CN117898433A - Synbiotic composition and application thereof - Google Patents

Synbiotic composition and application thereof Download PDF

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Publication number
CN117898433A
CN117898433A CN202410049024.XA CN202410049024A CN117898433A CN 117898433 A CN117898433 A CN 117898433A CN 202410049024 A CN202410049024 A CN 202410049024A CN 117898433 A CN117898433 A CN 117898433A
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China
Prior art keywords
lactobacillus
synbiotic composition
fermentation culture
present example
bifidobacterium
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Chinese (zh)
Inventor
银佳
冯立科
杨爱君
彭小霞
何瑛
纪坤发
林木娣
戴晓慧
黄娟
樊文博
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Guangdong Yantang Dairy Co ltd
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Guangdong Yantang Dairy Co ltd
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Priority to CN202410049024.XA priority Critical patent/CN117898433A/en
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Abstract

The invention discloses a synbiotics composition and application thereof, wherein the synbiotics composition comprises: bifidobacteria and/or lactobacilli, glucose or prebiotics; the prebiotic comprises isomaltooligosaccharide; inoculating the bifidobacterium or the lactobacillus into a fermentation substrate in the form of seed liquid, wherein the inoculating amount of the bifidobacterium or the lactobacillus is 1-5%, and the inoculating concentration is 1 multiplied by 10 7cfu/mL～1×1011 cfu/mL; the final concentration of the prebiotics is 10 g/L-50 g/L. The synbiotic composition provided by the invention can improve the yield of vitamin B7 synthesized by probiotics fermentation.

Description

Synbiotic composition and application thereof
Technical Field
The invention belongs to the technical field of microorganism culture, and particularly relates to a synbiotics composition and application thereof.
Background
Vitamin B7, also known as biotin, is abundant in eggs, fish, soybeans and nuts, and the world health organization WHO/United nations grain and agriculture organization FAO recommends that 30 μg vitamin B7 be taken by adults each day. Vitamin B7 plays an important role in gene regulation and cell signaling, and simultaneously participates in the metabolism of fatty acid, carbohydrate and amino acid of the organism, and the dietary supplement of vitamin B7 can accelerate the metabolism of the organism so as to reduce the risks of obesity and diabetes. Vitamin B7 can inhibit the expression of nuclear factor NF- κB related genes through histone binding mechanism, thereby inhibiting inflammation.
The result of metagenome research shows that actinomycetes such as bifidobacterium, such as ruminococcus, lactobacillus and the like lack genes necessary for synthesizing vitamin B7, and vitamin B7 needs to be obtained from the environment to meet the nutrition requirement. Studies have shown that a decrease in the pH level in the intestine increases vitamin B7 transport and absorption, whereas bifidobacteria, lactobacilli, enterococci and streptococci produce lactic acid etc. metabolites in the intestine, lowering the pH in the intestine, facilitating vitamin B7 absorption. At present, less researches are carried out on microorganisms for synthesizing vitamin B7, no report on the influence of probiotics on synthesizing vitamin B7 by probiotics is seen, and development of a novel synbiotics composition for improving the production efficiency of vitamin B7 by fermentation of probiotics is needed.
Disclosure of Invention
The present invention aims to solve at least one of the technical problems in the prior art described above. Therefore, the invention provides a synbiotic composition which can improve the yield of vitamin B7 synthesized by probiotics fermentation.
The invention also provides a method for promoting the fermentation and synthesis of vitamin B7 by probiotics.
The invention also provides a fermentation culture prepared by the method.
The invention also provides a food or medicine.
The invention also provides application of the synbiotics composition.
The invention also provides application of the fermentation culture.
According to a first aspect of the present invention, there is provided a synbiotic composition comprising:
(1) Bifidobacteria and/or lactobacillus, and
(2) Glucose or prebiotics;
the prebiotic comprises isomaltooligosaccharide;
Inoculating the bifidobacterium or the lactobacillus into a fermentation substrate in the form of seed liquid, wherein the inoculating amount of the bifidobacterium or the lactobacillus is 1-5%, and the inoculating concentration is 1 multiplied by 10 7cfu/mL～1×1011 cfu/mL;
The final concentration of the prebiotics is 10 g/L-50 g/L.
