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CN113355375B - Method for synthesizing B vitamins by fermenting lactobacillus paracasei K56 and application - Google Patents

Method for synthesizing B vitamins by fermenting lactobacillus paracasei K56 and application Download PDF

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Publication number
CN113355375B
CN113355375B CN202011355562.XA CN202011355562A CN113355375B CN 113355375 B CN113355375 B CN 113355375B CN 202011355562 A CN202011355562 A CN 202011355562A CN 113355375 B CN113355375 B CN 113355375B
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vitamin
fermentation
vitamins
lactobacillus paracasei
milk
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CN113355375A (en
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洪维鍊
赵雯
刘伟贤
侯保朝
马国文
吴超
倪丹
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Inner Mongolia Yili Industrial Group Co Ltd
Inner Mongolia Dairy Technology Research Institute Co Ltd
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Inner Mongolia Yili Industrial Group Co Ltd
Inner Mongolia Dairy Technology Research Institute Co Ltd
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    • C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
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    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
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    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
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    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
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    • A61K31/525—Isoalloxazines, e.g. riboflavins, vitamin B2
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    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
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    • A61K8/673—Vitamin B group
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
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    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/67—Vitamins
    • A61K8/673—Vitamin B group
    • A61K8/675—Vitamin B3 or vitamin B3 active, e.g. nicotinamide, nicotinic acid, nicotinyl aldehyde
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
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    • C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
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Abstract

The invention provides a method for synthesizing B vitamins by fermenting lactobacillus paracasei K56 and application thereof. The invention firstly provides a method for synthesizing B vitamins by fermenting Lactobacillus paracasei (Lactobacillus paracasei subsp. Paracasei) K56, which comprises the following steps: the fermentation product containing B vitamins is prepared by fermenting lactobacillus paracasei CGMCC No.15139 or DSM 27447. The research of the invention finds that the lactobacillus paracasei strain can be used for the production and application of foods, daily chemicals, feeds or medicines and the like rich in B vitamins.

Description

Method for synthesizing B vitamins by fermenting lactobacillus paracasei K56 and application
Technical Field
The invention relates to the technical field of microbial fermentation, in particular to a method for synthesizing B vitamins by fermenting Lactobacillus paracasei (Lactobacillus paracasei subsp. Paracasei) K56 (with the preservation number of CGMCC No.15139 or DSM 27447) and related application.
Background
The B vitamins are also called B vitamins (also called vitamin B, vitamin B group or vitamin B complex group), and are classified as "families" because of 8 types in total. The B vitamins are water-soluble vitamins which are indispensable for maintaining the normal functions and metabolic activities of human bodies, and can be quickly consumed in the human bodies in the tense life and working pressure, improper dietary habits or due to the use of certain specific medicines and the property of dissolving the B vitamins in water. The supplement of vitamin B can relieve emotion. Moreover, according to research reports, the B vitamins can help to maintain the functions of the heart and the nervous system, maintain the health of the digestive system and the skin, participate in energy metabolism, enhance physical strength, tonify and strengthen the body. If vitamin B is absent, cell function is immediately reduced, and metabolic disorder is caused, at which the human body suffers from malaise and inappetence. Furthermore, excessive drinking and the like cause liver damage, which is concurrent with vitamin B deficiency in many cases.
The B vitamins are widely present in rice bran, wheat bran, yeast, animal livers, coarse grain vegetables and other foods. Studies have shown that part of the B vitamins can be synthesized in the human gut.
Lactobacillus paracasei is a gram-positive facultative heterotrophic lactic acid bacterium commonly used in dairy fermentation and probiotics. Lactobacillus paracasei is commonly found in a variety of human habitats, such as the human intestinal tract and oral cavity, as well as silage and naturally fermented foods.
No relevant research report about the B vitamins produced by the fermentation of lactobacillus paracasei is found.
Disclosure of Invention
One object of the present invention is to provide a novel use of lactobacillus paracasei K56.
Lactobacillus paracasei (Lactobacillus paracasei subsp. Paracasei) K56 strain has been deposited in German Collection of Microorganisms and Cell Cultures (German Collection of Microorganisms and Cell Cultures) on 27.6.2013 under accession number DSM27447; in addition, lactobacillus paracasei (Lactobacillus paracasei subsp. Paracasei) K56 strain has also been stored in China General Microbiological Culture Collection Center (CGMCC) with the Collection number of CGMCC No.15139 in 29.12 months and 29.2017.
