CN106635912A - United fermentation process for compound lactic acid bacteria - Google Patents
United fermentation process for compound lactic acid bacteria Download PDFInfo
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- CN106635912A CN106635912A CN201611253562.2A CN201611253562A CN106635912A CN 106635912 A CN106635912 A CN 106635912A CN 201611253562 A CN201611253562 A CN 201611253562A CN 106635912 A CN106635912 A CN 106635912A
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Abstract
The invention provides a united fermentation process for compound lactic acid bacteria. The process comprises the steps that a seed culture medium is adopted to conduct separated culture on the four lactic acid bacteria including lactobacillus acidophilus, pediococcus acidilactici, enterococcus faecium and enterococcus faecalis for 10-25 hours, and a lactobacillus acidophilus seed solution, a pediococcus acidilactici seed solution, an enterococcus faecium seed solution and an enterococcus faecalis seed solution are prepared; 1%-2% of lactobacillus acidophilus seed solution, 1%-2% of pediococcus acidilactici seed solution, 1%-2% of enterococcus faecium seed solution and1%-2% of enterococcus faecalis seed solution are respectively weighed by mass and are transferred to a fermenting culture medium, united fermentation is performed under the conditions of the temperature of 37+/-2 DEG C, the pH value of 6+/-0.5 and the stirring speed of 80-200 r/min for 12-24 hours to obtain a compound lactic acid bacterium fermentation product. The compound lactic acid bacterium fermentation product prepared by the united fermentation process for compound lactic acid bacteria is good in stability, simple in process and convenient to use and has a wide application prospect.
Description
Technical field
The present invention relates to micro-ecology fermentation field, in particular it relates to a kind of combined ferment technique of compound lactobacillus.
Background technology
Lactic acid bacteria(Lactic acid bacteria, LAB)It is that a class can be produced in a large number using fermentable carbohydrate
The common name of the bacterium of lactic acid.It is a kind of probio being present in mankind's body.Species is enriched very much, common are Bifidobacterium,
Lactobacillus acidophilus, Pediococcus acidilactici, enterococcus faecalis, VREF etc..Lactic acid bacteria has splendid health-care effect to grind in a large number to human body
Study carefully and show, lactic acid bacteria can adjust body gastrointestinal tract normal flora, keep microecological balance, improve food digestion rate and biology
Valency, reduce serum cholesterol controls endotoxin, suppresses the generation of spoilage organisms growth and breeding and spoilage product in enteron aisle, manufacture battalion
Foster material, stimulate tissue development, so as to the nutritional status of body, physiological function, cell infection, drug effect, toxic reaction,
The generation effects such as immune response, tumour generation, aging course and unexpected emergency reaction.
But common lactic acid bacteria, vigor is extremely weak, they can only survive in relatively limited environment, existing lactic acid
In bacterium product, lactic acid bacteria bacterium work number is not generally high, and preservation condition is harsh, and stability is poor, therefore develops high bacterium work number, stablizes
The good lactic fermentation process of property is very necessary.
The content of the invention
In view of this, the present invention provides a kind of combined ferment technique of compound lactobacillus, to solve the above problems.
Specifically, the present invention is adopted the following technical scheme that:
A kind of combined ferment technique of compound lactobacillus, comprises the following steps:
Seed liquor is prepared using seed culture medium respectively to lactobacillus acidophilus, Pediococcus acidilactici, VREF, enterococcus faecalis four
Planting lactic acid bacteria carries out individually cultivating 10~25 h respectively, and lactobacillus acidophilus seed liquor, Pediococcus acidilactici seed liquor, dung intestines ball is obtained
Bacterium seed liquor and enterococcus faecalis seed liquor;
Combined ferment takes respectively the lactobacillus acidophilus seed liquor that mass fraction is 1%~2%, the Pediococcus acidilactici seed
Liquid, the VREF seed liquor and the enterococcus faecalis seed liquor are gone in fermentation medium, are 37 ± 2 DEG C, pH in temperature
It is under conditions of 80~200 r/min, to carry out h~24 h of combined ferment 12 to be worth for 6 ± 0.5, mixing speed, obtains Composite Milk
Acid bacteria fermentation thing;
Wherein, the seed culture medium includes following components:The g/L of glucose 5~20, the g/L of yeast extract 5~20, peptone 1~
15 g/L, the g/L of diammonium hydrogen citrate 1~10, the g/L of dipotassium hydrogen phosphate 1~2.5, the g/L of magnesium sulfate 0.01~0.34, manganese sulfate
0.01~0.1 g/L, the g/L of sodium acetate 1~9, the ml/L of Tween-80 0.5~2, solvent is sterilized water;
The fermentation medium includes following components:30~50g/L of glucose, the g/L of yeast extract 1~45, the g/ of peptone 5~35
L, 0.3~2.5g/L of diammonium hydrogen citrate, 1~8g/L of dipotassium hydrogen phosphate, the g/L of magnesium sulfate 0.1~2, manganese sulfate 0.05~1.5
G/L, the g/L of sodium acetate 0.1~6, the ml/L of Tween-80 0.5~5, solvent is sterilized water.
