Culture medium for promoting growth of bifidobacterium and application

Abstract

The invention discloses a culture medium for promoting growth of bifidobacteria and application of the culture medium. 1L of the culture medium is prepared from the following components: 6.2 to 7.4 g of # imgabs0 # peptone, 1.5 to 2.5 g of soy peptone, 3.0 to 3.8 g of yeast extract powder, 1.0 to 2g of beef extract powder, 8.5 to 9.5 g of digestive serum powder, 0.5 to 1.5 g of bovine liver extract powder, 1.5 to 2.0 g of monopotassium phosphate, 1.5 to 2.5 g of sodium chloride, 0.1 to 0.3 g of L-cysteine, 0.1 to 0.3 g of sodium thioglycolate, 0.05 to 0.12 g of growth promoting factors and the balance of water. The growth promoting factor is periplocin, syringaldehyde, 4-methoxysalicylaldehyde, astragaloside, calycosin or formononetin. The raw materials of the culture medium are easy to obtain, the growth promoting effect on bifidobacterium is good, and time and cost are saved. Bifidobacterium grows fast in the culture medium provided by the invention.

Description

Technical Field

[0001] The invention relates to the field of microorganisms, and in particular to a culture medium for promoting the growth of bifidobacteria and an application thereof. Background Art

[0002] Intestinal flora plays an important role in human health. The balance of intestinal flora is closely related to the digestion, metabolism, nervous and immune systems of the human body. Bifidobacterium is one of the most important probiotics in the intestine, which has many functions such as promoting digestion, enhancing immunity and preventing intestinal diseases. [1] In addition, in recent years, the research on bifidobacteria has become increasingly in-depth, and bifidobacteria are also present in cardiovascular and cerebrovascular diseases. [2] This suggests that as a relatively dominant probiotic in the human intestine, the physiological functions and disease-healing potential of bifidobacteria are still to be developed, which requires modern bacterial culture technology to be able to culture bifidobacteria quickly and in large quantities.

[0003] Bifidobacterium is a type of non-motile, strictly anaerobic Gram-positive bacteria. Its optimal growth temperature is 37-41°C and its optimal growth pH is 6.7 to 7.0. It will not grow in an acidic or alkaline environment. [3] Currently, in taxonomy, the genus Bifidobacterium includes 34 species, 12 of which are derived from humans, but only Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium bifidum and Bifidobacterium adolescentis can colonize the human intestine and have been approved for use in food processing. [4] .

[0004] At present, the bifidobacterium culture media commonly used in laboratories are mainly "TPY medium" and "Bifidobacterium medium". In the process of bifidobacterium cultivation, the culture medium determines the growth rate of bacteria. However, bifidobacteria grow relatively slowly in existing culture media. It takes a long time to cultivate bifidobacteria to achieve the maximum growth. [5] Moreover, due to their high salt content, both will produce precipitation after sterilization, affecting bacterial culture and observation, which obviously cannot meet the needs of modern experiments and production.

[0005] Existing finished culture media are expensive, have high salt content, are unstable after sterilization and may produce precipitation. The growth rate of bifidobacteria in them is slow, and it takes a long time (4-20h) to reach the logarithmic growth phase. In addition, the maximum number of viable bacteria that different bifidobacteria can reach in the finished culture media in the same time is small, and the OD 600 Not higher than 0.3 [5] , can no longer meet the needs of modern multi-omics research.

[0006] With the development of science and technology and the deepening of research, the connection between traditional Chinese medicine and intestinal probiotics has become closer and closer. Common traditional Chinese medicine and its ingredients are gradually becoming one of the important sources of prebiotics that promote the growth of probiotics. Therefore, there is an urgent need for a culture medium that effectively promotes the growth of bifidobacteria.

[0007] References:

[0008] [1]SJGavzy,A.Kensiski,ZLLee,et al.Bifidobacterium mechanisms ofimmune modulation and tolerance.Gut Microbes,2023,15(2):2291164.

[0009] [2] PMBock, AF Martins, BDSchaan. Understanding how pre-andprobiotics affect the gut microbiome and metabolic health. Am J PhysiolEndocrinol Metab, 2024, 327(1):E89-E102.

