Method for preparing rebaudioside A by fermentation and catalysis of bacillus subtilis
A Bacillus subtilis, catalyzed preparation technology, applied in the field of microbial fermentation enzyme preparations, can solve the problems of complex operation, high production cost, and inability to apply industrial production, and achieve low enzyme activity, reduce operation steps, and be conducive to industrial production. Effect
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Embodiment 1
[0020] (1) Seed preparation: link the synthesized glycosyltransferase UGT76G1 gene to the PBR322 plasmid vector, and transfer the PBR322 plasmid vector carrying the glycosyltransferase UGT76G1 gene into a competent Bacillus subtilis, and implant the The Bacillus subtilis with the glycosyltransferase UGT76G1 gene was inserted into the LB medium, the temperature of the LB medium was controlled at 34 °C, the rotation speed was 150 rpm, and the culture time was 22 h;
[0021] (2) Seed tank cultivation: The Bacillus subtilis obtained in step (1) was inserted into the seed tank containing LB medium according to the inoculation amount of 5% volume ratio, and the rotation speed of the seed tank was controlled at 300 rpm, and the ventilation ratio ( The aeration ratio is the volume ratio of air passing through a unit volume of culture solution per minute) is 0.5 V / V.min, cultivated at 35°C for 22 hours to obtain a seed culture solution;
[0022] (3) Fermentation tank production: put th...
Embodiment 2
[0028] (1) Seed preparation: link the synthesized glycosyltransferase UGT76G1 gene to the PBR322 plasmid vector, and transfer the PBR322 plasmid vector carrying the glycosyltransferase UGT76G1 gene into a competent Bacillus subtilis, and implant the The Bacillus subtilis with the glycosyltransferase UGT76G1 gene was inserted into the LB medium, the temperature of the LB medium was controlled at 30°C, the rotation speed was 200 rpm, and the culture time was 20 h;
[0029] (2) Seed tank cultivation: The Bacillus subtilis obtained in step (1) was inserted into the seed tank containing LB medium according to the inoculation amount of 5% volume ratio, and the rotation speed of the seed tank was controlled at 300 rpm, and the ventilation ratio ( The aeration ratio is the volume ratio of air passing through a unit volume of culture solution per minute) is 0.4 V / V.min, cultivated at 30°C for 20 hours to obtain a seed culture solution;
[0030] (3) Fermentation tank production: The see...
Embodiment 3
[0036] (1) Seed preparation: link the synthesized glycosyltransferase UGT76G1 gene to the PBR322 plasmid vector, and transfer the PBR322 plasmid vector carrying the glycosyltransferase UGT76G1 gene into a competent Bacillus subtilis, and implant the The Bacillus subtilis with the glycosyltransferase UGT76G1 gene was inserted into the LB medium, the temperature of the LB medium was controlled at 30°C, the rotation speed was 220 rpm, and the culture time was 24 h;
[0037] (2) Seed tank cultivation: The Bacillus subtilis obtained in step (1) was inserted into the seed tank containing LB medium according to the inoculation amount of 8% volume ratio, and the rotation speed of the seed tank was controlled at 400 rpm, and the ventilation ratio ( The aeration ratio is the volume ratio of air passing through a unit volume of culture solution per minute) is 0.3 V / V.min, cultivated at 30°C for 24 hours to obtain a seed culture solution;
[0038](3) Fermentation tank production: The seed...
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