Bacterial strain for producing gliotoxin and method for producing gliotoxin by adopting bacterial strain

Abstract

The invention belongs to the technical field of bioengineering and provides a high-yield bacterial strain for producing gliotoxin and a method for preparing the gliotoxin by adopting the bacterial strain for performing liquid fermentation. The bacterial strain for producing the gliotoxin related to the invention is a filamentous fungus, is identified as a Trichodermaviride through 16SrDNA, has a strain code of JBSH-002 and has a preservation number of CGMCC No.5611 at the Common Micro-organism Center of China Microbial Culture Preservation Commission. The bacterial strain can be applied to the production of the gliotoxin. The yield of the gliotoxin prepared by fermenting according to the method is above 500mg / L.

Description

technical field

[0001] The invention belongs to the technical field of bioengineering and provides a high-yield gliomycin microbial strain, in particular to a viride trichoderma and a method for producing gliomycin by using the strain. Background technique

[0002] Glitomycin was the first substance identified in the class of Diketopiperazines. Weidling and Emerson (1936) first separated this substance from Trichoderma lignorum, and later Weidling defined this substance as gliomycin, also known as gliotoxin. In 1958, the structure of gliomycin was identified by Bell et al.

[0003] Jacobus D M et al. studied the metabolic pathway of gliomycin in 1979, and found that peptide synthetase (gliP) catalyzes serine and phenylalanine to form a cyclic product, followed by thioredoxin reductase (gliT) Sulfurization and oxidation, and finally through the methylation of o-methyltransferase (gliM) and methyltransferase (gliN) to form gliomycin, but there has been no method to induce hi...

Examples

Embodiment 1

[0086] (1) Activation of strains: move the strains on the slant of the test tube stored on PDA medium at 4°C to room temperature (20°C-25°C) for activation for 4 hours;

[0087] (2) Preparation of liquid seeds: On an aseptic operating table, use 10 mL of sterilized distilled water to make a bacterial suspension from the activated test tube slant strain, and rinse it into a sterile medium containing sterilized seed medium. In the Erlenmeyer flask, one test tube strain was inoculated into one Erlenmeyer flask, the pH value was natural, the culture temperature was 24°C, the shaker speed was 200r / min, and the culture time was 48h;

[0088] The composition (w / w) of the seed medium is: by weight (w / w): 2% sucrose, 0.2% yeast extract powder, 0.05% magnesium sulfate, 0.025% potassium dihydrogen phosphate, add water to 1000mL; sterilization conditions Sterilize at 121°C, 0.15Mpa for 20min;

[0089] (3) Fermentation and regulation process: Receive the liquid seed into the fermentation ...

Embodiment 2

[0095] (1) Activation of strains: move the strains on the slant of the test tube stored on PDA medium at 4°C to room temperature (20°C-25°C) for activation for 6 hours;

[0096] (2) Preparation of liquid seeds: On an aseptic operating table, use 10 mL of sterilized distilled water to make a bacterial suspension from the activated test tube slant strain, and rinse it into a triangular flask containing sterilized seed medium. Inoculate 1 Erlenmeyer flask with 1 test tube strain, the pH value is natural, the culture temperature is 24°C, the shaker speed is 200r / min, and the culture time is 48h;

[0097] The composition (w / w) of the seed medium is: by weight (w / w): 2% sucrose, 0.6% yeast extract powder, 0.05% magnesium sulfate, 0.025% potassium dihydrogen phosphate, add water to 1000mL;

[0098] (3) Fermentation and regulation process: Inoculate liquid seeds with 10% (v / v) inoculum into 2L fermentation medium, culture at 26°C for 72 hours, add 1w / v% glycerin and 0.05w of the total...

Embodiment 3

[0104] (1) Activation of strains: move the strains on the slant of the test tube stored on PDA medium at 4°C to room temperature (20°C-25°C) for activation for 4 hours;

[0105] (2) Preparation of liquid seeds: On an aseptic operating table, use 10 mL of sterilized distilled water to make a bacterial suspension from the activated test tube slant strain, and rinse it into a triangular flask containing sterilized seed medium. Inoculate 1 Erlenmeyer flask with 1 test tube strain, the pH value is natural, the culture temperature is 24°C, the shaker speed is 200r / min, and the culture time is 48h;

