Bacterial strain for producing ergothioneine and screening method thereof

A technology of ergothioneine and screening method, applied in the field of microorganisms, can solve the problems of unpublished screening method of Pleurotus solani strains, affecting cell viability, low yield of ergothioneine, etc., achieving easy screening, low cost and high yield Effect

Active Publication Date: 2019-03-08
BLOOMAGE BIOTECHNOLOGY CORP LTD +1
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0005] Chinese patent application CN103184246A "The biosynthetic preparation method of ergothioneine" adopts the hyphae inoculation and fermentation of large-scale fungi (Pleurotus pleurotus and wild Pleurotus aurantii) to synthesize ergothioneine. The highest content of ergothioneine is 51 mg / L, 48 mg / L in Pleurotus pulmonata, and 37 mg / L in wild Pleurotus purpurea. The production of ergothioneine is still not high, and the growth cycle is long
[0006] Chinese patent application CN103734022A "The bacterial strain for producing ergothioneine and the method for preparing ergothioneine" has announced the steps of fermenting and synthesizing ergothioneine and extracting ergothioneine from the mycelium cells of Pleurotus officinalis. Screening method of Pleurotus dermoides strain
However, biotransformation will affect the viability of bacterial cells, and the transformation efficiency is low

Method used

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  • Bacterial strain for producing ergothioneine and screening method thereof
  • Bacterial strain for producing ergothioneine and screening method thereof
  • Bacterial strain for producing ergothioneine and screening method thereof

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specific Embodiment approach

[0037] As a specific embodiment of the present invention, Hericium erinaceus HT-3 CCTCC No: M 2018567 grows faster on potato dextrose agar medium (i.e. PDA solid medium), and grows at 25°C for 8 The diameter of the colony reaches 50-55mm, it is white, velvet-like, and radially extends around; the back is light brown; the colony morphology characteristics can be found in figure 1 . The mycelium of this strain is stout, composed of tubular cells, thin-walled, with septa and branches, and lock-like unions, see figure 2 .

[0038] Hericium erinaceus (Rull exF.) Pers. is a kind of saprophytic fungus and a famous edible fungus. The whole shape resembles a hedgehog or Hericium erinaceus, so it is commonly known as Hericium erinaceus or Hericium erinaceus. Among them, Hericium erinaceus is preferably Hericium erinaceus CCTCC No: M 2018567, which has been preserved in the China Center for Type Culture Collection (CCTCC) on August 23, 2018.

[0039] The 18s rRNA gene sequence of th...

Embodiment 1

[0065] Screening of Ergothioneine-producing Hericium Hericium Strains

[0066] (1) Cut internal tissue pieces of about 0.5 × 0.5 cm from the caps of the fruiting bodies of Hericium erinaceus from different origins, place them on PDA solid medium containing antibiotics, and cultivate them at 24°C, and transfer the obtained hyphae to PDA Carry out purification culture on solid medium, inoculate the purified bacteria on PDA slant, and store at 3-4°C for later use;

[0067] (2) Inoculate the solid strain obtained in step (1) to the fermentation medium, cultivate it at 24°C for 16 days, and obtain the mycelia fermentation liquid. The fermentation medium consists of 35g / L sucrose, 20g / L corn steep liquor, KH 2 PO 4 3 g / L, vitamin B 1 3mg / L, the rest is water. The precursor amino acids cysteine, methionine and histidine were supplemented at the concentration of 2mM respectively on the 10th day of fermentation.

[0068] (3) After the end of fermentation, homogenize the mycelia f...

Embodiment 2

[0070] Morphological Observation of Hericium erinaceus HT-3

[0071] The Hericium erinaceus HT-3 CCTCC NO: M 2018567 grows quickly on the potato dextrose agar medium, crawls and spreads close to the surface of the medium, grows at 25°C for 8 days, and the colony diameter is 50 ~55mm, white, velvet-like, extending radially around; the back is light brown; see the colony morphological characteristics figure 1 . The mycelium of this strain is stout, composed of tubular cells with thin cell walls, with septa and branches, and lock-like unions, see figure 2 .

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Abstract

The invention belongs to the technical field of microorganisms and specifically relates to a hericium erinaceus HT-3 with a preservation number of CCTCC No: M 2018567. The invention also relates to ascreening method of the hericium erinaceus HT-3. The screening method comprises the following steps: obtaining a purified strain containing a hericium erinaceus tissue block; performing fermental cultivation on the strain; extracting ergothioneine from cells of fermentation liquor mycelium; and determining the content of the ergothioneine in fermentation liquor. The hericium erinaceus strain screened by the invention has a high yield of the ergothioneine, and the adopted screening method is simple in process, convenient in operation and low in production cost.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to Hericium erinaceus HT-3 and a screening method thereof. Background technique [0002] The scientific name of ergothioneine is 2-mercapto-L-histidine trimethyl internal salt, which is a natural 2-thioimidazole amino acid. There are two tautomers of thiol and thione in the dissolved state. Under physiological pH conditions, it mainly exists in the form of thione. Ergothioneine was first isolated in 1909 by Tanret C from ergot (the sclerotia formed by a fungus parasitic on the grass rye). The pure product is a white crystal with good water solubility. The upper limit of dissolution is 0.9mol / L, and it is quite stable under physiological pH and strong alkali conditions. [0003] Ergothioneine has unique physiological functions in anti-oxidation, scavenging free radicals, chelating metal ions, preventing ultraviolet radiation damage, inhibiting cancer, etc., and in some...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N1/02C12P13/04C12R1/645
CPCC12N1/02C12N1/14C12P13/04C12R2001/645C12N1/145C12P17/10Y02A50/30C12Q1/04C12N1/005C12Q1/02G01N33/68G01N2333/37
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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