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Clostridium bifermentans for generating equol by degrading daidzein and bacteria agent and application thereof

A double-enzyme Clostridium and daidzein technology, applied in the field of microorganisms, can solve the problems of limiting the application of daidzein products, individual differences in clinical application effects, and high prices, so as to benefit the health of the host, ensure animal health, production and The effect of low cost of use

Inactive Publication Date: 2011-04-20
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, daidzein products for human use are pure extracts derived from leguminous plants. Not only are the yields low and the price is expensive, but also the individual differences in clinical application effects are due to the differences in the composition of daidzein-degrading bacteria in different populations. , limiting the application of daidzein products

Method used

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  • Clostridium bifermentans for generating equol by degrading daidzein and bacteria agent and application thereof
  • Clostridium bifermentans for generating equol by degrading daidzein and bacteria agent and application thereof
  • Clostridium bifermentans for generating equol by degrading daidzein and bacteria agent and application thereof

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Experimental program
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Effect test

Embodiment 1

[0044] The isolation and identification of embodiment 1 bacterial strain

[0045] Take fresh feces samples from healthy adult pigs and place them in sterile, anaerobic BHI-daidzein broth, mix them evenly with a vortex mixer, and carry out gradient dilution in an anaerobic workstation, and spread them on BHI-daidzein agar Plates were grown in an anaerobic workstation at 37°C until colonies formed. Single colonies with obvious morphological differences were picked and purified, and then inoculated in BHI-daidzein broth respectively. After culturing at 37°C, HPLC was used to detect the contents of daidzein and equol in the culture medium ( Figure 4 and Figure 5 ). The bacterial strain Clostridium bienzyme zx-7 (CGMCC NO.1995), which can degrade daidzein and produce equol, was screened out. The photos of its colony and Gram staining are shown in figure 1 , figure 2 , identified as Clostridium bifermentans , the main biological characteristic is G + , Strictly anaerobic g...

Embodiment 2

[0046] Embodiment 2 strain growth characteristics experiment

[0047] 2.1 Determination of growth characteristics

[0048] Clostridium bienzyme zx-7 (CGMCC NO.1995) was inoculated into the liquid fermentation medium, cultured anaerobically at 37 °C, and the growth curve was drawn by measuring the absorbance value of the bacterial culture solution at 660 nm with a 722 spectrophotometer ( Figure 6 ), use a pH meter to measure the pH value in the culture solution and draw the pH value change curve ( Figure 7 ).

[0049] The formula of liquid fermentation medium was: tryptone 20 g / L; beef extract 10 g / L; yeast extract 6 g / L; glucose 4 g / L; K 2 HPO 4 2 g / L; KH 2 PO 4 1 g / L; MgSO 4 0.4 g / L; CaCl 2 0.2 g / L; FeSO 4 0.1 g / L; cysteine ​​hydrochloride 0.5 g / L.

[0050] 2.2 Carbon source utilization experiment

[0051] Use carbon source utilization medium (medium formulation (g / L): (NH 4 ) SO 4 2.0; MgSO 4 .7H2O 0.2; NaH 2 PO 4 ;CaCl 2 ·H 2 O 0.1; K 2 HPO ...

Embodiment 3

[0052] Example 3 Acid-resistant and bile salt-resistant characteristic experiments of bacterial strains

[0053] 3.1 Effect of pH value on the survival of Clostridium bienzyme zx-7

[0054] Inoculate the zx-7 (CGMCC NO.1995) bacteria that entered the late logarithmic growth phase into three groups of culture solutions that have been adjusted to pH values ​​of 1.5, 2.5, 3.5 and 7.2 with 1mol / L HCL (the control group is the liquid fermentation medium , 10mg / L daidzein group (that is, add 10mg / L daidzein to the liquid fermentation medium, 50mg / L daidzein group (that is, add 50mg / L daidzein to the liquid fermentation medium), culture anaerobically at 37 ℃, at 0 and At the time point of 3h, the bacterial culture solution was taken on the plate (BHI agar solid medium) and counted ( Figure 9 ). The results showed that at pH 7.2, after 3 hours of treatment, the number of viable bacteria increased significantly, and the strain showed a normal growth trend; at pH 3.5, after 3 hou...

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Abstract

The invention belongs to the technical field of microorganism and discloses a clostridium bifermentans for generating equol by degrading daidzein and bacteria agent and application thereof. The clostridium bifermentans for generating equol by degrading daidzein zx-7 is collected in the China General Microbiological Culture Collection Center, and the collection number is CGMCC NO.1995. The clostridium bifermentans CGMCC NO.1995 can degrade daidzein to generate equol, the low density lipoprotein content of rat serum is remarkably reduced, the ratio of the high-density lipoprotein to the low-density lipoprotein is obviously increased, the elimination of cholesterol in rat is facilitated, and the host health is favored. The clostridium bifermentans can be used for preparing animal feed, a preparation for eliminating cholesterol and a fermented product of soy or bean pulp.

Description

technical field [0001] The invention belongs to the technical field of microbes, and relates to a strain of Clostridium bienzyme capable of degrading daidzein to produce equol, as well as its bacterial agent and application. Background technique [0002] Daidzein is a kind of soybean isoflavones (Isoflavones). Soy isoflavones include twelve components including daidzein, genistein, and glycitein. About 50%-60% of soybean isoflavones in soybean grains are genistein, 30%-35% are daidzein, and 5%-15% are glycitein. Since the 1980s, many research reports and animal experiments have shown that soybean isoflavones have weak estrogen activity, antioxidant activity, antihemolytic activity and antifungal activity, and can effectively prevent and control leukemia, osteoporosis, gastric cancer, The occurrence of various diseases such as breast cancer and prostate cancer, especially has positive preventive and therapeutic effects on breast cancer and prostate cancer. In animal produc...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P17/06A23K1/16C12R1/145A23K10/18
Inventor 姚文张逊郑卫江黄莎娜孙小燕
Owner NANJING AGRICULTURAL UNIVERSITY
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