In some embodiments of the invention, the bifidobacterium or lactobacillus is inoculated in an amount of 1%.
In some embodiments of the invention, the bifidobacterium or lactobacillus is inoculated at a concentration of 1 x 10 8cfu/mL～1×1010 cfu/mL.
In some preferred embodiments of the invention, the bifidobacterium or lactobacillus is inoculated at a concentration of 1X 10 9 cfu/mL.
In some embodiments of the invention, the final concentration of glucose or prebiotic is 15g/L to 30g/L.
In some preferred embodiments of the invention, the final concentration of glucose or prebiotic is 20g/L.
In some embodiments of the invention, the isomaltooligosaccharides meet the specifications of the national standard GB/T20885-2017.
In some embodiments of the invention, the bifidobacterium comprises at least one of bifidobacterium animalis subsp Bifidobacterium animalis subsp.
In some preferred embodiments of the present invention, the bifidobacterium animalis subspecies include at least one of bifidobacterium animalis subspecies BB-12, bifidobacterium animalis subspecies BL-04, bifidobacterium animalis subspecies B420, bifidobacterium animalis subspecies HN019, bifidobacterium animalis subspecies Y6, and bifidobacterium animalis subspecies CECT 8145.
In some preferred embodiments of the invention, the bifidobacterium longum subspecies longum comprises bifidobacterium subspecies longum W68.
In some preferred embodiments of the invention, the bifidobacterium bifidum comprises bifidobacterium bifidum Y22.
In some embodiments of the invention, the lactobacillus comprises at least one of lactobacillus plantarum Lactiplantibacillus plantarum, lactobacillus acidophilus Lactobacillus acidophilus, lactobacillus rhamnosus Lacticaseibacillus rhamnosus, lactobacillus paracasei Lacticaseibacillus paracasei, lactobacillus casei Lacticaseibacillus casei, lactobacillus gasseri Lactobacillus gasseri, and lactobacillus reuteri Limosilactobacillus reuteri.
In some preferred embodiments of the invention, the lactobacillus plantarum comprises at least one of lactobacillus plantarum FEED8, lactobacillus plantarum LP45 and lactobacillus plantarum CN 2018.
In some preferred embodiments of the present invention, the lactobacillus acidophilus comprises at least one of lactobacillus acidophilus K43, lactobacillus acidophilus NCFM and lactobacillus acidophilus LA-5.
In some preferred embodiments of the present invention, the lactobacillus rhamnosus comprises at least one of lactobacillus rhamnosus YGG and lactobacillus rhamnosus HN 001.
In some preferred embodiments of the invention, the lactobacillus paracasei includes at least one of lactobacillus paracasei K9 and lactobacillus paracasei Lpc-37.
In some preferred embodiments of the invention, the lactobacillus casei comprises lactobacillus casei K35.
In some preferred embodiments of the invention, the lactobacillus gasseri comprises lactobacillus gasseri BDUP.
In some preferred embodiments of the invention, the lactobacillus reuteri comprises lactobacillus reuteri K07.
In some embodiments of the invention, the synbiotic composition is bifidobacteria and glucose.
In some preferred embodiments of the invention, the synbiotic composition is glucose and bifidobacterium longum subspecies longum W68.
In some embodiments of the invention, the synbiotic composition is lactobacillus and isomaltooligosaccharide.
In some preferred embodiments of the invention, the synbiotic composition is isomaltooligosaccharide and lactobacillus acidophilus LA-5.
According to a second aspect of the present invention, there is provided a method of promoting the fermentative synthesis of microorganism B7 by probiotics, said method comprising the steps of: the synbiotic composition according to the first aspect of the invention is added into a culture medium and is cultivated for 18 to 30 hours at a temperature of between 30 and 40 ℃ to obtain a fermentation culture containing vitamin B7.
In some embodiments of the invention, the incubation time is 20 to 28 hours.
In some preferred embodiments of the invention, the time of the incubation is 24 hours.
In some embodiments of the invention, the medium is a carbon source free MRS medium supplemented with 0.03% -0.07% cysteine hydrochloride.
