Method for preparing low impurity acarbose
An acarbose and low impurity technology, applied in the field of preparing low impurity C acarbose, can solve the problems of difficulty in separation, high content of impurity component C, increased purification difficulty and production cost, etc.
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Embodiment 1
[0029] With reference to the medium and process of shake flask culture, seed culture and fermentation culture in Example 7 of Chinese patent 2016102923585, the seeds cultivated in the seed tank were inserted into 1m with an inoculum size of 10% (v / v). 3 The acclimatization tank is used for acclimation culture, and the culture medium of the acclimatization tank is as follows: in every 100ml of culture medium, sucrose 0.5g, glucose 2g, maltose 3g, soybean cake powder 2g, peptone 0.2g, ammonium sulfate 0.1g, sodium glutamate 0.5g, 0.3g of ferric chloride, 0.5g of calcium chloride, 0.5g of calcium carbonate, and the rest are water; the culture conditions are controlled as follows: the stirring speed is controlled at 200rpm, the tank pressure is controlled at 0.05MPa, and the ventilation rate is controlled at 1:1.5 (v / v ), the temperature is 28° C., and cultured for 72 hours. Subsequent access to 10m 3 Fermentation tanks for fermentation.
[0030] Take the cultivated domesticatio...
Embodiment 2-7
[0035] The following examples are all the same as in Example 1 except for the acclimatization medium and acclimatization culture conditions.
[0036] The cultured acclimatization medium was taken for microscopic examination to observe the growth status of the mycelia and the shape of the mycelium, and then the acclimatization medium was inserted into the fermentation medium, and the titer of acarbose was detected by HPLC. The feed liquid of the fermenter was purified by filtration, acidification, chromatography and other processes, and then tested by HPLC. The content of impurity component C was measured in Table 1.
[0037] Wherein embodiment 6 is that the same acclimatization medium is cultivated under different acclimatization conditions, and the impurity component C measured after the fermentation broth is purified is 0.53% (RRT24.7 is impurity C), see appendix image 3 .
[0038] Table: Effects of different acclimatization media and acclimatization conditions on the tite...
Embodiment 8
[0042] According to the culture medium and technology of shake flask culture, seed tank culture, fermentor tank of embodiment 7 of Chinese patent 2016102923585, carry out 60m 3 Fermentation tank enlargement.
[0043] The introduction of domestication culture between seed culture and fermentation culture is as follows:
[0044] 1. Seed culture:
[0045] Culture medium: 2% sucrose, 1% glycerin, 4.8% corn steep liquor powder, 0.5% calcium carbonate, 0.2% Soybean, pH 7.0, sterilized at 121°C for 30 minutes, and the volume of seed medium after sterilization is about 500L.
[0046] Seed culture method: 1m 3 Seed tank, put the cultivated shake flask seed solution into 1m with 0.2% (v / v) inoculum 3 The seed tank was cultured at a constant temperature for 48 hours at a stirring speed of 160 rpm, a tank pressure of 0.05 MPa, a ventilation rate of 1:1.1 (v / v), and a temperature of 28°C.
[0047] 2. Domestication and cultivation:
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