Penicillium notatum strain and application thereof in biotransformation mulberry leaf flavonoid glycoside
A technology for biotransformation and mulberry leaf flavonoids, applied in the field of microorganisms, can solve the problems of lack of basic data, unstable yield and conversion rate, and no research reports on the application of mulberry leaf flavonoid aglycones.
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Embodiment 1
[0015] This example illustrates the screening procedure for Penicillium notatum LJ-2.
[0016] Collect multiple soil samples, put them into a conical flask with 50mL of sterile water, add about 20 glass beads, shake vigorously for 20 minutes to completely mash the soil, and let it stand still. Add 1mL of soil sample suspension into each 50mL sterile water Erlenmeyer flask, put it into a constant temperature shaking incubator and cultivate it at 37°C for 2 days. Take the suspension and spread it on the primary screening plate medium to carry out the primary screening of the enzyme-producing strains. The quality composition of the primary screening medium is: dipotassium hydrogen phosphate 1.0%, potassium dihydrogen phosphate 0.5%, ammonium sulfate 0.1%, sodium nitrate 0.05%, magnesium sulfate 0.05%, ferrous sulfate 0.005%, zinc sulfate 0.005%, agar 5%, pH6.5, and add 1% rutin. The temperature is 35°C, and the cultivation time is 1 to 6 days. Select 20 strains that produce la...
Embodiment 2
[0022]This example illustrates the method of using Penicillium notatum LJ-2 to carry out fermentation and enzyme production to produce glycoside hydrolase and its biotransformation of mulberry flavone glycosides to prepare highly active aglycones.
[0023] The mass ratio composition of the medium: dipotassium hydrogen phosphate 0.1%, potassium dihydrogen phosphate 0.1%, ammonium sulfate 0.1%, sodium nitrate 0.01%, magnesium sulfate 0.01%, ferrous sulfate 0.001%, zinc sulfate 0.001%, pH5.0, And add 1% rutin.
[0024] The spore suspension (mass concentration) of Penicillium notatum LJ-2 was inoculated in the culture medium with an inoculum amount of 1%, and cultured aerobically at a temperature of 25° C. for 1 day, that is, the enzyme production ended.
[0025] The flavonoid glucoside of mulberry leaves is dissolved to prepare a solution with a mass concentration of 1%, and the pH is adjusted to 4.0 with NaOH or HCl solution, which is used as a substrate for the enzyme-catalyzed...
Embodiment 3
[0027] The method of this embodiment is the same as that of Embodiment 2, and the parameters of the method used are changed.
[0028] The mass ratio composition of the medium: dipotassium hydrogen phosphate 1.0%, potassium dihydrogen phosphate 0.5%, ammonium sulfate 0.1%, sodium nitrate 0.05%, magnesium sulfate 0.05%, ferrous sulfate 0.005%, zinc sulfate 0.005%, pH6.0.
[0029] The spore suspension (mass concentration) of Penicillium notatum LJ-2 was inoculated in the culture medium at an inoculum size of 2%, and cultured aerobically at a temperature of 35° C. for 4 days, that is, the enzyme production ended.
[0030] The solution is prepared into a solution with a mass concentration of 3%, which is used as the substrate of the enzyme-catalyzed reaction. Add the substrate solution to the crude enzyme solution or wet bacteria obtained by filtering the fermentation broth, and adjust the pH to 6.5 with NaOH or HCl solution, at 35°C, with a rotation speed of 150r / min, and react fo...
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