[0106] The composition (w / w) of the seed medium is: by weight (w / w): 2% sucrose, 0.6% yeast extract powder, 0.05% magnesium sulfate, 0.025% potassium dihydrogen phosphate, add water to 1000mL;

[0107] (3) Fermentation and regulation process: Inoculate liquid seeds with 10% (v / v) inoculation amount into 1L fermentation medium, culture at 26°C for 72 hours, add 1w / v% ethyl acetate and 0....

technical field

[0001] The invention belongs to the technical field of bioengineering and provides a high-yield gliomycin microbial strain, in particular to a viride trichoderma and a method for producing gliomycin by using the strain. Background technique

[0002] Glitomycin was the first substance identified in the class of Diketopiperazines. Weidling and Emerson (1936) first separated this substance from Trichoderma lignorum, and later Weidling defined this substance as gliomycin, also known as gliotoxin. In 1958, the structure of gliomycin was identified by Bell et al.

[0003] Jacobus D M et al. studied the metabolic pathway of gliomycin in 1979, and found that peptide synthetase (gliP) catalyzes serine and phenylalanine to form a cyclic product, followed by thioredoxin reductase (gliT) Sulfurization and oxidation, and finally through the methylation of o-methyltransferase (gliM) and methyltransferase (gliN) to form gliomycin, but there has been no method to induce hi...

Examples

Embodiment 1

[0086] (1) Activation of strains: move the strains on the slant of the test tube stored on PDA medium at 4°C to room temperature (20°C-25°C) for activation for 4 hours;

[0087] (2) Preparation of liquid seeds: On an aseptic operating table, use 10 mL of sterilized distilled water to make a bacterial suspension from the activated test tube slant strain, and rinse it into a sterile medium containing sterilized seed medium. In the Erlenmeyer flask, one test tube strain was inoculated into one Erlenmeyer flask, the pH value was natural, the culture temperature was 24°C, the shaker speed was 200r / min, and the culture time was 48h;

[0088] The composition (w / w) of the seed medium is: by weight (w / w): 2% sucrose, 0.2% yeast extract powder, 0.05% magnesium sulfate, 0.025% potassium dihydrogen phosphate, add water to 1000mL; sterilization conditions Sterilize at 121°C, 0.15Mpa for 20min;

[0089] (3) Fermentation and regulation process: Receive the liquid seed into the fermentation ...

Embodiment 2

[0095] (1) Activation of strains: move the strains on the slant of the test tube stored on PDA medium at 4°C to room temperature (20°C-25°C) for activation for 6 hours;

[0096] (2) Preparation of liquid seeds: On an aseptic operating table, use 10 mL of sterilized distilled water to make a bacterial suspension from the activated test tube slant strain, and rinse it into a triangular flask containing sterilized seed medium. Inoculate 1 Erlenmeyer flask with 1 test tube strain, the pH value is natural, the culture temperature is 24°C, the shaker speed is 200r / min, and the culture time is 48h;

[0097] The composition (w / w) of the seed medium is: by weight (w / w): 2% sucrose, 0.6% yeast extract powder, 0.05% magnesium sulfate, 0.025% potassium dihydrogen phosphate, add water to 1000mL;

[0098] (3) Fermentation and regulation process: Inoculate liquid seeds with 10% (v / v) inoculum into 2L fermentation medium, culture at 26°C for 72 hours, add 1w / v% glycerin and 0.05w of the total...

Embodiment 3

[0104] (1) Activation of strains: move the strains on the slant of the test tube stored on PDA medium at 4°C to room temperature (20°C-25°C) for activation for 4 hours;

[0105] (2) Preparation of liquid seeds: On an aseptic operating table, use 10 mL of sterilized distilled water to make a bacterial suspension from the activated test tube slant strain, and rinse it into a triangular flask containing sterilized seed medium. Inoculate 1 Erlenmeyer flask with 1 test tube strain, the pH value is natural, the culture temperature is 24°C, the shaker speed is 200r / min, and the culture time is 48h;

[0106] The composition (w / w) of the seed medium is: by weight (w / w): 2% sucrose, 0.6% yeast extract powder, 0.05% magnesium sulfate, 0.025% potassium dihydrogen phosphate, add water to 1000mL;

[0107] (3) Fermentation and regulation process: Inoculate liquid seeds with 10% (v / v) inoculation amount into 1L fermentation medium, culture at 26°C for 72 hours, add 1w / v% ethyl acetate and 0....

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