[0010] [3] Diao Zhimin, Zhang Wenjing, Ma Yonggui. Physiological effects and development prospects of Bifidobacterium. Qinghai Science and Technology, 2002, 6: 37-40.

[0011] [4] LAKSaleena, SKChang, K.Simarani, et al. A comprehensive review of Bifidobacterium spp: as a probiotic, application in the food and therapeutic, and forthcoming trends. Crit Rev Microbiol, 2024, 50(5): 581-597.

[0012] [5] Du Peng, Li Aili, Huo Guicheng. Optimization of Bifidobacterium culture medium. Industrial Microbiology, 2005, 3:14-18. Summary of the invention

[0013] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a culture medium for promoting the growth of bifidobacteria.

[0014] The second object of the present invention is to provide an application of a culture medium for promoting the growth of bifidobacteria.

[0015] The technical solution of the present invention is summarized as follows:

[0016] A culture medium for promoting the growth of bifidobacteria, wherein 1 L of the culture medium is prepared with the following components:

[0017] Peptone 6.2-7.4g, soy peptone 1.5-2.5g, yeast extract powder 3.0-3.8g, beef extract powder 1.0-2g, digested serum powder 8.5-9.5g, beef liver extract powder 0.5-1.5g, potassium dihydrogen phosphate 1.5-2.0g, sodium chloride 1.5-2.5g, L-cysteine ​​0.1-0.3g, sodium thioglycolate 0.1-0.3g, growth-promoting factor 0.05-0.12g, and the balance is water;

[0018] The growth-promoting factors are periplocoside, syringaldehyde, 4-methoxysalicylaldehyde, astragaloside IV, calycosin or formononetin.

[0019] Preferably, 1L of the culture medium is made with the following components: 6.8g peptone, 2.0g soy peptone, 3.4g yeast extract powder, 1.5g beef extract powder, 9.1g digested serum powder, 0.8g beef liver extract powder, 1.7g potassium dihydrogen phosphate, 2.0g sodium chloride, 0.2g L-cysteine, 0.2g sodium thioglycolate, 0.08g growth-promoting factor, and the balance is water.

[0020] Preferably, the bifidobacterium is Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium adolescentis, Bifidobacterium infantis or Bifidobacterium bifidum.

[0021] Application of the culture medium in promoting the growth of bifidobacteria.

[0022] Advantages of the present invention:

[0023] The culture medium raw materials of the present invention are easy to obtain, and the culture medium of the present invention has a good growth promotion effect on bifidobacteria, and the growth promotion efficiency of different species of bifidobacteria reaches 1.5 times and more than that of TPY culture medium, and reaches the logarithmic growth phase about 4 hours in advance, saving time cost. The growth rate of bifidobacteria in the culture medium of the present invention is fast, and the number of bifidobacteria more than 1.5-2.5 times that of TPY culture medium can be obtained in a relatively short time; more live bacteria can also be obtained in enterprise-scale production, increasing factory efficiency and increasing production. The culture medium preparation method of the present invention is simple, low in inorganic salt content, not easy to produce precipitation, low in cost, and high in the growth rate of different bifidobacteria therein. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 The effect of 4% dimethyl sulfoxide on the growth of five species of Bifidobacterium;

[0025] Figure 2 The effect of adding different growth-promoting factors to the growth-promoting medium (Example 2) on the growth of Bifidobacterium breve;

[0026] Figure 3 The effect of adding different growth-promoting factors to the growth-promoting medium (Example 2) on the growth of Bifidobacterium longum;

[0027] Figure 4 The effect of adding different growth-promoting factors to the growth-promoting medium (Example 2) on the growth of Bifidobacterium adolescentis;

[0028] Figure 5 The effect of adding different growth-promoting factors to the growth-promoting medium (Example 2) on the growth of Bifidobacterium infantis;

[0029] Figure 6 The effect of adding different growth-promoting factors to the growth-promoting medium (Example 2) on the growth of Bifidobacterium bifidum;

[0030] Figure 7 The effect of adding different growth-promoting factors to the growth-promoting medium (Example 3) on the growth of Bifidobacterium breve;

[0031] Figure 8 The effect of adding different growth-promoting factors to the growth-promoting medium (Example 4) on the growth of Bifidobacterium breve. DETAILED DESCRIPTION

[0032] The present invention will be further described below in conjunction with specific embodiments.

