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Bacterial strain producing L-erythrothioneine and method of preparing the L-erythrothioneine

A technology of ergothioneine and mycelium, which is applied in fermentation engineering, biosynthesis of natural products, and biological resources, can solve the problems of low product yield and achieve the effect of high content and cheap and easy-to-obtain raw materials

Active Publication Date: 2014-04-23
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the main problem of preparing ergothioneine by submerged fermentation is the low yield of the product, and the two main factors affecting the fermentation yield of ergothioneine are the production strain and the production process.

Method used

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  • Bacterial strain producing L-erythrothioneine and method of preparing the L-erythrothioneine
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  • Bacterial strain producing L-erythrothioneine and method of preparing the L-erythrothioneine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Determination of the Fatty Acid Components of Pleurotus pachyrhiza CGMCC No.6232 Bacteria

[0037] (1) Measurement method

[0038] The fatty acid composition of the experimental strains was analyzed by using the Sherolock automatic bacterial identification system of the American MIDI (Microbial Identification) company.

[0039] (2) Preparation of reagents

[0040] Solution I: Dissolve 45g of sodium hydroxide in 150mL of methanol and 150mL of distilled water.

[0041] Solution II: Dissolve 190mL of concentrated hydrochloric acid and 275mL of methanol in 135mL of distilled water.

[0042] Solution III: Mix 200mL n-hexane and 200mL methyl tert-butyl ether evenly.

[0043] Solution IV: Dissolve 10.8 g of sodium hydroxide in 900 mL of distilled water.

[0044] Solution V: saturated sodium chloride solution.

[0045] (3) Extraction method

[0046] Use an inoculation loop to scrape an appropriate amount of culture from the surface of the PDA medium, put it int...

Embodiment 2

[0055] Embodiment 2: Determination of the G+C mol% content of Pleurotus pachyrhiza CGMCC No.6232 genomic DNA

[0056] Detection method: the G+C mol% content determination of strain genomic DNA uses the melting temperature (Tm) method, with Escherichia coli (E.coli K12, CGMCC1.365) as a reference control, and the instrument used is Lambda35 of PerkinElmer Company UV / VIS Spectrometer; use PerkinElmer's PTP-1Peltier System digital temperature controller to control the temperature. Proceed as follows:

[0057] (1) Dilute the DNA sample to be tested to OD with 0.1×SSC 260nm The value is between 0.3 and 0.4;

[0058] (2) First record the OD value at 25°C at a wavelength of 260nm, and then set the temperature rise program, starting from 65°C to 95°C, during which the temperature rises by 1°C per minute;

[0059] (3) The rise of OD value indicates the beginning of denaturation, record the cuvette temperature and OD value until the OD value remains unchanged, indicating that the den...

Embodiment 3

[0065] Embodiment 3: the ergothioneine fermentation of Pleurotus pleurotus CGMCCNo.6232

[0066] Liquid seed medium: corn flour (Meihekou Xingda Rice Industry Co., Ltd.) 30g / L, soybean meal powder (Beijing Zhongmian Ziguang Biotechnology Co., Ltd.) 15g / L, α-amylase (Beijing Soleibao Technology Co., Ltd.) Company) 54U / L, KH 2 PO 4 3g / L, MgSO 4 1.5g / L, the rest is water, sterilized at 121°C for 20 minutes, and the liquid volume in a 500mL Erlenmeyer flask is 150mL.

[0067] Fermentation medium: dextrin (Tianjin North Tianyi Chemical Reagent Factory) 20g / L, yeast extract powder (Angel Yeast Co., Ltd.) 15g / L, KH 2 PO 4 3g / L, MgSO 4 1.5g / L, the rest is water, sterilized at 121°C for 20 minutes, and the liquid volume in a 500mL Erlenmeyer flask is 150mL.

[0068] Inoculate each bottle of liquid seed medium with about 1 cm of 2 CGMCC No.6232 bacterial lawn of Pleurotus rugosa CGMCC No. 6232 was cultured at 25° C. on a shaker at 150 rpm for 4 days to obtain seed liquid. The seed...

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Abstract

The invention relates to a strain Pleurotus ostreatus TIB.BPE.10010 with the accession number being CGMCC No.6232. The invention also relates to a method of preparing L-erythrothioneine. The method comprises steps of culturing and fermenting the Pleurotus ostreatus CGMCC No.6232 to perform biosynthesis to form the L-erythrothioneine and extracting the L-erythrothioneine from mycelium cells.

Description

technical field [0001] The invention belongs to the fields of biological resources, fermentation engineering and biosynthesis of natural products. More specifically, the present invention relates to a bacterial strain producing ergothioneine, Pleurotus otreatus, and a method for preparing ergothioneine by fermentation of the bacterial strain. Background technique [0002] Ergothioneine (L-Ergothioneine, EGT) is a rare natural chiral amino acid, an important natural active substance in the body, and has strong antioxidant activity: remove active oxygen and nitrogen compounds, activate antioxidant Inhibit superoxide dismutase (Aruoma OI, Whiteman M, England TG, Halliwell B.Antioxidant action ofergothioneine assessment of its ability to scavenge peroxynitrite.BBRC, 1997, 231:389~391.), inhibit various heme Oxidation reaction of protein, etc. (RomaroR, et al. The reactivity of thiols and disulfides with different redox states tomyoglobin [J]. Biol. Chem., 1992, 267: 1680-1688.)...

Claims

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Application Information

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IPC IPC(8): A01H15/00A01G1/04C12P13/04C05G1/00
Inventor 姜文侠刘琦周涛
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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