A kind of Paenibacillus polymyxa, its application in the preparation of genistein and the method for preparing genistein
A polymyx spore-like and genistein technology is applied in the field of research and development of new traditional Chinese medicines, which can solve problems such as undiscovered transformation research, and achieve the effects of strong transformation specificity, high transformation efficiency and abundant resources.
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[0061] The preparation method of the Sophora japonicus extract in the present invention: take the solid Sophora japonicus powder, fully extract with water according to the solid-liquid ratio, and then filter to obtain the extract. Taking 10% W / V Sophora japonica extract as an example, its preparation method is as follows: take 10 g of solid Sophora japonica powder, fully boil and extract it with 100 ml of water, filter and take the filtrate to dilute to 100 ml.
[0062] The preparation method of the Paenibacillus polymyxa bacteria liquid in the present invention: transfer the strain stored in the refrigerator to a PDA slope for activation, cultivate at 37°C for 36 hours, transfer to the fermentation medium, and shake and cultivate for 2 days at 37°C and 220r / min on a shaking table That is, the bacterial liquid is obtained.
[0063] The preparation method of Paenibacillus polymyxa bacterial powder in the present invention: transfer the bacterial strains stored in the refrigerat...
Embodiment 1
[0073] Prepare 10% W / V, pH5 Sophora japonica extract, inoculate the Paenibacillus polymyxa bacterial solution of the present invention into the above-mentioned transformation substrate according to the inoculum amount of 5% (V / V) for transformation.
[0074] Transformation method: Shake flask culture at 37°C for 4 days.
[0075] After the conversion, centrifuge at 8000rpm, take the supernatant, and obtain the solid product of genistein by conventional extraction methods.
[0076] The content of genistin and genistein before and after conversion was detected by HPLC method. The result is as image 3 and Figure 4 shown.
[0077] figure 2 It is the standard HPLC figure of genistin and genistein. Wherein A is genistin, the retention time is 10.797min, B is genistein, the retention time is 19.01min.
[0078] image 3 It is the HPLC chart of the conversion stock solution of the present invention.
[0079] Figure 4 It is the HPLC figure after conversion of the present inv...
Embodiment 2-19
[0086] The biotransformation process of Examples 2-19 is basically the same as Example 1.
[0087] The addition form, dosage, transformation substrate and reaction conditions of the transformation method of the microorganisms used are slightly different, as shown in the table below:
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[0089]
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