[0033] The bifidobacteria used in the following embodiments are five common bifidobacteria on the market, which were purchased from Beina Bio (Suzhou Beina Chuanglian Biotechnology Co., Ltd.) in January 2024.

[0034] Bifidobacterium breve / BNCC186529

[0035] Bifidobacterium longum / BNCC185354

[0036] Bifidobacterium adolescentis / BNCC185973

[0037] Bifidobacterium longum-infantis / BNCC356005

[0038] Bifidobacterium bifidum / BNCC186304

[0039] The above introduction of the sources of bifidobacteria is for a better understanding of the present invention, but does not limit the present invention.

[0040] The culture medium prepared from other components excluding growth-promoting factors in step (1) of Examples 2, 3, and 4 is referred to as the basal culture medium.

[0041] The culture medium for promoting the growth of bifidobacteria of the present invention is referred to as growth-promoting culture medium for short.

[0042] Peptone was purchased from Qingdao Hibo Biotechnology Co., Ltd., HB8277.

[0043] Example 1

[0044] Experiment on the effect of 4% dimethyl sulfoxide-water solution on the growth of Bifidobacterium.

[0045] (1) Weigh: Peptone 6.8g, soy peptone 2.0g, yeast extract powder 3.4g, beef extract powder 1.5g, digested serum powder 9.1g, beef liver extract powder 0.8g, potassium dihydrogen phosphate 1.7g, sodium chloride 2.0g, L-cysteine ​​0.2g, sodium thioglycolate 0.2g;

[0046] (2) Add water to 1 L of the components weighed in step (1), mix well, boil until dissolved, sterilize at 121° C. with high pressure wet heat for 15 minutes, and cool to room temperature to obtain a basal culture medium;

[0047] (3) Inoculate 10 mL of basal medium with 1% (v / v) of Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium adolescentis, Bifidobacterium infantis and Bifidobacterium bifidum, respectively; obtain:

[0048] Bifidobacterium breve + basic medium;

[0049] Bifidobacterium longum + basic medium;

[0050] Bifidobacterium adolescentis + basic culture medium;

[0051] Bifidobacterium infantis + basic culture medium;

[0052] Bifidobacterium bifidum + basic culture medium;

[0053] (4) Inoculate 10 mL of basal medium with 1% (v / v) of Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium adolescentis, Bifidobacterium infantis and Bifidobacterium bifidum, respectively; then add dimethyl sulfoxide to a final concentration of 4% (referred to as 4% dimethyl sulfoxide, the same below), and obtain:

[0054] Bifidobacterium breve + 4% dimethyl sulfoxide + basic medium;

[0055] Bifidobacterium longum + 4% dimethyl sulfoxide + basic medium;

[0056] Bifidobacterium adolescentis + 4% dimethyl sulfoxide + basal medium;

[0057] Bifidobacterium infantis + 4% dimethyl sulfoxide + basic medium;

[0058] Bifidobacterium bifidum + 4% dimethyl sulfoxide + basic medium;

[0059] The growth curves of the above groups were measured within 72 hours. The results were as follows: Figure 1 As shown, medium containing DMSO at a final concentration of 4% had almost no effect on the growth of different Bifidobacteria.

[0060] Example 2

[0061] A medium for promoting the growth of bifidobacteria (growth-promoting medium), prepared by the following method:

[0062] (1) Prepare a growth-promoting medium with a final volume of 1 L and weigh: Peptone 6.8g, soy peptone 2.0g, yeast extract powder 3.4g, beef extract powder 1.5g, digested serum powder 9.1g, beef liver extract powder 0.8g, potassium dihydrogen phosphate 1.7g, sodium chloride 2.0g, L-cysteine ​​0.2g, sodium thioglycolate 0.2g, growth-promoting factors (growth-promoting factors are periplocoside, syringaldehyde, 4-methoxysalicylaldehyde, astragaloside IV, calycosin or formononetin) 0.08g;

[0063] (2) Add water to 990 mL of the components except the growth-promoting factors in step (1), mix well, boil until dissolved, and package while hot into 10 bottles, each bottle containing 99 mL. Sterilize at 121° C. with high pressure wet heat for 15 minutes, cool to room temperature, and take 6 bottles and number them;

[0064] Add 1 mL of 8 mg / mL periplocoside solution (solvent: dimethyl sulfoxide) sterilized by membrane filtration to the first bottle, mix well, and obtain the 1-1 culture medium.