The invention discovers that the Lactobacillus paracasei (Lactobacillus paracasei subsp. Paracasei) K56 takes the material liquid (milk substrate) mainly containing milk components as the fermentation substrate, and the fermentation product has high yield of B vitamins (containing vitamin B) 1 Vitamin B 2 Vitamin B 3 Vitamin B 6 Vitamin B 9 ) Of the cell.
Thus, in one aspect, the present invention provides a method for the fermentative synthesis of B vitamins from Lactobacillus paracasei (Lactobacillus paracasei subsp. Paracasei), comprising: the fermentation product containing B vitamins is prepared by fermenting lactobacillus paracasei CGMCC No.15139 or DSM 27447.
According to a particular embodiment of the invention, the method of the invention further comprises: extracting purified B vitamins from the supernatant of the fermentation product.
According to a specific embodiment of the present invention, the B vitamins include vitamin B 1 Vitamin B 2 Vitamin B 3 Vitamin B 6 And vitamin B 9 One or more of (a).
According to a specific embodiment of the present invention, the B vitamins include vitamin B 1 Vitamin B 3 And/or vitamin B 6 。
According to a particular embodiment of the invention, the fermentation substrate used in the fermentation may be an emulsion. Preferably, the milk may be raw milk or reconstituted milk, and may be whole milk, low fat milk or skim milk. The emulsion may optionally contain 5% to 10% sucrose.
According to a particular embodiment of the invention, the fermentation conditions are: fermenting for 1-7 days at 35-45 ℃.
According to a specific embodiment of the invention, lactobacillus paracasei is inoculated in the fermentation substrate in the form of seed liquid, with an inoculation amount of 2% to 5%. The preparation of the seed liquid may be carried out according to conventional techniques in the field of probiotics.
According to the specific embodiment of the present invention, the milk substrate used as the fermentation substrate of lactobacillus paracasei K56 in the present invention may be milk (whole milk, low-fat milk or skim milk) with milk content of more than 80%, wherein auxiliary materials such as sweetener, stabilizer and the like may be optionally added according to the need, such as fermentation substrate commonly used in the preparation of fermented milk or milk beverage, and the milk substrate fermented by lactobacillus paracasei K56 of the present invention can achieve the effect of high yield of B vitamins, and the fermentation product is particularly rich in vitamin B 1 Vitamin B 2 Vitamin B 3 Vitamin B 6 Vitamin B 9 。
Vitamin B 1 The content in milk is 0.3 mg/L on average. Vitamin B in milk 1 Is not completely from feed, and can be synthesized by microorganism in rumen of milk cow, so that sour milk fermented by lactic acid bacteria is superior to normalTonghua milk vitamin B 1 The content is increased by about 30 percent. Children less than one week in age need vitamin B daily 1 5-10mg. If vitamin B is deficient 1 It can cause anorexia, slow growth, damage of nervous system activity, polyneuritis, milk regurgitation or constipation, dyspepsia, greenish stool, pale complexion, inappetence, muscle relaxation, frequent crying, and weight loss in children. According to some embodiments of the invention, lactobacillus paracasei K56 ferments vitamin B in the emulsion 1 The content can reach 30-70 mu g/mL. The yogurt product fermented by K56 is taken 100 mL/day, and can be supplemented with VB for human body intake 1 。
Vitamin B 2 In human milk, the content of vitamin B in cow milk is not high 2 The content can reach 1-2mg/L. The children are recommended to take 1-3mg of vitamin B every day. If vitamin B2 is lacked, the children will suffer from growth retardation, anorexia, inflammation of mouth corners and lips, red and swollen tongue, malnutrition, and asthenia. According to some embodiments of the invention, lactobacillus paracasei K56 ferments vitamin B in the emulsion 2 The content can reach 2.8-3 mug/mL.
Vitamin B 3 (nicotinic acid) is recommended to be taken by adults at 13-19mg per day (dietary guidelines of Chinese residents), 20mg per day is recommended for pregnant women, and 22mg per day is recommended for pregnant women in lactation. Vitamin B 3 It is mainly contained in livestock meat, fish and poultry, and plant foods such as mushroom, asparagus, sunflower seed, peanut, beans and grains. Vitamin B in ordinary yoghurt 3 Is present in an amount of about 0.2mg/100g. According to some embodiments of the invention, lactobacillus paracasei K56 ferments vitamin B in the emulsion 3 50-80 mug/mL, which is far higher than the content in the common yoghourt.
