Method for biologically synthesizing natural aromadendrin by escherichia coli through utilizing naringenin
A technology of Escherichia coli and naringenin is applied in the field of fermentation engineering to achieve the effects of simple production process, low production cost and simple composition
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Embodiment 1
[0030] Starting strain: the starting strain is Escherichia coli (E.coli) DHK-161114CCTCC NO:M 2016645.
[0031] Plate culture: Streak inoculate the Escherichia coli stored in the puncture tube onto the plate medium, and culture in a 30°C incubator for 15-24 hours.
[0032] Preparation of Escherichia coli CCTCC NO:M 2016645 strain bank: Pick a single colony, inoculate it into the seed medium, culture it on a shaker at 200 rpm for 13-17 hours at 30°C, absorb 0.5ml of the obtained seed culture solution and transfer it to 50% In the glycerol solution, prepare a seed glycerol tube with a final concentration of 25%, and store at -80°C.
[0033] Preparation of seed liquid: absorb 20 uL of seed glycerol tube, inoculate into seed medium, and culture at 30° C. with shaking at 200 rpm for 13 to 17 hours to obtain seed liquid.
[0034] Preparation of seed medium: peptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L, prepared from tap water, pH adjusted to 7.2, sterilized by high pres...
Embodiment 2
[0040] Embodiment 2 different peptone concentrations
[0041] Starting strain: the same as in Example 1.
[0042]Preparation of seed medium: same as in Example 1.
[0043] Preparation of fermentation medium: prepare peptone 0, 2.5, 5, 7.5, 10.0 and 12.5 g / L, yeast extract 5 g / L, sodium chloride 10 g / L, tap water, adjust pH to 7.2, and extinguish with high-pressure steam at 121 °C Bacteria 20min.
[0044] Pipette 20 μL seed glycerol tube into the seed shaker flask culture medium, the liquid volume is 50ml / 250ml, culture at 30°C, 200rpm shaker for 16h, and transfer to fermentation medium with different peptone concentrations according to 5% inoculum size (50ml / 250ml), under the condition of 37°C, 200rpm shaker shake culture for 3h, cool down to 30°C, add 1.5% lactose to induce 3h, maintain 30°C, add 2.7g / L substrate naringenin in total, fermentation time After 54h, the substrate and product content were measured after the fermentation was finished, and the results were shown ...
Embodiment 3
[0046] Embodiment 3 different yeast extract concentrations
[0047] Starting strain: the same as in Example 1.
[0048] Preparation of seed medium: same as in Example 1.
[0049] Preparation of fermentation medium: peptone 10g / L, yeast extract concentrations of 0, 2.5, 5.0 and 7.5g / L, sodium chloride 10g / L, tap water, pH adjusted to 7.2, sterilized by high pressure steam at 121°C for 20min .
[0050] Pipette 20 μL of seed glycerol tube into the seed shaker flask culture medium, the liquid volume is 50ml / 250ml, and culture at 30°C, 200rpm shaker for 16h, and transfer to fermentation culture with different yeast extract concentration according to 5% inoculum size Medium (50ml / 250ml), at 37°C, 200rpm shaker culture for 3h, cooled to 30°C, added 1.5% lactose for induction for 3h, maintained at 30°C, added 2.7g / L substrate naringenin in total, The fermentation time was 54h, and the substrate and product content were measured at the end of the fermentation. The results are shown ...
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