[0065] Add 1 mL of 8 mg / mL syringaldehyde solution (solvent: dimethyl sulfoxide) sterilized by membrane filtration to the second bottle, mix well, and obtain the 1-2 culture medium.

[0066] To the third bottle, add 1 mL of 8 mg / mL 4-methoxysalicylaldehyde solution (solvent: dimethyl sulfoxide) sterilized by membrane filtration, mix well, and obtain the 1-3 culture medium.

[0067] To the fourth bottle, add 1 mL of 8 mg / mL astragaloside IV solution (solvent: dimethyl sulfoxide) sterilized by membrane filtration, mix well, and obtain culture medium No. 1-4.

[0068] Add 1 mL of 8 mg / mL calycosin solution (solvent: dimethyl sulfoxide) sterilized by membrane filtration to the fifth bottle, mix well, and obtain medium No. 1-5.

[0069] To the sixth bottle, add 1 mL of 8 mg / mL formononetin solution (solvent: dimethyl sulfoxide) sterilized by membrane filtration, mix well, and obtain the 1-6 culture medium.

[0070] TPY medium: Weigh 26.4 g of finished TPY medium (Qingdao Haibo Biological HB8570-1), dissolve it in 1000 mL of distilled water by heating, sterilize it by high pressure at 121°C for 15 min, and return it to room temperature to obtain TPY medium.

[0071] The first negative control group is prepared by uniformly mixing the 1-1 culture medium, the 1-2 culture medium, the 1-3 culture medium, the 1-4 culture medium, the 1-5 culture medium, the 1-6 culture medium and the TPY culture medium in equal volumes to obtain the first negative control.

[0072] Example 3

[0073] A medium for promoting the growth of bifidobacteria (growth-promoting medium), prepared by the following method:

[0074] (1) Prepare a growth-promoting medium with a final volume of 1 L and weigh: Peptone 6.2g, soy peptone 2.5g, yeast extract powder 3.0g, beef extract powder 2g, digested serum powder 8.5g, beef liver extract powder 1.5g, potassium dihydrogen phosphate 1.5g, sodium chloride 2.5g, L-cysteine ​​0.1g, sodium thioglycolate 0.3g, growth-promoting factors (growth-promoting factors are periplocoside, syringaldehyde, 4-methoxysalicylaldehyde, astragaloside IV, calycosin or formononetin) 0.05g;

[0075] (2) Add water to 990 mL of the components except the growth-promoting factors in step (1), mix well, boil until dissolved, and package while hot into 10 bottles, each bottle containing 99 mL. Sterilize at 121°C for 15 min, cool to room temperature, and take 6 bottles and number them.

[0076] Add 1 mL of 5 mg / mL periplocoside solution (solvent: dimethyl sulfoxide) sterilized by membrane filtration to the first bottle, mix well, and obtain the 2-1 culture medium.

[0077] Add 1 mL of 5 mg / mL syringaldehyde solution (solvent: dimethyl sulfoxide) sterilized by membrane filtration to the second bottle, mix well, and obtain the 2-2 culture medium.

[0078] To the third bottle, add 1 mL of 5 mg / mL 4-methoxysalicylaldehyde solution (solvent: dimethyl sulfoxide) sterilized by membrane filtration, mix well, and obtain the 2-3 culture medium.

[0079] Add 1 mL of 5 mg / mL astragaloside IV solution (solvent: dimethyl sulfoxide) sterilized by membrane filtration to the fourth bottle, mix well, and obtain the second-fourth culture medium.

[0080] Add 1 mL of 5 mg / mL calycosin solution (solvent: dimethyl sulfoxide) sterilized by membrane filtration into the fifth bottle, mix well, and obtain the second-5 culture medium.

[0081] To the sixth bottle, add 1 mL of a 5 mg / mL formononetin solution (solvent: dimethyl sulfoxide) sterilized by membrane filtration, mix well, and obtain the 2-6 culture medium.

[0082] TPY medium, same as the TPY medium in Example 2.