Vitamin B 6 Also known as pyridoxine, which includes pyridoxine, pyridoxal and pyridoxamine, exists as phosphate esters in vivo, and is a water-soluble vitamin. Vitamin B 6 The functionality involves a number of aspects including: participating in protein synthesis and catabolism and participating in all amino acid metabolism; participating in gluconeogenesis and UFA generationMetabolism, associated with the metabolism of glycogen, sphingomyelin, and steroids; participate in the synthesis of certain neuromediators (5-hydroxytryptamine, taurine, dopamine, norepinephrine and gamma-aminobutyric acid); involving nucleic acid and DNA synthesis, and lacking vitamin B 6 DNA synthesis is impaired, a process which is important for maintaining proper immune function. Papastoyianidis et al (2006) use commercial ferments YO-1, YO-2 to ferment at 42 ℃ for 3.5h, then store for 1.5h in cold storage, detect vitamin B 6 The content of (A) is 1.37mg/kg and 1.50mg/kg respectively; vitamin B after fermentation assisted by Bifidobacterium BB-12 and Lactobacillus acidophilus LA-5 6 The contents of (A) are 1.37mg/kg and 1.31mg/kg, respectively. According to some embodiments of the invention, lactobacillus paracasei K56 ferments vitamin B in the emulsion 6 The content is 8-20 mug/mL.
Vitamin B 9 The content of the (folic acid) in the milk is generally 40 +/-10.00 mu g/L, and the content of the (folic acid) in the common yoghourt is 80 +/-20.00 mu g/L. Vitamin B 9 The main effects on organisms are as follows: are involved in the metabolism of genetic material and proteins; influence animal reproductive performance; affecting secretion of animal pancreas; promoting the growth of animals; improving immunity of organism. Vitamin B 9 As one of the essential vitamins for the growth and reproduction of body cells, the deficiency can affect the normal physiological activities of human bodies. Many reports in the literature of vitamin B deficiency 9 Has direct relation with diseases such as neural tube deformity, megaloblastic anemia, cleft lip and palate, depression, tumor and the like. In general, normal humans require vitamin B 9 The amount of the medicine is 100-200 mu g/d, and the recommended amount of the world health organization is as follows: adult 200 ug/d, pregnant woman and lactating mother 400 ug/d. The latest folic acid consumption standard of the FDA is as follows: between 25 and 50 years old (male) 240 μ g/d; 190 μ g/d between 25 and 50 years old (female); the concentration of the extract is 400 mug/d for the nannie and the pregnant woman; the infant is 200-400 mug/d. According to some embodiments of the invention, lactobacillus paracasei K56 ferments vitamin B in the emulsion 9 The content of (b) is 1.6-1.8 mu g/mL.
Therefore, in another aspect, the invention also provides the application of the lactobacillus paracasei in the preparation of the composition rich in B vitamins by fermentation, wherein the fermentation product rich in B vitamins is prepared by utilizing the fermentation of the lactobacillus paracasei with CGMCC No.15139 or DSM 27447.
According to a particular embodiment of the invention, the lactobacillus paracasei of the invention is used in the fermentative preparation of a composition enriched in B vitamins, which may be a food composition, a daily chemical composition or a pharmaceutical composition. Specifically, the fermentation product obtained by fermenting the milk substrate with lactobacillus paracasei K56 of the present invention can be directly used as a food (e.g., fermented milk, milk drink, milk powder, solid drink, etc.), a daily chemical (e.g., cosmetics), a feed (including pet food or probiotic products for livestock), a drug, etc., or further used for preparing such food, daily chemical, feed, or drug products. The process for preparing food, daily chemical, feed or pharmaceutical products from the fermentation product can be carried out with reference to the prior art in the field.
In a particular embodiment of the invention, the B vitamin containing fermentation product of the invention is used as or further for the preparation of a food product, preferably a fermented milk or fermented milk beverage. The emulsion used as a fermentation substrate may further comprise other materials commonly used in fermented milk or fermented milk drinks, such as appropriate amounts of sweeteners, stabilizers, nutritional additives, and the like.
According to a particular embodiment of the invention, the fermented milk matrix of lactobacillus paracasei of the invention results in a fermentation product containing B vitamins which is also enriched in conjugated linoleic acid and/or gamma-aminobutyric acid. Therefore, the invention also provides the application of the lactobacillus paracasei K56 in the preparation of a composition rich in conjugated linoleic acid and/or gamma-aminobutyric acid through fermentation. In the composition, the conjugated linoleic acid and/or the gamma-aminobutyric acid are mainly generated by fermenting the lactobacillus paracasei.
