proves that the antibacterial effect of the composite antibacterial peptide is greatly improved.
Test example 2 digestive absorption function test
1. Test samples: the biological bacteriostatic compositions prepared in example 3 and comparative examples 1 to 5; expanded feed purchased from jindingyi expanded feed ltd of coastal city;
2. test animals: 4 months after birth, 90 pigs with the weight of about 80 plus or minus 5kg, 10 pigs in each group are averagely divided into 9 groups, and the female and male groups are half each, and the requirements are as follows: each pig is healthy and has no hereditary diseases;
3. the test method comprises the following steps: the biological bacteriostatic compositions prepared in example 3 and comparative examples 1-5 are added to the expanded feed as feed additives, and the experimental components are three groups: respectively mixing the biological bacteriostatic composition of the group 3 in the example with commercial feed according to the weight ratio of 6: 94 (test group a), 8: 92 (test group B), 7: 93 (test group C), and comparing examples 1-5, respectively mixing the prepared biological antibacterial composition with commercial feed according to the weight ratio of 7: 93 (runs D-H, respectively); using commercially available puffed feed alone for a control group, feeding 110kg of test samples for each group per day, updating the feed and freely drinking water every day for 10 consecutive days, measuring the weight and the food intake for 2 times per day, and calculating the average weight, the weight gain, the total food intake and the food utilization rate of each group at the end of the test, wherein the average values are used as the final weight; daily gain and daily food intake were averaged over 10 animals per group.
4. And (3) test results: the specific test results are shown in Table 2.
TABLE 2 comparison of the average weight gain results for different test samples
As can be seen from Table 2, the daily gain of the experimental group to which the feed prepared in example 3 of the present invention was added was significantly increased and the average daily food intake was not greatly different from that of the control group, indicating that the pigs fed with the feed of the example of the present invention had a higher digestibility, particularly in test C group, and the daily gain of 942.16g, which is the best example of the present invention, whereas the groups of comparative examples 1 to 5 had a significantly decreased digestibility due to the deterioration of the mutual synergy between the components.
Test example 3 Immunometric assay
1. Test samples: the biological bacteriostatic compositions prepared in example 3 and comparative examples 1 to 5; expanded feed purchased from jindingyi expanded feed ltd of coastal city; alpha-hemolytic streptococcus, beta-hemolytic streptococcus available from Shanghai-Yizhao Biotechnology Ltd;
2. test animals: 4 months after birth, 90 pigs with the weight of about 80 plus or minus 5kg, 10 pigs in each group are averagely divided into 9 groups, and the female and male groups are half each, and the requirements are as follows: each pig is healthy and has no hereditary diseases;
3. the test method comprises the following steps: the biological bacteriostatic compositions prepared in example 3 and comparative examples 1-5 are added to the expanded feed as feed additives, and the experimental components are three groups: respectively mixing the biological bacteriostatic composition of the group 3 in the example with commercial feed according to the weight ratio of 6: 94 (test group a), 8: 92 (test group B), 7: 93 (test group C), and comparing examples 1-5, respectively mixing the prepared biological antibacterial composition with commercial feed according to the weight ratio of 7: 93 (runs D-H, respectively); mixing alpha-hemolytic streptococcus and beta-hemolytic streptococcus according to the weight ratio of 1: 1 mixing to obtain mixed bacterial powder.
The experiment group is divided into an experiment group and a control group, the experiment group is fed with the feed added with the experiment groups A to H every day, and enough feed is fed every day, so that the experiment group can drink water and eat food freely; meanwhile, each group of pigs were fed 200mg of the mixed bacterial powder twice a day for 10 consecutive days (all eaten). After continuous 3 months, the immunopotentiation effect on swine was observed. The immune enhancement index is determined by measuring leukocytes in blood collected from pigs before and after administration of the "mixed bacterial powder"; the average value of each group was taken as the final value.
White blood cell change value-final white blood cell number-initial white blood cell number.