[0083] The second negative control group is prepared by uniformly mixing the 2-1 culture medium, the 2-2 culture medium, the 2-3 culture medium, the 2-4 culture medium, the 2-5 culture medium, the 2-6 culture medium and the TPY culture medium in equal volumes to obtain the second negative control.

[0084] Example 4

[0085] A medium for promoting the growth of bifidobacteria (growth-promoting medium), prepared by the following method:

[0086] (1) Prepare a growth-promoting medium with a final volume of 1 L and weigh: Peptone 7.4g, soy peptone 1.5g, yeast extract powder 3.8g, beef extract powder 1g, digested serum powder 9.5g, beef liver extract powder 0.5g, potassium dihydrogen phosphate 2.0g, sodium chloride 1.5g, L-cysteine ​​0.3g, sodium thioglycolate 0.1g, growth-promoting factors (growth-promoting factors are periplocoside, syringaldehyde, 4-methoxysalicylaldehyde, astragaloside IV, calycosin or formononetin) 0.12g;

[0087] (2) Add water to 990 mL of the components except the growth-promoting factors in step (1), mix well, boil until dissolved, and package while hot into 10 bottles, each bottle containing 99 mL. Sterilize at 121°C for 15 min, cool to room temperature, and take 6 bottles and number them.

[0088] Add 1 mL of 12 mg / mL periplocoside solution (solvent: dimethyl sulfoxide) sterilized by membrane filtration to the first bottle, mix well, and obtain the 3-1 culture medium.

[0089] Add 1 mL of 12 mg / mL syringaldehyde solution (solvent: dimethyl sulfoxide) sterilized by membrane filtration to the second bottle, mix well, and obtain the 3-2 culture medium.

[0090] To the third bottle, add 1 mL of 12 mg / mL 4-methoxysalicylaldehyde solution (solvent: dimethyl sulfoxide) sterilized by membrane filtration, mix well, and obtain the 3-3 culture medium.

[0091] To the fourth bottle, add 1 mL of 12 mg / mL astragaloside IV solution (solvent: dimethyl sulfoxide) sterilized by membrane filtration, mix well, and obtain the 3-4 culture medium.

[0092] To the fifth bottle, add 1 mL of a 12 mg / mL solution of calycosin (solvent: dimethyl sulfoxide) sterilized by filtration through a filter membrane, mix well, and obtain the 3-5 culture medium.

[0093] To the sixth bottle, add 1 mL of a 12 mg / mL formononetin solution (solvent: dimethyl sulfoxide) sterilized by membrane filtration, mix well, and obtain the 3-6 culture medium.

[0094] TPY medium, same as the TPY medium in Example 2.

[0095] The third negative control group is prepared by uniformly mixing the 3-1 medium, the 3-2 medium, the 3-3 medium, the 3-4 medium, the 3-5 medium, the 3-6 medium and the TPY medium in equal volumes to obtain the third negative control.

[0096] Example 5

[0097] Application of a culture medium for promoting the growth of bifidobacteria in promoting the growth of bifidobacteria

[0098] The six growth-promoting culture media (medium 1-1 to medium 1-6) prepared in Example 2 were inoculated with 1% Bifidobacterium breve by volume, and the samples were repeated for three times, and cultured under the gas conditions of O2 2%, CO2 4%, and relative humidity 65%. OD was continuously measured. 600 The value is 72h, and a growth curve is drawn. The results are as follows Figure 2 shown.

[0099] The results showed that medium 1-2 significantly promoted the growth of Bifidobacterium breve (P<0.05), with the maximum OD 600 was 0.80, which was the TPY medium group (OD 600The TPY medium group reached the logarithmic growth phase within 4-14 hours, and the 1-2 medium group reached the logarithmic growth phase within 4-8 hours, significantly promoting the growth rate of Bifidobacterium breve (P<0.05) and shortening the experimental time. In addition, the 1-1, 1-3, 1-4, 1-5, and 1-6 mediums all had a growth-promoting effect on Bifidobacterium breve.

[0100] Example 6

[0101] Application of a culture medium for promoting the growth of bifidobacteria in promoting the growth of bifidobacteria

[0102] The six growth-promoting culture media (medium 1-1 to medium 1-6) prepared in Example 2 were inoculated with Bifidobacterium longum at a volume ratio of 1%, and the samples were repeated for three times, and cultured in an environment with a gas condition of O2 2%, CO2 4%, and a relative humidity of 65%. OD was continuously measured. 600 The value is 72h, and a growth curve is drawn. The results are as follows Figure 3 shown.