In conclusion, the invention provides the new application of the lactobacillus paracasei K56, and the invention discovers that the single-strain fermented milk substrate of the strain has good vitamin B-containing synthetic vitamin B 1 Vitamin B 2 Vitamin B 3 Vitamin B 6 Vitamin B 9 The ability of the cell to perform. The Lactobacillus paracasei K56 strain can be used for enriching B vitaminsFood, daily chemical products, feed or medicine, etc.
Detailed Description
For a more clear understanding of the technical features, objects and advantages of the present invention, reference is now made to the following detailed description taken in conjunction with the accompanying specific embodiments, and the technical solutions of the present invention are described, it being understood that these examples are intended to illustrate the present invention and are not intended to limit the scope of the present invention. In the examples, each raw reagent material is commercially available, and the experimental method not specifying the specific conditions is a conventional method and a conventional condition well known in the art, or a condition recommended by an instrument manufacturer.
Example 1: production of B vitamins from Lactobacillus paracasei fermented milk
1. Experimental strains
The lactobacillus paracasei K56 (CGMCC No. 15139) used in the technical proposal of the invention;
control strain 1: lactobacillus paracasei strain l.casei 431;
control strain 2: the commercial strain Lactobacillus paracasei LPC-37.
2. Activation of strains and liquid culture of seeds
(1) Strain activation: activating lactobacillus paracasei by using MRS culture medium, taking the strain growing to logarithmic phase, and preserving glycerol tubes for later use. Wherein, MRS culture medium (g/L): 10.0g of peptone; 5.0g of yeast extract; 20.0g of glucose; 5.0g of anhydrous sodium acetate; 2.0g of citric acid dihydroamine; tween-80.0 mL; dipotassium phosphate 2.0g; magnesium sulfate heptahydrate 0.2g; 0.05g of manganese sulfate; 10.0g of beef extract; 15g to 20g of agar.
(2) Determination of growth dynamics of strains
Dissolving 12% whole milk powder and 6% white sugar in water at normal temperature, sterilizing at 95 deg.C for 5min, cooling to 36 + -1 deg.C, inoculating strain as milk matrix, fermenting and culturing for 120h, sampling at different fermentation time periods, and measuring viable count and pH value.
And (3) detecting the viable count: after mixing 25ml of the sample with 75ml of physiological saline uniformly, counting by a plate dilution method.
And (3) acidity detection: the sample pH was measured using a pH meter and recorded. Taking 10g of the fermented dairy product, adding 2 drops of phenolphthalein solution, mixing uniformly, and titrating 0.1M standard NaOH solution to pink. The calculation formula of the titration acidity is as follows:
°T＝C×V×100/(m×0.1)。
the results of the measurements are shown in tables 1 and 2, respectively.
TABLE 1 colony count of Lactobacillus paracasei in milk matrix for different fermentation times
Figure BDA0002802513100000051
Figure BDA0002802513100000061
From the analysis of the test data, the pH value of K56 fermentation is rapidly increased after 28h, and then is reduced to be stable. The viable count of the control strain 1 and the control strain 2 increased rapidly after 36 hours, and then decreased steadily. The analysis of the K56, the control strain 1 and the control strain 2 shows that p is more than 0.05, and no significant difference exists; control strain 1 and control strain 2 analyzed p >0.05 with no significant difference. Wherein the blank group is the milk matrix without the addition of bacteria liquid.
TABLE 2 pH determination of Lactobacillus paracasei in milk substrates for different fermentation times
Figure BDA0002802513100000062
Figure BDA0002802513100000071
From the analysis of the test data, K56, the control strain 1 and the control strain 2 gradually decrease within 12-36 h. The p of K56 is less than 0.01 when being analyzed by a control strain 2, and the difference is very obvious; k56 was >0.05 as analyzed for p with control strain 1, with no significant difference; control strain 1 and control strain 2 analyzed p >0.05 with no significant difference. The presence of non-killed micro-organisms in the blank resulted in milk spoilage and a decrease in pH due to temperature. Wherein the blank group is an emulsion matrix without added bacteria liquid.
(3) Preparation of strain seed liquid: dissolving 12% whole milk powder and 6% white sugar in water at normal temperature, sterilizing at 95 deg.C for 5min, cooling to 36 + -1 deg.C, inoculating strain, fermenting at 36 + -1 deg.C for 48 + -2 h, and prolonging to 72 + -2 h to obtain seed liquid.
