Preparation method of stable isotop labelled 15N-L-asparagic acid
A stable isotope, aspartic acid technology, applied in the direction of fermentation, to achieve the effect of improving extraction process, improving market competitiveness and improving product quality
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Embodiment 1
[0031] The Escherichia coli cultured on the slant for 18 hours was inserted into the fermentation medium, filled with 50ml medium in a 500ml Erlenmeyer flask, and cultivated in a constant temperature shaker at 30°C for 20 hours with a rotation speed of 180rpm. Cultivate the obtained fermented liquid and centrifuge at 4800rpm for 30 minutes, pour off the supernatant, and add 0.5 g of the obtained wet thalline to the substrate concentration of 10% and the abundance of 10.39at%. 15 In 100 ml of a substrate solution of 7.7% by weight of N-ammonium sulfate and 0.01% by weight of magnesium sulfate, the pH value was adjusted to 7.9 with NaOH, and the reaction was carried out at a constant temperature of 35° C. for 23 hours. The obtained reaction solution was centrifuged, and after the supernatant was concentrated to pH 7.5 by ammonia, 0.2 gram of activated carbon was added to decolorize for 30 minutes to obtain a colorless solution, which was concentrated to 25 ml, and sulfuric acid w...
Embodiment 2
[0033] Get 2.0 grams of wet thalline cultured as in Example 1, add to the substrate concentration and be 11.6% by weight, containing abundance is 99.53at% 15 In 200 ml of a substrate solution of 7.4% by weight of N-ammonium chloride and 0.01% by weight of magnesium sulfate, the pH value was adjusted to 9.0 with 10M NaOH, and the reaction was carried out at a constant temperature of 39° C. for 27 hours. The obtained reaction solution was centrifuged, and the supernatant was concentrated to pH 6.5 with ammonia, and 0.6 g of active carbon was added to decolorize for 35 minutes to obtain a colorless solution, which was concentrated to 55 ml, and hydrochloric acid was added to adjust the pH to 3.00, and a large amount of white crystals were precipitated. Cooling, suction filtration and washing the next day, to obtain snow-white crystals, drying to obtain 23.65 grams of product, with an abundance of 99.18 at%, which is only 0.35 at% absolute and 0.35 at% relative to the raw material,...
Embodiment 3
[0035] The Escherichia coli cultured on the slant for 20 hours was inserted into the fermentation medium, 50ml of the medium was filled in a 500ml Erlenmeyer flask, and cultured on a constant temperature shaker at 32°C for 24 hours at a rotation speed of 200rpm. Cultivate the obtained fermented liquid and centrifuge at 4800rpm for 30 minutes, pour off the supernatant, and add 0.5 g of the obtained wet thalline to the substrate concentration of 10% and the abundance of 98.50at%. 15 In 100 ml of a substrate solution of 8% by weight of N-ammonium sulfate and 0.01% by weight of magnesium sulfate, the pH value was adjusted to 8.3 with NaOH, and the reaction was carried out at a constant temperature of 37° C. for 25 hours. The obtained reaction solution was centrifuged, and the supernatant was concentrated to pH 6.8 by catching up with ammonia, and after adding 0.25 g of active carbon for decolorization, a colorless solution was obtained for 40 minutes, concentrated to 20 ml, added s...
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