Lactobacillus fermentum 9-4 and application thereof

A technology for fermenting lactobacillus and lactic acid bacteria, applied in the application, bacteria, dairy products and other directions, can solve the problems of difficulty in determining the content of purine substances, difficulty in ingesting purines, and unfavorable health. Fermented product preparation, effect of good tolerance

Active Publication Date: 2020-01-14
GUANGXI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, because the content of purine substances in various foods is difficult to accurately determine, it is difficult to control purine intake through strict diet
A

Method used

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  • Lactobacillus fermentum 9-4 and application thereof
  • Lactobacillus fermentum 9-4 and application thereof
  • Lactobacillus fermentum 9-4 and application thereof

Examples

Experimental program
Comparison scheme
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Example Embodiment

[0052] Example 1

[0053] Isolation, purification and preliminary screening of lactic acid bacteria strains in raw rice flour

[0054] 1. Isolation and purification of lactic acid bacteria

[0055] Weigh 2.5g of each raw rice flour sample, and dilute it with sterilized normal saline at a rate of 1:9, which is 10 -1 Dilution, take the above diluent and dilute it 10 times with normal saline, which is 10 -2 Sparsity, and so on, for serial dilutions. take 10 -1 、10 -3 、10 -5 、10 -7 Dilution, draw 100 μL respectively and spread on MRS medium, do two repetitions for each dilution, place in a 37°C incubator, cultivate for 48 hours, wait for the colonies to grow, pick the characteristic colonies of lactic acid bacteria with calcium-dissolving circles on the plate ,like figure 1 . Pick 3-4 single colonies from different plates and further purify by streaking.

[0056] A total of 35 strains were isolated from ten samples in Nanning City. The purified strains were stored in 15%...

Example Embodiment

[0077] Example 2

[0078] Molecular biology identification of primary screening lactic acid bacteria

[0079] 1. Genome Extraction

[0080] Inoculate the 8 strains that have passed the preliminary screening in the MRS medium with 1% inoculum amount, and take 3 mL of the bacterial liquid when they are cultured to the logarithmic phase. Perform genomic DNA extraction. After extraction, agarose gel electrophoresis was carried out for verification, and the verification results were as follows: Image 6 shown.

[0081] 2. PCR amplification

[0082] Quantify the extracted gene DNA sample solution using an ultra-micro-volume full-wavelength spectrophotometer, and dilute it to 1 ng / μL, and use it as a template for PCR amplification. The specific sequences of primers 27F and 1492R are respectively SEQ ID No.2 and SEQ ID No.3 in the sequence listing. Construct a PCR reaction system according to Table 5, and use a PCR instrument for amplification. The amplification program is 94°C ...

Example Embodiment

[0091] Example 3

[0092] Screening of nornucleoside strains

[0093] Nucleosides in food are precursors for the formation of purines and are the main source of purines in the body. In order to screen out the strains with the function of reducing nucleoside, the metabolic ability of resting cells to inosine and guanosine was used as an index, and the inosine, guanosine, purine, etc. were detected by Waters e2695 high-performance liquid chromatography to understand each strain Ability to metabolize nucleosides.

[0094] Prepare 2mL HPLC inosine standard sample, guanosine standard sample and inosine-guanosine reaction solution system according to Table 7 below.

[0095] Table 7 HPLC 2mL solution system

[0096]

[0097] The 7 strains screened and identified were inoculated in MRS medium with 1% inoculum. After 16 hours, 10 mL of the bacterial liquid was taken at 4°C and centrifuged at 12,000 r / min for 3 minutes to collect the bacterial cells. The cells were washed with 1...

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Abstract

The invention discloses Lactobacillus fermentum 9-4 and application thereof. The strain is collected in China Center for Type Culture Collection (CCTCC) on August 12, 2019, with the collection numberof CCTCC NO:M 2019619. The Lactobacillus fermentum is a lactobacillus strain which is screened from traditional food raw-pressed rice flour in a Guangxi Zhuang nationality region and has purine reducing capacity, the strain grows well on an MRS agar culture medium, has high acid resistance and has survival rate of 34% after 3 h under the condition that pH is 1.5; and the strain can grow slowly incholate with concentration being 3% and has the survival rate of 65%. The strain has good acid resistance, cholate resistance and purine reducing capacity and is applicable to production of low-purinefood and fermented food with the purine reducing function.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to the field of development of functional lactic acid bacteria and production and development of products thereof. Specifically, it is a lactobacillus fermentum Lactobacillus fermentum 9-4 and an application thereof. Background technique [0002] Gout is a joint disease caused by urate deposition, which is directly related to hyperuricemia caused by purine metabolism disorder or decreased uric acid excretion. Gout patients often experience severe joint pain at night, and the pain lasts for several days or weeks, which can easily lead to redness, swelling and inflammation of the joints. After joint edema, the tissue becomes soft, which limits activities and eventually affects daily life. Gout is often accompanied by kidney disease, hyperlipidemia, hypertension, diabetes, arteriosclerosis and coronary heart disease. Therefore, gout is a metabolic disease that seriously endang...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/04A23L33/135A23C9/123C12R1/225
CPCA23C9/123A23L33/135C12N1/04C12N1/20C12N1/205C12R2001/225
Inventor 王成华卢良华刘田甜张冉朱春燕
Owner GUANGXI UNIV
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