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Genetically engineering lactobacillus and application thereof

A technology of Lactobacillus and preservation number, applied in the field of mutant Lactobacillus

Active Publication Date: 2014-08-27
THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the treatment of this part of the patients, the curative effect is recognized to be the use of angiotensin converting enzyme inhibitors (Angiotensin converting enzyme inhibitors, ACEI) and angiotensin II receptor antagonists (Angiotensin II receptor antagonists, ARB). The drug mainly delays the deterioration of renal function by reducing the high pressure in the glomerulus, high perfusion and high filtration, but ignores the damage and functional deterioration of all tissue cells including the kidney caused by the metabolites that have accumulated in the body

Method used

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  • Genetically engineering lactobacillus and application thereof
  • Genetically engineering lactobacillus and application thereof
  • Genetically engineering lactobacillus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] 1. Directional induction of Lactobacillus producing multiple uremic toxin-decomposing enzymes: inoculate activated L.B. in the serum of uremia patients at 4% (v / v), shake and mix well, and culture anaerobically for 24 hours to form the first generation. After mixing, take 1% and place it in the serum of a new uremia patient. After shaking and mixing, culture it anaerobically at 37° C. for 24 hours, which is the second generation. After repeated subculture to 25 generations, the bacteria were taken out to detect the ability of the bacteria to decompose creatinine and urea.

[0041] 2. Double and multiple mutagenesis of Lactobacillus producing multiple uremic toxin-decomposing enzymes

[0042] After the directional induction, the L.B inducer strain with the strongest ability to decompose urea toxin is selected as the starting strain, and double mutagenesis is carried out according to the mutagenesis scheme described in claim 4.

[0043] 1) Multiple UV mutagenesis: take t...

Embodiment 2

[0048] Determination of the ability of bacteria to decompose creatinine and urea: Activate the L.B obtained in Example 1 to make a bacterial solution, inoculate 4% in the MRS broth medium and mix well, then culture anaerobically at 37°C until the logarithmic phase Take it out, wash it repeatedly with sterile normal saline, centrifuge for 3 times, resuspend in normal saline, adjust the OD625 of the suspension in each tube to 0.08-0.1 by McFarland turbidimetry, about 1.5×108cfu / ml, take 200ul suspension bacteria each The solution was added to 800ul serum of uremic patients, cultured anaerobically at 37°C, taken out at 24h and 48h respectively, centrifuged at 40000rmp / min for 5min, and the supernatant was taken to detect the concentration of creatinine and urea nitrogen. Take 200ul of normal saline and add it to the above culture medium as a control group, and set 5 parallel tubes in each group. Jaffe's method and UV-GLDH method were used before and after cultivation, and the con...

Embodiment 3

[0072] Observation on the curative effect of mutagenic strains in the treatment of experimental renal failure

[0073] Nephrectomy model establishment: 30 male 6-week-old SD rats, weighing 170-190g, were fed adaptively for 1 week, and the feed supply at the beginning of the experiment was determined according to the feed consumption during the week. Eight rats were randomly selected as the sham operation group, and the remaining 32 rats underwent 5 / 6 nephrectomy. Preparation method of 5 / 6 nephrectomy model: ① The first step is 2 / 3 left nephrectomy: after weighing, anesthetize with 0.35mL / 100g of 10% chloral hydrate, disinfect the skin in the middle of the lower abdomen, and draw a 1mL skin test needle Pierce the abdominal wall vertically, withdraw blood and urine, that is, inject. After successful anesthesia, the limbs were fixed on the operating board in prone position. First touch the left costal ridge angle, then make an upward and downward incision at the center of 1 cm ...

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Abstract

The invention discloses a novel genetically engineering lactobacillus which is capable of decomposing a plurality of uremic toxins and is obtained by physical-chemical mutation breeding, characterized in that: the lactobacillus is obtained by carrying out directional induction on blood serums of uremic patients, and modifying the gene of the lactobacillus by dual repeated induction. The genetically engineering lactobacillus disclosed herein can be prepared into biological preparations for oral administration to plant the genetically engineering lactobacillus on intestinal mucosa for continuously decomposing uremic toxins, and the genetically engineering lactobacillus disclosed herein is used for treating uremia or chronic renal failure. Lactobacillus which is human intestinal probiotics and is planted on the inner layer of the intestinal mucosa has the functions of enhancing immunity, inhibiting pathogenic bacteria, synthesizing a plurality of vitamins, reducing cholesterol, etc. According to the invention, based on reducing a plurality of uremic toxins, the genetically engineering lactobacillus disclosed herein makes up the deficiencies of low immunity, vitamin deficiency, and fat metabolism disorder of people with chronic renal failure, and the invention provides chronic renal failure with a biological preparation for comprehensive treatment.

Description

technical field [0001] The invention relates to a mutated lactobacillus, in particular to a genetically engineered bacterium obtained by mutating the lactobacillus through human intervention. [0002] The invention also relates to the preparation process of the genetically engineered bacteria. [0003] The present invention further relates to the use of the genetically engineered bacteria. Background technique [0004] The key pathogenic link of chronic renal failure is the excessive retention of uremic toxins in the body. The urea toxins retained in the patient's body are mainly urea, creatinine, and uric acid, and other toxins are mostly the products of these three transformations, derivatives, and combinations. For example, urea can be derived into guanidines and carbamoyl compounds, and creatinine can be derived into hydrogen. Oxidized creatinine and methylguanidine. The accumulation of the three toxins and their derivatives in the body of patients with uremia can lea...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K35/74A61P13/12C12N15/01C12R1/225A61K35/747
Inventor 蒋云生王芳向大雄
Owner THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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