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Lactobacillus rhamnosus culture medium and culture method

A technology of Lactobacillus rhamnosus and culture method, which is applied in the field of culture medium and culture of Lactobacillus rhamnosus, can solve the problems of few studies on the proliferation and culture of strains, achieve cost reduction, increase the number of viable bacteria, and facilitate enrichment Effect

Active Publication Date: 2016-12-21
江西仁仁健康微生态科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, most of the studies on Lactobacillus rhamnosus focus on its functional characteristics, and there are not many studies on the propagation and cultivation of this strain

Method used

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  • Lactobacillus rhamnosus culture medium and culture method

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0042] 1. Preparation of strains for fermentation

[0043] 1) streak culture: after the Lactobacillus rhamnosus bacteria powder is diluted with sterile water, streak four areas on the MC medium, after the streak is completed, the plate is inverted, and cultured at a constant temperature of 37 ° C in an incubator for 48 to 72 hours;

[0044] 2) Primary purification culture: pick a single colony with a large calcium-dissolving circle on the plate and inoculate it into a test tube containing 5 ml of MRS liquid medium, seal the test tube, and culture at a constant temperature in a 37°C incubator for 17 to 24 hours;

[0045] 3) Secondary purification culture: according to the inoculum amount of 3 to 5%, inoculate the cultured primary bacterial suspension into the triangular flask containing the MRS liquid medium, and the volume of the medium filling accounts for 40% of the volume of the triangular flask, The triangular flask is sealed, and the incubator is kept at a constant temperat...

Embodiment 1

[0056] Embodiment 1 Lactobacillus rhamnosus culture medium and culture method

[0057] 1. Lactobacillus rhamnosus medium, including basal medium and optimized medium.

[0058] Formulation and preparation of basal medium: yeast peptone 8.0g / L, glucose 25.0g / L, yeast extract 3.0g / L, sodium acetate 6.0g / L, magnesium sulfate 0.5g / L, Tween 80 0.5g / L , Isomalt oligosaccharide 1.0g / L, potassium dihydrogen phosphate 3.0g / L, the balance is sterile water, pH value 6.20; weigh each component according to the formula ratio, mix, heat and dissolve, use 1mol / L food grade The pH value of the medium was adjusted to 6.20 with NaOH solution, and sterilized at 115 °C for 30 min.

[0059] The formulation and preparation of the optimized medium: yeast peptone 14.0g / L, glucose 30.0g / L, yeast extract 6.0g / L, sodium acetate 3.0g / L, magnesium sulfate 0.10g / L, Tween 80 0.5g / L , isomalt oligosaccharide 2.0g / L, potassium dihydrogen phosphate 3.0g / L, the balance is sterile water, pH value 6.80; weigh ea...

Embodiment 2

[0076] Embodiment 2 is used for the selection experiment of the strain preservation mode of fermentation

[0077] After Lactobacillus rhamnosus is cultured by streaking, primary and secondary purification, the bacterial suspension is frozen and stored in the slant, bacterial suspension and the preservation mode of the centrifuged bacteria mud in Example 1, and then the three The strains frozen in different preservation methods were fermented under the same culture conditions, and the viable count and yield of the bacterial suspension were determined respectively after the fermentation.

[0078] Determination method of the number of viable bacteria: plate counting method;

[0079] Yield determination method:

[0080]

[0081] Table 1 The effect of fermentation on yield and viable count of bacteria using different preservation methods

[0082] storage method

[0083] From the data in Table 1, it is shown that the bacterial species preserved by the centrifugal bact...

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Abstract

The invention discloses a lactobacillus rhamnosus culture medium. The lactobacillus rhamnosus culture medium and culture method has a basic culture medium and an optimized culture medium, wherein the basic culture medium is prepared from the following components: yeast peptone, glucose, yeast extract, sodium acetate, magnesium sulfate, tween 80, isomaltooligosaccharide, potassium dihydrogen phosphate and the balance of sterile water, and the pH value is 6.20 to 6.80; and the optimized culture medium is prepared from the following components: yeast peptone, glucose, yeast extract, sodium acetate, magnesium sulfate, tween 80, isomaltooligosaccharide, potassium dihydrogen phosphate and the balance the sterile water, and the pH value is 6.20 to 6.80. The culture medium is suitable for proliferation culture of lactobacillus rhamnosus. By utilizing the method for culturing the lactobacillus rhamnosus by virtue of the culture medium, not only can the culture of the lactobacillus rhamnosus be effectively proliferated, but also the industrialized mass production can be facilitated.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a Lactobacillus rhamnosus culture medium and a culture method. Background technique [0002] Lactobacillus rhamnosus belongs to the genus Lactobacillus, is a Gram-positive facultative anaerobic bacterium without plasmids; it cannot utilize lactose, but can metabolize monosaccharides; it can grow well under anaerobic conditions, and it can grow well in the presence of CO 2 Can grow even if it exists. [0003] Lactobacillus rhamnosus is one of the normal flora of the human body, with high intestinal adhesion rate and strong colonization ability. It is one of the most widely studied probiotics in human beings. Bifidobacteria and Lactobacillus acidophilus grow and function, prevent and help treat diarrhea, prevent respiratory infections, expel toxins, prevent dental caries, prevent allergies, etc. [0004] At present, most of the research on Lactobacillus rhamnosus focuses ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/225
Inventor 余萍张春宇闵祥博
Owner 江西仁仁健康微生态科技有限公司
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