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Ethionine resistance Candida utilis and application thereof

A technology of Candida utilis and ethionine, applied in fermentation, biochemical equipment and methods, microorganisms, etc., to achieve high yield, strong protein synthesis ability, and broad growth nutrition spectrum

Inactive Publication Date: 2012-04-04
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, research at home and abroad is mainly focused on the fermentation of single-purpose products such as SAM or GSH. So far, no Candida utilis has been selected for the co-production fermentation of SAM and GSH.

Method used

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  • Ethionine resistance Candida utilis and application thereof
  • Ethionine resistance Candida utilis and application thereof
  • Ethionine resistance Candida utilis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Co-production and fermentation of SAM and GSH, and determining the optimal concentration of ethionine

[0030] (1) Strain

[0031] The starting strain is Candida utilis SZU 07-01, preserved by the Industrial Microbiology Laboratory of Soochow University.

[0032] (2) Medium

[0033] Slope and seed culture medium (g / L): glucose 20, peptone 20, yeast extract 10, pH 6.0;

[0034] Screening medium: Add 0~0.7g / L ethionine to the seed medium;

[0035] Fermentation medium before optimization (g / L): glucose 30, ammonium sulfate 8, potassium dihydrogen phosphate 3, magnesium sulfate 0.25, pH 5.5.

[0036] (3) UV mutagenesis

[0037] The irradiation was carried out under a Philips ZWSSJD-30 ultraviolet lamp, the irradiation power and wavelength were 30W, 253.7nm, and the irradiation distance was 30cm. The specific steps are as follows: aspirate 1 mL of seed culture solution after 20 hours of culture, centrifuge for 2 min at 10000 r / min, discard the supernatant, add sterile saline ...

Embodiment 2

[0064] Example 2: Determine the best carbon source

[0065] The operation was carried out according to Example 1, but the difference was that the fermentation medium before optimization was used as a benchmark to investigate the effects of different carbon sources (carbon sources were glucose, sucrose, glycerol, ethanol, sodium acetate, sodium citrate and starch) on SAM and GSH. The impact of co-production fermentation, on this basis, the experimental results of co-production fermentation to prepare SAM and GSH, the results are shown in Table 2:

[0066] SAM output: 0~173.1mg / L; GSH output: 6.5~193.5mg / L;

[0067] SAM and GSH co-production total 6.5~366.6mg / L;

[0068] Among them, the best carbon source is sucrose.

[0069] Table 2 The effect of carbon source types on the co-production and fermentation of SAM and GSH

[0070]

Embodiment 3

[0071] Example three, determine the best nitrogen source

[0072] The operation was carried out according to the first embodiment, the difference is: the fermentation medium before optimization was used as a benchmark to investigate the effect of different nitrogen sources (the nitrogen sources are yeast extract, beef extract, peptone, potassium nitrate, ammonium sulfate, urea and ammonium nitrate) on SAM The effect of co-production and fermentation with GSH. On this basis, the experimental results of co-production and fermentation to prepare SAM and GSH are shown in Table 3:

[0073] SAM output: 41~146mg / L; GSH output: 57.5~167.7mg / L;

[0074] The combined production of SAM and GSH is 90.5~308.7mg / L;

[0075] Among them, the best nitrogen source is ammonium sulfate.

[0076] Table 3 The effect of nitrogen sources on the co-production and fermentation of SAM and GSH

[0077]

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Abstract

The invention discloses an ethionine resistance Candida utilis and application thereof, wherein the Candida utilis SUGS-01 is preserved in China Center for Type Culture Collection (CCTCC) on December 9th, 2009, and the preservation number is CCTCC M 209298. The Candida utilis SUGS-01 is applied to produce SAM and GSH by co-production and fermentation, and the application comprises the following steps: taking the Candida utilis SUGS-01 as the original strain, and after slant culturing and seed culturing, fermenting for at least 24 hours; one litre of fermentation medium contains 35-36g of canesugar, 10-12g of ammonium sulfate, 12-13g of monopotassium phosphate, 0.05-0.06g of magnesium sulfate and 4-5g of L-methionine; and the pH value of the fermentation medium is 5.5-6.0. Because adopting the combined action of UV mutagenesis and gamma-ray mutagenesis, the invention obtains the Candida utilis mutant strain which can co-produce SAM and GSH, and the obtained Candida utilis is used as production strains, has the advantages of wide growth nutrition spectrum, less production of ethanol during the fermentation process, strong capability to synthesize protein and the like, and has good industrial application prospect.

Description

Technical field [0001] The invention belongs to microbial fermentation technology, and specifically relates to an ethionine-resistant Candida utilis and its application in the co-production and fermentation of S-adenosylmethionine and glutathione. Background technique [0002] S-adenosylmethionine (SAM) is an important small molecule substance widely found in biological cells, discovered by Cantoni in 1953. Studies have found that SAM, as a group provider and enzyme inducer, participates in many key biochemical reactions, such as the methylation of lipids, proteins, nucleic acids, transsulfurization reactions, and polyamine synthesis. The physiological functions of SAM in cells involve: methyl donor in methyl reaction; production of methylthioadenosine and homoserine; donor of aminobutyl of tRNA; donor of amino acid chain in the synthesis of vitamin H; gland Donor of glycoside part; inhibition of cytolysin-2,3-amino allosteric enzyme, threonine synthase, pyruvate formate lyase, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/16C12P19/40C12P21/02C12R1/72
Inventor 王大慧邵娜卫功元
Owner SUZHOU UNIV
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