Enterobacter SYA2 and method for preparing lactase with same
A technology of lactase and Enterobacteriaceae, applied in the field of bioengineering, to achieve the effects of fast reproduction, easy fermentation process, and small size
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Embodiment 1
[0046] Enterobacteriaceae SYA2 was inoculated in the seed medium, and cultured on a shaker at 37 °C and 150 r / min for 24 hours to obtain the seed liquid. The composition of the seed medium was: lactose 10 g / L, protein powder 10 g / L, yeast extract 3 g / L, sodium chloride 3 g / L, pH 7.0;
[0047] Add the seed liquid to the liquid fermentation medium at an inoculum amount of 3%, and culture it with shaking at 37°C and 180r / min for 24 hours to obtain a fermentation liquid. The composition of the liquid fermentation medium is: lactose 15g / L, protein age 20 g / L , Yeast extract 3 g / L, sodium chloride 3 g / L, dipotassium hydrogen phosphate 1 g / L, manganese dioxide 0.1 g / L, pH 7.0;
[0048]Centrifuge the fermentation broth at 4°C and 6000r / min for 20min, collect the cells and wash them twice with 0.1mol / L phosphate buffer solution (pH 6.5), sonicate the cells for 15min, and centrifuge at 12000r / min for 20min at 4°C. Obtain the supernatant.
[0049] Dry and grind the ammonium sulfate, ta...
Embodiment 2
[0054] Enterobacteriaceae SYA2 was inoculated in the seed medium, and cultured on a shaker at 37 °C and 150 r / min for 24 hours to obtain the seed liquid. The composition of the seed medium was: lactose 10 g / L, protein powder 10 g / L, yeast extract 3 g / L, sodium chloride 3 g / L, pH 7.0;
[0055] Add the seed liquid to the liquid fermentation medium at an inoculum amount of 3%, and culture it with shaking at 37°C and 190r / min for 24 hours to obtain a fermentation liquid. The composition of the liquid fermentation medium is: lactose 15g / L, protein age 20 g / L , Yeast extract 3 g / L, sodium chloride 3 g / L, dipotassium hydrogen phosphate 1 g / L, manganese dioxide 0.1 g / L, pH 7.0;
[0056] Centrifuge the fermentation broth at 4°C and 6000r / min for 20min, collect the cells and wash them twice with 0.1mol / L phosphate buffer solution (pH 6.5), sonicate the cells for 15min, and centrifuge at 12000r / min for 20min at 4°C. Obtain the supernatant.
[0057] Dry and grind the ammonium sulfate, t...
Embodiment 3
[0060] Enterobacteriaceae SYA2 was inoculated in the seed medium, and cultured on a shaker at 37°C and 150 r / min for 24 hours to obtain the seed liquid. The composition of the seed medium was: lactose 10 g / L, protein powder 10 g / L, yeast extract 3 g / L, sodium chloride 3 g / L, pH 7.0;
[0061] Add the seed liquid into the liquid fermentation medium at an inoculum amount of 3%, and culture it with shaking at 37°C and 200r / min for 24 hours to obtain a fermentation liquid. The composition of the liquid fermentation medium is: lactose 15g / L, protein age 20 g / L , Yeast extract 3 g / L, sodium chloride 3 g / L, dipotassium hydrogen phosphate 1 g / L, manganese dioxide 0.1 g / L, pH 7.0;
[0062] The fermentation broth was centrifuged at 4°C and 6000r / min for 20min, the cells were collected and washed twice with 0.1mol / L phosphate buffer solution (pH 6.5), the cells were sonicated for 15min, centrifuged at 12000r / min for 20min, 4°C, Obtain the supernatant.
[0063] Dry and grind the ammonium...
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