Trichosporon asahii 2-1 and application thereof in preparing acrylic acid
A technology of Trichosporium and Trichosporium, applied in the application field of preparing acrylic acid, can solve the problems of easy pollution, high reaction energy consumption and high equipment requirements
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Embodiment 1
[0074] Seed medium formula: 1.0g acrylic acid, 15.0g sucrose, 5.0g glucose, 10g yeast extract, 5g NaCl, KH 2 PO 4 1.0g, MgSO 4 .7H 2 O 0.2g, FeSO 4 0.01g, prepared with deionized water, adjusted the pH to 7.0 with ammonia water, the total volume is 1L.
[0075] Fermentation medium: acrylic acid 10.0g, sucrose 20.0g, yeast extract 5.0g, NaCl 1.0g, KH 2 PO 4 1.0g, MgSO4.7H 2 O 0.2g, FeSO 4 0.1g, prepared with deionized water, adjusted the pH to 7.0 with ammonia water, the total volume is 1L.
[0076] Take 120mL of the above-mentioned seed culture medium, evenly distribute it in four 250mL Erlenmeyer flasks, and sterilize. Insert Trichosporon assahii 2-1, a slant fungus, at a shaker speed of 150r / min, and culture on a shaker at 30°C for 20h as a seed solution for later use.
[0077] Take 1L of the above-mentioned fermentation medium, evenly distribute it into ten 500mL shake flasks, and sterilize. The seed liquid was inserted, the inoculum amount was 2% (V / V), and c...
Embodiment 2
[0080] The composition of the seed medium and the cultivation of the seeds are the same as in Example 1.
[0081] Fermentation enzyme production medium formula: acrylic acid 5.0g, sucrose 15.0g, yeast extract 5.0g, urea 1.0g, NaCl 1.0g, KH 2 PO 4 0.5g, MgSO 4 .7H 2 O 0.5g, FeSO 4 0.1g, prepared with deionized water, adjusted the pH to 7.0 with ammonia water, the total volume is 1L;
[0082] Take 1L of the above-mentioned fermentation medium, evenly distribute it into ten 500mL shake flasks, and sterilize. The seed liquid was inserted, the inoculum amount was 2% (V / V), and culture was carried out, the culture temperature was 30° C., the culture time was 20 h, and the rotation speed of the shaker was 150 r / min.
[0083] After the cultivation was completed, the fermentation broth was centrifuged at 5000 r / min at 15° C. for 5 minutes to collect the bacterial cells, and the bacterial cells were suspended in phosphate buffer for transformation reaction. The reaction system i...
Embodiment 3
[0085] The composition of the seed medium and the cultivation of the seeds are the same as in Example 1.
[0086] Fermentation enzyme production medium formula: 3.0g acrylic acid, 15g sucrose, 5.0g yeast extract, 2.0g urea, 1.0g NaCl, KH 2 PO 4 0.5g, MgSO 4 .7H 2 O 0.5g, FeSO 4 0.2g, prepared with deionized water, adjusted the pH to 8.0 with ammonia water, the total volume is 1L.
[0087] Take 1L of the above-mentioned fermentation medium, evenly distribute it into ten 500mL shake flasks, and sterilize. The seed liquid was inserted, the inoculum amount was 2% (V / V), and culture was carried out, the culture temperature was 30° C., the culture time was 20 h, and the rotation speed of the shaker was 150 r / min.
[0088] After the cultivation is completed, the fermentation broth is centrifuged at 5000 r / min at 15° C. for 5 minutes to collect the bacterial cells, and the bacterial cells are suspended in the buffer for transformation reaction. The reaction system is 100mL, in...
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