3. Synthesis of B vitamins from lactobacillus paracasei fermented milk
Preparing fermented milk: adding sucrose accounting for 6.5% of the total weight of raw milk into raw milk, stirring, heating to 95 ℃, sterilizing for 5min, cooling the sterilized emulsion to 37 ℃, respectively adding various bacterial seed liquids (the inoculation amount of lactobacillus paracasei K56 is 3%, the inoculation amount of the control bacteria is equivalent to K56 according to the conversion of viable count), respectively fermenting each sample at 37 ℃ for 24, 48, 72, 96 and 120h, centrifuging the fermented emulsion at 8000r/min for 15min, collecting fermented supernatant, filtering through a 0.22 mu m filter membrane, and measuring the content of B vitamins in the filtrate.
The content of B vitamins is determined by high performance liquid chromatography, and the specific operation conditions refer to HPLC detection of water-soluble vitamins (Liyan, food research and development, 2018) in Collybia nigricans.
Blank milk base (without added probiotics) was used as a control, and blank control supernatant could not be obtained by centrifugation due to the inability to curd during the same fermentation time. Thus, the following data are the directly measured vitamin content in the supernatant of the fermentation broth, without subtracting the vitamin content in the blank milk matrix.
(1) Vitamin B in fermentation samples 1 In an amount of
Vitamin B in probiotic fermentation supernatant with different fermentation times 1 See table 3 for the contents of (d).
TABLE 3 fermentation supernatant vitamin B 1 In an amount of
Figure BDA0002802513100000081
From the analysis of the above test data, it can be seen that vitamin B is present in the fermentation supernatant of each strain 1 The content shows a clear rising trend along with the time. Differences were found between K56, control strain 1, and control strain 2, but were not significant.
(2) Fermentation sample vitamin B 2 In a content of
Vitamin B in probiotic fermentation supernatant with different fermentation times 2 See table 4 for the content of (d).
TABLE 4 vitamin B in fermentation supernatants 2 In a content of
Figure BDA0002802513100000082
From the analysis of the above test data, it can be seen that each strain produces vitamin B in the supernatant of the fermentation at different times 2 The contents are all close and the difference is not significant. Wherein vitamin B in the supernatant in the first 72h 2 The content shows a clear ascending trend, and the content is basically unchanged after 48 h.
(3) Fermentation sample vitamin B 3 In a content of
Vitamin B in probiotic fermentation supernatant with different fermentation times 3 See table 5 for the content of (d).
TABLE 5 vitamin B in fermentation supernatants 3 In a content of
Figure BDA0002802513100000083
The analysis of the test data shows that the vitamin B in the K56 120h fermentation supernatant 3 The content was about 1.4 times that of the control strain 2. Vitamin B in fermentation supernatant of each strain 3 The content showed a clear ascending trend with time, and the ascending trend of the control strain 2 was relatively slow. Vitamin B in supernatant of K56 and control Strain 1 when fermented for 120h 3 The content difference is obvious (p is less than 0.05). K56 and vitamin B in the fermentation supernatant of control Strain 2 3 The content difference is very obvious (p is less than 0.01).
(4) Fermentation sample vitamin B 6 In an amount of
Vitamin B in probiotic fermentation supernatant with different fermentation times 6 See table 6 for the contents of (d).
TABLE 6 HairVitamin B in fermentation supernatant 6 In a content of
Figure BDA0002802513100000091
The analysis of the test data shows that the vitamin B in the K56 120h fermentation supernatant 6 The content was about 1.4 times that of the control strain 2. Vitamin B in fermentation supernatant of each strain 6 The content showed a clear rising trend with time, the rising trend of the control strain 2 was relatively slow. Vitamin B in supernatant after 72h fermentation of K56 and control Strain 1 6 The content difference is obvious (p is less than 0.05). Vitamin B in supernatant after 72h fermentation of K56 and control Strain 2 6 The content difference is obvious (p is less than 0.05).
(5) Fermentation sample vitamin B 9 In a content of
Vitamin B in probiotic fermentation supernatant with different fermentation times 9 See table 7 for the contents of (d).
TABLE 7 vitamin B 9 In an amount of
Figure BDA0002802513100000092
From the analysis of the above test data, it can be seen that vitamin B is present in the fermentation supernatant of each strain 9 The content of vitamin B is substantially constant with time 9 The contents were all at a lower level.
