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Biosynthesis preparation method for ergothioneine, and fermentation culture medium

A fermentation medium and ergothioneine technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, fermentation, etc., can solve the problems of difficult purification treatment, increased fermentation by-products, low fermentation yield, etc.

Active Publication Date: 2019-03-08
BLOOMAGE BIOTECHNOLOGY CORP LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] 1. The fermentation yield is low, the production cost is high, and it cannot be produced and utilized on a large scale;
[0011] 2. The method of judging the end point of fermentation is not clear
If regular sampling is adopted, the method of liquid phase testing to determine whether the content of ergothioneine continues to increase after treatment is complicated, and the end point judgment is not timely enough.
If the fermentation time is prolonged blindly in order to make ergothionein fully synthesized, other fermentation by-products will definitely be increased, which will bring difficulties to the later stage purification treatment;
[0012] 3. Stirring and heating are often used to leach ergothioneine from the inside of mycelium cells to the outside of the cells. The cell breakdown may not be complete, and the ergothioneine cannot be completely leached, resulting in waste, and the energy consumption of stirring and heating is high. , which increases the production cost

Method used

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  • Biosynthesis preparation method for ergothioneine, and fermentation culture medium
  • Biosynthesis preparation method for ergothioneine, and fermentation culture medium
  • Biosynthesis preparation method for ergothioneine, and fermentation culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0137] Embodiment 1: Ergothioneine fermentation of Hericium erinaceus CCTCC NO: M 2018567

[0138] Liquid seed medium: 4% (w / v) sucrose, 1.5% (w / v) bean cake powder, 0.2% (w / v) sodium dihydrogen phosphate, 0.1% (w / v) sodium sulfate, The rest is water, pH 4.0-4.5, sterilized at 121°C for 20 minutes.

[0139] Fermentation medium: 3% (w / v) maltose, 2% (w / v) corn steep liquor, 0.05% (w / v) sodium dihydrogen phosphate, the rest is water, pH 4.0-4.5, at 121°C Sterilize for 20 minutes.

[0140] Each bottle of liquid seed medium was inoculated with a lawn of Hericium erinaceus CCTCCNo: M 2018567 picked from the slant of the strain of about 1 × 1 cm in size, and cultivated in a shaker at 20°C at 150rpm for 7 days to obtain a seed solution. The seed liquid is inserted into the fermentation medium with an inoculation amount of 5% by volume, cultivated on a shaker at 23°C at 180rpm, and supplemented with 5mM each of the precursor substances cysteine, betaine, and methionine when fermente...

Embodiment 2

[0141] Example 2: Extraction of Ergothioneine Using Helicase and Collapsing Enzymes

[0142] After fermentation, adjust the pH of the fermentation broth to 6.5±0.1, raise the temperature to 40±2°C, add 0.005% (w / v) helicase and 0.005% (w / v) collapse enzyme, and incubate at 40±2°C for 2.5 h, after the end of the enzymolysis, the temperature of the fermentation broth was raised to 90° C. and kept for 20 minutes to inactivate the enzyme. The ergothioneine extract is obtained by centrifuging the fermented liquid after enzymolysis, taking the supernatant, and then filtering and sterilizing through a 1.2 μm fine filter paper board and a 0.22 μm filter element.

Embodiment 3

[0143] Embodiment 3: the influence of different fermentation carbon sources on the output of ergothioneine

[0144] Replace the carbon source of the above-mentioned fermentation medium with: soluble starch, glycerol, glucose, sucrose, fructose, maltose, lactose, mannitol, maltitol, corn flour, galactose, maltodextrin, according to the method of Example 1 Fermentation culture, measure the content of ergothioneine (EGT) in the fermented liquid after the end of fermentation, the results are shown in Table 1 and figure 1 . When glucose was used as the carbon source, the content of ergothioneine in the fermented liquid was 151.7mg / L, and the yield increase was relatively large. It can be used as the preferred carbon source for the production of ergothioneine by the liquid fermentation of Hericium erinaceus mycelia, followed by maltose (ergothioneine). Thioneine content is 135.7mg / L) and sucrose (ergothioneine content is 132.4mg / L).

[0145] Table 1

[0146] carbon sourc...

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Abstract

The invention relates to a biosynthesis preparation method for ergothioneine, and a fermentation culture medium. The method comprises the following steps of S1: inoculating Hericium erinaceus myceliumslant cultures to a liquid seed culture medium to be cultured to obtain seed liquid; S2: inoculating the obtained seed liquid to the fermentation culture medium to be fermented, adding precusor substances for culturing, and fermenting to a fermentation destination through pH (Potential of Hydrogen) judging a fermentation liquor; S3: after fermentation finishes, adding enzyme, carrying out enzymolysis to the destination, raising the temperature to kill enzyme, and lixiviating the ergothioneine in the cell of the mycelium to the fermentation liquor out of the cell. By use of the method disclosed by the invention, the yield of the ergothioneine can be improved, the fermentation destination can be quickly and simply judged, fermentation byproducts are reduced, the enzyme is used for breakingmycelium cells, and ergothioneine is safe to be lixiviated.

Description

technical field [0001] The invention relates to a method for biosynthesizing ergothioneine by using Hericium erinaceus mycelium and a fermentation medium used therefor, belonging to the field of microbial fermentation engineering. Background technique [0002] Ergothioneine (EGT), scientifically named as 2-mercapto-L-histidine trimethyl inner salt, contains imidazole-2-thione group shown in the following general formula (1) in its molecular structure. Ergothioneine is a natural amino acid that exists in many animals and plants and is rich in content. It cannot be synthesized by the animal body itself and can only be ingested from food. It is a rare amino acid. [0003] [0004] Studies have shown that ergothioneine has strong antioxidant effects and can protect cells from cell damage or even death caused by ultraviolet rays and gamma rays. Therefore, it has broad application prospects in pharmaceutical, food and cosmetic industries. However, the content of ergothioneine ...

Claims

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Application Information

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IPC IPC(8): C12P13/04C12R1/645
CPCC12P13/04
Inventor 魏玉洁陆震贾玉倩孙元军陈雯雯石艳丽栾贻宏郭学平
Owner BLOOMAGE BIOTECHNOLOGY CORP LTD
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