Based on above-mentioned, the seed culture medium includes following components:The g/L of glucose 10~15, the g/ of yeast extract 10~15
L, the g/L of peptone 5~10, the g/L of diammonium hydrogen citrate 3~7, the g/L of dipotassium hydrogen phosphate 1.5~2.0, magnesium sulfate 0.22~
0.30 g/L, the g/L of manganese sulfate 0.44~0.62, the g/L of sodium acetate 3~7, the ml/L of Tween-80 1.0~1.5, solvent is aseptic
Water;
The fermentation medium includes following components:The g/L of glucose 35~45, the g/L of yeast extract 15~25, peptone 15~25
G/L, 1.0~2.0g/L of diammonium hydrogen citrate, the g/L of dipotassium hydrogen phosphate 3~6, the g/L of magnesium sulfate 1.0~1.5, manganese sulfate 0.75
~1.35 g/L, the g/L of sodium acetate 2.1~4.2, the ml/L of Tween-80 1.0~3.0, solvent is sterilized water.
Compared with prior art, the combined ferment technique of the compound lactobacillus that the present invention is provided, is matched somebody with somebody using two-step method and is closed
Seed culture medium and above-mentioned fermentation medium are stated, while to lactobacillus acidophilus, Pediococcus acidilactici, VREF, enterococcus faecalis four
Planting lactic acid bacteria carries out combined ferment, and these four lactic acid bacterias act synergistically, mutually promote, and greatly improves obtained compound lactobacillus
In bacterium live number and stability;It is easy to operate with wide and the combined ferment technology process of the compound lactobacillus is simple
Application prospect.
Specific embodiment
Below by specific embodiment, technical scheme is described in further detail.
Embodiment 1
A kind of combined ferment technique of compound lactobacillus, comprises the following steps:
Seed liquor is prepared using seed culture medium respectively to lactobacillus acidophilus, Pediococcus acidilactici, VREF, enterococcus faecalis four
Planting lactic acid bacteria carries out individually cultivating 10~25 h respectively, and lactobacillus acidophilus seed liquor, Pediococcus acidilactici seed liquor, dung intestines ball is obtained
Bacterium seed liquor and enterococcus faecalis seed liquor;
Combined ferment takes respectively the lactobacillus acidophilus seed liquor that mass fraction is 1%, the Pediococcus acidilactici seed liquor, institute
State VREF seed liquor and the enterococcus faecalis seed liquor gone in fermentation medium, temperature be 37 ± 2 DEG C, pH value be 6
± 0.5, mixing speed is under conditions of 80~200 r/min, to carry out combined ferment, obtains compound lactobacillus-fermencucumber thing;
Wherein, the seed culture medium includes following components:The g/L of glucose 10, the g/L of yeast extract 10, the g/L of peptone 7, lemon
The g/L of two ammonium of lemon acid hydrogen 5, the g/L of dipotassium hydrogen phosphate 2.0, the g/L of magnesium sulfate 0.22, the g/L of manganese sulfate 0.52, the g/ of sodium acetate 4.5
L, the ml/L of Tween-80 1.2, solvent is sterilized water;
The fermentation medium includes following components:The g/L of glucose 40, the g/L of yeast extract 22, the g/L of peptone 18, citric acid
The g/L of two ammonium of hydrogen 1.4, the g/L of dipotassium hydrogen phosphate 4, the g/L of magnesium sulfate 1.2, the g/L of manganese sulfate 0.58, the g/L of sodium acetate 3.2, tell
- 80 2.6 ml/L of temperature, solvent is sterilized water.