4. And (3) test results: the specific test results are shown in Table 3.
TABLE 3 comparison of the number of leukocytes in pigs fed with different test samples
As can be seen from table 3, the experimental groups fed with the feed containing the bacteriostatic composition prepared in example 3 of the present invention all had higher white blood cell number reduction values, indicating that the resistance was strong, especially the resistance was strongest in the experimental group C, which is the best example of the present invention, while the resistance of the pigs fed with the feed of the comparative examples 1 to 5 was reduced to different degrees, which laterally reflects the synergistic effect among the components of the present invention.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting the protection scope of the present invention, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.
Claims (10)
1. A biological bacteriostatic composition is characterized by comprising the following components in parts by weight: 10-20 parts of composite antibacterial peptide, 3-5 parts of soybean protein powder, 6-10 parts of maltose, 5-8 parts of II type collagen, 1-3 parts of vitamin C, 1-2 parts of glycerol and 40-50 parts of water.
2. The biological bacteriostatic composition as claimed in claim 1, which comprises the following components in parts by weight: 15 parts of composite antibacterial peptide, 4 parts of soybean protein powder, 8 parts of maltose, 7 parts of II type collagen, 2 parts of vitamin C, 1.5 parts of glycerol and 45 parts of water.
3. The biological bacteriostatic composition according to claim 1 or 2, wherein the composite antibacterial peptide is prepared from cecropin, porcine beta-defensin and nisin in a weight ratio of 1-2: 8-10: 2 to 5.
4. The biological bacteriostatic composition according to claim 3, wherein the complex antibacterial peptide is prepared from cecropin, porcine beta-defensin and nisin in a weight ratio of 1.5: 9: 4.
5. A method of preparing a bacteriostatic composition according to any one of claims 1 to 4, comprising the steps of:
s1, heating water to 40-50 ℃, adding maltose, vitamin C and glycerol, and mixing and stirring uniformly to obtain a mixture I;
s2, cooling the mixture I to 20-25 ℃, adding the composite antibacterial peptide, and mixing and stirring uniformly to prepare a mixture II;
s3, adding the soybean protein powder and the II-type collagen into the mixture II obtained in the step S2, and uniformly mixing and stirring to prepare a mixture III;
s4, freeze-drying the mixture III prepared in the step S3, and collecting dried powder to obtain the compound III.
6. The method for preparing a bacteriostatic composition according to claim 5, wherein the mixing and stirring process in steps S1, S2 and S3 is to stir at 100-200 rpm for 30-40 min.
7. The method for preparing a biological bacteriostatic composition according to claim 5, wherein the specific operation process of the freeze drying in step S4 is as follows: and (3) placing the mixture III under a vacuum condition, and freezing at a temperature of between 15 ℃ below zero and 20 ℃ below zero for 20 to 30min to obtain the compound III.
8. Use of a bacteriostatic biological composition according to claim 1 in the preparation of a feed additive.
9. The application of claim 8, wherein the addition ratio of the biological bacteriostatic composition to a commercial feed is 6-8: 92-94.
10. The use of claim 9, wherein the ratio of the bacteriostatic biological composition to the commercial feed is 7: 93 of the composition.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103734550A (en) * | 2014-01-29 | 2014-04-23 | 北京劲能生物科技股份公司 | Nursing pig concentrated feed |
| CN105558353A (en) * | 2016-02-04 | 2016-05-11 | 王植 | Feed additive and preparation method thereof |
| CN109511783A (en) * | 2018-12-29 | 2019-03-26 | 上海邦成生物工程有限公司 | A kind of bacillus subtilis feed addictive and preparation method thereof of the peptide of antibacterial protein containing feature |
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- 2020-11-06 CN CN202011230168.3A patent/CN112369510A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103734550A (en) * | 2014-01-29 | 2014-04-23 | 北京劲能生物科技股份公司 | Nursing pig concentrated feed |
| CN105558353A (en) * | 2016-02-04 | 2016-05-11 | 王植 | Feed additive and preparation method thereof |
| CN109511783A (en) * | 2018-12-29 | 2019-03-26 | 上海邦成生物工程有限公司 | A kind of bacillus subtilis feed addictive and preparation method thereof of the peptide of antibacterial protein containing feature |
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Application publication date: 20210219 |