Method for improving yield of endophytic fungus fermentation product 4-hydroxyphenethyl alcohol through epigenetics modification
A technology of epigenetics and hydroxyphenylethyl alcohol, applied in the field of biological fermentation, can solve the problems of unfriendly environment, low efficiency, long reaction steps, etc., and achieve the effect of reducing production cost, low economic cost and simple operation
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Embodiment 1
[0024] Prepare medium: 35g PDB medium dry powder, sodium pyruvate 200μM, distilled water 1L, adjust pH to 6.5, autoclave at 121°C for 20min.
[0025] Preparation of seed liquid: Add 100 mL of culture medium to a 500 mL Erlenmeyer flask, add activated strains, culture at 28°C and 180 rpm / min for 3 days to obtain seed liquid.
[0026] Large-scale fermentation: Use 500mL Erlenmeyer flasks to ferment 10 bottles in total, each bottle contains 150mL of culture medium, and inoculate the seed liquid into PDB medium containing sodium pyruvate according to the inoculum amount of 10%, and cultivate at 28°C and 180rpm / min 7 days.
[0027] Extraction and separation; combine 3 bottles of fermentation broth, use ethyl acetate to extract the fermentation broth 3 times, combine the ethyl acetate extracts, spin dry to obtain 630 mg of crude extract, use silica gel column separation to obtain 183 mg of the target product, and use nuclear magnetic resonance analysis to identify the target product...
Embodiment 2
[0030] Prepare medium: 35g PDB medium dry powder, sodium pyruvate 100μM, distilled water 1L, adjust pH to 7.0, autoclave at 121°C for 20min.
[0031] Preparation of seed liquid: Add 100 mL of culture medium to a 500 mL Erlenmeyer flask, add activated strains, culture at 26°C, 150 rpm / min, for 5 days to obtain seed liquid.
[0032] Large-scale fermentation: Use 500mL Erlenmeyer flasks to ferment 10 bottles in total, each bottle contains 150mL of culture medium, and inoculate the seed liquid into PDB medium containing sodium pyruvate according to the inoculum size of 10%, and cultivate at 26°C and 150rpm / min 7 days.
[0033] Extraction and separation; combine 3 bottles of fermentation broth, extract the fermentation broth 3 times with ethyl acetate, combine the ethyl acetate extracts, spin dry to obtain 603 mg of crude extract, and use silica gel column to separate to obtain 149 mg of the target product. The target product was identified as 4-hydroxyphenethyl alcohol by NMR ana...
Embodiment 3
[0035] Prepare medium: 35g PDB medium dry powder, sodium pyruvate 500μM, distilled water 1L, adjust pH to 7.5, autoclave at 121°C for 20min.
[0036] Preparation of seed solution: add 100mL of culture medium to a 500mL Erlenmeyer flask, add activated strains, culture at 30°C and 170rpm / min for 2 days to obtain seed solution.
[0037] Large-scale fermentation: use 500mL Erlenmeyer flasks to ferment 10 bottles in total, each bottle contains 150mL of culture medium, and inoculate the seed liquid into PDB medium containing sodium pyruvate according to the inoculum amount of 15%, and cultivate at 30°C and 170rpm / min 7 days.
[0038] Extraction and separation: combine 3 bottles of fermentation broth, extract the fermentation broth 3 times with ethyl acetate, combine the ethyl acetate extracts, spin dry to obtain 579 mg of crude extract, and use silica gel column separation to obtain 140 mg of the target product. The target product was identified as 4-hydroxyphenethyl alcohol by NMR...
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