Contrast experiment
Control group:Temperature be 37 ± 2 DEG C, pH be 6 ± 0.5, mixing speed be 80~200 r/min under the conditions of, using tradition
Lactic acid bacteria culturing medium be MRS medium culture lactobacillus acidophilus, obtain bacillus acidophilus fermented product.
The compound lactobacillus-fermencucumber thing and the bacillus acidophilus fermented product for taking the offer of embodiment 1 of equivalent continues in temperature
For 37 ± 2 DEG C, pH be 6 ± 0.5, mixing speed be to be fermented under the conditions of 80~200 r/min, determine respectively its bacterium live number with
Fermentation time changes, and test result is as shown in table 1;The compound lactobacillus-fermencucumber of the h of fermentation 24 of the offer of embodiment 1 of equivalent is provided
Bacillus acidophilus fermented product's normal temperature of 24 h of thing and fermentation is preserved, and its bacterium work number is determined respectively and is changed over, test knot
Fruit is as shown in table 2.
The bacterium of table 1 lives number with fermentation time change
Bacterium work number is changed under the normal temperature of table 2
Note:" --- " in table 2 represents zymotic fluid viable count seldom without measure meaning
As shown in Table 1, bacterium lives number as fermentation time increases in experimental group and control group, first increases, and then reduces, in 24 h
When, reach maximum;And the bacterium in experimental group lives number much larger than control group;As shown in Table 2, normal temperature is preserved, and bacterium lives in control group
Number is rapid to be reduced, and bacterium work number still keeps certain numerical value in experimental group.
Embodiment 2
The present embodiment provides a kind of combined ferment technique of compound lactobacillus, and it is with the difference of embodiment 1:
The seed culture medium includes following components:The g/L of glucose 5, the g/L of yeast extract 20, the g/L of peptone 15, hydrogen citrate
The g/L of two ammonium 10, the g/L of dipotassium hydrogen phosphate 2.5, the g/L of magnesium sulfate 0.01, the g/L of manganese sulfate 0.01, the g/L of sodium acetate 9, tween-
80 2 ml/L, solvent is sterilized water;
The fermentation medium includes following components:The g/L of glucose 30, the g/L of yeast extract 1, the g/L of peptone 35, hydrogen citrate
The g/L of two ammonium 0.3, the g/L of dipotassium hydrogen phosphate 1, the g/L of magnesium sulfate 2, the g/L of manganese sulfate 1.5, the g/L of sodium acetate 0.1, Tween-80 5
Ml/L, solvent is sterilized water;
In the step of the combined ferment, the lactobacillus acidophilus seed liquor, the lactic acid that mass fraction is 2% are taken respectively
Piece coccus seed liquor, the VREF seed liquor and the enterococcus faecalis seed liquor are gone in fermentation medium, are in temperature
37 ± 2 DEG C, pH value be 6 ± 0.5, mixing speed be 80~200 r/min under conditions of, carry out the h of combined ferment 12, answered
Close lactobacillus fermentation;
The compound lactobacillus-fermencucumber thing carries out normal temperature preservation, and after 30 d, bacterium lives number for 1.42 × 108;After 60 d, bacterium work number is
9.13×106;After 90 d, bacterium lives number for 5.04 × 105。
Embodiment 3
The present embodiment provides a kind of combined ferment technique of compound lactobacillus, and it is with the difference of embodiment 1:
The seed culture medium includes following components:The g/L of glucose 20, the g/L of yeast extract 5, the g/L of peptone 1, hydrogen citrate
The g/L of two ammonium 11, the g/L of dipotassium hydrogen phosphate 1, the g/L of magnesium sulfate 0.1, the g/L of manganese sulfate 1, the g/L of sodium acetate 1, Tween-80 0.5