In some preferred embodiments of the present invention, the medium is a carbon-free MRS medium supplemented with 0.05% cysteine hydrochloride. In some preferred embodiments of the present invention, the carbon-free MRS medium includes beef extract, tryptone, yeast extract, dipotassium hydrogen phosphate, tri-ammonium citrate, magnesium sulfate heptahydrate, manganese sulfate tetrahydrate, sodium acetate, tween-80.
In some more preferred embodiments of the present invention, the carbon source-free MRS medium includes 8 to 12g/L of beef extract powder, 8 to 12g/L of tryptone, 4 to 6g/L of yeast extract powder, 1 to 3g/L of dipotassium hydrogen phosphate, 1 to 3g/L of tri-ammonium citrate, 0.4 to 0.6g/L of magnesium sulfate heptahydrate, 0.2 to 0.3g/L of manganese sulfate tetrahydrate, 4 to 6g/L of sodium acetate, and 0.05v/v% to 0.2v/v% of tween-80.
According to a third aspect of the present invention there is provided a fermentation culture of a probiotic, the fermentation culture being prepared using a synbiotic composition according to the first aspect of the present invention and/or a method according to the second aspect of the present invention.
In some embodiments of the invention, the fermentation culture contains 4 to 25 μg/L vitamin B7.
In some preferred embodiments of the invention, the fermentation culture contains 10-20 μg/L vitamin B7.
In some embodiments of the invention, the synbiotic composition is glucose and bifidobacterium longum subspecies longum W68, and the fermentation culture contains 19.78 μg/L vitamin B7.
In some embodiments of the invention, the synbiotic composition is isomaltooligosaccharide and lactobacillus acidophilus LA-5, and the fermentation culture contains 10.67 μg/L vitamin B7.
According to a fourth aspect of the present invention, there is provided a food or pharmaceutical product comprising a synbiotic composition according to the first aspect of the present invention and/or a fermentation culture according to the third aspect of the present invention.
In some embodiments of the invention, the food product comprises at least one of a dairy product, a cereal product, a soy product, a condiment, an ice product, and a beverage.
In some preferred embodiments of the present invention, the dairy product is selected from at least one of sterilized milk, reconstituted milk, yogurt, cheese milk, milk powder, formula milk powder, condensed milk, cheese, casein, whey powder, and milk fat.
According to a fifth aspect of the present invention there is provided the use of a synbiotic composition according to the first aspect of the present invention, to facilitate the fermentative synthesis of vitamin B7 by probiotics.
According to a sixth aspect of the present invention there is provided the use of a synbiotic composition according to the first aspect of the present invention and/or a fermentation culture according to the third aspect of the present invention in any one of (1) to (4):
(1) Preparing a food;
(2) Preparing an anti-inflammatory drug;
(3) Preparing a medicament for preventing or treating obesity;
(4) Preparing the medicine for preventing or treating diabetes.
Detailed Description
The conception and the technical effects produced by the present invention will be clearly and completely described in conjunction with the embodiments below to fully understand the objects, features and effects of the present invention. It is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments, and that other embodiments obtained by those skilled in the art without inventive effort are within the scope of the present invention based on the embodiments of the present invention.
Example 1
The present example provides a synbiotic composition comprising bifidobacterium animalis subspecies lactis BB-12 and glucose; the embodiment also utilizes the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific steps are as follows:
1) Inoculating a small amount of lyophilized powder of Bifidobacterium animalis subspecies BB-12 in MRSc liquid culture medium in an ultra-clean bench, culturing in a 37 deg.C incubator for 24 hr, mixing the cultured bacterial liquid with 50% glycerol (obtained by mixing pure glycerol with physiological saline in equal volume) in equal volume, and preserving at-80deg.C; specific information of the freeze-dried bacterial powder used in the embodiment of the invention is shown in table 1;
2) Streaking the frozen strain in the step 1) on MRSc plates, culturing at 37 ℃ for 48 hours, and then picking a single colony to be cultured in MRSc liquid medium for 24 hours;
3) Transferring the bacterial liquid obtained by culturing in the step 2) into a fresh MRSc liquid culture medium with an inoculum size of 1%, culturing for 24 hours at 37 ℃, and transferring into a MRSc culture medium with glucose as a carbon source (glucose adding amount is 20 g/L) (the formula of the MRSc culture medium comprises: weighing 10g of beef extract powder, 10g of tryptone, 5g of yeast extract powder, 2g of dipotassium hydrogen phosphate, 2g of triammonium citrate, 0.5g of magnesium sulfate heptahydrate, 0.25g of manganese sulfate tetrahydrate, 5g of sodium acetate, 1mL of Tween-80 and 0.5g of cysteine hydrochloride, dissolving in 980mL of distilled water, sterilizing for 15min at 121 ℃ after constant volume is 1L, cooling), and culturing for 24h at 37 ℃;
4) Collecting the bacterial liquid obtained by culturing in the step 3), centrifuging at 4 ℃ and 8000rpm for 10min, collecting supernatant, filtering with a sterile 0.22 μm filter membrane, packaging in a sterile freezing tube, and freezing at-80 ℃ for standby.
