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Method and fermentation medium for biosynthetic preparation of ergothioneine

A fermentation medium, ergothioneine technology, applied in the directions of microorganism-based methods, biochemical equipment and methods, fermentation, etc., can solve problems such as increasing fermentation by-products, difficulty in purification and processing, and inability to produce and utilize on a large scale

Active Publication Date: 2021-01-26
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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  • Method and fermentation medium for biosynthetic preparation of ergothioneine
  • Method and fermentation medium for biosynthetic preparation of ergothioneine
  • Method and fermentation medium for biosynthetic preparation of ergothioneine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0137] Example 1: Fermentation of Ergothioneine by Hericium erinaceus CCTCC NO: M 2018567

[0138] Liquid seed medium: 4% (w / v) sucrose, 1.5% (w / v) bean cake flour, 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 20min.

[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 Hericium erinaceus CCTCCNo: M 2018567 bacterial lawn of about 1 × 1 cm in size picked from the slanted surface of the strain, and cultured at 20°C for 7 days on a shaker at 150 rpm to obtain a seed solution. The seed liquid was inserted into the fermentation medium at an inoculum of 5% by volume, cultivated at 180 rpm on a shaker at 23°C, and 5 mM of the precursors cysteine, betaine, and methionine were supplemented during fermentation for 3 day...

Embodiment 2

[0141] Example 2: Extraction of ergothioneine using helicase and crash enzymes

[0142] After the fermentation, the pH of the fermentation broth was adjusted to 6.5±0.1, the temperature was raised to 40±2°C, 0.005% (w / v) helicase and 0.005% (w / v) collapse enzyme were added, and the temperature was kept at 40±2°C for 2.5 h, after the enzymatic hydrolysis, the temperature of the fermentation broth was raised to 90° C. and kept for 20 min to inactivate the enzyme. The ergothioneine extract is obtained by centrifuging the enzymatically hydrolyzed fermentation broth, taking the supernatant, and then filtering and sterilizing it through a 1.2 μm fine filter cardboard and a 0.22 μm filter element.

Embodiment 3

[0143] Example 3: Effects of different fermentation carbon sources on ergothioneine yield

[0144] The carbon sources of the above-mentioned fermentation medium were respectively replaced with: soluble starch, glycerol, glucose, sucrose, fructose, maltose, lactose, mannitol, maltitol, corn flour, galactose, maltodextrin, and carried out according to the method of Example 1 Fermentation culture, measure the content of ergothioneine (EGT) in the fermentation broth after the fermentation finishes, the results are shown in Table 1 and figure 1 . When glucose was made carbon source, the content of ergothioneine in the fermented liquid was 151.7mg / L, and the output improvement rate was larger, which could be used as the preferred carbon source of Hericium erinaceus mycelium liquid fermentation to produce ergothioneine, followed by maltose (ergot). Thioneine content was 135.7 mg / L) and sucrose (ergothioneine content was 132.4 mg / L).

[0145] Table 1

[0146] carbon sourc...

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Abstract

The invention relates to a biosynthetic preparation method and a fermentation medium of ergothioneine. The method comprises the following steps: Step 1: inoculating Hericium erinaceus (Hericium erinaceus) mycelium slants into a liquid seed culture medium for cultivation to obtain seeds liquid; step 2: inoculate the obtained seed liquid into the fermentation medium for fermentation and add precursor substances for cultivation, judge the fermentation to the end point of fermentation by pH, and obtain the fermentation liquid; step 3: after the end of fermentation, add enzymes, and enzymatically hydrolyze to the end point , the temperature is raised to inactivate the enzyme, and the ergothioneine in the cells of the mycelium is leached into the extracellular fermentation broth. The method of the invention can increase the yield of ergothioneine, quickly and easily judge the fermentation end point, reduce the generation of fermentation by-products, use enzymes to break mycelia cells, and completely extract ergothioneine.

Description

technical field [0001] The invention relates to a method for biosynthesizing ergothioneine by utilizing Hericium erinaceus mycelium and a fermentation medium therefor, belonging to the field of microbial fermentation engineering. Background technique [0002] Ergothioneine (EGT), scientific name is 2-mercapto-L-histidine trimethyl inner salt, and its molecular structure contains an imidazole-2-thione group represented by the following general formula (1). 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, but 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 also protect cells from cell damage and even death caused by ultraviolet rays and gamma rays, so it has broad application prospects in medicine, food and cosmetics industries. However, the content of ergothioneine in...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12R1/645
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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