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A method for high-yield exopolysaccharide production by Lactobacillus plantarum

A technology of Lactobacillus plantarum and exopolysaccharide, which is applied in the field of high production of exopolysaccharide by Lactobacillus plantarum, can solve the problems of low yield, different EPS yield and performance, etc.

Active Publication Date: 2020-03-17
健码制药(广东)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, the production of microbial exopolysaccharides is still low at present, and it is necessary to further improve the production capacity of strains if it is to be put into large-scale production, and the structure and characteristics of EPS vary with changes in strains and fermentation conditions, such as carbon source and fermentation time. and medium composition, EPS yield and performance will also vary

Method used

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  • A method for high-yield exopolysaccharide production by Lactobacillus plantarum
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  • A method for high-yield exopolysaccharide production by Lactobacillus plantarum

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Embodiment 1 Fermentation and extraction and purification of exopolysaccharide of Lactobacillus plantarum La-10

[0078] Lactobacillus plantarum La-10 was inoculated in the improved MRS liquid medium at an inoculum size of 4% volume fraction, and the fermentation temperature was 30°C, and it was cultured statically for 30 hours, wherein the sucrose content in the improved MRS liquid medium was 30g / L.

[0079] Table 2 The highest extraction amount of each single factor

[0080] single factor EPS(g / L) Inoculation amount 3.1756±0.02 b

Sucrose content 2.8255±0.03 a

fermentation temperature 2.9417±0.09 a

fermentation time 2.6694±0.001 a

[0081] The composition of the improved MRS liquid medium: soybean peptone 10.0g / L, beef extract 10.0g / L, yeast extract 5.0g / L, sucrose 30g / L, K 2 PO 4 2.0g / L, sodium acetate 5.0g / L, triammonium citrate 2.0g / L, MgSO 4 ·7H 2 O is 0.58g / L, MnSO 4 4H 2 O is 0.25g / L, Tween-80 is 1.0mL / L, ...

Embodiment 2

[0092] The cholesterol-lowering qualitative method of embodiment 2 Lactobacillus plantarum La-10 exopolysaccharide

[0093] When the inoculum size was 4%, the sucrose content was 30g / L, the fermentation time was 30h, and the fermentation temperature was 30°C, La-10 was cultured statically in the improved MRS liquid medium to extract purified and freeze-dried EPS powder.

[0094] Weigh 0.8g / L of EPS solid powder into a 15mL clean centrifuge tube, and add 10mL of cholesterol standard solution (1mg / mL), and use no exopolysaccharide as a control. Under the condition of constant temperature, oscillating and adsorbing at 30°C for 15 hours, centrifuging at 10000rpm for 6 minutes, taking the supernatant, measuring the cholesterol content of the supernatant by the o-phthalaldehyde method, and using the above formula to calculate the cholesterol adsorption rate of La-10 exopolysaccharide is 41.45 %±1.3%.

[0095] Phthalaldehyde method: absorb 500 μL of supernatant, dilute the solution ...

Embodiment 3

[0097] Embodiment 3 Lactobacillus plantarum La-10 liquid static fermentation produces EPS

[0098] Lactobacillus plantarum La-10 was inoculated in the improved MRS liquid medium at an inoculum size of 5% by volume, at a fermentation temperature of 31° C., and cultured statically for 35 hours, wherein the sucrose content in the improved MRS liquid medium was 34 g / L.

[0099] MRS liquid medium composition: soybean peptone 10.0g / L, beef extract 10.0g / L, yeast extract 5.0g / L, sucrose 34g / L, K 2 PO 4 2.0g / L, sodium acetate 5.0g / L, triammonium citrate 2.0g / L, MgSO 4 ·7H 2 O is 0.58g / L, MnSO 4 4H 2 O is 0.25g / L, Tween-80 is 1.0mL / L, pH6.2~6.4.

[0100] EPS extraction method is:

[0101] (1) Centrifuge at 4°C and 8000r / min for 15min, take the supernatant and discard the bacterial precipitate.

[0102] (2) Add trichloroacetic acid (TCA) to the supernatant to a final mass fraction of 5%, and let stand overnight at 4° C. to remove proteins.

[0103] (3) Centrifuge at 4°C and 8000...

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Abstract

The invention discloses a method for high yield of exopolysaccharides from lactobacillus plantarum. EPS (Exopolysaccharides) generated from a strain used in the method under a fermentation condition has a function of efficiently reducing cholesterol, that is: 1) the fermentation condition for optimal production of exopolysaccharides is obtained through single factor optimization tests, and the yield of the exopolysaccharides is as high as 2.67-3.18g / L; 2) through comparison of the cholesterol reduction performance of the EPS before optimization and after optimization, the cholesterol reduction performance of the EPS can be remarkably improved; 3) single factor in-vitro qualitative experiment shows that the EPS has an efficient cholesterol elimination property which can be up to 59% at most; 4) tests show that the EPS which is secreted outside cells and is not extracted and purified still has a certain cholesterol elimination property. By adopting the fermentation condition and the preparation method disclosed by the invention, the microorganism EPS yield can be increased, the obtained EPS has an efficient cholesterol reduction property, and wide development and application of fat reduction medicines and healthcare products can be achieved.

Description

technical field [0001] The invention belongs to the field of exopolysaccharides of lactic acid bacteria, and in particular relates to a method for high-production of exopolysaccharides by Lactobacillus plantarum. Background technique [0002] Bacterial exopolysaccharides (Exopolysaccharides, EPS) are a class of microorganisms that use the carbon source in the culture medium to go through a series of phosphorylation, lipidation, linkage, polymerization and transportation to the outside of the cell, and then attach to the cell wall or directly penetrate into the culture medium. Among them, macromolecular polysaccharides are polymerized from oligosaccharides through lipid carriers. Among them, polysaccharides formed by one type of oligosaccharides are homopolysaccharides, otherwise they are heteropolysaccharides. The polysaccharides after extraction, purification and lyophilization are white. [0003] The physiological and biochemical functions of exopolysaccharides are mainly ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/04C12R1/25
CPCC12P19/04
Inventor 林俊芳唐灿叶志伟郭丽琼郭天芬
Owner 健码制药(广东)有限公司
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