TABLE 1 seed name, number, procurement Source and corresponding examples
Example 2
The present example provides a synbiotic composition comprising bifidobacterium animalis subspecies lactis BB-12 and fructooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 3
The present example provides a synbiotic composition comprising bifidobacterium animalis subspecies lactis BB-12 and galactooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 4
The present example provides a synbiotic composition comprising bifidobacterium animalis subspecies lactis BB-12 and isomaltooligosaccharide; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 5
The embodiment provides a synbiotic composition, which comprises bifidobacterium animalis subspecies BL-04 and glucose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 6
The embodiment provides a synbiotic composition, which comprises bifidobacterium animalis subspecies lactis BL-04 and fructooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 7
The present example provides a synbiotic composition comprising bifidobacterium animalis subspecies lactis BL-04 and galactooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 8
The present example provides a synbiotic composition comprising bifidobacterium animalis subspecies lactis BL-04 and isomaltooligosaccharide; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 9
The present example provides a synbiotic composition comprising bifidobacterium animalis subspecies B420 and glucose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 10
The present example provides a synbiotic composition comprising bifidobacterium animalis subspecies B420 and fructooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 11
The present example provides a synbiotic composition comprising bifidobacterium animalis subspecies B420 and galactooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 12
The present example provides a synbiotic composition comprising bifidobacterium animalis subspecies B420 and isomaltooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 13
The present example provides a synbiotic composition comprising bifidobacterium animalis subspecies lactis HN019 and glucose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 14
The present example provides a synbiotic composition comprising bifidobacterium animalis subspecies lactis HN019 and fructooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 15
The present example provides a synbiotic composition comprising bifidobacterium animalis subspecies lactis HN019 and galactooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 16
The present example provides a synbiotic composition comprising bifidobacterium animalis subspecies lactis HN019 and isomaltooligosaccharide; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 17
The present example provides a synbiotic composition comprising bifidobacterium animalis subspecies Y6 and glucose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 18
The present example provides a synbiotic composition comprising bifidobacterium animalis subspecies Y6 and fructooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 19
The present example provides a synbiotic composition comprising bifidobacterium animalis subspecies Y6 and galactooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 20
The present example provides a synbiotic composition comprising bifidobacterium animalis subspecies Y6 and isomaltooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 21
The present example provides a synbiotic composition comprising bifidobacterium animalis subspecies CECT8145 and glucose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 22
The present example provides a synbiotic composition comprising bifidobacterium animalis subspecies CECT8145 and fructooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 23
The present example provides a synbiotic composition comprising bifidobacterium animalis subspecies CECT8145 and galactooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 24
The present example provides a synbiotic composition comprising bifidobacterium animalis subspecies CECT8145 and isomaltooligosaccharide; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 25
The present example provides a synbiotic composition comprising bifidobacterium longum subspecies longum W68 and glucose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 26
The present example provides a synbiotic composition comprising bifidobacterium longum subspecies longum W68 and fructooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 27
The present example provides a synbiotic composition comprising bifidobacterium longum subspecies longum W68 and galactooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 28
The present example provides a synbiotic composition comprising bifidobacterium longum subspecies longum W68 and isomaltooligosaccharide; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 29
The present example provides a synbiotic composition comprising bifidobacterium bifidum Y22 and glucose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 30
