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Method for controlling pH of fermentation liquor of strain capable of producing aflatoxin B1 degrading enzyme

An aflatoxin and enzyme-degrading technology, applied in the field of microorganisms, can solve the problems of toxin regeneration, increased enzyme activity, unable to change the physical and sensory state components of products, etc., and achieves the effects of low cost, easy availability, dosage, and convenient regulation.

Inactive Publication Date: 2014-11-19
HUBEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] (3) The physical and sensory state of the product cannot be changed
[0008] (5) It must be able to destroy the spores and mycelia of aflatoxin-producing strains, because if they remain in the product, under suitable conditions, they may cause regeneration of the toxin
Therefore, in the process of producing aflatoxin detoxification enzyme by microbial fermentation, the pH value of the fermentation broth should be controlled, in case the aflatoxin B1 degrading enzyme enzyme activity in the detection fermentation broth, after adding the fermentation broth to the aflatoxin B1 sample The pH of the reaction system deviates from neutral, which will lead to the degradation of aflatoxin B1 due to the pH deviation, rather than the result of enzymatic reaction, resulting in the illusion of increased enzyme activity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1: adding glucose shake flask fermentation to fermentation pH control

[0028] (1) strain

[0029] bacteria Sinomonas sp.HSD8, CCTCC NO: M2014109.

[0030] (2) Medium

[0031] The slant medium per 1 L contains: beef extract 3g, peptone 10g, NaCl 5g, agar 20g, and the pH of the slant medium is natural;

[0032] The seed culture medium per 1 L contains: beef extract 3g, peptone 10g, NaCl 5g, and the pH of the seed culture medium is natural;

[0033] The fermentation medium (basic medium) on each 1 L shake flask contains: beef extract 3g, peptone 10g, NaCl 8.5g, KH 2 PO 4 1g, K 2 HPO 4 ·3H 2 O 1g, MgSO 4 1g, coumarin 1g, the pH of the fermentation medium is 5.0-5.5;

[0034] (3) The solution used in the experiment

[0035] PBS buffer (pH 7.0), composed of 0.2mol / L Na 2 HPO 4 ﹒ 12H 2 O solution and 0.2mol / L NaH 2 PO 4 ﹒ 2H 2 O solution prepared.

[0036] (4) Add different amounts of glucose to control the pH of the fermentation broth

[...

Embodiment 2

[0046] Example 2: Adding KH 2 PO 4 Shake Flask Fermentation vs Fermentation pH Control

[0047] (1) strain

[0048] Same as Example 1.

[0049] (2) Medium

[0050] The slant medium per 1 L is the same as in Example 1.

[0051] The seed medium per 1 L is the same as in Example 1.

[0052] The fermentation medium (basic medium) on each 1 L shake flask contains: beef extract 3g, peptone 10g, glucose 4g, NaCl 8.5g, K 2 HPO 4 ·3H 2 O 1g, MgSO 4 1g, coumarin 1g, the pH of the fermentation medium is 5.0-5.5;

[0053] (3) The solution used in the experiment

[0054] Same as Example 1.

[0055] (4) Add different amounts of KH 2 PO 4 pH control of fermentation broth

[0056] In the fermentation medium (basic medium) (basic medium formula (g / L): beef extract 3, peptone 10, NaCl 8.5, glucose 4g, K 2 HPO 4 ·3H 2 O 1, MgSO 4 1. Add 0% (W / V), 0.1% (W / V), 0.15% (W / V), 0.2% (W / V), 0.25% (W / V) on the basis of coumarin 1) , 0.3% (W / V) KH 2 PO 4 As a medium f...

Embodiment 3

[0065] Example 3: Adding Glucose and KH 2 PO 4 50L Fermentation Tank Fermentation Control of Fermentation Broth pH

[0066] (1) strain

[0067] Same as Example 1.

[0068] (2) Medium

[0069] The slant culture medium described in every 1 L is the same as in Example 1;

[0070] The seed culture medium described in every 1 L is the same as in Example 1;

[0071] The fermentation medium on the level of the fermentor per 1 L contains: corn steep liquor 13g, glucose 2g, NaCl 8.5g, KH 2 PO 4 2.5g, K 2 HPO 4 ·3H 2 O 1g, MgSO 4 1g, coumarin 1g, the pH of the fermentation medium is 4.5-5.0;

[0072] (3) Seed cultivation

[0073]After the slant seeds are activated for 24-36 hours, insert them into a 250 mL Erlenmeyer flask filled with 50 mL of seed medium and cultivate them for 12-14 hours to obtain first-grade seeds; Put the primary seeds into the same fermentation medium and cultivate them for 12-14 hours to prepare the secondary seeds. The cultivation tem...

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PUM

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Abstract

The invention discloses a method for controlling pH of fermentation liquor in a fermentation process by utilizing bacteria Sinomonas atrocyanea capable of producing aflatoxin detoxifying enzyme and belongs to the technical field of microorganism. On the basis of a fermentation culture medium, the mixture of 0.4% (w / v) of glucose and 0.25%(w / v) of monopotassium phosphate is added, so that the pH value of fermentation supernate is controlled to be about 7.0, and the problem that enzyme activity detection is influenced by pH can be effectively solved.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, in particular to a bacterium producing aflatoxin detoxification enzyme Sinomonas sp.HSD8 is a method for controlling the pH of the fermentation broth during the fermentation process. technical background [0002] Aflatoxin is a compound with strong biological toxicity, often produced by Aspergillus flavus and several other molds in moldy grains, such as rice, beans, peanuts, etc. It is by far the strongest carcinogen , and its toxicity is much higher than that of cyanide, arsenic and organic pesticides. When people ingest a large amount, acute poisoning can occur, and acute hepatitis, hemorrhagic necrosis, fatty degeneration of liver cells and hyperplasia of bile ducts can occur. When ingested in small amounts continuously, it can cause chronic poisoning, growth retardation, fibrous lesions, and fibrous tissue proliferation. The main aflatoxins are B 1 , B 2 , G 1 with G 2 Wait for...

Claims

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Application Information

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IPC IPC(8): C12N9/00C12R1/01
CPCC12N9/00C12Q3/00
Inventor 蔡俊王常高戴军杜馨周安盛
Owner HUBEI UNIV OF TECH
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