[0103] The results showed that the 1st to 6th culture medium groups significantly promoted the growth of Bifidobacterium longum (P<0.05), reaching the maximum OD at 20h. 600 value (0.36), which was 1.80 times that of the TPY medium group under the same conditions, reaching the maximum OD 600 The time to reach the maximum OD value was shortened by 24 hours, and the logarithmic growth phase was reached in 4-8 hours, which shortened the logarithmic growth period by 4 hours, saving experimental time, and the maximum OD was reached within 12 hours after reaching the logarithmic growth phase. 600 The value was 48h earlier than that of the TPY medium group. In addition, the 1-1, 1-2, 1-3, 1-4, and 1-5 medium groups all promoted the growth of Bifidobacterium longum.

[0104] Example 7

[0105] Application of a culture medium for promoting the growth of bifidobacteria in promoting the growth of bifidobacteria

[0106] Six growth-promoting culture media (medium 1-1 to medium 1-6) prepared in Example 2 were inoculated with Bifidobacterium adolescentis at a volume ratio of 1%, and three samples were repeated. The culture was placed in an environment with a gas condition of O2 2%, CO2 4%, and a relative humidity of 65%. OD was continuously measured. 600 The value is 72h, and a growth curve is drawn. The results are as follows Figure 4 shown.

[0107] The results showed that the 1st to 6th culture medium groups significantly promoted the growth of Bifidobacterium adolescentis (P<0.05), with the maximum OD 600The maximum OD value of the 1st to 5th culture medium groups was 0.32, which was 1.6 times that of the TPY culture medium group. The logarithmic growth phase was reached within 4-8 hours, which was 4 hours earlier than that of the TPY culture medium group. 600 The value can reach 0.30, which is 1.5 times that of the TPY medium group. The logarithmic growth phase is reached within 4-8 hours, which is 4 hours shorter than the TPY medium group, saving experimental time. In addition, the 1-1, 1-2, 1-3, and 1-4 medium groups all have a promoting effect on the growth of Bifidobacterium adolescentis.

[0108] Example 8

[0109] Application of a culture medium for promoting the growth of bifidobacteria in promoting the growth of bifidobacteria

[0110] The six growth-promoting culture media (medium 1-1 to medium 1-6) prepared in Example 2 were inoculated with 1% of infantis Bifidobacterium by volume, and the samples were repeated for three times, and cultured under the gas conditions of O2 2%, CO2 4%, and relative humidity 65%. OD was continuously measured. 600 The value is 72h, and a growth curve is drawn. The results are as follows Figure 5 shown.

[0111] The results showed that the 1st to 5th culture medium groups significantly promoted the growth of Bifidobacterium infantis (P<0.05), with the maximum OD 600 The value can reach 0.35, which is 2.19 times that of the TPY medium group. The logarithmic growth period is reached within 0-8h, which is 8h shorter than the TPY medium group, saving experimental time. In addition, the 1-1, 1-2, 1-3, 1-4, and 1-6 medium groups all have a growth-promoting effect on infant Bifidobacterium.

[0112] Example 9

[0113] Application of a culture medium for promoting the growth of bifidobacteria in promoting the growth of bifidobacteria

[0114] The six growth-promoting culture media (medium 1-1 to medium 1-6) prepared in Example 2 were inoculated with 1% of Bifidobacterium bifidum by volume, and the samples were repeated for three times, and the culture was placed in an environment with a gas condition of O2 2%, CO2 4%, and a relative humidity of 65%. OD was continuously measured. 600 The value is 72h, and a growth curve is drawn. The results are as follows Figure 6 shown.

[0115] The results showed that the 1st to 6th culture medium groups significantly promoted the growth of Bifidobacterium bifidum (P<0.05), with the maximum OD 600The value can reach 0.33, which is 1.65 times that of the TPY group. The logarithmic growth phase is reached within 4-8 hours, which is 4 hours shorter than the TPY culture medium group, saving experimental time. Moreover, the 1-1, 1-2, 1-3, 1-4, and 1-5 culture medium groups all have different degrees of promoting effects on Bifidobacterium bifidum.