According to the experimental result of the viable count of each strain, the lactobacillus paracasei reaches the maximum value of the viable count within about 48 hours, namely the stable period, and the viable counts are kept unchanged after 48 hours. After 24h fermentation, fermentation supernatant can not be obtained to detect the content of folic acid, but folic acid is supposed to be metabolized and generated in the process of strain growth. There was no significant difference in the folate content over time during the subsequent fermentation time, a possible reason: the folic acid content reaches the maximum value in the stable growth period (48 h) of the strain, and the thallus is aged and the folic acid synthesis amount is reduced along with the continuous metabolism of the strain; the influence of environmental factors such as light, heat, temperature and the like on the folic acid content in the experimental process.
(6) Content of Conjugated Linoleic Acid (CLA) and gamma-aminobutyric acid in fermentation sample
In the invention, the beneficial substances such as Conjugated Linoleic Acid (CLA) and gamma-aminobutyric acid are generated in the K56 fermented milk. See tables 8 and 9 for relevant test results. Wherein, the content determination method of the conjugated linoleic acid is based on a gas chromatography NY/T1671-2008 for determining the content of the Conjugated Linoleic Acid (CLA) in milk and dairy products; the content determination method of the gamma-aminobutyric acid is based on QB/T4587-2013 gamma-aminobutyric acid.
TABLE 8 content of conjugated linoleic acid in K56 fermentation supernatant
Figure BDA0002802513100000101
TABLE 9 content of gamma-aminobutyric acid in fermentation supernatant
Figure BDA0002802513100000102
Example 2 fermentation of Lactobacillus paracasei K56 milk beverage enriched in vitamin B group
Heating fresh milk to more than 50 ℃, adding 6% of white granulated sugar, stirring until the white granulated sugar is completely dissolved, preheating to 60-65 ℃, homogenizing under 20Mpa pressure, carrying out heat treatment for 5-8 minutes at about 90 ℃, cooling to 37 ℃, and inoculating lactobacillus paracasei K56 seed liquid with the inoculation amount of 2-3%. Fermenting at 37 deg.C for 72-96 h. Diluting the fermented product with sterilized and cooled mixture of water, syrup, emulsion stabilizer, etc. by 2-4 times, and making into directly drinkable lactobacillus beverage. The lactobacillus beverage has sour and sweet taste, good flavor, and high vitamin B content 1 Vitamin B 3 Vitamin B 6 Conjugated linoleic acid and gamma-aminobutyric acid content.

Claims (10)

1. A method for synthesizing B vitamins by fermenting Lactobacillus paracasei (Lactobacillus paracasei subsp. Paracasei), which comprises the following steps: the lactobacillus paracasei CGMCC No.15139 or DSM27447 is used for fermentation to prepare the product containingA fermentation product with B vitamins; wherein, the fermentation substrate used in the fermentation is emulsion, and the emulsion is fresh milk or reconstituted milk; the B vitamins are selected from vitamin B 1 Vitamin B 2 Vitamin B 3 Vitamin B 6 And vitamin B 9 One or more of (a).
2. The method of claim 1, further comprising: extracting purified B vitamins from the supernatant of the fermentation product.
3. The method of claim 1 wherein the B vitamins are vitamin B 1 Vitamin B 3 And/or vitamin B 6 。
4. The method of claim 1, wherein the emulsion comprises 5% to 10% sucrose.
5. The process according to claim 1 or 4, wherein the fermentation is carried out under the following conditions: fermenting for 1-7 days at 35-45 ℃.
6. The method according to claim 1 or 4, wherein the Lactobacillus paracasei is inoculated in the fermentation substrate in the form of seed liquid, in an amount of 2% to 5%.
7. The application of the lactobacillus paracasei in the preparation of the composition rich in B vitamins by fermentation is characterized in that the lactobacillus paracasei is fermented by CGMCC No.15139 or DSM27447 to prepare a fermentation product rich in B vitamins; the obtained fermentation product containing the B vitamins is used as or further used for preparing food, daily chemicals, feed or medicines; wherein, the fermentation substrate used in the fermentation is emulsion, and the emulsion is fresh milk or reconstituted milk; the B vitamins are selected from vitamin B 1 Vitamin B 2 Vitamin B 3 Vitamin B 6 And vitamin B 9 One or more of (a).
8. Use according to claim 7, wherein the food product is a fermented milk, a fermented milk drink or a milk powder.
9. Use according to claim 7, wherein the food product is a solid beverage.
10. Use according to claim 7 or 8 or 9, wherein the composition is further enriched in conjugated linoleic acid and/or gamma-aminobutyric acid, the conjugated linoleic acid and/or gamma-aminobutyric acid being produced by fermentation of the lactobacillus paracasei.
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