Method for producing homoeriodictyol by fermentation of recombinant escherichia coli
A technology for recombining Escherichia coli and Escherichia coli, which is applied in the field of bioengineering, can solve the problems of harsh transformation conditions, difficulty in large-scale production, low yield of high eriodictyol, etc., and achieve the effect of simple and easy production process
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Embodiment 1
[0033] Medium:
[0034] 1. Seed medium: peptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L, magnesium sulfate heptahydrate 0.5g / L, mixed vitamin solution 10ml / L, tap water, kanamycin 50mg / L, the pH was adjusted to 7.2, the medium was sterilized at 121°C for 20 minutes, and the vitamin mixture and kanamycin were filtered through a 0.22 μm filter membrane and added.
[0035] 2. Fermentation medium: glucose 4g / L, lactose 10g / L, yeast extract 5g / L, disodium hydrogen phosphate 3g / L, sodium dihydrogen phosphate 1.2g / L, ammonium sulfate 0.5g / L, chloride Sodium 0.5g / L, magnesium sulfate heptahydrate 0.5g / L, calcium chloride 0.05g / L, mixed vitamin solution 15ml / L, kanamycin 30mg / L, prepared with tap water, pH adjustment 7.2, culture medium 121℃ Bacteria for 20 minutes, in which glucose and lactose were sterilized separately at 115°C for 20 minutes, glucose was added before inoculation, lactose was added later as feeding sugar, and vitamin mixture and kanamycin were added after ...
Embodiment 2
[0043] Medium:
[0044] 1. Plate medium: peptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L, agar 20g / L, kanamycin 50mg / L. Among them, kanamycin was added after filtering through a 0.22 μm filter membrane.
[0045] 2. Primary seed medium: consistent with the seed medium in Example 1.
[0046] 3. Secondary seed culture: glucose 4g / L, yeast extract 5g / L, disodium hydrogen phosphate 3g / L, sodium dihydrogen phosphate 1.2g / L, ammonium sulfate 0.5g / L, sodium chloride 0.5g / L L, Magnesium Sulfate Heptahydrate 0.5g / L, Calcium Chloride 0.05g / L, Mixed Vitamin Solution 15ml / L, Kanamycin 50mg / L, prepared from tap water, pH adjusted to 7.2, medium sterilized at 121°C for 20min, of which Glucose and lactose were sterilized separately at 115°C for 20 minutes. Glucose was added before inoculation, and vitamin mixture and kanamycin were added after filtering through a 0.22 μm filter membrane.
[0047] 4. Fermentation medium: glucose 4g / L, lactose 15g / L, yeast extract 5g / L, disodium hyd...
Embodiment 3
[0057] Medium:
[0058] The primary seed and secondary seed medium are the same as in Example 2 in this embodiment. The initial concentration of glucose in the fermentation medium continued to maintain 4g / L, and an additional 8g / L was added in stage 3. 4g / L other ingredients are the same as in Example 2.
[0059] Training method:
[0060] The stage 1 and stage 2 in the cultivation method in this embodiment are consistent with the embodiment 2. In stage 3, the addition of eriodictyol was changed to 0.4g / L at 4.5h, 16h, 28h and 40h of fermentation, and 2g / L of glucose was added at the same time as eriodictyol was added. In the third stage, a total of 1.6g / L eriodictyol and 8g / L glucose were added to ferment for 72 hours, and the yield of homo eriodictyol was 1.17g / L. About 73.1%.
[0061] Compared with Example 2, Example 3 increased the glucose supplementation process in the conversion stage, and this adjustment improved the yield of homo-eriodicty by about 21.8%.
[0062]...
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