Ml/L, solvent is sterilized water;
The fermentation medium includes following components:The g/L of glucose 50, the g/L of yeast extract 45, the g/L of peptone 5, hydrogen citrate
The g/L of two ammonium 2.5, the g/L of dipotassium hydrogen phosphate 8, the g/L of magnesium sulfate 0.1, the g/L of manganese sulfate 0.05, the g/L of sodium acetate 6, tween -8
0.5 ml/L, solvent is sterilized water;
In the step of the combined ferment, the lactobacillus acidophilus seed liquor, the lactic acid that mass fraction is 1% are taken respectively
Piece coccus seed liquor, the VREF seed liquor and the enterococcus faecalis seed liquor are gone in fermentation medium, are in temperature
37 ± 2 DEG C, pH value be 6 ± 0.5, mixing speed be 80~200 r/min under conditions of, carry out the h of combined ferment 24, answered
Close lactobacillus fermentation;
The compound lactobacillus-fermencucumber thing carries out normal temperature preservation, and after 30 d, bacterium lives number for 1.52 × 108;After 60 d, bacterium work number is
9.01×106;After 90 d, bacterium lives number for 4.96 × 105。
Embodiment 4
The present embodiment provides a kind of combined ferment technique of compound lactobacillus, and it is with the difference of embodiment 1:The kind
Sub- culture medium includes following components:The g/L of glucose 13, the g/L of yeast extract 11, the g/L of peptone 5, the g/L of diammonium hydrogen citrate 7,
The g/L of dipotassium hydrogen phosphate 2.0, the g/L of magnesium sulfate 0.22, the g/L of manganese sulfate 0.62, the g/L of sodium acetate 3, the ml/L of Tween-80 1.0,
Solvent is sterilized water;
The fermentation medium includes following components:The g/L of glucose 35, the g/L of yeast extract 15, the g/L of peptone 25, citric acid
The g/L of two ammonium of hydrogen 1.0, the g/L of dipotassium hydrogen phosphate 3, the g/L of magnesium sulfate 1.5, the g/L of manganese sulfate 0.75, the g/L of sodium acetate 4.2, tell
- 8 1.2 ml/L of temperature, solvent is sterilized water;
In the step of the combined ferment, the lactobacillus acidophilus seed liquor, the lactic acid that mass fraction is 2% are taken respectively
Piece coccus seed liquor, the VREF seed liquor and the enterococcus faecalis seed liquor are gone in fermentation medium, are in temperature
37 ± 2 DEG C, pH value be 6 ± 0.5, mixing speed be 80~200 r/min under conditions of, carry out the h of combined ferment 12, answered
Close lactobacillus fermentation;
The compound lactobacillus-fermencucumber thing carries out normal temperature preservation, and after 30 d, bacterium lives number for 1.55 × 108;After 60 d, bacterium work number is
9.17×106;After 90 d, bacterium lives number for 5.12 × 105。
Therefore, the combined ferment technique of above-mentioned compound lactobacillus provided in an embodiment of the present invention can be effectively improved and is obtained again
The bacterium work number in lactic acid bacteria is closed, and bacterium work number has good stability, and has broad application prospects.
Finally it should be noted that:Above example is only to illustrate technical scheme rather than a limitation;To the greatest extent
Pipe has been described in detail with reference to preferred embodiment to the present invention, and those of ordinary skill in the art should be understood:Still
The specific embodiment of the present invention can be modified or equivalent is carried out to some technical characteristics;Without deviating from this
The spirit of bright technical scheme, it all should cover in the middle of the technical scheme scope being claimed in the present invention.