The present example provides a synbiotic composition comprising bifidobacterium bifidum Y22 and fructooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 31
The present example provides a synbiotic composition comprising bifidobacterium bifidum Y22 and galactooligosaccharide; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 32
The present example provides a synbiotic composition comprising bifidobacterium bifidum Y22 and isomaltooligosaccharide; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 33
The present example provides a synbiotic composition comprising lactobacillus plantarum FEED8 and glucose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 34
The present example provides a synbiotic composition comprising lactobacillus plantarum FEED8 and fructooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 35
The present example provides a synbiotic composition comprising lactobacillus plantarum FEED8 and galacto-oligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 36
The present example provides a synbiotic composition comprising lactobacillus plantarum FEED8 and isomaltooligosaccharide; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 37
The present example provides a synbiotic composition comprising lactobacillus plantarum LP45 and glucose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 38
The present example provides a synbiotic composition comprising lactobacillus plantarum LP45 and fructooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 39
The present example provides a synbiotic composition comprising lactobacillus plantarum LP45 and galacto-oligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 40
The present example provides a synbiotic composition comprising lactobacillus plantarum LP45 and isomaltooligosaccharide; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 41
The embodiment provides a synbiotic composition, which comprises lactobacillus plantarum CN2018 and glucose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 42
The embodiment provides a synbiotic composition, which comprises lactobacillus plantarum CN2018 and fructooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 43
The present example provides a synbiotic composition comprising lactobacillus plantarum CN2018 and galacto-oligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 44
The present example provides a synbiotic composition comprising lactobacillus plantarum CN2018 and isomaltooligosaccharide; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 45
The embodiment provides a synbiotic composition, which comprises lactobacillus acidophilus K43 and glucose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 46
The embodiment provides a synbiotic composition, which comprises lactobacillus acidophilus K43 and fructo-oligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 47
The present example provides a synbiotic composition comprising lactobacillus acidophilus K43 and galacto-oligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 48
The present example provides a synbiotic composition comprising lactobacillus acidophilus K43 and isomaltooligosaccharide; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 49
The present example provides a synbiotic composition comprising lactobacillus acidophilus NCFM and glucose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 50
The present example provides a synbiotic composition comprising lactobacillus acidophilus NCFM and fructooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 51
The present example provides a synbiotic composition comprising lactobacillus acidophilus NCFM and galacto-oligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 52
The present example provides a synbiotic composition comprising lactobacillus acidophilus NCFM and isomaltooligosaccharide; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 53
The embodiment provides a synbiotic composition, which comprises lactobacillus acidophilus LA-5 and glucose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 54
The embodiment provides a synbiotic composition, which comprises lactobacillus acidophilus LA-5 and fructooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 55
The embodiment provides a synbiotic composition, which comprises lactobacillus acidophilus LA-5 and galacto-oligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 56
The present example provides a synbiotic composition comprising lactobacillus acidophilus LA-5 and isomaltooligosaccharide; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 57
The embodiment provides a synbiotic composition, which comprises lactobacillus rhamnosus YGG and glucose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 58
The embodiment provides a synbiotic composition, which comprises lactobacillus rhamnosus YGG and fructo-oligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 59
The embodiment provides a synbiotic composition, which comprises lactobacillus rhamnosus YGG and galacto-oligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 60
The present embodiment provides a synbiotic composition comprising lactobacillus rhamnosus YGG and isomaltooligosaccharide; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 61
