Method for promoting microbes to synthesize ascomycin by using exogenous additive and preparation of seed liquor and culture medium
A technology of exogenous additives and microbial synthesis, applied in the field of microbial fermentation, can solve problems such as gaps, achieve low additional investment, increase the amount of ascomycin synthesis, and simple operation
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Embodiment 1
[0017] Take the mature slant spores of Streptomyces hygroscopicus subspecies (ATCC14891) at room temperature to make a spore suspension with sterile water; Glucose 32g / L, peptone 5g / L, yeast powder 5g / L, calcium carbonate 1g / L, initial pH 7.0) in a 250mL shaker flask, cultured in a shaker at 30°C and 180rpm for 48h; , added to the fermentation medium (composition and content: starch 22g / L, dextrin 40g / L, gluten powder 3g / L, peptone 4g / L, yeast powder 12g / L, corn steep liquor 4g / L, cold Pressed bean cake powder 4g / L, dipotassium hydrogen phosphate 0.5g / L, ammonium sulfate 1g / L, magnesium sulfate 0.5g / L, calcium carbonate 0.5g / L, shikimic acid 0.5g / L, initial pH7.0) 7.5LDE fermenter; fermentation conditions: 30°C, controlled ventilation 0.6v / v / m, rotating speed 500rpm, pH 7.0, fermented for 192h. The concentration of ascomycin in the fermentation broth was detected by HPLC. Ascomycin fermentation unit increased from 170mg / L to 265mg / L, an increase of 55.9%.
Embodiment 2
[0019]Take the mature slant spores of Streptomyces hygroscopicus subspecies (ATCC14891) at room temperature to make a spore suspension with sterile water; Glucose 32g / L, peptone 5g / L, yeast powder 7g / L, calcium carbonate 2g / L, initial pH 7.0) in a 250mL shake flask, cultured in a shaker at 30°C and 180rpm for 48h; , added to the fermentation medium (composition and content: starch 18g / L, dextrin 42g / L, gluten powder 2g / L, peptone 6g / L, yeast powder 10g / L, corn steep liquor 5g / L, cold Pressed bean cake powder 4g / L, dipotassium hydrogen phosphate 1.5g / L, ammonium sulfate 0.5g / L, magnesium sulfate 0.8g / L, calcium carbonate 1.2g / L, initial pH7.0) automatic 7.5LDE fermentation tank. Add 0.5g / L shikimic acid after 36h of fermentation. Fermentation conditions: 30°C, controlled ventilation 0.6v / v / m, rotation speed 500rpm, pH 7.0, fermented for 180h. The concentration of ascomycin in the fermentation broth was detected by HPLC. Ascomycin fermentation unit increased from 170mg / L to 2...
Embodiment 3
[0021] Take the mature slant spores of Streptomyces hygroscopicus subspecies (ATCC14891) at room temperature to make a spore suspension with sterile water; Glucose 29g / L, peptone 4.5g / L, yeast powder 6.5g / L, calcium carbonate 1.5g / L, initial pH7.0) in a 250mL shaker flask, cultured in a shaker at 26°C and 220rpm for 48h; with 10% The amount of inoculum was added to the fermentation medium (composition and content: starch 21g / L, dextrin 41g / L, gluten powder 2.2g / L, peptone 5.5g / L, yeast powder 11g / L, corn steep liquor 5.5g / L, cold-pressed bean cake powder 5.5g / L, dipotassium hydrogen phosphate 0.8g / L, ammonium sulfate 0.8g / L, magnesium sulfate 0.6g / L, calcium carbonate 0.7g / L, isoleucine 2g / L L, in an automatic 7.5LDE fermenter with an initial pH of 7.0). Fermentation conditions: 26°C, 1.0v / v / m aeration, 500rpm rotation speed, pH 7.0, 144h fermentation. The concentration of ascomycin in the fermentation broth was detected by HPLC. Ascomycin fermentation unit increased from 1...
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