[0116] Example 10

[0117] Application of a culture medium for promoting the growth of bifidobacteria in promoting the growth of bifidobacteria

[0118] The six growth-promoting culture media (medium 2-1 to medium 2-6) prepared in Example 3 were inoculated with 1% Bifidobacterium breve by volume, and the samples were repeated for three times, and the culture was placed in an environment with a gas condition of O2 2%, CO2 4%, and a relative humidity of 65%. OD was continuously measured. 600 The value is 72h, and a growth curve is drawn. The results are as follows Figure 7 shown.

[0119] The results showed that the 2-2 medium group significantly promoted the growth of Bifidobacterium breve (P<0.05), with the maximum OD 600 The value can reach 0.63, which is 1.97 times that of the TPY medium group. The logarithmic growth phase is reached within 4-8 hours, which is 4 hours shorter than the TPY medium group, saving experimental time. The 2-1, 2-3, 2-4, 2-5, and 2-6 medium groups all have different degrees of promoting effects on Bifidobacterium breve, and the effects are better than those of the TPY medium group.

[0120] Experiments have shown that when Bifidobacterium longum, Bifidobacterium adolescentis, Bifidobacterium infantis or Bifidobacterium bifidum are used to replace Bifidobacterium breve of this embodiment, the effects are better than those of the TPY medium group, while the other aspects are the same as those of this embodiment.

[0121] Embodiment 11

[0122] Application of a culture medium for promoting the growth of bifidobacteria in promoting the growth of bifidobacteria

[0123] The six growth-promoting culture media (medium 3-1 to medium 3-6) prepared in Example 4 were inoculated with 1% Bifidobacterium breve by volume, and the samples were repeated for three times, and cultured in an environment with a gas condition of 2% O2, 4% CO2, and a relative humidity of 65%. OD was continuously measured. 600 The value is 72h, and a growth curve is drawn. The results are as follows Figure 8 shown.

[0124] The results showed that the 3-2 medium group significantly promoted the growth of Bifidobacterium breve (P<0.05), with the maximum OD 600The value can reach 0.62, which is 1.94 times that of the TPY medium group. The logarithmic growth phase is reached within 4-10 hours, which is 4 hours shorter than the TPY medium group, saving experimental time. The 3-1, 3-3, 3-4, 3-5, and 3-6 medium groups all have different degrees of promotion effects on Bifidobacterium breve, and the effects are better than those of the TPY medium group.

[0125] Experiments have shown that when Bifidobacterium longum, Bifidobacterium adolescentis, Bifidobacterium infantis or Bifidobacterium bifidum are used to replace Bifidobacterium breve of this embodiment, the effects are better than those of the TPY medium group, while the other aspects are the same as those of this embodiment.

Claims

1. A culture medium for promoting the growth of bifidobacteria, characterized in that 1L of the culture medium is prepared with the following components: Peptone 6.2-7.4g, soy peptone 1.5-2.5g, yeast extract powder 3.0-3.8g, beef extract powder 1-2g, digested serum powder 8.5-9.5g, beef liver extract powder 0.5-1.5g, potassium dihydrogen phosphate 1.5-2.0g, sodium chloride 1.5-2.5g, L-cysteine ​​0.1-0.3g, sodium thioglycolate 0.1-0.3g, growth-promoting factor 0.05-0.12g, and the balance is water; The growth-promoting factors are periplocoside, syringaldehyde, 4-methoxysalicylaldehyde, astragaloside IV, calycosin or formononetin.

2. The culture medium according to claim 1, characterized in that 1L of the culture medium is prepared with the following components: 6.8g peptone, 2.0g soy peptone, 3.4g yeast extract powder, 1.5g beef extract powder, 9.1g digested serum powder, 0.8g beef liver extract powder, 1.7g potassium dihydrogen phosphate, 2.0g sodium chloride, 0.2g L-cysteine, 0.2g sodium thioglycolate, 0.08g growth-promoting factor, and the balance is water.

3. The culture medium according to claim 1, characterized in that The bifidobacterium is Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium adolescentis, Bifidobacterium infantis or Bifidobacterium bifidum.

4. Use of the culture medium according to any one of claims 1 to 3 in promoting the growth of bifidobacteria.

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