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Trichoderma reesei for producing ergothioneine

A technology of ergothioneine and Trichoderma reesei, which is applied in the field of microorganisms, can solve the problems of long fermentation period, slow growth of mushroom mycelium, and high production cost, and achieve fast growth, simple post-treatment process, and no product inhibition sexual effect

Active Publication Date: 2020-12-01
ZHEJIANG HUARUI BIOTECHNOLOGY CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, there are two important defects in mushroom fermentation. One is that the growth of mushroom mycelium is relatively slow, resulting in a longer fermentation period, generally 7-10 days; the other is that most of the ergothioneine synthesized by mushroom fermentation In the filament body, the cumbersome post-treatment process of crushing and extracting the mycelium is required, resulting in relatively high production costs

Method used

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  • Trichoderma reesei for producing ergothioneine
  • Trichoderma reesei for producing ergothioneine
  • Trichoderma reesei for producing ergothioneine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1: Screening of filamentous fungi resistant to ergothioneine

[0028] Collect food waste samples from 30 areas in Huzhou City, Zhejiang Province, resuspend them with sterile water, filter them with sterile absorbent cotton, and coat the suspension with solids containing penicillin, kanamycin, chloramphenicol and ergothioneine Medium (glucose 20g / L, peptone 10g / L, yeast extract 5g / L, penicillin (100μg / ml), kanamycin (50μg / ml), chloramphenicol (25μg / ml), ergothioneine 1g / L, agar 20g / L, pH7.0), cultivated at 28-30°C for 4-5 days. Pick vigorously growing filamentous microorganisms and transfer them to new solid plates for pure culture. After more than two months, a total of more than 600 strains of microorganisms with different shapes were obtained, most of which were molds, which were used for the next step of shake flask fermentation screening.

Embodiment 2

[0029] Embodiment 2: the microbial screening of producing ergothioneine

[0030] More than 600 strains of microorganisms obtained in Example 1 were inoculated into test tubes of liquid medium (glucose 20g / L, peptone 10g / L, yeast extract 5g / L, pH7.0) respectively, and each bacterial strain was inoculated with 3 filling volumes for 4ml test tubes. 30°C, 230rpm shaker culture for 72h.

[0031] After the cultivation, the three test tube culture fluids of each bacterial strain were combined, centrifuged to get the supernatant, and the content of ergothioneine was detected by HPLC. The production of ergothioneine could be detected in the fermentation broth of 19 strains of microorganisms. The ergothioneine content in two of the microorganisms was relatively high, and they were named SH444 and SH445 respectively.

Embodiment 3

[0032] Embodiment 3: bacterial strain identification

[0033] The bacterial strains SH444 and SH445 obtained in Example 2 were sent to the Institute of Microbiology, Chinese Academy of Sciences to carry out strain identification, and according to the data such as cultivation, i.e. cell micromorphological characteristics, physiological and biochemical characteristics and rRNA gene sequence, it was determined that SH444 was Rhizopus oryzae; Determine SH445 as Trichoderma reesei. Specifically

[0034] 3.1 Strain SH444

[0035] The colony is sparse and fluffy, the hyphae are white, the rhizomes are slightly dark brown, the stolons crawl, the rhizoids are well developed, chemo-heterotrophic, aerobic. The bacterial strain was observed under a microscope, and the shape was strip-shaped, colorless and transparent.

[0036] 3.2 Strain SH445

[0037] The colony is in the form of cotton wool widely spread, white dense flat hyphae at first, slightly raised in the middle, sticky when w...

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Abstract

The invention discloses trichoderma reesei which is preserved in China General Microbiological Culture Collection Center with the preservation number of CGMCC No. 17667. The trichoderma reesei CGMCC No.17667 disclosed by the invention can be used for producing ergothioneine through fermentation, and has prospect in industrial development and application.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and relates to Trichoderma reesei and its application for producing ergothioneine. Background technique [0002] Ergothioneine (L-ergothionine, EGT), the chemical name is 2-mercaptohistidine trimethyl inner salt, and its structural formula is as follows: [0003] [0004] Ergothioneine is the only known natural 2-thioimidazole amino acid. Ergothioneine has various physiological effects such as anti-oxidation, anti-inflammation, prolonging cell survival cycle or anti-aging activity, and improving nerve cell generation. At the same time, in a variety of disease models, including Alzheimer's, diabetes and other complications, it has a good effect on protecting cells and resisting damage. It has a good market application prospect. [0005] At present, ergothioneine can be obtained through chemical synthesis, edible fungus extraction and microbial fermentation. Among them, a variety of m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P13/04C12R1/885
CPCC12P13/04C12R2001/885C12N1/145
Inventor 范文超高书良王金刚梁岩任亮
Owner ZHEJIANG HUARUI BIOTECHNOLOGY CO LTD