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Lactobacillus fermentum and application thereof in preparation of conjugated fatty acid

A technology for fermenting lactobacillus and fatty acid, applied in the biological field, can solve the problems of far different isomer content, complex oil composition, low yield and the like

Active Publication Date: 2020-09-18
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, conjugated linoleic acid is mainly derived from natural conjugated linoleic acid and artificially synthesized conjugated linoleic acid, because natural conjugated linoleic acid mainly exists in milk fat and meat products of rumen animals such as cattle and sheep Among them, the CLA content per gram of milk fat ranges from 2 mg to 25 mg, and the CLA content increases with the age of dairy cows, and its sources are limited, so it is not suitable for large-scale industrial production
However, in the current method of artificially synthesizing CLA, the content of each isomer in the obtained product is very different because of its different raw materials and synthetic methods.
[0005] Since natural conjugated linolenic acid exists in some plant seeds in nature, such as pomegranate seeds, tung seeds, bitter gourd seeds, calendula seeds, Trichosanthes and Jacaranda seeds, etc.; however, many plants containing conjugated linolenic acid Among plant seeds, only Trichosanthes can be eaten directly, and the oil composition in plant seeds is very complex, so it is very difficult to separate and purify conjugated linolenic acid from plant seed oils
Although conjugated linolenic acid can also be produced by isomerization of linolenic acid by alkali treatment, its yield is low and there are reagent residues, so the industrial production of conjugated linolenic acid has not been realized so far.

Method used

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  • Lactobacillus fermentum and application thereof in preparation of conjugated fatty acid
  • Lactobacillus fermentum and application thereof in preparation of conjugated fatty acid
  • Lactobacillus fermentum and application thereof in preparation of conjugated fatty acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Collection of samples and isolation and identification of Lactobacillus

[0042] 1. Sample collection and separation

[0043] (1) The samples were collected from the feces of a 90-year-old man in Rugao City, Nantong City, Jiangsu Province.

[0044] (2) Take 1g of feces sample, spread the sample on MRS solid medium after serial dilution, place it in an anaerobic environment, and culture it at 37°C for 72 hours, observe and record the colony shape; Cultivate in liquid medium at 37°C for 48 hours, perform Gram staining on the obtained colonies and record the strain morphology; discard the Gram-negative strains and Gram-positive cocci in the colonies, and select the Gram-positive bacilli; After the obtained Gram-positive bacilli were analyzed by catalase, the catalase-positive strains were discarded, and the catalase-negative strains were retained; the resulting strain was identified as Lactobacillus fermentum through 16S rDNA sequencing, and named as Lactobacil...

Embodiment 2

[0060] Embodiment 2: the preparation of the microbial preparation containing lactobacillus fermentum (Lactobacillus fermentum) CCFM1116

[0061] The glycerin tube containing Lactobacillus fermentum CCFM1116 was taken out from the -80°C refrigerator, and 200-600 μL of Lactobacillus fermentum CCFM1116 bacterial liquid was inoculated into 10-30 mL of MRS liquid medium. Under anaerobic environment, 37 Activate 2 to 3 generations at ℃ until Lactobacillus fermentum (Lactobacillusfermentum) CCFM1116 reaches 1×10 8 When the number of viable bacteria is above cfu / mL, centrifuge at 5000-10000rpm for 10-20min, remove the supernatant, and then add buffer (normal saline or 0.2M phosphate buffer with a pH value of 7) and Cryoprotectant (15%-20% (w / v) sucrose solution), the concentration of the cells should not be lower than 1×10 9 cfu / mL, vacuum freeze-drying to obtain solid bacterial agent.

Embodiment 3

[0062] Example 3: Application of Lactobacillus fermentum CCFM1116 in the preparation of conjugated linoleic acid

[0063] Specific steps are as follows:

[0064] (1) Strain activation

[0065] Take out the glycerin tube containing Lactobacillus fermentum (Lactobacillus fermentum) CCFM1116 from the -80°C refrigerator, take out the bacterial solution and streak it on the MRS solid medium, and culture it at 37°C for 48h in an anaerobic environment; pick the grown single colony and inoculate it on In the MRS liquid medium, cultured at 37° C. for 48 hours under anaerobic environment, and continuously activated for 3 generations to obtain the bacterial liquid.

[0066] (2) Preparation of linoleic acid mother liquor

[0067] Weigh 300mg of linoleic acid (LA) and 200mg of Tween-80, dissolve them in water and dilute to 10mL, stir and emulsify fully, filter and sterilize through a 0.45μm sterile filter membrane to obtain linoleic acid with a concentration of 0.48mg / mL Acid mother liq...

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Abstract

The invention relates to Lactobacillus fermentum and application thereof in preparation of conjugated fatty acid, and belongs to the technical field of biology. The invention provides Lactobacillus fermentum CCFM1116 capable of producing conjugated linoleic acid and conjugated linolenic acid. The conversion rate of converting linoleic acid into conjugated linoleic acid by fermentation supernatantcan reach 26.19%, wherein c9, t11-CLA isomer accounts for 63.65%; and the conversion rate of converting linolenic acid into conjugated linolenic acid by fermentation supernatant can reach 41.79%, wherein c9, t11 and c15-CLANA accounts for 95.01%.

Description

technical field [0001] The invention relates to a lactobacillus fermentum and its application in the preparation of conjugated fatty acids, belonging to the field of biotechnology. Background technique [0002] Conjugated linoleic acid (CLA) is the general term for octadecadienoic acid containing conjugated double bonds, and is the positional isomer and geometric isomer of linoleic acid (Linoleic acid, 18:2) . The most common isomer is cis 9, trans 11-CLA (c9, t11-CLA), also known as rumenic acid. In addition, trans 10, cis 12-CLA (t10, c12-CLA) is also an isomer with relatively high content in nature. Conjugated linoleic acid has attracted attention because of its biological functions. Different conjugated linoleic acid isomers have different physiological functions, among which c9,t11-CLA (CLA1) and t10,c12-CLA are recognized as the most physiological The main functions of the active conjugated linoleic acid isomers, c9, t11-CLA are anti-cancer, anti-inflammation and im...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P7/64C12R1/225
CPCC12N1/20C12P7/6427C12R2001/225C12N1/205
Inventor 陈卫杨波夏嘉祎王欣陈科学沈慧敏嵇熠彬赵建新张灏
Owner JIANGNAN UNIV
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