Preparation method and application of Bacillus methylotrophicus and tobacco angelica sinensis extract thereof
A technology of eating methanol spores and angelica extract, which is applied in the field of tobacco flavors, can solve the problems of dry throat, many by-products, and large irritation, and achieve the effect of wide sources of raw materials, wide sources, and simple process
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Embodiment 1
[0033] The present invention utilizes the microorganism Bacillus methylotrophicus VJ4-1CCTCC NO: M 2012004 to biotransform the preparation method of Angelica extract for tobacco as follows:
[0034] 1) Crush the Chinese herbal medicine Angelica sinensis to 30 mesh, then reflux extract the coarse powder of Angelica sinensis and water at a mass ratio of 1:20 for 3 hours, then filter, repeat the reflux extraction filtrate once, combine the filtrates and sterilize at 115°C for 25 minutes;
[0035] 2) Slant culture: inoculate Bacillus methylotrophicus VJ4-1CCTCC NO: M 2012004 on the potato dextrose agar slant medium, and culture it statically for 72 hours at 25°C and pH 12, then place it in a refrigerator at 4°C;
[0036] Slant medium: potato infusion 200mL / L, glucose 20g / L, agar 13g / L;
[0037] 3) Preparation of seed culture medium: use the slant culture obtained in the above step 2) to inoculate the seed culture medium, vibrate for 12 hours at 25°C and rotate at 150 rpm, and tra...
Embodiment 2
[0042] The present invention utilizes the microorganism Bacillus methylotrophicus VJ4-1CCTCC NO: M 2012004 to biotransform the preparation method of angelica extract for tobacco as follows:
[0043] 1) Crush the Chinese herbal medicine Angelica sinensis to 50 mesh, then reflux extract the coarse powder of Angelica sinensis and water at a mass ratio of 1:40 for 2 hours, then filter, repeat the reflux extraction filtrate twice, combine the filtrates and sterilize at 115°C for 25 minutes;
[0044] 2) Slant culture: inoculate Bacillus methylotrophicus VJ4-1CCTCC NO: M 2012004 on the potato dextrose agar slant medium, and culture it statically for 36 hours at 30°C and pH 10, then place it in a refrigerator at 4°C;
[0045] Slant medium: potato infusion 200mL / L, glucose 20g / L, agar 13g / L;
[0046] 3) Preparation of seed culture solution: use the slant culture obtained in the above step 2) to inoculate the seed medium, vibrate and culture at 30°C and 120 rpm for 24 hours, and make t...
Embodiment 3
[0051] The present invention utilizes the microorganism Bacillus methylotrophicus VJ4-1 CCTCC NO: M 2012004 to biotransform the preparation method of Angelica sinensis extract for tobacco as follows:
[0052] 1) Crush the Chinese herbal medicine Angelica sinensis to 80 meshes, then reflux extract the coarse powder of Angelica sinensis and water at a mass ratio of 1:60 for 1 hour, then filter, repeat the reflux extraction filtrate 3 times, combine the filtrates and sterilize;
[0053] 2) Slant culture: inoculate Bacillus methylotrophicus VJ4-1CCTCC NO: M 2012004 on the potato dextrose agar slant medium, and culture it statically for 24 hours at 35°C and pH 8, then place it in a 4°C refrigerator;
[0054] Slant medium: potato infusion 200mL / L, glucose 20g / L, agar 13g / L;
[0055] 3) Preparation of seed culture medium: use the slant culture obtained in the above step 2) to inoculate the seed culture medium, vibrate and culture at 35°C and 100 rpm for 36 hours, and make the seed c...
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