The present example provides a synbiotic composition comprising lactobacillus rhamnosus YGG and 2' -fucosyllactose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 62
The embodiment provides a synbiotic composition, which comprises lactobacillus rhamnosus HN001 and glucose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 63
The embodiment provides a synbiotic composition, which comprises lactobacillus rhamnosus HN001 and fructo-oligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 64
The embodiment provides a synbiotic composition, which comprises lactobacillus rhamnosus HN001 and galacto-oligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 65
The embodiment provides a synbiotic composition, which comprises lactobacillus rhamnosus HN001 and isomaltooligosaccharide; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 66
The present example provides a synbiotic composition comprising lactobacillus rhamnosus HN001 and 2' -fucosyllactose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 67
The embodiment provides a synbiotic composition, which comprises lactobacillus paracasei K9 and glucose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 68
The embodiment provides a synbiotic composition, which comprises lactobacillus paracasei K9 and fructooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 69
The present example provides a synbiotic composition comprising lactobacillus paracasei K9 and galacto-oligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 70
The present example provides a synbiotic composition comprising lactobacillus paracasei K9 and isomaltooligosaccharide; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 71
The present example provides a synbiotic composition comprising lactobacillus paracasei K9 and 2' -fucosyllactose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 72
The present example provides a synbiotic composition comprising lactobacillus paracasei Lpc-37 and glucose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 73
The present example provides a synbiotic composition comprising lactobacillus paracasei Lpc-37 and fructooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 74
The present example provides a synbiotic composition comprising lactobacillus paracasei Lpc-37 and galacto-oligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 75
The present example provides a synbiotic composition comprising lactobacillus paracasei Lpc-37 and isomaltooligosaccharide; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 76
The present example provides a synbiotic composition comprising lactobacillus paracasei Lpc-37 and 2' -fucosyllactose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 77
The present example provides a synbiotic composition comprising lactobacillus casei K35 and glucose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 78
The present example provides a synbiotic composition comprising lactobacillus casei K35 and fructooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 79
The present example provides a synbiotic composition comprising lactobacillus casei K35 and galacto-oligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 80
The present example provides a synbiotic composition comprising lactobacillus casei K35 and isomaltooligosaccharide; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 81
The present example provides a synbiotic composition comprising lactobacillus casei K35 and 2' -fucosyllactose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 82
The present example provides a synbiotic composition comprising lactobacillus gasseri BDUP and glucose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 83
The present example provides a synbiotic composition comprising lactobacillus gasseri BDUP and fructooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 84
The present example provides a synbiotic composition comprising lactobacillus gasseri BDUP and galacto-oligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 85
The present example provides a synbiotic composition comprising lactobacillus gasseri BDUP and isomaltooligosaccharide; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 86
The present example provides a synbiotic composition comprising lactobacillus reuteri K07 and glucose; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 87
The present example provides a synbiotic composition comprising lactobacillus reuteri K07 and fructooligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 88
The present example provides a synbiotic composition comprising lactobacillus reuteri K07 and galacto-oligosaccharides; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Example 89
The present example provides a synbiotic composition comprising lactobacillus reuteri K07 and isomaltooligosaccharide; the present example also used the synbiotic composition to prepare a fermentation culture containing vitamin B7, and the specific procedure was as in example 1.
Test examples
The present test example tested the vitamin B7 content of the fermentation cultures prepared in examples 1 to 89, and the specific test procedure and test results were as follows:
1) Sample pretreatment: taking out a sample of the fermentation culture in a refrigerator at-80 ℃, freeze-drying 1mL of the sample, adding 10. Mu.L of an internal standard and 400. Mu.L of a pre-cooled methanol/acetonitrile (1:1 mixture, v/v) solution containing 0.3% formic acid, adding black ceramic beads, homogenizing for 2 times (20 s each), vortex mixing for 30s, and incubating at 4 ℃ for 10min to precipitate proteins; centrifuging at 4deg.C and 14000rcf for 10min, collecting 400 μl of supernatant, adding 400 μl of pre-cooled mass spectrometry grade ultrapure water, mixing by vortex for 30s, sucking 800 μl of supernatant, performing solid phase extraction, blow drying filtrate, and preserving at-80deg.C to be detected; the internal standard is a mixture of ：Pyridoxine-(methyl-d3)hydrochloride(Sigma,809659,1mg)、Nicotinamide-13C6solution(Sigma,V-034-1ML,1ML)、Thiamine-(4-methyl-13C-thiazol-5-yl-13C3)hydrochloride(Vitamin B1-13C4)(Sigma,731188,2mg)、biotin(3',3',4',4'-d4)(CIL,DLM-9751-PK,1mg)、Vitamin B5(di-β-alanine-13C6,15N2)calcium salt(Sigma,705837,5mg)、Riboflavin-(dioxopyrimidine-13C4,15N2)(Sigma,705292,1mg)、Folic acid-(glutamic acid-13C5)(Sigma,803049,1mg)、Nicotinic acid-13C6 solution(Sigma,V-035-1ML,1ML);
2) Vitamin B7 content determination:
Separating the sample by adopting an Agilent 1290 Infinicity LC ultra-high performance liquid chromatography system; placing the sample in an automatic sampler at 4 ℃, wherein the column temperature is 45 ℃, the mobile phase A is 5mM ammonium formate and 0.3% formic acid aqueous solution (315.28 mg ammonium formate and 3mL formic acid are dissolved in 900mL mass spectrum grade ultrapure water, after all the ammonium formate and 3mL formic acid are dissolved, the volume is fixed to 1L, the mobile phase B is pure methanol, the flow rate is 300 mu L/min, and the sample injection amount is 5 mu L;
The relevant liquid phase gradients are as follows: 0-2min, wherein the liquid B is 0%, and the liquid B is linearly changed from 0% to 60% in 2-3 min; 3-4.5min, the liquid B is linearly changed from 60% to 100%;4.5-6min, wherein the liquid B is linearly changed from 100% to 5%, and 6-11min, and the liquid B is 5%; setting a QC sample for detecting and evaluating the stability and repeatability of the system every a certain number of experimental samples in the sample queue; a standard mixture of set target substances of the sample queue for correction of chromatographic retention times;
Adopting 5500QTRAP mass spectrometer (AB SCIEX) to carry out mass spectrometry in positive ion mode, wherein the detection mode is multi-reaction monitoring (MRM); the specific conditions include: ion source temperature: 550 ℃; atomizing Gas (Gas 1): 55psi; assist Gas (Gas 2): 55psi; curtain gas (CUR): 40psi; electrospray gas pressure (ISVF): +4500v; after the measurement result is calculated, performing inter-group T-test significance analysis on the data; the results are shown in tables 2 to 3.
TABLE 2 content of vitamin B7 in fermentation supernatants of bifidobacteria (examples 1 to 32) (. Mu.g/L)
Strain Glucose Fructooligosaccharides Galacto-oligosaccharides Oligomeric isomaltose
BB12 8.89±0.71 8.38±0.60 9.06±0.27 9.94±0.24
BB04 8.23±0.30 8.74±0.63 8.67±0.35 8.81±0.32
BB420 8.51±0.43 9.23±0.40 8.61±0.31 9.25±0.29
BB019 9.20±0.06 9.41±0.58 9.31±0.63 9.56±1.06
BB6 9.39±0.34 9.89±1.01 9.89±0.20 9.65±0.36
BB8145 7.91±0.16 8.35±0.30 8.38±1.07 8.51±0.52
BB68 19.78±1.21 13.89±1.02 11.70±1.52 16.45±1.22
BB22 8.45±0.50 9.06±0.38 9.10±0.61 9.12±0.27
TABLE 3 vitamin B7 content (μg/L) in fermentation supernatants of Lactobacillus (examples 33-89)
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As can be seen from Table 2, when the probiotic bacteria in the synbiotic composition are bifidobacteria, a higher vitamin B7 yield (19.78. Mu.g/L) can be obtained using a composition of glucose and Bifidobacterium longum subspecies W68 (example 25).
As can be seen from Table 3, when the probiotic bacteria in the synbiotic composition were Lactobacillus, a higher yield of microorganism B7 (10.67. Mu.g/L) was obtained using the composition of isomaltooligosaccharide and Lactobacillus acidophilus LA-5 (example 56).
While the embodiments of the present invention have been described in detail, the present invention is not limited to the above embodiments, and various changes can be made without departing from the spirit of the present invention within the knowledge of those skilled in the art. Furthermore, embodiments of the invention and features of the embodiments may be combined with each other without conflict.

Claims (10)

1. A synbiotic composition, characterized in that the synbiotic composition comprises:
(1) Bifidobacteria and/or lactobacillus, and
(2) Glucose or prebiotics;
the prebiotic comprises isomaltooligosaccharide;
Inoculating the bifidobacterium or the lactobacillus into a fermentation substrate in the form of seed liquid, wherein the inoculating amount of the bifidobacterium or the lactobacillus is 1-5%, and the inoculating concentration is 1 multiplied by 10 7cfu/mL～1×1011 cfu/mL;
The final concentration of the prebiotics is 10 g/L-50 g/L.