Claims (2)
1. a kind of combined ferment technique of compound lactobacillus, comprises the following steps:
Seed liquor is prepared using seed culture medium respectively to lactobacillus acidophilus, Pediococcus acidilactici, VREF, enterococcus faecalis four
Planting lactic acid bacteria carries out individually cultivating 10~25 h respectively, and lactobacillus acidophilus seed liquor, Pediococcus acidilactici seed liquor, dung intestines ball is obtained
Bacterium seed liquor and enterococcus faecalis seed liquor;
Combined ferment takes respectively the lactobacillus acidophilus seed liquor that mass fraction is 1%~2%, the Pediococcus acidilactici seed
Liquid, the VREF seed liquor and the enterococcus faecalis seed liquor are gone in fermentation medium, are 37 ± 2 DEG C, pH in temperature
It is under conditions of 80~200 r/min, to carry out h~24 h of combined ferment 12 to be worth for 6 ± 0.5, mixing speed, obtains Composite Milk
Acid bacteria fermentation thing;
Wherein, the seed culture medium includes following components:The g/L of glucose 5~20, the g/L of yeast extract 5~20, peptone 1~
15 g/L, the g/L of diammonium hydrogen citrate 1~10, the g/L of dipotassium hydrogen phosphate 1~2.5, the g/L of magnesium sulfate 0.01~0.34, manganese sulfate
0.01~0.1 g/L, the g/L of sodium acetate 1~9, the ml/L of Tween-80 0.5~2, solvent is sterilized water;
The fermentation medium includes following components:30~50g/L of glucose, the g/L of yeast extract 1~45, the g/ of peptone 5~35
L, 0.3~2.5g/L of diammonium hydrogen citrate, 1~8g/L of dipotassium hydrogen phosphate, the g/L of magnesium sulfate 0.1~2, manganese sulfate 0.05~1.5
G/L, the g/L of sodium acetate 0.1~6, the ml/L of Tween-80 0.5~5, solvent is sterilized water.
2. the combined ferment technique of compound lactobacillus according to claim 1, it is characterised in that the seed culture medium
Including following components:The g/L of glucose 10~15, the g/L of yeast extract 10~15, the g/L of peptone 5~10, diammonium hydrogen citrate 3
~7 g/L, the g/L of dipotassium hydrogen phosphate 1.5~2.0, the g/L of magnesium sulfate 0.22~0.30, the g/L of manganese sulfate 0.44~0.62, acetic acid
The g/L of sodium 3~7, the ml/L of Tween-80 1.0~1.5, solvent is sterilized water;
The fermentation medium includes following components:The g/L of glucose 35~45, the g/L of yeast extract 15~25, peptone 15~25
G/L, 1.0~2.0g/L of diammonium hydrogen citrate, the g/L of dipotassium hydrogen phosphate 3~6, the g/L of magnesium sulfate 1.0~1.5, manganese sulfate 0.75
~1.35 g/L, the g/L of sodium acetate 2.1~4.2, the ml/L of Tween-80 1.0~3.0, solvent is sterilized water.
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Cited By (6)
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CN111728081A (en) * | 2020-04-03 | 2020-10-02 | 河南金百合生物科技股份有限公司 | Compound bacterium fermentation liquor for feed additive and preparation method thereof |
CN111961631A (en) * | 2020-09-04 | 2020-11-20 | 国科智盛(沁阳市)生物科技有限公司 | Fermentation production process of lactobacillus fermentum |
CN112175851A (en) * | 2019-07-01 | 2021-01-05 | 中国科学院上海有机化学研究所湖州生物制造创新中心 | High-density fermentation of lactobacillus mixture and preparation of lactobacillus composite bacteria preparation |
CN115354005A (en) * | 2022-09-26 | 2022-11-18 | 福建益昕葆生物制药有限公司 | Compound microbial preparation and preparation method and storage method thereof |
CN116585255A (en) * | 2023-04-24 | 2023-08-15 | 广州领衔生物科技有限公司 | Composition containing lactobacillus fermentation product and preparation method and application thereof |
WO2023226430A1 (en) * | 2022-05-25 | 2023-11-30 | 江苏大学 | Method for preparing shrimp paste on basis of fast fermentation strengthened with combined strains |
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CN112175851A (en) * | 2019-07-01 | 2021-01-05 | 中国科学院上海有机化学研究所湖州生物制造创新中心 | High-density fermentation of lactobacillus mixture and preparation of lactobacillus composite bacteria preparation |
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CN115354005A (en) * | 2022-09-26 | 2022-11-18 | 福建益昕葆生物制药有限公司 | Compound microbial preparation and preparation method and storage method thereof |
CN116585255A (en) * | 2023-04-24 | 2023-08-15 | 广州领衔生物科技有限公司 | Composition containing lactobacillus fermentation product and preparation method and application thereof |
CN116585255B (en) * | 2023-04-24 | 2024-05-03 | 广州领衔生物科技有限公司 | Composition containing lactobacillus fermentation product and preparation method and application thereof |
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Application publication date: 20170510 |
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RJ01 | Rejection of invention patent application after publication |