2. The synbiotic composition according to claim 1, wherein the bifidobacteria comprise at least one of bifidobacterium animalis subsp Bifidobacterium animalis subsp.
Preferably, the bifidobacterium animalis subspecies include at least one of bifidobacterium animalis subspecies BB-12, bifidobacterium animalis subspecies BL-04, bifidobacterium animalis subspecies B420, bifidobacterium animalis subspecies HN019, bifidobacterium animalis subspecies Y6 and bifidobacterium animalis subspecies CECT 8145;
Preferably, the bifidobacterium longum subspecies longum comprises bifidobacterium subspecies longum W68;
preferably, the bifidobacterium bifidum comprises bifidobacterium bifidum Y22.
3. The synbiotic composition according to claim 1, wherein the lactobacillus comprises at least one of lactobacillus plantarum Lactiplantibacillus plantarum, lactobacillus acidophilus Lactobacillus acidophilus, lactobacillus rhamnosus Lacticaseibacillus rhamnosus, lactobacillus paracasei Lacticaseibacillus paracasei, lactobacillus casei Lacticaseibacillus casei, lactobacillus gasseri Lactobacillus gasseri and lactobacillus reuteri Limosilactobacillus reuteri;
Preferably, the lactobacillus plantarum comprises at least one of lactobacillus plantarum FEED8, lactobacillus plantarum LP45 and lactobacillus plantarum CN 2018;
preferably, the lactobacillus acidophilus comprises at least one of lactobacillus acidophilus K43, lactobacillus acidophilus NCFM and lactobacillus acidophilus LA-5;
preferably, the lactobacillus rhamnosus comprises at least one of lactobacillus rhamnosus YGG and lactobacillus rhamnosus HN 001;
Preferably, the lactobacillus paracasei comprises at least one of lactobacillus paracasei K9 and lactobacillus paracasei Lpc-37;
Preferably, the lactobacillus casei comprises lactobacillus casei K35;
preferably, the lactobacillus gasseri comprises lactobacillus gasseri BDUP;
preferably, the lactobacillus reuteri comprises lactobacillus reuteri K07.
4. The synbiotic composition according to claim 1, wherein the synbiotic composition is bifidobacteria and glucose;
And/or, the synbiotic composition is lactobacillus and isomaltooligosaccharide.
5. A method for promoting the fermentation and synthesis of vitamin B7 by probiotics, said method comprising the steps of: the synbiotic composition according to any one of claims 1-4, added to a medium, and incubated at 30 ℃ -40 ℃ for 18-30 hours, resulting in a fermentation culture containing vitamin B7.
6. The method according to claim 5, wherein the medium is MRS medium supplemented with 0.03% -0.07% cysteine hydrochloride;
Preferably, the carbon-free MRS medium comprises beef extract powder, tryptone, yeast extract powder, dipotassium hydrogen phosphate, tri-ammonium citrate, magnesium sulfate heptahydrate, manganese sulfate tetrahydrate, sodium acetate, and tween-80.
7. A fermentation culture of a probiotic, characterized in that it is prepared by using a synbiotic composition according to any one of claims 1 to 4 and/or a method according to any one of claims 5 to 6.
8. A food or pharmaceutical product, characterized in that it comprises a synbiotic composition according to any one of claims 1 to 4 and/or a fermentation culture according to claim 7;
preferably, the food product comprises at least one of a dairy product, a cereal product, a soy product, a condiment, an ice product, and a beverage;
More preferably, the dairy product is selected from at least one of sterilized milk, reconstituted milk, yogurt, cheese milk, milk powder, formula milk powder, condensed milk, cheese, casein, whey powder, and milk fat.
9. Use of a synbiotic composition as claimed in any one of claims 1 to 4 to facilitate the fermentative synthesis of vitamin B7 by probiotics.
10. Use of a synbiotic composition according to any one of claims 1 to 4 and/or a fermentation culture according to claim 7 in any one of (1) to (4):
(1) Preparing a food;
(2) Preparing an anti-inflammatory drug;
(3) Preparing a medicament for preventing or treating obesity;
(4) Preparing the medicine for preventing or treating diabetes.
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